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Estradiol inhibits the activity of proton-coupled amino acid transporter PAT1 expressed in Xenopus oocytes.
Estrogen has great potential as a therapeutic agent in focal ischemic brain injury. Amino acids as energy resources and neurotransmitters in the central nervous system are crucial for proper neuronal function and excitability. The proton-coupled amino acid transporter PAT1 has clear potential in drug absorption. In this paper, human brain PAT1 was cloned and expressed in Xenopus oocytes. The effects of estradiol on the activity of PAT1 were investigated. Glycine-induced membrane currents mediated by PAT1 were measured using the two-electrode voltage clamp technique. The amplitude of the glycine-elicited current was decreased progressively with increasing concentrations of ?-estradiol. A concentration-dependent outwards current of PAT1 was also detected by the presence of ?-estradiol. We conclude that estrogen attenuates the activity of PAT1 by directly closing PAT1 channel. Our results may provide an additional mechanism for estrogen on neurotransmission and neuronal metabolism during ischemic injury. PMID:22975709
Shan, Linlin; Yang, Yujie; Wang, Jin; Zuo, Ji; Dong, Xianhong; Li, Chaokun; Li, Dongliang
Influence of a proton gradient on the transport kinetics of the H +\\/amino acid cotransporter PAT1 in Caco-2 cells
Microsoft Academic Search
The recently cloned proton-coupled amino acid transporter 1 (PAT1) not only accepts several amino acids as substrates but also pharmaceutically relevant l-proline or GABA derivatives such as cis-4-hydroxy-l-proline, l-azetidine-2-carboxylic acid (LACA), 3-amino-1-propanesulfonic acid, nipecotic acid, and the antituberculotic agent d-cycloserine. Because human intestine expresses hPAT1 at the brush border membrane, the transporter may serve as a new oral drug delivery
Linda Metzner; Matthias Brandsch
Influence of a proton gradient on the transport kinetics of the H+/amino acid cotransporter PAT1 in Caco-2 cells.
The recently cloned proton-coupled amino acid transporter 1 (PAT1) not only accepts several amino acids as substrates but also pharmaceutically relevant L-proline or GABA derivatives such as cis-4-hydroxy-L-proline, L-azetidine-2-carboxylic acid (LACA), 3-amino-1-propanesulfonic acid, nipecotic acid, and the antituberculotic agent D-cycloserine. Because human intestine expresses hPAT1 at the brush border membrane, the transporter may serve as a new oral drug delivery route. Using the human intestinal cell line Caco-2, we have investigated the influence of an inwardly directed proton gradient on the kinetic parameters of L-proline uptake. H+ altered only the apparent affinity of L-proline transport and not the maximal transport velocity. Similarly, treatment of the cells with diethylpyrocarbonate (DEPC), known to chemically modify histidyl residues and block their function, affected only the Kt value of L-proline transport. Both increasing pH and DEPC treatment strongly increased the inhibition constants (Ki) of several drugs at hPAT1. It is concluded that H+ stimulates hPAT1 primarily by increasing the substrate affinity with no detectable influence on the maximal transport velocity of the transporter. PMID:16531028
Metzner, Linda; Brandsch, Matthias
Role of Rck-Pat1b binding in assembly of processing-bodies
PubMed Central
The DEAD box RNA helicase Rck and the scaffold protein Pat1b participate in controlling gene expression at the post-transcriptional level by suppressing mRNA translation and promoting mRNA decapping. In addition, both proteins are required for the assembly of processing (P)-bodies, cytoplasmic foci that contain stalled mRNAs and numerous components of the mRNA decay machinery. The C-terminal RecA-like domain of Rck interacts with the N-terminal acidic domain of Pat1b. Here, we identified point mutations in human Rck and Pat1b that prevent the two proteins from binding to each other. By analyzing interaction-deficient mutants in combination with knockdown and rescue strategies in human HeLa cells, we found that Pat1b assembles P-bodies and suppresses expression of tethered mRNAs in the absence of Rck binding. In contrast, Rck requires the Pat1b-binding site in order to promote P-body assembly and associate with the decapping enzyme Dcp2 as well as Ago2 and TNRC6A, two core components of the RNA-induced silencing complex. Our data indicate that P-body assembly occurs in a step-wise manner, where Rck participates in the initial suppression of mRNA translation, whereas Pat1b in a second step triggers P-body assembly and promotes mRNA decapping.
Ozgur, Sevim; Stoecklin, Georg
Identification of homologues to the pathogenicity factor Pat-1, a putative serine protease of Clavibacter michiganensis subsp. michiganensis.
Hybridization of Clavibacter michiganensis subsp. michiganensis total DNA against the pathogenicity gene pat-1 indicated the presence of pat-1 homologous nucleotide sequences on the chromosome and on plasmid pCM2. Isolation of the corresponding DNA fragments and nucleotide sequence determination showed that there are three pat-1 homologous genes: chpA (chromosome) and phpA and phpB (plasmid pCM2). The gene products share common characteristics, i.e. a signal sequence for Sec-dependent secretion, a serine protease motif, and six cysteine residues at conserved positions. Gene chpA located on the chromosome is a pseudogene since it contains a translational stop codon after 97 of 280 amino acids. In contrast to pat-1, cloning of the plasmid encoded homologs phpA and phpB into the avirulent plasmid free Cmm strain CMM100 did not result in a virulent phenotype. So far, no proteolytic activity could be demonstrated for Pat-1, however, site specific mutagenesis of pat-1 showed that the serine residue in the motif GDSGG is required for the virulent phenotype of pat-1 and thus Pat-1 could be a functional protease. PMID:16255147
Burger, Annette; Gr�fen, Ines; Engemann, Jutta; Niermann, Erik; Pieper, Martina; Kirchner, Oliver; Gartemann, Karl-Heinz; Eichenlaub, Rudolf
PAT-1 safety analysis report addendum.
SciTech Connect
The Plutonium Air Transportable Package, Model PAT-1, is certified under Title 10, Code of Federal Regulations Part 71 by the U.S. Nuclear Regulatory Commission (NRC) per Certificate of Compliance (CoC) USA/0361B(U)F-96 (currently Revision 9). The purpose of this SAR Addendum is to incorporate plutonium (Pu) metal as a new payload for the PAT-1 package. The Pu metal is packed in an inner container (designated the T-Ampoule) that replaces the PC-1 inner container. The documentation and results from analysis contained in this addendum demonstrate that the replacement of the PC-1 and associated packaging material with the T-Ampoule and associated packaging with the addition of the plutonium metal content are not significant with respect to the design, operating characteristics, or safe performance of the containment system and prevention of criticality when the package is subjected to the tests specified in 10 CFR 71.71, 71.73 and 71.74.
Weiner, Ruth F.; Schmale, David T.; Kalan, Robert J.; Akin, Lili A.; Miller, David Russell; Knorovsky, Gerald Albert; Yoshimura, Richard Hiroyuki; Lopez, Carlos; Harding, David Cameron; Jones, Perry L.; Morrow, Charles W.
Conserved charged amino acid residues in the extracellular region of sodium\\/iodide symporter are critical for iodide transport activity
BACKGROUND: Sodium\\/iodide symporter (NIS) mediates the active transport and accumulation of iodide from the blood into the thyroid gland. His-226 located in the extracellular region of NIS has been demonstrated to be critical for iodide transport in our previous study. The conserved charged amino acid residues in the extracellular region of NIS were therefore characterized in this study. METHODS: Fourteen
Chia-Cheng Li; Tin-Yun Ho; Chia-Hung Kao; Shih-Lu Wu; Ji-An Liang; Chien-Yun Hsiang
Functional sodium iodide symporter expression in breast cancer xenografts in�vivo after systemic treatment with retinoic acid and dexamethasone
Context��The sodium iodide symporter (NIS) mediates iodide uptake in the thyroid gland as well as in lactating breast, and is also\\u000a expressed in the majority of breast cancers. Recently, we have reported stimulation of all-trans retinoic acid (atRA)-induced NIS expression in the human breast cancer cell line MCF-7 by dexamethasone (Dex), resulting\\u000a in an enhanced therapeutic effect of 131I in�vitro.
Michael J. Willhauck; Bibi Sharif-Samani; Reingard Senekowitsch-Schmidtke; Nathalie Wunderlich; Burkhard G�ke; John C. Morris; Christine Spitzweg
Transport of the photodynamic therapy agent 5-aminolevulinic acid by distinct H+-coupled nutrient carriers coexpressed in the small intestine.
5-Aminolevulinic acid (ALA) is a prodrug used in photodynamic therapy, fluorescent diagnosis, and fluorescent-guided resection because it leads to accumulation of the photosensitizer protoporphyrin IX (PpIX) in tumor tissues. ALA has good oral bioavailability, but high oral doses are required to obtain selective PpIX accumulation in colonic tumors because accumulation is also observed in normal gut mucosa. Structural similarities between ALA and GABA led us to test the hypothesis that the H(+)-coupled amino acid transporter PAT1 (SLC36A1) will contribute to luminal ALA uptake. Radiolabel uptake and electrophysiological measurements identified PAT1-mediated H(+)-coupled ALA symport after heterologous expression in Xenopus oocytes. The selectivity of the nontransported inhibitors 5-hydroxytryptophan and 4-aminomethylbenzoic acid for, respectively, PAT1 and the H(+)-coupled di/tripeptide transporter PepT1 (SLC15A1) were examined. 5-Hydroxytryptophan selectively inhibited PAT1-mediated amino acid uptake across the brush-border membrane of the human intestinal (Caco-2) epithelium whereas 4-aminomethylbenzoic acid selectively inhibited PepT1-mediated dipeptide uptake. The inhibitory effects of 5-hydroxytryptophan and 4-aminomethylbenzoic acid were additive, demonstrating that both PAT1 and PepT1 contribute to intestinal transport of ALA. This is the first demonstration of overlap in substrate specificity between these distinct transporters for amino acids and dipeptides. PAT1 and PepT1 expression was monitored by reverse transcriptase-polymerase chain reaction using paired samples of normal and cancer tissue from human colon. mRNA for both transporters was detected. PepT1 mRNA was increased 2.3-fold in cancer tissues. Thus, increased PepT1 expression in colonic cancer could contribute to the increased PpIX accumulation observed. Selective inhibition of PAT1 could enhance PpIX loading in tumor tissue relative to that in normal tissue. PMID:19789362
Anderson, Catriona M H; Jevons, Mark; Thangaraju, Muthusamy; Edwards, Noel; Conlon, Nichola J; Woods, Steven; Ganapathy, Vadivel; Thwaites, David T
Synergy and specificity of two Na+-aromatic amino acid symporters in the model alimentary canal of mosquito larvae
SUMMARY The nutrient amino acid transporter (NAT) subfamily is the largest subdivision of the sodium neurotransmitter symporter family (SNF; also known as SLC6; HUGO). There are seven members of the NAT population in the African malaria mosquito Anopheles gambiae, two of which, AgNAT6 and AgNAT8, preferably transport indole- and phenyl-branched substrates, respectively. The relative expression and distribution of these aromatic NATs were examined with transporter-specific antibodies in Xenopus oocytes and mosquito larval alimentary canal, representing heterologous and tissue expression systems, respectively. NAT-specific aromatic-substrate-induced currents strongly corresponded with specific accumulation of both transporters in the plasma membrane of oocytes. Immunolabeling revealed elevated expressions of both transporters in specific regions of the larval alimentary canal, including salivary glands, cardia, gastric caeca, posterior midgut and Malpighian tubules. Differences in relative expression densities and spatial distribution of the transporters were prominent in virtually all of these regions, suggesting unique profiles of the aromatic amino acid absorption. For the first time reversal of the location of a transporter between apical and basal membranes was identified in posterior and anterior epithelial domains corresponding with secretory and absorptive epithelial functions, respectively. Both aromatic NATs formed putative homodimers in the larval gut whereas functional monomers were overexpressed heterologously in Xenopus oocytes. The results unequivocally suggest functional synergy between substrate-specific AgNAT6 and AgNAT8 in intracellular absorption of aromatic amino acids. More broadly, they suggest that the specific selectivity, regional expression and polarized membrane docking of NATs represent key adaptive traits shaping functional patterns of essential amino acid absorption in the metazoan alimentary canal and other tissues.
Okech, Bernard A.; Meleshkevitch, Ella A.; Miller, Melissa M.; Popova, Lyudmila B.; Harvey, William R.; Boudko, Dmitri Y.
Retinoic acid induces sodium/iodide symporter gene expression and radioiodide uptake in the MCF-7 breast cancer cell line
The sodium/iodide symporter (NIS) stimulates iodide uptake in normal lactating breast, but is not known to be active in nonlactating breast or breast cancer. We studied NIS gene regulation and iodide uptake in MCF-7 cells, an estrogen receptor (ER)-positive human breast cancer cell line. All-trans retinoic acid (tRA) treatment stimulated iodide uptake in a time- and dose-dependent fashion up to ?9.4-fold above baseline. Stimulation with selective retinoid compounds indicated that the induction of iodide uptake was mediated by retinoic acid receptor. Treatment with tRA markedly stimulated NIS mRNA and immunoreactive protein (?68 kDa). tRA stimulated NIS gene transcription ?4-fold, as shown by nuclear run-on assay. No induction of iodide uptake was observed with RA treatment of an ER-negative human breast cancer cell line, MDA-MB 231, or a normal human breast cell line, MCF-12A. The iodide efflux rate of tRA-treated MCF-7 cells was slow (t1/2 = 24 min), compared with that in FRTL-5 thyroid cells (t1/2 = 3.9 min), favoring iodide retention in MCF-7 cells. An in vitro clonogenic assay demonstrated selective cytotoxicity with 131I after tRA stimulation of MCF-7 cells. tRA up-regulates NIS gene expression and iodide uptake in an ER-positive breast cancer cell line. Stimulation of radioiodide uptake after systemic retinoid treatment may be useful for diagnosis and treatment of some differentiated breast cancers.
Kogai, Takahiko; Schultz, James J.; Johnson, Laura S.; Huang, Min; Brent, Gregory A.
Crystal structure of a bacterial homologue of the bile acid sodium symporter ASBT
High cholesterol levels greatly increase the risk of cardiovascular disease. About 50 per cent of cholesterol is eliminated from the body by its conversion into bile acids. However, bile acids released from the bile duct are constantly recycled, being reabsorbed in the intestine by the apical sodium-dependent bile acid transporter (ASBT, also known as SLC10A2). It has been shown in
Nien-Jen Hu; So Iwata; Alexander D. Cameron; David Drew
Synchronized fission yeast meiosis using an ATP analog-sensitive Pat1 protein kinase
Synchronous cultures are often indispensable for studying meiosis. Here, we present an optimized protocol for induction of synchronous meiosis in the fission yeast Schizosaccharomyces pombe. Chemical inactivation of an ATP analog-sensitive form of the Pat1 kinase (pat1-as2) by adding the ATP-analog 1-NM-PP1 in G1-arrested cells allows induction of synchronous meiosis at optimal temperature (25 �C). Importantly, this protocol eliminates detrimental effects of elevated temperature (34 �C) which is required to inactivate the commonly used temperature-sensitive Pat1 kinase mutant (pat1-114). Addition of the mat-Pc gene to a mat1-M strain further improves chromosome segregation and spore viability. Thus, our protocol offers highly synchronous meiosis at optimal temperature with most characteristics similar to those of wild-type meiosis. The synchronization protocol can be completed in 5 days.
Cipak, Lubos; Polakova, Silvia; Hyppa, Randy W.; Smith, Gerald R.; Gregan, Juraj
Air transport of plutonium metal : content expansion initiative for the Plutonium Air Transportable (PAT-1) packaging.
The National Nuclear Security Administration (NNSA) has submitted an application to the Nuclear Regulatory Commission (NRC) for the air shipment of plutonium metal within the Plutonium Air Transportable (PAT-1) packaging. The PAT-1 packaging is currently authorized for the air transport of plutonium oxide in solid form only. The INMM presentation will provide a limited overview of the scope of the plutonium metal initiative and provide a status of the NNSA application to the NRC.
Mann, Paul T. (National Nuclear Security Administration); Caviness, Michael L. (Los Alamos National Laboratory); Yoshimura, Richard Hiroyuki
Crystal structure of a bacterial homologue of the bile acid sodium symporter ASBT.
High cholesterol levels greatly increase the risk of cardiovascular disease. About 50 per cent of cholesterol is eliminated from the body by its conversion into bile acids. However, bile acids released from the bile duct are constantly recycled, being reabsorbed in the intestine by the apical sodium-dependent bile acid transporter (ASBT, also known as SLC10A2). It has been shown in animal models that plasma cholesterol levels are considerably lowered by specific inhibitors of ASBT, and ASBT is thus a target for hypercholesterolaemia drugs. Here we report the crystal structure of a bacterial homologue of ASBT from Neisseria meningitidis (ASBT(NM)) at 2.2?�. ASBT(NM) contains two inverted structural repeats of five transmembrane helices. A core domain of six helices harbours two sodium ions, and the remaining four helices pack in a row to form a flat, 'panel'-like domain. Overall, the architecture of the protein is remarkably similar to the sodium/proton antiporter NhaA, despite having no detectable sequence homology. The ASBT(NM) structure was captured with the substrate taurocholate present, bound between the core and panel domains in a large, inward-facing, hydrophobic cavity. Residues near this cavity have been shown to affect the binding of specific inhibitors of human ASBT. The position of the taurocholate molecule, together with the molecular architecture, suggests the rudiments of a possible transport mechanism. PMID:21976025
Hu, Nien-Jen; Iwata, So; Cameron, Alexander D; Drew, David
Pat1 Contains Distinct Functional Domains That Promote P-Body Assembly and Activation of Decapping?
The control of mRNA degradation and translation are important aspects of gene regulation. Recent results suggest that translation repression and mRNA decapping can be intertwined and involve the formation of a quiescent mRNP, which can accumulate in cytoplasmic foci referred to as P bodies. The Pat1 protein is a key component of this complex and an important activator of decapping, yet little is known about its function. In this work, we analyze Pat1 in Saccharomyces cerevisiae function by deletion and functional analyses. Our results identify two primary functional domains in Pat1: one promoting translation repression and P-body assembly and a second domain promoting mRNA decapping after assembly of the mRNA into a P-body mRNP. In addition, we provide evidence that Pat1 binds RNA and has numerous domain-specific interactions with mRNA decapping factors. These results indicate that Pat1 is an RNA binding protein and a multidomain protein that functions at multiple stages in the process of translation repression and mRNA decapping.
Pilkington, Guy R.; Parker, Roy
All-trans-retinoic acid promotes iodine uptake via up- regulating the sodium iodide symporter in medullary thyroid cancer stem cells.
Recently, the main therapy of medullary thyroid cancer (MTC) is surgical, but by which way there is a poor prognosis with a mean survival of only 5 years. In some cases, some researchers found that it is the medullary thyroid cancer stem cells (MTCSCs) that cause metastasis and recurrence. This study aimed to eradicate MTCSCs through administration of all-trans-retinoic acid (ATRA). Here we demonstrate that MTCSCs possess stem- like properties in serum-free medium. The ABCG2, OCT4 and sodium iodide symporter (NIS) were changed by ATRA. Additionally, we found that ATRA can increase the expression of NIS in vivo. All the data suggested that ATRA could increase the iodine uptake of MTCSCs through NIS. PMID:24641421
Tang, Min; Hou, Yan-Li; Kang, Qiang-Qiang; Chen, Xing-Yue; Duan, Li-Qun; Shu, Jin; Li, Shao-Lin; Hu, Xiao-Li; Peng, Zhi-Ping
His-65 in the proton-sucrose symporter is an essential amino acid whose modification with site-directed mutagenesis increases transport activity
The proton�sucrose symporter that mediates phloem loading is a key component of assimilate partitioning in many higher plants. Previous biochemical investigations showed that a diethyl pyrocarbonate-sensitive histidine residue is at or near the substrate-binding site of the symporter. Among the proton�sucrose symporters cloned to date, only the histidine residue at position 65 of AtSUC1 from Arabidopsis thaliana is conserved across species. To test whether His-65 is involved in the transport reaction, we have used site-directed mutagenesis and functional expression in yeast to determine the significance of this residue in the reaction mechanism. Symporters with mutations at His-65 exhibited a range of activities; for example, the H65C mutant resulted in the complete loss of transport capacity, whereas H65Q was almost as active as wild type. Surprisingly, the H65K and H65R symporters transport sucrose at significantly higher rates (increased Vmax) than the wild-type symporter, suggesting His-65 may be associated with a rate-limiting step in the transport reaction. RNA gel blot and protein blot analyses showed that, with the exception of H65C, the variation in transport activity was not because of alterations in steady-state levels of mRNA or symporter protein. Significantly, those symporters with substitutions of His-65 that remained transport competent were no longer sensitive to inactivation by diethyl pyrocarbonate, demonstrating that this is the inhibitor-sensitive histidine residue. Taken together with our previous results, these data show that His-65 is involved in sucrose binding, and increased rates of transport implicate this region of the protein in the transport reaction.
Lu, Jade M.-Y.; Bush, Daniel R.
PAT-1 safety analysis report addendum author responses to request for additional information.
The Plutonium Air Transportable Package, Model PAT-1, is certified under Title 10, Code of Federal Regulations Part 71 by the U.S. Nuclear Regulatory Commission (NRC) per Certificate of Compliance (CoC) USA/0361B(U)F-96 (currently Revision 9). The National Nuclear Security Administration (NNSA) submitted SAND Report SAND2009-5822 to NRC that documented the incorporation of plutonium (Pu) metal as a new payload for the PAT-1 package. NRC responded with a Request for Additional Information (RAI), identifying information needed in connection with its review of the application. The purpose of this SAND report is to provide the authors responses to each RAI. SAND Report SAND2010-6106 containing the proposed changes to the Addendum is provided separately.
Interactions between Upf1 and the Decapping Factors Edc3 and Pat1 in Saccharomyces cerevisiae
In Saccharomyces cerevisiae, mRNA transcripts with premature termination codons are targeted for deadenylation independent decapping and 5? to 3? decay in a quality control pathway termed nonsense-mediated decay (NMD). Critical factors in NMD include Upf1, Upf2, and Upf3, as well as the decapping enzyme, Dcp2/Dcp1. Loss of Upf2 or Upf3 leads to the accumulation of not only Upf1 and Dcp2 in P-bodies, but also of the decapping-activators Pat1, Dhh1, and Lsm1. An interaction between Upf1 and Dcp2 has been identified, which might recruit Dcp2 to the NMD decapping complex. To determine the nature and significance of the Dcp2-Upf1 interaction, we utilized the yeast two-hybrid assay to assess Upf1 interactions with various mRNA decapping factors. We find that although Dcp2 can interact with Upf1, this interaction is indirect and is largely dependent on the Edc3 protein, which interacts with the N-terminal domain of Upf1 at an overlapping, but not identical, site as Upf2. We also found that Pat1 has an independent two-hybrid interaction with the N-terminus of Upf1. Assessment of both reporter and endogenous NMD transcripts suggest that the decapping stimulators, including Edc3 and Pat1, as well as Edc1 and Edc2, are not essential for NMD under normal conditions. This work defines a larger decapping complex involved in NMD, but indicates that components of that complex are not required for general NMD and might either regulate a subset of NMD transcripts or be essential for proper NMD under different environmental conditions.
Swisher, Kylie D.; Parker, Roy
Taurine uptake across the human intestinal brush-border membrane is via two transporters: H+-coupled PAT1 (SLC36A1) and Na+- and Cl(-)-dependent TauT (SLC6A6).
Taurine is an essential amino acid in some mammals and is conditionally essential in humans. Taurine is an abundant component of meat and fish-based foods and has been used as an oral supplement in the treatment of disorders such as cystic fibrosis and hypertension. The purpose of this investigation was to identity the relative contributions of the solute transporters involved in taurine uptake across the luminal membrane of human enterocytes. Distinct transport characteristics were revealed following expression of the candidate solute transporters in Xenopus laevis oocytes: PAT1 (SLC36A1) is a H(+)-coupled, pH-dependent, Na(+)- and Cl(-)-independent, low-affinity, high-capacity transporter for taurine and beta-alanine; TauT (SLC6A6) is a Na(+)- and Cl(-)-dependent, high-affinity, low-capacity transporter of taurine and beta-alanine; ATB(0,+) (SLC6A14) is a Na(+)- and Cl(-)-dependent, high-affinity, low-capacity transporter which accepts beta-alanine but not taurine. Taurine uptake across the brush-border membrane of human intestinal Caco-2 cell monolayers showed characteristics of both PAT1- and TauT-mediated transport. Under physiological conditions, Cl(-)-dependent TauT-mediated uptake predominates at low taurine concentrations, whereas at higher concentrations typical of diet, Cl(-)-independent PAT1-mediated uptake is the major absorptive mechanism. Real-time PCR analysis of human duodenal and ileal biopsy samples demonstrates that PAT1, TauT and ATB(0,+) mRNA are expressed in each tissue but to varying degrees. In conclusion, this study is the first to demonstrate both taurine uptake via PAT1 and functional coexpression of PAT1 and TauT at the apical membrane of the human intestinal epithelium. PAT1 may be responsible for bulk taurine uptake during a meal whereas TauT may be important for taurine supply to the intestinal epithelium and for taurine capture between meals. PMID:19074966
Anderson, Catriona M H; Howard, Alison; Walters, Julian R F; Ganapathy, Vadivel; Thwaites, David T
Analysis of Human Sodium Iodide Symporter Gene Expression in Extrathyroidal Tissues and Cloning of Its Complementary Deoxyribonucleic Acids from Salivary Gland, Mammary Gland, and Gastric Mucosa
The ability to concentrate iodide is a fundamental property of normally functioning thyroid tissue and represents the first step in the production of thyroid hormones. Iodide uptake has been demon- strated in various extrathyroidal tissues, including salivary gland, gastric mucosa, and lactating mammary gland. Recently, cloning and molecular characterization of the human sodium iodide symporter (hNIS) have been reported; however,
C. Spitzweg; W. JOBA; W. EISENMENGER; A. E. HEUFELDER
Arabidopsis Bile Acid:Sodium Symporter Family Protein 5 is Involved in Methionine-Derived Glucosinolate Biosynthesis
Glucosinolates (GSLs) are a group of plant secondary metabolites that have repellent activity against herbivore insects and pathogens, and anti-carcinogenic activity in humans. They are produced in plants of the Brassicaceae and other related families. Biosynthesis of GSLs from precursor amino acids takes place in two subcellular compartments; amino acid biosynthesis and side chain elongation occur mainly in the chloroplast,
Yuji Sawada; Kiminori Toyooka; Ayuko Kuwahara; Akane Sakata; Mutsumi Nagano; Kazuki Saito; Masami Yokota Hirai
The C-Terminal Domain from S. cerevisiae Pat1 Displays Two Conserved Regions Involved in Decapping Factor Recruitment
Eukaryotic mRNA decay is a highly regulated process allowing cells to rapidly modulate protein production in response to internal and environmental cues. Mature translatable eukaryotic mRNAs are protected from fast and uncontrolled degradation in the cytoplasm by two cis-acting stability determinants: a methylguanosine (m7G) cap and a poly(A) tail at their 5? and 3? extremities, respectively. The hydrolysis of the m7G cap structure, known as decapping, is performed by the complex composed of the Dcp2 catalytic subunit and its partner Dcp1. The Dcp1-Dcp2 decapping complex has a low intrinsic activity and requires accessory factors to be fully active. Among these factors, Pat1 is considered to be a central scaffolding protein involved in Dcp2 activation but also in inhibition of translation initiation. Here, we present the structural and functional study of the C-terminal domain from S. cerevisiae Pat1 protein. We have identified two conserved and functionally important regions located at both extremities of the domain. The first region is involved in binding to Lsm1-7 complex. The second patch is specific for fungal proteins and is responsible for Pat1 interaction with Edc3. These observations support the plasticity of the protein interaction network involved in mRNA decay and show that evolution has extended the C-terminal alpha-helical domain from fungal Pat1 proteins to generate a new binding platform for protein partners.
Back, Regis; Keller, Jenny; Charenton, Clement; Taverniti, Valerio; Plesse, Claudine Gaudon; Lazar, Noureddine; Durand, Dominique; van Tilbeurgh, Herman; Seraphin, Bertrand; Graille, Marc
The 3' overhangs at Tetrahymena thermophila telomeres are packaged by four proteins, Pot1a, Tpt1, Pat1, and Pat2.
Although studies with the ciliate Tetrahymena thermophila have played a central role in advancing our understanding of telomere biology and telomerase mechanisms and composition, the full complement of Tetrahymena telomere proteins has not yet been identified. Previously, we demonstrated that in Tetrahymena, the telomeric 3' overhang is protected by a three-protein complex composed of Pot1a, Tpt1, and Pat1. Here we show that Tpt1 and Pat1 associate with a fourth protein, Pat2 (Pot1 associated Tetrahymena 2). Mass spectrometry of proteins copurifying with Pat1 or Tpt1 identified peptides from Pat2, Pot1a, Tpt1, and Pat1. The lack of other proteins copurifying with Pat1 or Tpt1 implies that the overhang is protected by a four-protein Pot1a-Tpt1-Pat1-Pat2 complex. We verified that Pat2 localizes to telomeres, but we were unable to detect direct binding to telomeric DNA. Cells depleted of Pat2 continue to divide, but the telomeres exhibit gradual shortening. The lack of growth arrest indicates that, in contrast to Pot1a and Tpt1, Pat2 is not required for the sequestration of the telomere from the DNA repair machinery. Instead, Pat2 is needed to regulate telomere length, most likely by acting in conjunction with Pat1 to allow telomerase access to the telomere. PMID:24297442
Premkumar, Vidjaya Letchoumy; Cranert, Stacey; Linger, Benjamin R; Morin, Gregg B; Minium, Sasha; Price, Carolyn
About P systems with symport\\/antiport
It is proved that four membranes su-ce to P systems with minimal symport\\/antiport to generate all recursively enumerable sets of numbers. It is also proved that P systems with symport\\/antiport without maximal par- allelism are equivalent to partially blind counter automata.
Pierluigi Frisco
Lactose Permease H+-Lactose Symporter: Mechanical Switch or Brownian Ratchet?
Lactose permease structure is deemed consistent with a mechanical switch device for H+-coupled symport. Because the crystallography-assigned docking position of thiodigalactoside (TDG) does not make close contact with several amino acids essential for symport; the switch model requires allosteric interactions between the proton and sugar binding sites. The docking program, Autodock 3 reveals other lactose-docking sites. An alternative cotransport mechanism is proposed where His-322 imidazolium, positioned in the central pore equidistant (5�7 �) between six charged amino acids, Arg-302 and Lys-319 opposing Glu-269, Glu-325, Asp-237, and Asp-240, transfers a proton transiently to an H-bonded lactose hydroxyl group. Protonated lactose and its dissociation product H3O+ are repelled by reprotonated His-322 and drift in the electrostatic field toward the cytosol. This Brownian ratchet model, unlike the conventional carrier model, accounts for diminished symport by H322N mutant; how H322 mutants become uniporters; why exchanging Lys-319 with Asp-240 paradoxically inactivates symport; how some multiple mutants become revertant transporters; the raised export rate and affinity toward lactose of uncoupled mutants; the altered specificity toward lactose, melibiose, and galactose of some mutants, and the proton dissociation rate of H322 being 100-fold faster than the symport turnover rate.
Naftalin, Richard J.; Green, Nicholas; Cunningham, Philip
Parsing apical oxalate exchange in Caco-2BBe1 monolayers: siRNA knockdown of SLC26A6 reveals the role and properties of PAT-1
The purpose of this investigation was to quantitate the contribution of the anion exchanger PAT-1 (putative anion transporter-1), encoded by SLC26A6, to oxalate transport in a model intestinal epithelium and to discern some characteristics of this exchanger expressed in its native environment. Control (Con) Caco-2 BBe1 monolayers, 6�8 days postseeding, were compared with those transfected with a small interfering RNA targeted to SLC26A6 (A6KD). Radiotracer and Ussing chamber techniques were used to determine the transepithelial unidirectional fluxes of Ox2?, Cl?, and SO42? whereas fluorometric/BCECF measurements of intracellular pH were used to assess HCO3? exchange. PAT-1 was functionally targeted to the apical membrane, and SLC26A6 knockdown reduced PAT-1 protein (>60%) and mRNA (>75%) expression in A6KD. No net flux of Ox2?, Cl?, or SO42? was detected in Con or A6KD monolayers, yet the unidirectional fluxes in A6KD were reduced 50, 35, and 15%, respectively. Cl?-dependent HCO3? efflux from A6KD was reduced 50% compared with Con. The difference between Con and A6KD properties represents that mediated solely by PAT-1, and by this approach we found that PAT-1-mediated oxalate influx and efflux are inhibited equally by mucosal DIDS (EC50 ?5 ?M) and that mucosal Cl? inhibits oxalate uptake with an EC50 < 20 mM. Transepithelial Cl? gradients supported large, DIDS-sensitive net absorptive or secretory fluxes of oxalate in a direction opposite that of the imposed Cl? gradient. The overall symmetry of PAT-1-mediated oxalate exchange suggests that vectorial oxalate transport observed in vivo is principally dependent on the magnitude and direction of counterion gradients.
Morozumi, Makoto; Hatch, Marguerite
Chloride binding site of neurotransmitter sodium symporters
Neurotransmitter:sodium symporters (NSSs) play a critical role in signaling by reuptake of neurotransmitters. Eukaryotic NSSs are chloride-dependent, whereas prokaryotic NSS homologs like LeuT are chloride-independent but contain an acidic residue (Glu290 in LeuT) at a site where eukaryotic NSSs have a serine. The LeuT-E290S mutant displays chloride-dependent activity. We show that, in LeuT-E290S cocrystallized with bromide or chloride, the anion is coordinated by side chain hydroxyls from Tyr47, Ser290, and Thr254 and the side chain amide of Gln250. The bound anion and the nearby sodium ion in the Na1 site organize a connection between their coordinating residues and the extracellular gate of LeuT through a continuous H-bond network. The specific insights from the structures, combined with results from substrate binding studies and molecular dynamics simulations, reveal an anion-dependent occlusion mechanism for NSS and shed light on the functional role of chloride binding.
Kantcheva, Adriana K.; Quick, Matthias; Shi, Lei; Winther, Anne-Marie Lund; Stolzenberg, Sebastian; Weinstein, Harel; Javitch, Jonathan A.; Nissen, Poul
Activation of Hormone-sensitive Lipase Requires Two Steps, Protein Phosphorylation and Binding to the PAT-1 Domain of Lipid Droplet Coat Proteins*
Lipolysis is an important metabolic pathway controlling energy homeostasis through degradation of triglycerides stored in lipid droplets and release of fatty acids. Lipid droplets of mammalian cells are coated with one or more members of the PAT protein family, which serve important functions in regulating lipolysis. In this study, we investigate the mechanisms by which PAT family members, perilipin A, adipose differentiation-related protein (ADFP), and LSDP5, control lipolysis catalyzed by hormone-sensitive lipase (HSL), a major lipase in adipocytes and several non-adipose cells. We applied fluorescence microscopic tools to analyze proteins in situ in cultured Chinese hamster ovary cells using fluorescence recovery after photobleaching and anisotropy Forster resonance energy transfer. Fluorescence recovery after photobleaching data show that ADFP and LSDP5 exchange between lipid droplet and cytoplasmic pools, whereas perilipin A does not. Differences in protein mobility do not correlate with PAT protein-mediated control of lipolysis catalyzed by HSL or endogenous lipases. Forster resonance energy transfer and co-immunoprecipitation experiments reveal that each of the three PAT proteins bind HSL through interaction of the lipase with amino acids within the highly conserved amino-terminal PAT-1 domain. ADFP and LSDP5 bind HSL under basal conditions, whereas phosphorylation of serine residues within three amino-terminal protein kinase A consensus sequences of perilipin A is required for HSL binding and maximal lipolysis. Finally, protein kinase A-mediated phosphorylation of HSL increases lipolysis in cells expressing ADFP or LSDP5; in contrast, phosphorylation of perilipin A exerts the major control over HSL-mediated lipolysis when perilipin is the main lipid droplet protein.
Wang, Hong; Hu, Liping; Dalen, Knut; Dorward, Heidi; Marcinkiewicz, Amy; Russell, Deanna; Gong, Dawei; Londos, Constantine; Yamaguchi, Tomohiro; Holm, Cecilia; Rizzo, Mark A.; Brasaemle, Dawn; Sztalryd, Carole
Substrate-Na{sup +} complex formation: Coupling mechanism for {gamma}-aminobutyrate symporters
Crystal structures of transmembrane transport proteins belonging to the important families of neurotransmitter-sodium symporters reveal how they transport neurotransmitters across membranes. Substrate-induced structural conformations of gated neurotransmitter-sodium symporters have been in the focus of research, however, a key question concerning the mechanism of Na{sup +} ion coupling remained unanswered. Homology models of human glial transporter subtypes of the major inhibitory neurotransmitter {gamma}-aminobutyric acid were built. In accordance with selectivity data for subtype 2 vs. 3, docking and molecular dynamics calculations suggest similar orthosteric substrate (inhibitor) conformations and binding crevices but distinguishable allosteric Zn{sup 2+} ion binding motifs. Considering the occluded conformational states of glial human {gamma}-aminobutyric acid transporter subtypes, we found major semi-extended and minor ring-like conformations of zwitterionic {gamma}-aminobutyric acid in complex with Na{sup +} ion. The existence of the minor ring-like conformation of {gamma}-aminobutyric acid in complex with Na{sup +} ion may be attributed to the strengthening of the intramolecular H-bond by the electrostatic effect of Na{sup +} ion. Coupling substrate uptake into cells with the thermodynamically favorable Na{sup +} ion movement through substrate-Na{sup +} ion complex formation may be a mechanistic principle featuring transmembrane neurotransmitter-sodium symporter proteins.
Pallo, Anna; Simon, Agnes [Department of Neurochemistry, Institute of Biomolecular Chemistry, Chemical Research Center, Hungarian Academy of Sciences (Hungary)] [Department of Neurochemistry, Institute of Biomolecular Chemistry, Chemical Research Center, Hungarian Academy of Sciences (Hungary); Bencsura, Akos [Department of Theoretical Chemistry, Institute of Structural Chemistry, Chemical Research Center, Hungarian Academy of Sciences, Budapest (Hungary)] [Department of Theoretical Chemistry, Institute of Structural Chemistry, Chemical Research Center, Hungarian Academy of Sciences, Budapest (Hungary); Heja, Laszlo [Department of Neurochemistry, Institute of Biomolecular Chemistry, Chemical Research Center, Hungarian Academy of Sciences (Hungary)] [Department of Neurochemistry, Institute of Biomolecular Chemistry, Chemical Research Center, Hungarian Academy of Sciences (Hungary); Kardos, Julianna, E-mail: firstname.lastname@example.com [Department of Neurochemistry, Institute of Biomolecular Chemistry, Chemical Research Center, Hungarian Academy of Sciences (Hungary)] [Department of Neurochemistry, Institute of Biomolecular Chemistry, Chemical Research Center, Hungarian Academy of Sciences (Hungary)
The importance of being aromatic: ? interactions in sodium symporters.
In the LeuT family of sodium solute symporters, 13-17% of the residues in transmembrane domains are aromatic. The unique properties of aromatic amino acids allow them to play specialized roles in proteins, but their function in membrane transporters is underappreciated. Here we analyze the ? bonding pattern in the LeuT (5TMIR) family and then describe the role of a triad of aromatic residues in sodium-dependent sugar cotransporters (SGLTs). In SLC5 symporters, three aromatic residues in TM6 (SGLT1 W289, Y290, and W291) are conserved in only those transporting sugars and inositols. We used biophysical analysis of mutants to discover their functional roles, which we have interpreted in terms of CH-?, ?-?, and cation-? bonding. We discovered that (1) glucose binding involves CH-? stacking with Y290, (2) ? T-stacking interactions between Y290 and W291 and H-bonding between Y290 and N78 (TM1) are essential to form the sodium and sugar binding sites, (3) the Na(+):sugar stoichiometry is determined by these residues, and (4) W289 may be important in stabilizing the structure through H-bonding to TM3. We also find that the WYW triad plays a role in Na(+) coordination at the Na1 site, possibly through cation-? interactions. Surprisingly, this Na(+) is not necessarily coupled to glucose translocation. Our analysis of ? interactions in other LeuT proteins suggests that they also contribute to the structure and function in this whole family of transporters. PMID:23116249
Jiang, Xuan; Loo, Donald D F; Hirayama, Bruce A; Wright, Ernest M
XYLH encodes a xylose/H+ symporter from the highly related yeast species Debaryomyces fabryi and Debaryomyces hansenii.
The closely related yeasts Debaryomyces fabryi and Debaryomyces hansenii are excellent xylose consumers. We previously described the activity of a high-affinity xylose/H(+) symport from an industrial strain of D.�hansenii subsequently reclassified as D.�fabryi. We now report the identification of the gene encoding this permease, AY347871.2. This was retrieved from D.�fabryi gDNA using a degenerate primer PCR strategy, based on conserved regions from the amino acid sequences of three well-characterized bacterial xylose/H(+) symporters. This sequence is 86% identical to another, DEHA2C11374p from D.�hansenii type strain. DEHA2C11374p was conceptually ascribed to the major facilitator superfamily. The putative amino acid sequence of AY347871.2 and DEHA2C11374p presented a hydrophobicity pattern compatible with plasma membrane proteins. The last was functionally expressed in Saccharomyces cerevisiae. The sensitivity of transport activity to a protonophore confirmed its dependence on proton motive force, as expected from a symporter. We named D.�fabryi AY347871.2 and D.�hansenii DEHA2C11374p as XYLH from Xylose/H(+) symport. Based on the very high similarity, we suggested that Scheffersomyces stipitis Xut3 and Aspergillus nidulans AN8400.2 may also encode xylose high-affinity permeases. PMID:23809840
Ferreira, Danielly; Nobre, Alexandra; Silva, Marta Luisa; Faria-Oliveira, F�bio; Tulha, Joana; Ferreira, C�lia; Lucas, C�ndida
The decapping activator Lsm1p-7p-Pat1p complex has the intrinsic ability to distinguish between oligoadenylated and polyadenylated RNAs
Decapping is a critical step in mRNA decay. In the 5?-to-3? mRNA decay pathway conserved in all eukaryotes, decay is initiated by poly(A) shortening, and oligoadenylated mRNAs (but not polyadenylated mRNAs) are selectively decapped allowing their subsequent degradation by 5? to 3? exonucleolysis. The highly conserved heptameric Lsm1p-7p complex (made up of the seven Sm-like proteins, Lsm1p�Lsm7p) and its interacting partner Pat1p activate decapping by an unknown mechanism and localize with other decapping factors to the P-bodies in the cytoplasm. The Lsm1p-7p�Pat1p complex also protects the 3?-ends of mRNAs in vivo from trimming, presumably by binding to the 3?-ends. In order to determine the intrinsic RNA-binding properties of this complex, we have purified it from yeast and carried out in vitro analyses. Our studies revealed that it directly binds RNA at/near the 3?-end. Importantly, it possesses the intrinsic ability to distinguish between oligoadenylated and polyadenylated RNAs such that the former are bound with much higher affinity than the latter. These results indicate that the intrinsic RNA-binding characteristics of this complex form a critical determinant of its in vivo interactions and functions.
Chowdhury, Ashis; Mukhopadhyay, Jaba; Tharun, Sundaresan
Structure-based mechanism for Na+/melibiose symport by MelB
The bacterial melibiose permease (MelB) belongs to the glycoside�pentoside�hexuronide:cation symporter family, a part of the major facilitator superfamily (MFS). Structural information regarding glycoside�pentoside�hexuronide:cation symporter family transporters and other Na+-coupled permeases within MFS has been lacking, although a wealth of biochemical and biophysical data are available. Here we present the three-dimensional crystal structures of Salmonella typhimurium MelBSt in two conformations, representing an outward partially occluded and an outward inactive state of MelBSt. MelB adopts a typical MFS fold and contains a previously unidentified cation-binding motif. Three conserved acidic residues form a pyramidal-shaped cation-binding site for Na+, Li+ or H+, which is in close proximity to the sugar-binding site. Both cosubstrate-binding sites are mainly contributed by the residues from the amino-terminal domain. These two structures and the functional data presented here provide mechanistic insights into Na+/melibiose symport. We also postulate a structural foundation for the conformational cycling necessary for transport catalysed by MFS permeases in general.
Ethayathulla, Abdul S.; Yousef, Mohammad S.; Amin, Anowarul; Leblanc, Gerard; Kaback, H. Ronald; Guan, Lan
Effect of Ligands of Nuclear Hormone Receptors on Sodium\\/Iodide Symporter Expression and Activity in Breast Cancer Cells
Iodide uptake by normal and cancerous thyroid cells is an active process mediated by the sodium\\/iodide symporter (NIS). Using quantitative real-time RT-PCR, we found that all 22 fresh human breast cancer samples had very low NIS expression similar to levels in untreated MCF-7 breast cancer cells. 9-cis retinoic acid (9-cis RA), a ligand for both retinoic acid receptor (RAR)\\/retinoic X
Sakae Tanosaki; Takayuki Ikezoe; Anthony Heaney; Jonathan W. Said; Kazuo Dan; Makoto Akashi; H. Phillip Koeffler
Substrate�Na + complex formation: Coupling mechanism for ?-aminobutyrate symporters
Crystal structures of transmembrane transport proteins belonging to the important families of neurotransmitter-sodium symporters reveal how they transport neurotransmitters across membranes. Substrate-induced structural conformations of gated neurotransmitter-sodium symporters have been in the focus of research, however, a key question concerning the mechanism of Na+ ion coupling remained unanswered. Homology models of human glial transporter subtypes of the major inhibitory neurotransmitter
Anna Pall�; �gnes Simon; �kos Bencsura; L�szl� H�ja; Julianna Kardos
Expression of the sodium iodide symporter in human kidney
Expression of the sodium iodide symporter in human kidney.BackgroundThe human sodium iodide symporter (hNIS) is a transmembrane protein that mediates the active transport of iodide in the thyroid gland. Following cloning of NIS, NIS expression has been detected in a broad range of nonthyroidal tissues, suggesting that iodide transport in these tissues is conferred by the expression of functional NIS
Christine Spitzweg; Charyl M Dutton; Maria R Castro; Elizabeth R Bergert; John R Goellner; Armin E Heufelder; John C Morris
Differential Regulation of Sodium\\/Iodide Symporter Gene Expression by Nuclear Receptor Ligands in MCF7 Breast Cancer Cells
The sodium\\/iodide symporter (NIS) mediates iodide uptake in lactating breast tissue and is expressed in some breast can- cers. We have previously demonstrated that all-trans retinoic acid (tRA) stimulates NIS gene expression and the selective cytotoxic effect of -emitting radioiodide-131 (131I) in both in vitro and in vivo MCF-7 breast cancer cell systems. We studied the ability of natural and
Takahiko Kogai; Yoko Kanamoto; Andrew I. Li; Lisa H. Che; Emi Ohashi; Katsumi Taki; Roshantha A. Chandraratna; Tsukasa Saito; Gregory A. Brent
Anionic selectivity sequence of the Cl(-)-H+ symporter in the synaptosomal preparation from rat brain cortex.
The Na(+)/H(+) exchanger has been the only unequivocally demonstrated H(+)-transport mechanism in the synaptosomal preparation. We had previously suggested that a Cl(-)-H(+) symporter (in its acidifying mode) is involved in cytosolic pH regulation in the synaptosomal preparation. Supporting this suggestion, we now show that: (1) when synaptosomes are transferred from PSS to either gluconate or sulfate solutions, the Fura-2 ratio remains stable instead of increasing as it does in 50 mM K solution. This indicates that these anions do not promote a plasma membrane depolarization. (2) Based in the recovery rate from the cytosolic alkalinization, the anionic selectivity of the Cl(-)-H(+) symporter is NO(3)(-) > Br(-) > Cl(-) > I(-) = isethionate = sulfate = methanesulfonate = gluconate. (3) PCMB 10 muM inhibits the gluconate-dependent alkalinization by 30 +/- 6%. (4) Neither Niflumic acid, 9AC, Bumetanide nor CCCP inhibits the recovery from the cytosolic alkalinization. PMID:18404376
Torres, M L; Ortega, F; Cuaranta, I; Gonz�lez, J; Sanchez-Armass, S
P systems with symport\\/antiport simulating counter automata
The generative capability of several variants of P systems with symport\\/antiport is studied via the simulation of counter automata. This leads to the reduction of the complexity, expressed in number of membranes and weight of rules, of P systems generating recursively enu- merable sets.
Pierluigi Frisco; Hendrik Jan Hoogeboom
Simulating Counter Automata by P Systems with Symport\\/Antiport
The complexity, expressed in number of membranes and weight of rules, of P systems with symport\\/antiport generating recur- sively enumerable sets is reduced if counter automata instead of matrix grammars are simulated. We consider both subsets of N obtained by counting objects in a designated membrane, and string languages ob- tained by following the traces of a designated object.
Unexpected universality results for three classes of P systems with symport\\/antiport
Symport and antiport are biological ways of transporting molecules through membranesin ``collaborating'' pairs; in the case\\u000a of symport the two molecules pass in the same direction, in the case of antiport the two molecules pass in opposite directions.\\u000a Here we first survey the results about the computing power of membrane systems (P systems) using only symport\\/antiport rules\\u000a (hence these systems
Mihai Ionescu; Carlos Mart�n-vide; Andrei Paun; Gheorghe Paun
Unexpected Universality Results for Three Classes of P Systems with Symport\\/Antiport
Symport and antiport are biological ways of transporting molecules through membranes in �collaborating� pairs; in the case\\u000a of symport the two molecules pass in the same direction, in the case of antiport the two molecules pass in opposite directions.\\u000a Here we first survey the results about the computing power of membrane systems (P systems) using only symport\\/antiport rules\\u000a (hence these
Breast cancer brain metastases express the sodium iodide symporter
Breast cancer brain metastases are on the rise and their treatment is hampered by the limited entry and efficacy of anticancer\\u000a drugs in this sanctuary. The sodium iodide symporter, NIS, actively transports iodide across the plasma membrane and is exploited\\u000a clinically to deliver radioactive iodide into cells. As in thyroid cancers, NIS is expressed in many breast cancers including\\u000a primary
Corinne Renier; Hannes Vogel; Onyinye Offor; Chen Yao; Irene Wapnir
The sodium\\/substrate symporter family: structural and functional features
Members of the sodium\\/substrate symporter family (SSSF, TC 2.A.21) catalyze the uptake of a wide variety of solutes including sugars, proline, pantothenate, and iodide into cells of pro- and eukaryotic origin. Extensive analyses of the topology of different SSSF proteins suggest an arrangement of 13 transmembrane domains as a common topological motif. Regions involved in sodium and\\/or substrate binding were
Heinrich Jung
The sodium iodide symporter: its emerging relevance to clinical thyroidology
The sodium iodide symporter (NIS), an integral basolateral cell membrane protein of thyroid follicular cells, is responsible for active accumulation of iodide within the thyroid gland, a critical step in the biosynthesis of thyroid hormones. After cloning of rat NIS cDNA and of its human counterpart (hNIS), molecular characterization of the human NIS gene has revealed that the hNIS coding
Christine Spitzweg; Armin E Heufelder
Sodium Iodide Symporter and the Radioiodine Treatment of Thyroid Carcinoma
Since the specific accumulation of iodide in thyroid was found in 1915, radioiodine has been widely applied to diagnose and\\u000a treat thyroid cancer. Iodide uptake occurs across the membrane of the thyroid follicular cells and cancer cells through an\\u000a active transporter process mediated by the sodium iodide symporter (NIS). The NIS coding genes were cloned and identified\\u000a from rat and
June-Key Chung; Hye Won Youn; Joo Hyun Kang; Ho Young Lee; Keon Wook Kang
The sodium/substrate symporter family: structural and functional features.
Members of the sodium/substrate symporter family (SSSF, TC 2.A.21) catalyze the uptake of a wide variety of solutes including sugars, proline, pantothenate, and iodide into cells of pro- and eukaryotic origin. Extensive analyses of the topology of different SSSF proteins suggest an arrangement of 13 transmembrane domains as a common topological motif. Regions involved in sodium and/or substrate binding were identified. Furthermore, protein chemical and spectroscopic studies reveal ligand-induced structural alterations which are consistent with close interactions between the sites of sodium and substrate binding, thereby supporting an ordered binding mechanism for transport. PMID:12354616
Jung, Heinrich
Ontogenic changes in human placental sodium iodide symporter expression.
The human fetus requires a maternal supply of iodide to synthesize thyroid hormone from 16 weeks gestation. Placental iodide transport is regulated by the sodium iodide symporter (NIS). We studied the ontogeny of NIS in placentas from surgically terminated pregnancies and from normal term pregnancies. NIS mRNA was low at 6 weeks gestation and peaked at 12 weeks gestation. Placental NIS protein levels are significantly correlated with gestational age during early pregnancy and increase with increased placental vascularization. This would lead to increased iodide supply to meet increased fetal requirements for thyroid hormone synthesis as the pregnancy progresses. PMID:22954554
Li, H; Patel, J; Mortimer, R H; Richard, K
The major amino acid transporter superfamily has a similar core structure as Na+-galactose and Na+-leucine transporters
The sodium solute symporters (SSS) and neurotransmitter sodium symporters (NSS) are two families of secondary transporters that are not related in amino acid sequence. Nonetheless, recent crystal structures showed that the Na+\\/galactose (SSS) and Na+\\/leucine (NSS) transporters have similar core structures. The structural relatedness highlights the need for classification methods for membrane protein structures based on other criteria than amino
Juke S. Lolkema; Dirk-Jan Slotboom
Sodium Solute Symporter and Cadherin Proteins Act as Bacillus thuringiensis Cry3Ba Toxin Functional Receptors in Tribolium castaneum*
Understanding how Bacillus thuringiensis (Bt) toxins interact with proteins in the midgut of susceptible coleopteran insects is crucial to fully explain the molecular bases of Bt specificity and insecticidal action. In this work, aminopeptidase N (TcAPN-I), E-cadherin (TcCad1), and sodium solute symporter (TcSSS) have been identified by ligand blot as putative Cry3Ba toxin-binding proteins in Tribolium castaneum (Tc) larvae. RNA interference knockdown of TcCad1 or TcSSS proteins resulted in decreased susceptibility to Cry3Ba toxin, demonstrating the Cry toxin receptor functionality for these proteins. In contrast, TcAPN-I silencing had no effect on Cry3Ba larval toxicity, suggesting that this protein is not relevant in the Cry3Ba toxin mode of action in Tc. Remarkable features of TcSSS protein were the presence of cadherin repeats in its amino acid sequence and that a TcSSS peptide fragment containing a sequence homologous to a binding epitope found in Manduca sexta and Tenebrio molitor Bt cadherin functional receptors enhanced Cry3Ba toxicity. This is the first time that the involvement of a sodium solute symporter protein as a Bt functional receptor has been demonstrated. The role of this novel receptor in Bt toxicity against coleopteran insects together with the lack of receptor functionality of aminopeptidase N proteins might account for some of the differences in toxin specificity between Lepidoptera and Coleoptera insect orders.
Contreras, Estefania; Schoppmeier, Michael; Real, M. Dolores; Rausell, Carolina
Crystal structure of a glucose/H+ symporter and its mechanism of action.
Glucose transporters are required to bring glucose into cells, where it is an essential energy source and precursor in protein and lipid synthesis. These transporters are involved in important common diseases such as cancer and diabetes. Here, we report the crystal structure of the Staphylococcus epidermidis glucose/H(+) symporter in an inward-facing conformation at 3.2-� resolution. The Staphylococcus epidermidis glucose/H(+) symporter is homologous to human glucose transporters, is very specific and has high avidity for glucose, and is inhibited by the human glucose transport inhibitors cytochalasin B, phloretin, and forskolin. On the basis of the crystal structure in conjunction with mutagenesis and functional studies, we propose a mechanism for glucose/H(+) symport and discuss the symport mechanism versus facilitated diffusion. PMID:24127585
Iancu, Cristina V; Zamoon, Jamillah; Woo, Sang Bum; Aleshin, Alexander; Choe, Jun-yong
Sodium Iodide Symporter Gene Transfer for Imaging and Ablation of Prostate Cancer.
National Technical Information Service (NTIS)
The sodium iodide symporter (NIS) mediates iodide uptake in thyroid follicular cells and provides a mechanism for effective radioiodide treatment of residual, recurrent, and metastatic thyroid cancers. The objective of the proposed research is to test the...
S. M. Jhiang
Modified Sodium Iodide Symporter Proteins and Genes for Imaging and Cancer Therapy.
The invention provides modified sodium iodide symporter (NIS) proteins, and polynucleotides encoding modified NIS proteins. The modified NIS proteins have a net electrostatic charge more positive than that of corresponding wild-type NIS proteins. Expressi...
D. H. Shen S. M. Jhiang X. Lin
Crystal structure of a glucose/H+ symporter and its mechanism of action
Glucose transporters are required to bring glucose into cells, where it is an essential energy source and precursor in protein and lipid synthesis. These transporters are involved in important common diseases such as cancer and diabetes. Here, we report the crystal structure of the Staphylococcus epidermidis glucose/H+ symporter in an inward-facing conformation at 3.2-� resolution. The Staphylococcus epidermidis glucose/H+ symporter is homologous to human glucose transporters, is very specific and has high avidity for glucose, and is inhibited by the human glucose transport inhibitors cytochalasin B, phloretin, and forskolin. On the basis of the crystal structure in conjunction with mutagenesis and functional studies, we propose a mechanism for glucose/H+ symport and discuss the symport mechanism versus facilitated diffusion.
Iancu, Cristina V.; Zamoon, Jamillah; Woo, Sang Bum; Aleshin, Alexander; Choe, Jun-yong
The Crystal Structure of a Sodium Galactose Transporter Reveals Mechanistic Insights into Na+\\/Sugar Symport
Membrane transporters that use energy stored in sodium gradients to drive nutrients into cells constitute a major class of proteins. We report the crystal structure of a member of the solute sodium symporters (SSS), the Vibrio parahaemolyticus sodium\\/galactose symporter (vSGLT). The ~3.0 angstrom structure contains 14 transmembrane (TM) helices in an inward-facing conformation with a core structure of inverted repeats
Salem Faham; Akira Watanabe; Gabriel Mercado Besserer; Duilio Cascio; Alexandre Specht; Bruce A. Hirayama; Ernest M. Wright; Jeff Abramson
Radioisotope Concentrator Gene Therapy Using the Sodium\\/Iodide Symporter Gene
We demonstrate a novel method of concentrating radiation for tumor imaging or killing. The rat sodium\\/iodide symporter gene (rNIS) was cloned into a retroviral vector for transfer into cancer cells to mimic the iodide uptake of thyroid follicular cells. In vitro iodide transport shows that the symporter functions similarly in rNIS-transduced tumor cells and rat thyroid follicular cells. rNIS-transduced and
Robert B. Mandell; Leisa Z. Mandell; Charles J. Link
Identification of in vivo phosphorylation sites and their functional significance in the sodium iodide symporter.
The Na+/I- symporter (NIS)-mediated iodide uptake activity is the basis for targeted radioiodide ablation of thyroid cancers. Although it has been shown that NIS protein is phosphorylated, neither the in vivo phosphorylation sites nor their functional significance has been reported. In this study, Ser-43, Thr-49, Ser-227, Thr-577, and Ser-581 were identified as in vivo NIS phosphorylation sites by mass spectrometry. Kinetic analysis of NIS mutants of the corresponding phosphorylated amino acid residue indicated that the velocity of iodide transport of NIS is modulated by the phosphorylation status of Ser-43 and Ser-581. We also found that the phosphorylation status of Thr-577 may be important for NIS protein stability and that the phosphorylation status of Ser-227 is functionally silent. Thr-49 appears to be critical for proper local structure/conformation of NIS because mutation of Thr-49 to alanine, aspartic acid, or serine results in reduced NIS activity without alterations in total or cell surface NIS protein levels. Taken together, we showed that NIS protein levels and functional activity could be modulated by phosphorylation through distinct mechanisms. PMID:17913707
Vadysirisack, Douangsone D; Chen, Eric S-W; Zhang, Zhaoxia; Tsai, Ming-Daw; Chang, Geen-Dong; Jhiang, Sissy M
Intestinal absorption of the antiepileptic drug substance vigabatrin in G�ttingen mini-pigs is unaffected by co-administration of amino acids.
The anti-epileptic drug substance vigabatrin is used against infantile spasms. In vitro evidence suggests that vigabatrin is transported via the proton coupled amino acid transporter (PAT1). The aim of the present study was to investigate whether the intestinal absorption of vigabatrin in vivo was mediated via PAT1 in non-rodents. This was investigated by oral co-administration of vigabatrin and PAT1-ligands to G�ttingen mini-pigs. Vigabatrin had an oral absorption fraction (Fabs) of 75-80%, and the maximal plasma concentration (Cmax) was reached within 0.5-1.0 h (tmax). Co-administration of vigabatrin and amino acids generally did not significantly affect Fa, Tmax or Cmax. However, co-administration with sarcosine prolonged the time to reach Cmax. After co-administration with amino acids, vigabatrin absorption showed a slightly lowered onset. This may indicate an effect of amino acids on either the rate of gastric emptying or an effect directly on the absorption of vigabatrin, possibly via inhibition of PAT1 or another drug transporter. In conclusion, co-administration of PAT1-ligands together with vigabatrin did not significantly alter the pharmacokinetic profile of vigabatrin. PMID:24607206
N�hr, Martha Kampp; Holm, Ren�; Thale, Zia Irene; Nielsen, Carsten Uhd
The Sodium Iodide Symporter (NIS): Regulation and Approaches to Targeting for Cancer Therapeutics
Expression of the sodium iodide symporter (NIS) is required for efficient iodide uptake in thyroid and lactating breast. Since most differentiated thyroid cancer expresses NIS, ?-emitting radioactive iodide is routinely utilized to target remnant thyroid cancer and metastasis after total thyroidectomy. Stimulation of NIS expression by high levels of thyroid-stimulating hormone is necessary to achieve radioiodide uptake into thyroid cancer that is sufficient for therapy. The majority of breast cancer also expresses NIS, but at a low level insufficient for radioiodine therapy. Retinoic acid is a potent NIS inducer in some breast cancer cells. NIS is also modestly expressed in some non-thyroidal tissues, including salivary glands, lacrimal glands and stomach. Selective induction of iodide uptake is required to target tumors with radioiodide. Iodide uptake in mammalian cells is dependent on the level of NIS gene expression, but also successful translocation of NIS to the cell membrane and correct insertion. The regulatory mechanisms of NIS expression and membrane insertion are regulated by signal transduction pathways that differ by tissue. Differential regulation of NIS confers selective induction of functional NIS in thyroid cancer cells, as well as some breast cancer cells, leading to more efficient radioiodide therapy for thyroid cancer and a new strategy for breast cancer therapy. The potential for systemic radioiodide treatment of a range of other cancers, that do not express endogenous NIS, has been demonstrated in models with tumor-selective introduction of exogenous NIS.
Kogai, Takahiko; Brent, Gregory A.
Phosphate-responsive promoter of a Pichia pastoris sodium phosphate symporter.
To develop a functional phosphate-regulated promoter in Pichia pastoris, a phosphate-responsive gene, PHO89, which encodes a putative sodium (Na(+))-coupled phosphate symporter, was isolated. Sequencing analyses revealed a 1,731-bp open reading frame encoding a 576-amino-acid polypeptide with 12 putative transmembrane domains. The properties of the PHO89 promoter (P(PHO89)) were investigated using a bacterial lipase gene as a reporter in 5-liter jar fermentation experiments. P(PHO89) was tightly regulated by phosphate and was highly activated when the cells were grown in a phosphate-limited external environment. Compared to translation elongation factor 1alpha and the glyceraldehyde-3-phosphate dehydrogenase promoter, P(PHO89) exhibited strong transcriptional activity with higher specific productivity (amount of lipase produced/cell/h). Furthermore, a cost-effective and simple P(PHO89)-based fermentation process was developed for industrial application. These results demonstrate the potential for efficient use of P(PHO89) for controlled production of recombinant proteins in P. pastoris. PMID:19329662
Ahn, Jungoh; Hong, Jiyeon; Park, Myongsoo; Lee, Hyeokweon; Lee, Eungyo; Kim, Chunsuk; Lee, Joohwan; Choi, Eui-sung; Jung, Joon-ki; Lee, Hongweon
The sodium iodide symporter (NIS): regulation and approaches to targeting for cancer therapeutics.
Expression of the sodium iodide symporter (NIS) is required for efficient iodide uptake in thyroid and lactating breast. Since most differentiated thyroid cancer expresses NIS, ?-emitting radioactive iodide is routinely utilized to target remnant thyroid cancer and metastasis after total thyroidectomy. Stimulation of NIS expression by high levels of thyroid-stimulating hormone is necessary to achieve radioiodide uptake into thyroid cancer that is sufficient for therapy. The majority of breast cancer also expresses NIS, but at a low level insufficient for radioiodine therapy. Retinoic acid is a potent NIS inducer in some breast cancer cells. NIS is also modestly expressed in some non-thyroidal tissues, including salivary glands, lacrimal glands and stomach. Selective induction of iodide uptake is required to target tumors with radioiodide. Iodide uptake in mammalian cells is dependent on the level of NIS gene expression, but also successful translocation of NIS to the cell membrane and correct insertion. The regulatory mechanisms of NIS expression and membrane insertion are regulated by signal transduction pathways that differ by tissue. Differential regulation of NIS confers selective induction of functional NIS in thyroid cancer cells, as well as some breast cancer cells, leading to more efficient radioiodide therapy for thyroid cancer and a new strategy for breast cancer therapy. The potential for systemic radioiodide treatment of a range of other cancers, that do not express endogenous NIS, has been demonstrated in models with tumor-selective introduction of exogenous NIS. PMID:22750642
Kogai, Takahiko; Brent, Gregory A
Synthetic K(+)/Cl(-)-Selective Symporter across a Phospholipid Membrane.
Synthetic molecules which selectively transport sodium or potassium chloride across a lipid membrane have been prepared. The salt carriers consist of two heteroditopic binding sites, an anion-binding cavity with three hydrogen bond donors and an azacrown ether for binding an alkali metal cation. The association constants between the carriers and chloride ion have been enhanced by 1 order of the magnitude in the presence of sodium or potassium ion in 10% (v/v) CD3OH/CD3CN, due to the formation of a contact ion-pair between the bound cation and chloride as demonstrated by the single-crystal X-ray structure of a sodium chloride complex. A series of transport experiments have demonstrated that the synthetic molecule functions as a mobile carrier of transporting salts via M(+)/Cl(-) symport. Among alkali metal chlorides, the carrier with an 18-azacrown-6 exhibits a strong selectivity toward potassium chloride, while the carrier with a 15-azacrown-5 displays a moderate selectivity for sodium chloride. PMID:24971563
Lee, Jung Ha; Lee, Ji Hyun; Choi, Ye Rin; Kang, Philjae; Choi, Moon-Gun; Jeong, Kyu-Sung
Intracellular anion fluorescence assay for sodium/iodide symporter substrates.
The sodium/iodide symporter (NIS) is primarily responsible for iodide accumulation in the thyroid gland for the synthesis of thyroid hormones; however, it can also transport other lyotropic anions in the thyroid gland and nonthyroid tissues. Some NIS substrates have important physiological or clinical roles, and others are environmental contaminants with health-related consequences. The aim of this study was to assess the utility of a yellow fluorescent protein variant, YFP-H148Q/I152L, as a biosensor to monitor the cellular uptake of NIS substrates, including thiocyanate (SCN(-)), nitrate (NO(3)(-)), chlorate (ClO(3)(-)), perchlorate (ClO(4)(-)), and perrhenate (ReO(4)(-)). The fluorescence of purified YFP-H148Q/I152L was suppressed by anions with an order of potency of ReO(4)(-)>ClO(4)(-)=I(-)=SCN(-)=ClO(3)(-)>NO(3)(-)?Cl(-). Anions also suppressed the fluorescence of YFP-H148Q/I152L expressed in FRTL-5, a thyroid cell line with high NIS expression. Quantitation of intracellular concentrations revealed differences among anions in the affinity and maximal velocity of NIS-mediated uptake as well as in the rate constant for passive efflux. These results suggest that YFP-H148Q/I152L can serve as an intracellular biosensor of NIS-transported anions and may be useful to study the physiology of endogenous anions as well as the health-related consequences of environmental anions. PMID:21545783
Di Bernardo, Julie; Iosco, Carmela; Rhoden, Kerry J
Chloride and membrane potential dependence of sodium ion-proline symport.
Proline accumulation by renal proximal tubule brush border membrane vesicles is Na+ dependent, but little is known about the role of anions or membrane potential on proline uptake. Recent studies in a variety of transport systems, including rat renal brush border membrane vesicles, indicate that halide anions chloride (Cl-) and bromide (Br-) are essential for glycine, beta-alanine, gamma-aminobutyric acid, and taurine uptake, so the possibility that Na(+)-proline symport is Cl- dependent was explored. Also, the role of membrane potential on transport was assessed by determining the effect of external anions with different membrane permeabilities. The ratio of initial rate Cl- stimulated to thiocyanate (SCN)(-)-stimulated uptake values serves to measure Cl- dependence. The initial rate of proline uptake to equilibrium value was 3.11 +/- 0.5 (SE) in the presence of Cl- versus SCN-. The ratio for D-glucose, whose uptake is governed only by electrogenic status of the membrane, was 0.61 +/- 0.47 (P less than 0.001 versus proline). In another series of experiments, uptake values for various anions as a percent of equilibrium (I/E x 100) were: SCN-, 84.9 +/- 10.9; NO3, 49.9 +/- 11.0; SO4(2-), 27.3 +/- 4.4; F-, 68.5 +/- 18.3; Cl-, 164.1 +/- 44.6; Br-, 150.6 +/- 30.2; I-, 56.7 +/- 13.5. The stoichiometry of uptake by Hill plot analysis of proline uptake in the presence of varying concentrations of Na+ (0 to 100 mM) and Cl- (0 to 100 mM) was 2Na+:1Cl-:1 proline.(ABSTRACT TRUNCATED AT 250 WORDS) PMID:1751792
Chesney, R W; Zelikovic, I; Budreau, A; Randle, D
Prodigiosins uncouple lysosomal vacuolar-type ATPase through promotion of H+/Cl- symport.
We reported previously [Kataoka, Muroi, Ohkuma, Waritani, Magae, Takatsuki, Kondo, Yamasaki and Nagai (1995) FEBS Lett. 359, 53-59] that prodigiosin 25-C (one of the red pigments of the prodigiosin group produced by micro-organisms like Streptomyces and Serratia) uncoupled vacuolar H+-ATPase, inhibited vacuolar acidification and affected glycoprotein processing. In the present study we show that prodigiosin, metacycloprodigiosin and prodigiosin 25-C, all raise intralysosomal pH through inhibition of lysosomal acidification driven by vacuolar-type (V-)ATPase without inhibiting ATP hydrolysis in a dose-dependent manner with IC50 values of 30-120 pmol/mg of protein. The inhibition against lysosomal acidification was quick and reversible, showing kinetics of simple non-competitive (for ATP) inhibition. However, the prodigiosins neither raised the internal pH of isolated lysosomes nor showed ionophoric activity against H+ or K+ at concentrations where they strongly inhibited lysosomal acidification. They required Cl- for their acidification inhibitory activity even when driven in the presence of K+ and valinomycin, suggesting that their target is not anion (chloride) channel(s). In fact, the prodigiosins inhibited acidification of proteoliposomes devoid of anion channels that were reconstituted from lysosomal vacuolar-type (V-)ATPase and Escherichia coli phospholipids. However, they did not inhibit the formation of an inside-positive membrane potential driven by lysosomal V-ATPase. Instead, they caused quick reversal of acidified pH driven by lysosomal V-ATPase and, in acidic buffer, produced quick acidification of lysosomal pH, both only in the presence of Cl-. In addition, they induced swelling of liposomes and erythrocytes in iso-osmotic ammonium salt of chloride but not of gluconate, suggesting the promotion of Cl- entry by prodigiosins. These results suggest that prodigiosins facilitate the symport of H+ with Cl- (or exchange of OH- with Cl-) through lysosomal membranes, resulting in uncoupling of vacuolar H+-ATPase.
Ohkuma, S; Sato, T; Okamoto, M; Matsuya, H; Arai, K; Kataoka, T; Nagai, K; Wasserman, H H
Sodium iodide symporter-mediated radioiodide imaging and therapy of ovarian tumor xenografts in mice
Ovarian cancer represents the fifth leading cause of cancer death among women in the United States, with >16 000 deaths expected this year. This study was carried out to investigate the potential of sodium iodide symporter (NIS)-mediated radioiodide therapy as a novel approach for ovarian cancer treatment. Radioiodide is routinely and effectively used for the treatment of benign and malignant
R M Dwyer; E R Bergert; M K O'Connor; S J Gendler; J C Morris
A Homozygous Missense Mutation of the Sodium\\/Iodide Symporter Gene Causing Iodide Transport Defect
Iodide transport defect is a disorder characterized by an inability of the thyroid to maintain an iodide concentration difference between the plasma and the thyroid. The recent cloning of the sodium\\/iodide symporter (NIS) gene enabled us to characterize the NIS gene in this disorder. We identified a homozygous missense mutation of A3 Ca t nucleotide 11060 in NIS complementary DNA
Failure of Membrane Targeting Causes the Functional Defect of Two Mutant Sodium Iodide Symporters
Molecular cloning of the sodium\\/iodide symporter (NIS) allowed identification of NIS gene mutations in patients with iodide trapping defect. Whereas various mutant human (h) NIS molecules display loss of function when expressed by transfection in mammalian cells, the precise mechanism(s) responsible for the functional abnormality of these proteins remains unknown. With the aim to explore these mech- anisms in three
?-Fetoprotein promoter-targeted sodium iodide symporter gene therapy of hepatocellular carcinoma
Due to limited treatment options the prognosis of patients with advanced hepatocellular cancer (HCC) has remained poor. To investigate an alternative therapeutic approach, we examined the feasibility of radioiodine therapy of HCC following human sodium iodide symporter (NIS) gene transfer using a mouse ?-fetoprotein (AFP) promoter construct to target NIS expression to HCC cells. For this purpose, the murine Hepa
M J Willhauck; B R Sharif Samani; K Klutz; N Cengic; I Wolf; L Mohr; M Geissler; R Senekowitsch-Schmidtke; B G�ke; J C Morris; C Spitzweg
Analysis of Human Sodium Iodide Symporter Immunoreactivity in Human Exocrine Glands
The human sodium iodide symporter (hNIS) is an intrinsic trans- membrane protein that mediates the active transport of iodide across the basolateral membrane of thyroid follicular cells. In addition to normally functioning thyroid tissue, various extrathyroidal tissues, including sal- ivary gland, lacrimal gland, gastric mucosa, choroid plexus, and lactat- ing mammary gland, have been demonstrated to accumulate iodide. After cloning
C. Spitzweg; W. JOBA; K. SCHRIEVER; J. R. GOELLNER; J. C. MORRIS; A. E. HEUFELDER
The crystal structure of a sodium galactose transporter reveals mechanistic insights into Na[superscript +]\\/sugar symport
Membrane transporters that use energy stored in sodium gradients to drive nutrients into cells constitute a major class of proteins. We report the crystal structure of a member of the solute sodium symporters (SSS), the Vibrio parahaemolyticus sodium\\/galactose symporter (vSGLT). The -3.0 angstrom structure contains 14 transmembrane (TM) helices in an inward-facing conformation with a core structure of inverted repeats
S. Faham; A. Watanabe; G. M. Besserer; D. Cascio; A. Specht; B. A. Hirayama; E. M. Wright; J. Abramson
Implications of the molecular characterization of the sodium-iodide symporter (NIS).
The recently cloned sodium-iodide symporter (NIS) represents a key molecule for thyroid function by efficiently accumulating iodide from the circulation into the thyrocyte against an electrochemical gradient. This uptake requires energy, is coupled to the action of Na+/K+-ATPase, and stimulated by TSH, the main hormone regulating thyroid-specific functions. NIS mutations are found in congenital hypothyroidism, and potential defects in the NIS gene, its expression, or function of the NIS protein are currently under investigation in various thyroid diseases. Increased NIS expression has been found in autonomous adenoma and Graves' disease, decreased levels of NIS protein and/or mRNA were observed in Hashimoto's disease, cold nodules, most thyroid cancers and cell lines derived therefrom. Autoantibodies directed against NIS have been identified in autoimmune thyroid disease and blocking antibodies isolated from sera of patients with Hashimoto's disease inhibit NIS function in NIS-transfected CHO cells. NIS mRNA expression can be up-regulated by retinoic acid in human thyroid carcinoma cell lines whereas retinoic acid treatment decreases NIS expression and function in differentiated rat thyroid FRTL-5 cells. Apart from thyrocytes, NIS is also expressed in other tissues known to transiently accumulate radioiodide, albeit at much lower levels, requiring RT-PCR for detection of the transcript. Diagnostic and therapeutic implications of the recent cloning of the human NIS gene such as development of NIS-directed drugs, ligands, antibodies, vaccines, gene therapeutic approaches combining NIS targeting and expression together with the long-established, efficient and safe method of radioiodide therapy are discussed both for application to thyroid related diseases and carcinoma, and non-thyroid benign and malignant diseases. Apart from these therapeutic and diagnostic perspectives the availability of the NIS gene will also open new opportunities to develop sensitive and homologous diagnostic test systems to identify factors involved in autoimmune thyroid disease, evolution of goitre, adenoma and thyroid cancer as well as NIS-directed new drugs. Advanced and sophisticated molecular diagnostic approaches (RT-PCR from fine needle aspirations, screening for mutations, analysis of gene defects) are already developed for NIS and will complement or overcome some established procedures in thyroid diagnostics. PMID:9865544
Schmutzler, C; K�hrle, J
Relationship between the Na+/H+ antiporter and Na+/substrate symport in Bacillus alcalophilus.
The Na+/H+ antiporter of the obligate alkalophile Bacillus alcalophilus facilitates growth at alkaline pH and precludes growth below pH 8.5. Thus, nonalkalophilic mutant strains do not exhibit Na+/H+ antiport activity and, interestingly, such strains concomitantly lose the ability to catalyze Na+-dependent accumulation of alpha-aminoisobutyrate [Krulwich, T. A., Mandel, D. G. Bornstein, R. F. & Guffanti, A. A. (1979) Biochem. Biophys. Res. Commun. 91, 58-62]. Several other Na+-dependent transport systems are now documented in vesicles from the wild-type strain, and it is demonstrated that these systems are defective in vesicles from the nonalkalophilic mutant KM23. Surprisingly, the defect seems to result not from the loss of Na+/H+ antiport activity per se but from a pleiotropic defect in the Na+/substrate symporters themselves. Monensin, an ionophore that catalyzes Na+/H+ exchange, does not restore respiration-driven Na+/substrate symport in KM23 vesicles. Moreover, with KM23 vesicles, efflux of alpha-aminoisobutyrate, L-malate, and L-aspartate down their respective concentration gradients is not stimulated by Na+, in contrast to the observations with wild-type vesicles. Because monensin should ameliorate a simple defect in Na+/H+ antiport activity and the antiporter should not be required for Na+/substrate symport down a concentration gradient, the results suggest that there may be a direct relationship between the antiporter and various Na+/substrate symporters. One possibility is that the systems share a Na+-translocating subunit.
Guffanti, A A; Cohn, D E; Kaback, H R; Krulwich, T A
MEK signaling modulates sodium iodide symporter at multiple levels and in a paradoxical manner
The NaC\\/IK symporter (NIS)-mediated iodide uptake is the basis for targeted radioiodine ablation of thyroid cancers. However, NIS-mediated radioiodide uptake (RAIU) activity is often reduced in thyroid cancers. As mitogen activated protein kinase (MAPK) signaling pathway is activated in about 70% of papillary thyroid carcinoma, we investigated whether MEK (MAPK kinase) inhibition will restore NIS protein levels and NIS-mediated RAIU
Douangsone D Vadysirisack; Anjli Venkateswaran; Zhaoxia Zhang; Sissy M Jhiang
Immunohistochemical Expression of the Human Sodium\\/Iodide Symporter Distinguishes Malignant From Benign Gastric Lesions
Aim. Sodium\\/iodide symporter (NIS) is a transmembrane protein that mediates the transport of I?. The aim was to evaluate the immunohistochemical expression of the human homolog of NIS (hNIS) in a wide spectrum of gastric lesions. Materials and methods. Seventy-seven samples were stained immunohistochemically with a monoclonal antibody for hNIS, including 14 with normal gastric mucosa, 14 with chronic atrophic
Anna Farnedi; Leonardo H. Eusebi; Francesca Poli; Maria P. Foschini
Radioiodine therapy of colon cancer following tissue-specific sodium iodide symporter gene transfer
We investigated the feasibility of using radioiodine therapy in colon carcinoma cells (HCT 116) following tumor-specific expression of the human sodium iodide symporter (hNIS) using the carcinoembryonic antigen (CEA) promoter. HCT 116 cells were stably transfected with an expression vector, in which hNIS cDNA has been coupled to a CEA promoter fragment. This promoter is responsible for tissue-specific expression of
I V Scholz; N Cengic; C H Baker; K J Harrington; K Maletz; E R Bergert; R Vile; B G�ke; J C Morris; C Spitzweg
Radioiodine Therapy of Hepatoma Using Targeted Transfer of the Human Sodium\\/Iodide Symporter Gene
We investigated the feasibility of radioiodine therapy targeting hepatoma cells (MH3924A) by tissue-specific expression of the human sodium\\/iodide symporter (hNIS) gene directed by the murine albumin enhancer and promoter (mAlb). Methods: The cell-specific transcriptional activity of mAlb was examined by a luciferase assay in several transiently transfected cell lines. MH3924A cells were stably transfected with the recombinant retroviral vector, in
Libo Chen; Annette Altmann; Walter Mier; Helmut Eskerski; Karin Leotta; Lihe Guo; Ruisen Zhu; Uwe Haberkorn
Update on the thyroid sodium iodide symporter: a novel thyroid antigen emerging on the horizon
The ability to transport and concentrate iodide, a fundamental property of normally functioning thyroid epithelial cells, represents a key step in the production of the iodine-containing thyroid hormones. Iodide uptake across the basolateral membrane of thyroid follicular cells is made possible by the Na?\\/I? symporter (NIS), an active co-transport mechanism that is driven by an inwardly directed Na? gradient. In
Detection of mRNA of sodium iodide symporter in benign and malignant human thyroid tissues
Human sodium iodide symporter (hNIS) is an intrinsic membrane protein with 12 transmembrane regions, which shows homology to other sodium-dependent transporters. There is controversy as to the amount of hNIS expression in different kinds of human thyroid cancer tissues and cell lines. In this study, reverse transcription-polymerase chain reaction (RT-PCR) was used to detect mRNA of hNIS in various fresh
Miaw-Jene Liou; Jen-Der Lin; Err-Cheng Chan; Feng-Hsuan Liu; Tzu-Chieh Chao; Hsiao-Fen Weng
On the Universality of P Systems with Minimal Symport\\/Antiport Rules
\\u000a P systems with symport\\/antiport rules of a minimal size (only one object passes in any direction in a communication step)\\u000a were recently proven to be computationally universal. The proof from [2] uses systems with nine membranes. In this paper we\\u000a improve this results, by showing that six membranes suffice. The optimality of this result remains open (we believe that the
Lila Kari; Carlos Mart�n-vide; Andrei Paun
Electrogenicity, pH-Dependence, and Stoichiometry of the Proton-Sucrose Symport
The electrogenicity, pH-dependence, and stoichiometry of the proton-sucrose symport were examined in plasma membrane vesicles isolated from sugar beet (Beta vulgaris L. cv Great Western) leaves. Symport mediated sucrose transport was electrogenic as demonstrated by the effect of membrane potential on ?pH-dependent flux. In the absence of significant charge compensation, a low rate of sucrose transport was observed. When membrane potential was clamped at zero with symmetric potassium concentrations and valinomycin, the rate of sucrose flux was stimulated fourfold. In the presence of a negative membrane potential, transport increased six-fold. These results are consistent with electrogenic sucrose transport which results in a net flux of positive charge into the vesicles. The effect of membrane potential on the kinetics of sucrose transport was on Vmax only with no apparent change in Km. Sucrose transport rates driven by membrane potential only, i.e. in the absence of ?pH, were comparable to ?pH-driven flux. Both membrane potential and ?pH-driven sucrose transport were used to examine proton binding to the symport and the apparent Km for H+ was 0.7 micromolar. The kinetics of sucrose transport as a function of proton concentration exhibited a simple hyperbolic relationship. This observation is consistent with kinetic models of ion-cotransport systems when the stoichiometry of the system, ion:substrate, is 1:1. Quantitative measurements of proton and sucrose fluxes through the symport support a 1:1 stoichiometry. The biochemical details of protoncoupled sucrose transport reported here provide further evidence in support of the chemiosmotic hypothesis of nutrient transport across the plant cell plasma membrane.
Bush, Daniel R.
KT5823 differentially modulates sodium iodide symporter expression, activity, and glycosylation between thyroid and breast cancer cells.
Na(+)/I(-) symporter (NIS)-mediated iodide uptake into thyroid follicular cells serves as the basis of radioiodine therapy for thyroid cancer. NIS protein is also expressed in the majority of breast tumors, raising potential for radionuclide therapy of breast cancer. KT5823, a staurosporine-related protein kinase inhibitor, has been shown to increase thyroid-stimulating hormone-induced NIS expression, and thus iodide uptake, in thyroid cells. In this study, we found that KT5823 does not increase but decreases iodide uptake within 0.5 h of treatment in trans-retinoic acid and hydrocortisone-treated MCF-7 breast cancer cells. Moreover, KT5823 accumulates hypoglycosylated NIS, and this effect is much more evident in breast cancer cells than thyroid cells. The hypoglycosylated NIS is core glycosylated, has not been processed through the Golgi apparatus, but is capable of trafficking to the cell surface. KT5823 impedes complex NIS glycosylation at a regulatory point similar to brefeldin A along the N-linked glycosylation pathway, rather than targeting a specific N-glycosylated site of NIS. KT5823-mediated effects on NIS activity and glycosylation are also observed in other breast cancer cells as well as human embryonic kidney cells expressing exogenous NIS. Taken together, KT5823 will serve as a valuable pharmacological reagent to uncover mechanisms underlying differential NIS regulation between thyroid and breast cancer cells at multiple levels. PMID:21209020
Beyer, Sasha; Lakshmanan, Aparna; Liu, Yu-Yu; Zhang, Xiaoli; Wapnir, Irene; Smolenski, Albert; Jhiang, Sissy
Molecular Characterization of V59E NIS, a Na+/I? Symporter Mutant that Causes Congenital I? Transport Defect
I? is actively transported into thyrocytes via the Na+/I? symporter (NIS), a key glycoprotein located on the basolateral plasma membrane. The cDNA encoding rat NIS was identified in our laboratory, where an extensive structure/function characterization of NIS is being conducted. Several NIS mutants have been identified as causes of congenital I? transport defect (ITD), including V59E NIS. ITD is characterized by low thyroid I? uptake, low saliva/plasma I? ratio, hypothyroidism, and goiter and may cause mental retardation if untreated. Studies of other ITD-causing NIS mutants have revealed valuable information regarding NIS structure/function. V59E NIS was reported to exhibit as much as 30% of the activity of wild-type NIS. However, this observation was at variance with the patients� phenotype of total lack of activity. We have thoroughly characterized V59E NIS and studied several amino acid substitutions at position 59. We demonstrated that, in contrast to the previous report, V59E NIS is inactive, although it is properly targeted to the plasma membrane. Glu and all other charged amino acids or Pro at position 59 also yielded nonfunctional NIS proteins. However, I? uptake was rescued to different degrees by the other substitutions. Although the Km values for Na+ and I? were not altered in these active mutants, we found that the structural requirement for NIS function at position 59 is a neutral, helix-promoting amino acid. This result suggests that the region that contains V59 may be involved in intramembrane helix-helix interactions during the transport cycle without being in direct contact with the substrates.
Reed-Tsur, Mia D.; De la Vieja, Antonio; Ginter, Christopher S.; Carrasco, Nancy
Construction of an MUC1 promoter driven, conditionally replicating adenovirus that expresses the sodium iodide symporter for gene therapy of breast cancer
INTRODUCTION: The sodium iodide symporter (NIS) directs the uptake and concentration of iodide in thyroid cells. This in turn allows radioiodine imaging and therapy for thyroid cancer. To extend the use of NIS-mediated radioiodine therapy to other types of cancer, we successfully transferred and expressed the sodium-iodide symporter (NIS) gene in prostate, colon, and breast cancer cells both in vivo
Miguel A Trujillo; Michael J Oneal; Julia Davydova; Elizabeth Bergert; Masato Yamamoto; John C Morris
The amino acid\\/auxin:proton symport permease family 1 The accompanying review paper �Phylogenetic characterization of novel transport protein families revealed by genome analysis� by M.H. Saier Jr. et al. will be published in Biochim. Biophys. Acta, Vol. 1422\\/1, February 1999 issue. 1
Amino acids and their derivatives are transported into and out of cells by a variety of permease types which comprise several distinct protein families. We here present a systematic analysis of a group of homologous transport proteins which together comprise the eukaryotic-specific amino acid\\/auxin permease (AAAP) family (TC #2.18). In characterizing this family, we have (1) identified all sequenced members
Gregory B. Young; Donald L. Jack; Douglas W. Smith; Milton H. Saier
The iodide-transport-defect-causing mutation R124H: a ?-amino group at position 124 is critical for maturation and trafficking of the Na+/I- symporter.
Na(+)/I(-) symporter (NIS)-mediated active accumulation of I(-) in thyrocytes is a key step in the biosynthesis of the iodine-containing thyroid hormones T3 and T4. Several NIS mutants have been identified as a cause of congenital I(-) transport defect (ITD), and their investigation has yielded valuable mechanistic information on NIS. Here we report novel findings derived from the thorough characterization of the ITD-causing mutation R124H, located in the second intracellular loop (IL-2). R124H NIS is incompletely glycosylated and colocalizes with endoplasmic reticulum (ER)-resident protein markers. As a result, R124H NIS is not targeted to the plasma membrane and therefore does not mediate any I(-) transport in transfected COS-7 cells. Strikingly, however, the mutant is intrinsically active, as revealed by its ability to mediate I(-) transport in membrane vesicles. Of all the amino acid substitutions we carried out at position 124 (K, D, E, A, W, N and Q), only Gln restored targeting of NIS to the plasma membrane and NIS activity, suggesting a key structural role for the ?-amino group of R124 in the transporter's maturation and cell surface targeting. Using our NIS homology model based on the structure of the Vibrio parahaemolyticus Na(+)/galactose symporter, we propose an interaction between the ?-amino group of either R or Q124 and the thiol group of C440, located in IL-6. We conclude that the interaction between IL-2 and IL-6 is critical for the local folding required for NIS maturation and plasma membrane trafficking. PMID:23690546
Paroder, Viktoriya; Nicola, Juan P; Ginter, Christopher S; Carrasco, Nancy
MEK inhibition leads to lysosome-mediated Na+/I- symporter protein degradation in human breast cancer cells.
The Na(+)/I(-) symporter (NIS (SLC5A5)) is a transmembrane glycoprotein that mediates active iodide uptake into thyroid follicular cells. NIS-mediated iodide uptake in thyroid cells is the basis for targeted radionuclide imaging and treatment of differentiated thyroid carcinomas and their metastases. Furthermore, NIS is expressed in many human breast tumors but not in normal non-lactating breast tissue, suggesting that NIS-mediated radionuclide uptake may also allow the imaging and targeted therapy of breast cancer. However, functional cell surface NIS expression is often low in breast cancer, making it important to uncover signaling pathways that modulate NIS expression at multiple levels, from gene transcription to posttranslational processing and cell surface trafficking. In this study, we investigated NIS regulation in breast cancer by MAPK/extracellular signal-regulated kinase (ERK) kinase (MEK) signaling, an important cell signaling pathway involved in oncogenic transformation. We found that MEK inhibition decreased NIS protein levels in all-trans retinoic acid/hydrocortisone-treated MCF-7 cells as well as human breast cancer cells expressing exogenous NIS. The decrease in NIS protein levels by MEK inhibition was not accompanied by a decrease in NIS mRNA or a decrease in NIS mRNA export from the nucleus to the cytoplasm. NIS protein degradation upon MEK inhibition was prevented by lysosome inhibitors but not by proteasome inhibitors. Interestingly, NIS protein level was correlated with MEK/ERK activation in human breast tumors from a tissue microarray. Taken together, MEK activation appears to play an important role in maintaining NIS protein stability in human breast cancers. PMID:23404856
Zhang, Zhaoxia; Beyer, Sasha; Jhiang, Sissy M
MEK Inhibition Leads To Lysosome-Mediated Na+/I- Symporter Protein Degradation In Human Breast Cancer Cells
The Na+/I- symporter (NIS) is a transmembrane glycoprotein that mediates active iodide uptake into thyroid follicular cells. NIS-mediated iodide uptake in thyroid cells is the basis for targeted radionuclide imaging and treatment of differentiated thyroid carcinomas and their metastases. Furthermore, NIS is expressed in many human breast tumors but not in normal non-lactating breast tissue, suggesting that NIS-mediated radionuclide uptake may also allow the imaging and targeted therapy of breast cancer. However, functional cell surface NIS expression is often low in breast cancer, making it important to uncover signaling pathways that modulate NIS expression at multiple levels, from gene transcription to post-translational processing and cell surface trafficking. In this study, we investigated NIS regulation in breast cancer by MEK (MAPK/ERK kinase) signaling, an important cell signaling pathway involved in oncogenic transformation. We found that MEK inhibition decreased NIS protein levels in all-trans retinoic acid (tRA)/hydrocortisone treated MCF-7 cells as well as human breast cancer cells expressing exogenous NIS. The decrease in NIS protein levels by MEK inhibition was not accompanied by a decrease in NIS mRNA or a decrease in NIS mRNA export from the nucleus to the cytoplasm. NIS protein degradation upon MEK inhibition was prevented by lysosome inhibitors, but not by proteasome inhibitors. Interestingly, NIS protein level was correlated with MEK/ERK activation in human breast tumors from a tissue microarray. Taken together, MEK activation appears to play an important role in maintaining NIS protein stability in human breast cancers.
Regulated exocytosis of an H+/myo-inositol symporter at synapses and growth cones
Phosphoinositides, synthesized from myo-inositol, play a critical role in the development of growth cones and in synaptic activity. As neurons cannot synthesize inositol, they take it up from the extracellular milieu. Here, we demonstrate that, in brain and PC12 cells, the recently identified H+/myo-inositol symporter HMIT is present in intracellular vesicles that are distinct from synaptic and dense-core vesicles. We further show that HMIT can be triggered to appear on the cell surface following cell depolarization, activation of protein kinase C or increased intracellular calcium concentrations. HMIT cell surface expression takes place preferentially in regions of nerve growth and at varicosities and leads to increased myo-inositol uptake. The symporter is then endocytosed in a dynamin-dependent manner and becomes available for a subsequent cycle of stimulated exocytosis. HMIT is thus expressed in a vesicular compartment involved in activity-dependent regulation of myo-inositol uptake in neurons. This may be essential for sustained signaling and vesicular traffic activities in growth cones and at synapses.
Uldry, Marc; Steiner, Pascal; Zurich, Marie-Gabrielle; Beguin, Pascal; Hirling, Harald; Dolci, Wanda; Thorens, Bernard
Mitogen-Activated Protein Kinase Signaling Enhances Sodium Iodide Symporter Function and Efficacy of Radioiodide Therapy in Nonthyroidal Cancer Cells
Although the success of sodium\\/iodide symporter (NIS) gene- based cancer therapy is critically dependent on the level of radio- iodide accumulation attained, recent evidence indicates that successful therapy relies not solely on NIS amount but also cru- cially on its functional activity. In this study, we investigated the role of kinase-linked signaling on the regulation of NIS function in cancer
Kyung-Ho Jung; Jin-Young Paik; Bong-Ho Ko; Kyung-Han Lee
FSY1, a Novel Gene Encoding a Specific Fructose/H+ Symporter in the Type Strain of Saccharomyces carlsbergensis
A novel gene, FSY1, encoding a permease involved in active fructose uptake by a proton symport mechanism in the type strain of Saccharomyces carlsbergensis has been isolated. Fsy1p is only distantly related to the Hxt proteins that mediate facilitated diffusion of glucose and fructose in Saccharomyces cerevisiae and related species.
Goncalves, Paula; Rodrigues de Sousa, Helena; Spencer-Martins, Isabel
Effect of thyroid statuses on sodium\\/iodide symporter (NIS) gene expression in the extrathyroidal tissues in mice
BACKGROUND: Iodide that is essential for thyroid hormone synthesis is actively transported into the thyroid follicular cells via sodium\\/iodide symporter (NIS) protein in vertebrates. It is well known that NIS expression in thyroid is regulated by the thyroid statuses mainly through thyroid stimulating hormone (TSH). Although NIS mRNA expressions in extrathyroidal tissues have been qualitatively reported, their regulation by thyroid
Masato Asai; Xiao-yang Sun; Yoshitaka Hayashi; Junichi Sakamoto; Yoshiharu Murata
Treatment of Prostate Cancer by Radioiodine Therapy after Tissue-specific Expression of the Sodium Iodide Symporter1
Causing prostate cancer cells to express functionally active sodium iodide symporter (NIS) by targeted NIS gene transfer might offer the possibility of radioiodine therapy of prostate cancer. Therefore, we inves- tigated radioiodine accumulation and therapeutic effectiveness of 131 Ii n NIS-transfected prostate cancer cells in vitro and in vivo. The human prostatic adenocarcinoma cell line LNCaP was stably transfected with
Christine Spitzweg; Michael K. O'Connor; Elizabeth R. Bergert; Donald J. Tindall; Charles Y. F. Young; John C. Morris
A Novel Peculiar Mutation in the Sodium\\/Iodide Symporter Gene in Spanish Siblings with Iodide Transport Defect
Previously, we reported two Spanish siblings with congenital hypothyroidism due to total failure of iodide transport. These were the only cases reported to date who received long-term iodide treatment over 10 yr. We examined the sodium\\/iodide symporter (NIS) gene of these patients. A large deletion was observed by long and accurate PCR using primers derived from introns 2 and 7
Effects of Histone Acetylation on Sodium Iodide Symporter Promoter and Expression of Thyroid-Specific Transcription Factors
Inhibitors of histone deacetylases (HDACs) activate the so- dium iodide symporter (NIS) expression in thyroid tumor cells. In this study, mechanisms accounting for these effects were investigated. Various human thyroid tumor cell lines (ARO, BCPAP, FRO, TPC-1) were treated with the HDAC in- hibitors Na butyrate (NaB) and tricostatin A (TSA), and the effects on the expression of NIS and
Cinzia Puppin; Federica D'Aurizio; Angela V. D'Elia; Laura Cesaratto; Gianluca Tell; Diego Russo; Sebastiano Filetti; Elisabetta Ferretti; Emanuele Tosi; Tiziana Mattei; Annalisa Pianta; Lucia Pellizzari; Giuseppe Damante
Characterisation of the purified human sodium\\/iodide symporter reveals that the protein is mainly present in a dimeric form and permits the detailed study of a native C-terminal fragment
The sodium\\/iodide symporter is an intrinsic membrane protein that actively transports iodide into thyroid follicular cells. It is a key element in thyroid hormone biosynthesis and in the radiotherapy of thyroid tumours and their metastases. Sodium\\/iodide symporter is a very hydrophobic protein that belongs to the family of sodium\\/solute symporters. As for many other membrane proteins, particularly mammalian ones, little
Sylvaine Huc-Brandt; Didier Marcellin; Fanny Graslin; Olivier Averseng; Laurent Bellanger; Patrick Hivin; Eric Quemeneur; C�cile Basquin; Val�rie Navarro; Thierry Pourcher; Elisabeth Darrouzet
Sodium\\/iodide-symporter: distribution in different mammals and role in entero-thyroid circulation of iodide
The sodium (Na+)\\/iodide (I-)-symporter (NIS) is abundantly expressed and accumulates iodide in thyroid follicular cells. The NIS is also found in extrathyroidal tissues, particularly gastric mucosa. Controversies exist on the localization of extrathyroidal NIS. We have studied the presence of both NIS peptide and NIS messenger RNA (mRNA) in the digestive tract and thyroid from different mammals. The role of
Visualization of Hypoxia-Inducible Factor1 Transcriptional Activation in C6 Glioma Using Luciferase and Sodium Iodide Symporter Genes
Hypoxia-inducible factor-1 (HIF-1) is a transcription factor of hy- poxic response in cancer cells and is associated with tumor pro- gression, angiogenesis, metastasis, and resistance to therapy. We assessed whether the human sodium iodide symporter (NIS)reportersystemscanbeusedtovisualizetranscriptionalac- tivation of HIF-1 in C6 glioma. Methods: Two types of plasmid- expressing human NIS or luciferase (Luc) genes, controlled by 5copiesofhypoxiaresponseelement(5HRE),wereconstructed: p5HRE-NIS or p5HRE-Luc.
Chan Joo Yeom; June-Key Chung; Joo Hyun Kang; Yong Hyun Jeon; Kwang Il Kim; Yong Nan Jin; You Mie Lee; Jae Min Jeong; Dong Soo Lee
Comparison of expressed human and mouse sodium\\/iodide symporters reveals differences in transport properties and subcellular localization
The active transport of iodide from the bloodstream into thyroid follicular cells is mediated by the NaC\\/IK symporter (NIS). We studied mouse NIS (mNIS) and found that it catalyzes iodide transport into transfected cells more efficiently than human NIS (hNIS). To further characterize this difference, we compared 125I uptake in the transiently transfected human embryonic kidney (HEK) 293 cells. We
Manal Dayem; Cecile Basquin; Valerie Navarro; Patricia Carrier; Robert Marsault; Patrick Chang; Sylvaine Huc; Elisabeth Darrouzet; Sabine Lindenthal; Thierry Pourcher
Targeting of tumor radioiodine therapy by expression of the sodium iodide symporter under control of the survivin promoter
To test the feasibility of using the survivin promoter to induce specific expression of sodium\\/iodide symporter (NIS) in cancer cell lines and tumors for targeted use of radionuclide therapy, a recombinant adenovirus, Ad-SUR-NIS, that expressed the NIS gene under control of the survivin promoter was constructed. Ad-SUR-NIS mediating iodide uptake and cytotoxicity was performed in vitro. Scintigraphic, biodistribution and radioiodine
R Huang; Z Zhao; X Ma; S Li; R Gong; A Kuang
Development of Monoclonal Antibodies Against the Human Sodium Iodide Symporter: Immunohistochemical Characterization of This Protein in Thyroid Cells
The thyroid sodium-iodide symporter (NIS) is responsible for io- dide concentrating ability within thyroid follicular cells. We sought to develop monoclonal antibodies against human NIS (hNIS) for use as reagents in structure-function studies of the protein, as well as po- tential tools in the assessment of NIS expression in benign and ma- lignant thyroid tissues. Synthetic peptides corresponding to the
PI3K activation is associated with intracellular sodium\\/iodide symporter protein expression in breast cancer
Background��The sodium\\/iodide symporter (NIS) is a membrane glycoprotein mediating active iodide uptake in the thyroid gland and is the\\u000a molecular basis for radioiodide imaging and therapeutic ablation of thyroid carcinomas. NIS is expressed in the lactating\\u000a mammary gland and in many human breast tumors, raising interest in similar use for diagnosis and treatment. However, few human\\u000a breast tumors have clinically
Katherine AB Knostman; James A McCubrey; Carl D Morrison; Zhaoxia Zhang; Charles C Capen; Sissy M Jhiang
Feasibility of sodium\\/iodide symporter gene as a new imaging reporter gene: comparison with HSV1-tk
Positron emission tomography (PET) imaging reporter genes, such as HSV1-tk and D 2 receptor genes, make it possible to visualise gene expression non-invasively and repetitively in vivo. However, these systems require the synthesis of complicated substrates and the availability of expensive PET equipment. Expression of the sodium\\/iodide symporter ( NIS) gene can be easily monitored with radioiodines and technetium-99m using
JaeHoon Shin; June-Key Chung; JooHyun Kang; YongJin Lee; KwangIl Kim; ChulWoo Kim; JaeMin Jeong; DongSoo Lee; MyungChul Lee
Do cell surface trafficking impairments account for variable cell surface sodium iodide symporter levels in breast cancer?
The Na+\\/I? symporter (NIS) is a transmembrane glycoprotein that mediates iodide uptake into thyroid follicular cells and serves as the\\u000a molecular basis of radioiodine imaging and therapy for thyroid cancer patients. The finding that NIS protein is present in\\u000a 80�90% of breast tumors suggests that breast cancer patients may also benefit from NIS-mediated radionuclide imaging and targeted\\u000a therapy. However, only
S. J. Beyer; R. E. Jimenez; C. L. Shapiro; J. Y. Cho; S. M. Jhiang
AtDUR3 encodes a new type of high-affinity urea\\/H+ symporter in Arabidopsis
Urea is the major nitrogen form supplied as fertilizer in agricultural plant production but also an important nitrogen metabolite in plants. We report the cloning and functional characterization of AtDUR3, a high-affinity urea transporter in plants. AtDUR3 contains 14 putative transmembrane-spanning domains and represents an individual member in Arabidopsis that belongs to a superfamily of sodium-solute symporters. Heterologous expression in
Lai-Hua Liu; Uwe Ludewig; Wolf B. Frommer; N. von Wiren
Low Frequency of Autoantibodies to the Human Na1\\/I2 Symporter in Patients with Autoimmune Thyroid Disease
Several studies suggest that the sodium-iodide symporter (NIS) may represent a major autoantigen in autoimmune diseases of the thyroid. The aim of the present paper was to investigate the impor- tance of autoantibodies to human NIS (hNIS-Ab) in patients suffering from Hashimoto's thyroiditis (HT) and Graves' disease (GD). Full- length human NIS (hNIS) was cloned from thyroid tissue, expressed by
Radioiodide Treatment after Sodium Iodide Symporter Gene Transfer Is a Highly Effective Therapy in Neuroendocrine Tumor Cells1
This study evaluates the possibility of treating Bon1 and QGP pancre- atic neuroendocrine tumor cells with radioactive iodide (131I) after stable transfection with the thyroid sodium iodide symporter (NIS). NIS expres- sion was driven either by the strong viral cytomegalovirus promoter or by the tissue-specific chromogranin A promoter. Using either approach, NIS expression was confirmed by reverse transcription-PCR and Western
Meike L. Schipper; Alexander Weber; Martin Behe; Rudiger Goke; Werner Joba; Harald Schmidt; Till Bert; Babette Simon; Rudolf Arnold; Armin E. Heufelder; Thomas M. Behr
Phosphoinositide3-kinase inhibition induces sodium\\/iodide symporter expression in rat thyroid cells and human papillary thyroid cancer cells
TSH stimulation of sodium iodide symporter (NIS) expression in thyroid cancer promotes radioiodine uptake and is required to deliver an effective treatment dose. Activation of the insulin\\/phosphoinositide-3-kinase (PI3K) signaling pathway in TSH-stimulated thyroid cells reduces NIS expression at the transcriptional level. We, therefore, investigated the effects of PI3K pathway inhibition on iodide uptake and NIS expression in rat thyroid cell
Takahiko Kogai; Saima Sajid-Crockett; Lynell S Newmarch; Yan-Yun Liu; Gregory A Brent
State-dependent conformations of the translocation pathway in the tyrosine transporter Tyt1, a novel neurotransmitter:sodium symporter from Fusobacterium nucleatum.
The gene of a novel prokaryotic member (Tyt1) of the neurotransmitter:sodium symporter (NSS) family has been cloned from Fusobacterium nucleatum. In contrast to eukaryotic and some prokaryotic NSSs, which contain 12 transmembrane domains (TMs), Tyt1 contains only 11 TMs, a characteristic shared by approximately 70% of prokaryotic NSS homologues. Nonetheless upon heterologous expression in an engineered Escherichia coli host, Tyt1 catalyzes robust Na+-dependent, highly selective l-tyrosine transport. Genetic engineering of Tyt1 variants devoid of cysteines or with individually retained endogenous cysteines at positions 18 or 238, at the cytoplasmic ends of TM1 and TM6, respectively, preserved normal transport activity. Whereas cysteine-less Tyt1 was resistant to the inhibitory effect of sulfhydryl-alkylating reagents, N-ethylmaleimide inhibited transport by Tyt1 variants containing either one or both of the endogenous cysteines, and this inhibition was altered by the substrates sodium and tyrosine, consistent with substrate-induced dynamics in the transport pathway. Our findings support a binding model of Tyt1 function in which an ordered sequence of substrate-induced structural changes reflects distinct conformational states of the transporter. This work identifies Tyt1 as the first functional bacterial NSS member putatively consisting of only 11 TMs and shows that Tyt1 is a suitable model for the study of NSS dynamics with relevance to structure/function relationships of human NSSs, including the dopamine, norepinephrine, serotonin, and gamma-aminobutyric acid transporters. PMID:16798738
Quick, Matthias; Yano, Hideaki; Goldberg, Naomi R; Duan, Lihua; Beuming, Thijs; Shi, Lei; Weinstein, Harel; Javitch, Jonathan A
Structural basis for dynamic mechanism of proton-coupled symport by the peptide transporter POT
Proton-dependent oligopeptide transporters (POTs) are major facilitator superfamily (MFS) proteins that mediate the uptake of peptides and peptide-like molecules, using the inwardly directed H+ gradient across the membrane. The human POT family transporter peptide transporter 1 is present in the brush border membrane of the small intestine and is involved in the uptake of nutrient peptides and drug molecules such as ?-lactam antibiotics. Although previous studies have provided insight into the overall structure of the POT family transporters, the question of how transport is coupled to both peptide and H+ binding remains unanswered. Here we report the high-resolution crystal structures of a bacterial POT family transporter, including its complex with a dipeptide analog, alafosfalin. These structures revealed the key mechanistic and functional roles for a conserved glutamate residue (Glu310) in the peptide binding site. Integrated structural, biochemical, and computational analyses suggested a mechanism for H+-coupled peptide symport in which protonated Glu310 first binds the carboxyl group of the peptide substrate. The deprotonation of Glu310 in the inward open state triggers the release of the bound peptide toward the intracellular space and salt bridge formation between Glu310 and Arg43 to induce the state transition to the occluded conformation.
Doki, Shintaro; Kato, Hideaki E.; Solcan, Nicolae; Iwaki, Masayo; Koyama, Michio; Hattori, Motoyuki; Iwase, Norihiko; Tsukazaki, Tomoya; Sugita, Yuji; Kandori, Hideki; Newstead, Simon; Ishitani, Ryuichiro; Nureki, Osamu
Dynamics of the l-fucose/H+ symporter revealed by fluorescence spectroscopy
FucP of Escherichia coli catalyzes l-fucose/H+ symport, and a crystal structure in an outward-facing conformation has been reported. However, nothing is known about FucP conformational dynamics. Here, we show that addition of l-fucose to purified FucP in detergent induces ?20% quenching of Trp fluorescence in a concentration-dependent manner without a shift in ?max. Quenching is essentially abolished when both Trp38 and Trp278, which are positioned on opposing faces of the outward-facing cavity walls, are replaced with Tyr or Phe, and reduced quenching is observed when either Trp is mutated. Therefore, both Trp residues are involved in the phenomenon. Furthermore, replacement of either Trp38 or Trp278, predominantly Trp38, causes decreased quenching, decreased apparent affinity for l-fucose, and significant inhibition of active l-fucose transport, indicating that the two residues are likely involved directly in sugar binding. It is proposed that sugar binding induces a conformational change in which the outward-facing cavity in FucP closes, thereby bringing Trp38 and Trp278 into close proximity around the bound sugar to form an �occluded� intermediate. The location of these two Trp residues provides a unique method for analyzing structural dynamics in FucP.
Sugihara, Junichi; Sun, Linfeng; Yan, Nieng; Kaback, H. Ronald
The Na+/I- symporter mediates active iodide uptake in the intestine.
Absorption of dietary iodide, presumably in the small intestine, is the first step in iodide (I(-)) utilization. From the bloodstream, I(-) is actively taken up via the Na(+)/I(-) symporter (NIS) in the thyroid for thyroid hormone biosynthesis and in such other tissues as lactating breast, which supplies I(-) to the newborn in the milk. The molecular basis for intestinal I(-) absorption is unknown. We sought to determine whether I(-) is actively accumulated by enterocytes and, if so, whether this process is mediated by NIS and regulated by I(-) itself. NIS expression was localized exclusively at the apical surface of rat and mouse enterocytes. In vivo intestine-to-blood transport of pertechnetate, a NIS substrate, was sensitive to the NIS inhibitor perchlorate. Brush border membrane vesicles accumulated I(-) in a sodium-dependent, perchlorate-sensitive manner with kinetic parameters similar to those of thyroid cells. NIS was expressed in intestinal epithelial cell line 6, and I(-) uptake in these cells was also kinetically similar to that in thyrocytes. I(-) downregulated NIS protein expression and its own NIS-mediated transport both in vitro and in vivo. We conclude that NIS is functionally expressed on the apical surface of enterocytes, where it mediates active I(-) accumulation. Therefore, NIS is a significant and possibly central component of the I(-) absorption system in the small intestine, a system of key importance for thyroid hormone biosynthesis and thus systemic intermediary metabolism. PMID:19052257
Nicola, Juan Pablo; Basquin, C�cile; Portulano, Carla; Reyna-Neyra, Andrea; Paroder, Monika; Carrasco, Nancy
A novel mechanism of sodium iodide symporter repression in differentiated thyroid cancer.
Differentiated thyroid cancers and their metastases frequently exhibit reduced iodide uptake, impacting on the efficacy of radioiodine ablation therapy. PTTG binding factor (PBF) is a proto-oncogene implicated in the pathogenesis of thyroid cancer. We recently reported that PBF inhibits iodide uptake, and have now elucidated a mechanism by which PBF directly modulates sodium iodide symporter (NIS) activity in vitro. In subcellular localisation studies, PBF overexpression resulted in the redistribution of NIS from the plasma membrane into intracellular vesicles, where it colocalised with the tetraspanin CD63. Cell-surface biotinylation assays confirmed a reduction in plasma membrane NIS expression following PBF transfection compared with vector-only treatment. Coimmunoprecipitation and GST-pull-down experiments demonstrated a direct interaction between NIS and PBF, the functional consequence of which was assessed using iodide-uptake studies in rat thyroid FRTL-5 cells. PBF repressed iodide uptake, whereas three deletion mutants, which did not localise within intracellular vesicles, lost the ability to inhibit NIS activity. In summary, we present an entirely novel mechanism by which the proto-oncogene PBF binds NIS and alters its subcellular localisation, thereby regulating its ability to uptake iodide. Given that PBF is overexpressed in thyroid cancer, these findings have profound implications for thyroid cancer ablation using radioiodine. PMID:19706688
Smith, Vicki E; Read, Martin L; Turnell, Andrew S; Watkins, Rachel J; Watkinson, John C; Lewy, Greg D; Fong, Jim C W; James, Sally R; Eggo, Margaret C; Boelaert, Kristien; Franklyn, Jayne A; McCabe, Christopher J
Decreased Expression of Thyroglobulin and Sodium Iodide Symporter Genes in Hashimoto's Thyroiditis.
Aim. The aim of the study was to compare the expression of sodium iodide symporter (NIS), thyroglobulin (Tg), tumor necrosis factor- ? (TNF ? ), and interleukin-1 ? genes in patients with Hashimoto's thyroiditis (HT) and healthy individuals. Subjects and Methods. Thyroid cells were obtained from 39 patients with HT and 15 controls by an ultrasound guided fine needle aspiration biopsy. Results. The patients with HT had lower Tg and NIS mRNA (P = 0.002 and P = 0.001, resp.), as well as higher TNF ? mRNA expression (P = 0.049) than the controls. In the HT group Tg mRNA expression correlated positively with NIS mRNA expression (R = 0.739, P = 0.0001) and thyroid volume (R = 0.465, P = 0.0005), as well as negatively with TNF ? mRNA expression (R = -0.490, P = 0.001) and anti-peroxidase antibodies (TPOAb) level (R = -0.482, P = 0.0002), whereas NIS mRNA expression correlated positively with thyroid volume (R = 0.319, P = 0.02), as well as negatively with TNF ? mRNA expression (R = -0.529, P = 0.0006) and TPOAb level (R = -0.422, P = 0.001). Conclusions. Our results suggest that decreased Tg and NIS expression in thyroid cells may result in reduced active iodide transport and reduced thyroid volume in patients with HT. PMID:24723949
Pop?awska-Kita, Anna; Telejko, Beata; Siewko, Katarzyna; Ko?ciuszko-Zdrodowska, Maria; Wawrusewicz-Kurylonek, Natalia; Kr?towski, Adam; Hryniewicka, Justyna; Dzi?cio?, Janusz; Bauer, Witold; Milewski, Robert; Szelachowska, Ma?gorzata; G�rska, Maria
Sodium iodide symporter for nuclear molecular imaging and gene therapy: from bedside to bench and back.
Molecular imaging, defined as the visual representation, characterization and quantification of biological processes at the cellular and subcellular levels within intact living organisms, can be obtained by various imaging technologies, including nuclear imaging methods. Imaging of normal thyroid tissue and differentiated thyroid cancer, and treatment of thyroid cancer with radioiodine rely on the expression of the sodium iodide symporter (NIS) in these cells. NIS is an intrinsic membrane protein with 13 transmembrane domains and it takes up iodide into the cytosol from the extracellular fluid. By transferring NIS function to various cells via gene transfer, the cells can be visualized with gamma or positron emitting radioisotopes such as Tc-99m, I-123, I-131, I-124 and F-18 tetrafluoroborate, which are accumulated by NIS. They can also be treated with beta- or alpha-emitting radionuclides, such as I-131, Re-186, Re-188 and At-211, which are also accumulated by NIS. This article demonstrates the diagnostic and therapeutic applications of NIS as a radionuclide-based reporter gene for trafficking cells and a therapeutic gene for treating cancers. PMID:22539935
Ahn, Byeong-Cheol
Physiological sodium concentrations enhance the iodide affinity of the Na(+)/I(-) symporter.
The Na(+)/I(-) symporter (NIS) mediates active I(-) transport-the first step in thyroid hormonogenesis-with a 2Na(+):1I(-) stoichiometry. NIS-mediated (131)I(-) treatment of thyroid cancer post-thyroidectomy is the most effective targeted internal radiation cancer treatment available. Here to uncover mechanistic information on NIS, we use statistical thermodynamics to obtain Kds and estimate the relative populations of the different NIS species during Na(+)/anion binding and transport. We show that, although the affinity of NIS for I(-) is low (Kd=224??M), it increases when Na(+) is bound (Kd=22.4??M). However, this Kd is still much higher than the submicromolar physiological I(-) concentration. To overcome this, NIS takes advantage of the extracellular Na(+) concentration and the pronounced increase in its own affinity for I(-) and for the second Na(+) elicited by binding of the first. Thus, at physiological Na(+) concentrations, ~79% of NIS molecules are occupied by two Na(+) ions and ready to bind and transport I(-). PMID:24888603
Nicola, Juan P; Carrasco, Nancy; Mario Amzel, L
Sodium iodide symporter (NIS) in extrathyroidal malignancies: focus on breast and urological cancer
Background Expression and function of sodium iodide symporter (NIS) is requisite for efficient iodide transport in thyrocytes, and its presence in cancer cells allows the use of radioiodine as a diagnostic and therapeutic tool in thyroid neoplasia. Discovery of NIS expression in extrathyroidal tissues, including transformed cells, has opened a novel field of research regarding NIS-expressing extrathyroidal neoplasia. Indeed, expression of NIS may be used as a biomarker for diagnostic, prognostic, and therapeutic purposes. Moreover, stimulation of endogenous NIS expression may permit the radioiodine treatment of extrathyroidal lesions by concentrating this radioisotope. Results This review describes recent findings in NIS research in extrathyroidal malignancies, focusing on breast and urological cancer, emphasizing the most relevant developments that may have clinical impact. Conclusions Given the recent progress in the study of NIS regulation as molecular basis for new therapeutic approaches in extrathyroidal cancers, particular attention is given to studies regarding the relationship between NIS and clinical-pathological aspects of the tumors and the regulation of NIS expression in the experimental models.
Decreased Expression of Thyroglobulin and Sodium Iodide Symporter Genes in Hashimoto's Thyroiditis
Aim. The aim of the study was to compare the expression of sodium iodide symporter (NIS), thyroglobulin (Tg), tumor necrosis factor-? (TNF?), and interleukin-1? genes in patients with Hashimoto's thyroiditis (HT) and healthy individuals. Subjects and Methods. Thyroid cells were obtained from 39 patients with HT and 15 controls by an ultrasound guided fine needle aspiration biopsy. Results. The patients with HT had lower Tg and NIS mRNA (P = 0.002 and P = 0.001, resp.), as well as higher TNF? mRNA expression (P = 0.049) than the controls. In the HT group Tg mRNA expression correlated positively with NIS mRNA expression (R = 0.739, P = 0.0001) and thyroid volume (R = 0.465, P = 0.0005), as well as negatively with TNF? mRNA expression (R = ?0.490, P = 0.001) and anti-peroxidase antibodies (TPOAb) level (R = ?0.482, P = 0.0002), whereas NIS mRNA expression correlated positively with thyroid volume (R = 0.319, P = 0.02), as well as negatively with TNF? mRNA expression (R = ?0.529, P = 0.0006) and TPOAb level (R = ?0.422, P = 0.001). Conclusions. Our results suggest that decreased Tg and NIS expression in thyroid cells may result in reduced active iodide transport and reduced thyroid volume in patients with HT.
Poplawska-Kita, Anna; Telejko, Beata; Siewko, Katarzyna; Kosciuszko-Zdrodowska, Maria; Wawrusewicz-Kurylonek, Natalia; Kretowski, Adam; Hryniewicka, Justyna; Dzieciol, Janusz; Bauer, Witold; Milewski, Robert; Szelachowska, Malgorzata; Gorska, Maria
MEK signaling modulates sodium iodide symporter at multiple levels and in a paradoxical manner.
The Na(+)/I(-) symporter (NIS)-mediated iodide uptake is the basis for targeted radioiodine ablation of thyroid cancers. However, NIS-mediated radioiodide uptake (RAIU) activity is often reduced in thyroid cancers. As mitogen activated protein kinase (MAPK) signaling pathway is activated in about 70% of papillary thyroid carcinoma, we investigated whether MEK (MAPK kinase) inhibition will restore NIS protein levels and NIS-mediated RAIU activity in RET/PTC oncogene-transformed thyroid cells. We found that MEK inhibitor PD98059 increased NIS protein levels within 30 min of treatment. However, the increase of NIS protein level was not accompanied with an increase in NIS-mediated RAIU activity, particularly at early time points of PD98059 treatment. PD98059 also decreased RAIU activity mediated by exogenous NIS in non-thyroid cells. The transient decrease of RAIU activity by PD98059 in thyroid cells was not due to decreased NIS cell surface level, decreased NIS binding affinity for I(-) , or increased iodide efflux. While PD98059 moderately decreased Na(+)/K(+)-ATPase activity, ouabain titration indicates that the extent of decrease in Na(+)/K(+)-ATPase activity is much greater than the extent of decrease in RAIU activity. Additionally, a decrease of Na(+)/K(+)-ATPase activity was not accompanied with a decrease of biotin uptake activity mediated by Na(+)-dependent multivitamin transporter. Since PD98059 reduced V(max)- I(-) without decreasing NIS cell surface levels, it is most likely that PD98059 decreases the turnover rate of iodide transport with an yet to be identified mechanism. PMID:17639055
Vadysirisack, Douangsone D; Venkateswaran, Anjli; Zhang, Zhaoxia; Jhiang, Sissy M
Expression of Thyrotropin Receptor, Thyroglobulin, Sodium-Iodide Symporter, and Thyroperoxidase by Fibrocytes Depends on AIRE.
Context: CD34(+) fibrocytes, bone marrow-derived progenitor cells, infiltrate orbital connective tissue in thyroid-associated ophthalmopathy, a manifestation of Graves' disease. In the orbit, they become CD34(+) fibroblasts and coexist with native CD34(-) fibroblasts. Fibrocytes have been shown to express TSH receptor and thyroglobulin. Objective: The objective of the study was to determine whether a broader repertoire of thyroid protein expression can be detected in fibrocytes and whether a common factor is responsible. Design/Setting/Participants: Fibrocytes and fibroblasts were collected and analyzed from healthy individuals and those with Graves' disease in an academic clinical practice. Main Outcome Measures: Real-time PCR, Western blot analysis, gene promoter analysis, cell transfections, and flow cytometric cell sorting were performed. Results: We detect two additional thyroid proteins expressed by fibrocytes, namely sodium-iodide symporter and thyroperoxidase. The autoimmune regulator (AIRE) protein appears necessary for this expression. AIRE expression in fibrocytes results from an active AIRE gene promoter and stable AIRE mRNA. Knocking down AIRE with a targeting small interfering RNA reduces the expression of these thyroid proteins in fibrocytes as well as the transcription factors paired box-8 and thyroid transcription factor-1. When compared with an unaffected first-degree relative, levels of these proteins are substantially reduced in fibrocytes from an individual with an inactivating AIRE mutation. Levels of AIRE and the thyroid proteins are lower in orbital fibroblasts from patients with thyroid-associated ophthalmopathy than in fibrocytes. However, when mixed fibroblast populations are sorted into pure CD34(+) and CD34(-) subsets, the levels of these proteins are dramatically increased selectively in CD34(+) fibroblasts. Conclusions: Fibrocytes express four proteins, the aggregate expression of which was previously thought to be restricted to thyroid epithelium. These proteins represent the necessary molecular biosynthetic machinery necessary for thyroid hormone production. Our findings implicate AIRE in the promiscuous expression of thyroid proteins in fibrocytes. PMID:24708100
Fernando, Roshini; Lu, Ying; Atkins, Stephen J; Mester, Tunde; Branham, Kari; Smith, Terry J
Sodium iodide symporter (NIS)-mediated radiovirotherapy of hepatocellular cancer using a conditionally replicating adenovirus.
In this study, we determined the in vitro and in vivo efficacy of sodium iodide symporter (NIS) gene transfer and the therapeutic potential of oncolytic virotherapy combined with radioiodine therapy using a conditionally replicating oncolytic adenovirus. For this purpose, we used a replication-selective adenovirus in which the E1a gene is driven by the mouse alpha-fetoprotein (AFP) promoter and the human NIS gene is inserted in the E3 region (Ad5-E1/AFP-E3/NIS). Human hepatocellular carcinoma cells (HuH7) infected with Ad5-E1/AFP-E3/NIS concentrated radioiodine at a level that was sufficiently high for a therapeutic effect in vitro. In vivo experiments demonstrated that 3 days after intratumoral (i.t.) injection of Ad5-E1/AFP-E3/NIS HuH7 xenograft tumors accumulated approximately 25% ID g(-1) (percentage of the injected dose per gram tumor tissue) (123)I as shown by (123)I gamma camera imaging. A single i.t. injection of Ad5-E1/AFP-E3/NIS (virotherapy) resulted in a significant reduction of tumor growth and prolonged survival, as compared with injection of saline. Combination of oncolytic virotherapy with radioiodine treatment (radiovirotherapy) led to an additional reduction of tumor growth that resulted in markedly improved survival as compared with virotherapy alone. In conclusion, local in vivo NIS gene transfer using a replication-selective oncolytic adenovirus is able to induce a significant therapeutic effect, which can be enhanced by additional (131)I application. PMID:23038026
Gr�nwald, G K; Klutz, K; Willhauck, M J; Schwenk, N; Senekowitsch-Schmidtke, R; Schwaiger, M; Zach, C; G�ke, B; Holm, P S; Spitzweg, C
The Na+/I? symporter (NIS) mediates electroneutral active transport of the environmental pollutant perchlorate
The Na+/I? symporter (NIS) is a key plasma membrane protein that mediates active I? uptake in the thyroid, lactating breast, and other tissues with an electrogenic stoichiometry of 2 Na+ per I?. In the thyroid, NIS-mediated I? uptake is the first step in the biosynthesis of the iodine-containing thyroid hormones, which are essential early in life for proper CNS development. In the lactating breast, NIS mediates the translocation of I? to the milk, thus supplying this essential anion to the nursing newborn. Perchlorate (ClO4?) is a well known competitive inhibitor of NIS. Exposure to food and water contaminated with ClO4? is common in the U.S. population, and the public health impact of such exposure is currently being debated. To date, it is still uncertain whether ClO4? is a NIS blocker or a transported substrate of NIS. Here we show in vitro and in vivo that NIS actively transports ClO4?, including ClO4? translocation to the milk. A simple mathematical fluxes model accurately predicts the effect of ClO4? transport on the rate and extent of I? accumulation. Strikingly, the Na+/ ClO4? transport stoichiometry is electroneutral, uncovering that NIS translocates different substrates with different stoichiometries. That NIS actively concentrates ClO4? in maternal milk suggests that exposure of newborns to high levels of ClO4? may pose a greater health risk than previously acknowledged because ClO4? would thus directly inhibit the newborns' thyroidal I? uptake.
Dohan, Orsolya; Portulano, Carla; Basquin, Cecile; Reyna-Neyra, Andrea; Amzel, L. Mario; Carrasco, Nancy
1014. Adenovirus-Mediated MUC1Driven Expression of the Sodium Iodide Symporter (NIS) in Pancreatic Tumors Provides for In Vivo Radioiodide Imaging and Therapy
Pancreatic cancer is the fourth leading cause of cancer death in the United States. It is highly aggressive with no effective chemotherapy available for metastatic disease. The Sodium Iodide Symporter (NIS) is a transmembrane glycoprotein responsible for uptake of iodide into cells. The presence of NIS in thyroid cells has permitted non-invasive diagnostic imaging and ablation of thyroid tumors using
Roisin M. Dwyer; Elizabeth R. Bergert; Michael K. O'Connor; Sandra J. Gendler; John C. Morris
Neurotransmitter/sodium symporter orthologue LeuT has a single high-affinity substrate site
Neurotransmitter/sodium symporters (NSSs) couple the uptake of neurotransmitter with one or more sodium ions1�3, removing neurotransmitter from the synaptic cleft. NSSs are essential to the function of chemical synapses, are associated with multiple neurological diseases and disorders4, and are the targets of therapeutic and illicit drugs5. LeuT, a prokaryotic orthologue of the NSS family, is a model transporter for understanding the relationships between molecular mechanism and atomic structure in a broad range of sodium-dependent and sodium-independent secondary transporters6�13. At present there is a controversy over whether there are one or two high-affinity substrate binding sites in LeuT. The first-reported crystal structure of LeuT, together with subsequent functional and structural studies, provided direct evidence for a single, high-affinity, centrally located substrate-binding site, defined as the S1 site14,15. Recent binding, flux and molecular simulation studies, however, have been interpreted in terms of a model where there are two high-affinity binding sites: the central, S1, site and a second, the S2 site, located within the extracellular vestibule16. Furthermore, it was proposed that the S1 and S2 sites are allosterically coupled such that occupancy of the S2 site is required for the cytoplasmic release of substrate from the S1 site16. Here we address this controversy by performing direct measurement of substrate binding to wild-type LeuT and to S2 site mutants using isothermal titration calorimetry, equilibrium dialysis and scintillation proximity assays. In addition, we perform uptake experiments to determine whether the proposed allosteric coupling between the putative S2 site and the S1 site manifests itself in the kinetics of substrate flux. We conclude that LeuT harbours a single, centrally located, high-affinity substrate-binding site and that transport is well described by a simple, single-substrate kinetic mechanism.
Piscitelli, Chayne L.; Krishnamurthy, Harini; Gouaux, Eric
The invertebrate B 0 system transporter, D. melanogaster NAT1, has unique d-amino acid affinity and mediates gut and brain functions
The CG3252 gene product, DmNAT1, represents the first Nutrient Amino acid Transporter cloned from Drosophila. It absorbs a broader set of neutral amino acids versus earlier characterized insect NATs and mammalian NATs-B0 system transporters from the Sodium Neurotransmitter symporter Family (SNF, a.k.a. solute carrier family 6, SLC6). In addition to B0-specific l-substrates, DmNAT1 equally or more effectively transports d-amino acids
Melissa M. Miller; Lyudmila B. Popova; Ella A. Meleshkevitch; Philip V. Tran; Dmitri Y. Boudko
Synthesis and evaluation of 3,4-dihydropyrimidin-2(1H)-ones as sodium iodide symporter inhibitors.
The sodium iodide symporter (NIS) is responsible for the accumulation of iodide in the thyroid gland. This transport process is involved in numerous thyroid dysfunctions and is the basis for human contamination in the case of exposure to radioactive iodine species. 4-Aryl-3,4-dihydropyrimidin-2(1H)-ones were recently discovered by high-throughput screening as the first NIS inhibitors. Described herein are the synthesis and evaluation of 115 derivatives with structural modifications at five key positions on the pyrimidone core. This study provides extensive structure-activity relationships for this new class of inhibitors that will serve as a basis for further development of compounds with in?vivo efficacy and adequate pharmacokinetic properties. In addition, the SAR investigation provided a more potent compound, which exhibits an IC(50) value of 3.2?nM in a rat thyroid cell line (FRTL5). PMID:23132843
Lacotte, Pierre; Puente, Celine; Ambroise, Yves
Transport of beta-lactam antibiotics in kidney brush border membrane. Determinants of their affinity for the oligopeptide/H+ symporter.
This study was designed to determine whether beta-lactam antibiotics (cephalosporins and penicillins) are all substrates for the renal oligopeptide/H+ symporter and, if so, whether the transport system discriminates among the numerous beta-lactam antibiotics. We used [3H]glycylglutamine, [3H]cephalexin, and [3H]-ampicillin as probes for the transport of oligopeptides, cephalosporins, and penicillins in kidney brush border membrane vesicles, respectively. Among the beta-lactam antibiotics, only those with an alpha-amino group in the phenylacetamido moiety were found to interact with the oligopeptide/H+ symporter. Aminocephalosporins displayed high affinities (KiS generally < 250 microM), whereas aminopenicillins displayed low affinities (Ki 0.78-3.03 mM). These differences in affinities appeared to be a consequence of conformational features of the substrates, especially the sterical location of the carboxy group. The affinities of aminolactams for the oligopeptide/H+ symporter were, furthermore, related to the hydrophobicity of the phenylglycyl chains and the substituents attached to the thiazolidine and dihydrothiazine ring. In sharp contrast to the uptake of [3H]glycylglutamine and [3H]cephalexin, the uptake of [3H]ampicillin was not dependent on a pH gradient and was inhibited by various beta-lactam antibiotics, whether or not they contained an alpha-amino group. Our data suggest that: (a) the transport of aminocephalosporins is largely mediated by the oligopeptide/H+ symporter, which is highly influenced by the substrate structure; and (b) penicillins are transported by another system, which is less discriminative with respect to substrate structure.
Daniel, H; Adibi, S A
The pan-DAC inhibitor LBH589 is a multi-functional agent in breast cancer cells: cytotoxic drug and inducer of sodium-iodide symporter (NIS)
New drugs with anti-tumor activity, also able to modify the expression of selected molecules, are under evaluation in breast\\u000a cancer which is becoming resistant to conventional treatment, or in metastatic disease. The sodium-iodide symporter (NIS),\\u000a which mediates iodide uptake into thyroid cells, and is the molecular basis of radioiodine imaging and therapy in thyroid\\u000a cancer, is also expressed in a
N. Fortunati; M. G. Catalano; F. Marano; V. Mugoni; M. Pugliese; O. Bosco; F. Mainini; G. Boccuzzi
Epidermal Growth Factor Receptor-targeted 131I-therapy of Liver Cancer Following Systemic Delivery of the Sodium Iodide Symporter Gene
We recently demonstrated tumor-selective iodide uptake and therapeutic efficacy of radioiodine in neuroblastoma tumors after systemic nonviral polyplex-mediated sodium iodide symporter (NIS) gene delivery. In the present study, we used novel polyplexes based on linear polyethylenimine (LPEI), polyethylene glycol (PEG), and the synthetic peptide GE11 as an epidermal growth factor receptor (EGFR)-specific ligand to target a NIS-expressing plasmid to hepatocellular
Kathrin Klutz; David Schaffert; Michael J Willhauck; Geoffrey K Gr�nwald; Rudolf Haase; Nathalie Wunderlich; Christian Zach; Franz J Gildehaus; Reingard Senekowitsch-Schmidtke; Burkhard G�ke; Ernst Wagner; Manfred Ogris; Christine Spitzweg
Expression of cAMP response element-binding protein and sodium iodide symporter in benign non-functioning and malignant thyroid tumours
Objective: Reduced expression or defective targeting of the sodium\\/iodide symporter (NIS) to the cell membrane in thyroid tumours has been reported. The expression of the NIS gene is up-regulated by TSH through the cAMP pathway and the characterization of the promoter region of the rat NIS gene revealed the existence of a degenerate cAMP response element (CRE) sequence. The cAMP-
Paola Luciani; Lisa Buci; Barbara Conforti; Massimo Tonacchera; Patrizia Agretti; Rossella Elisei; Agnese Vivaldi; Federica Cioppi; Giancarlo Biliotti; Giuseppe Manca; Paolo Vitti; Mario Serio; Alessandro Peri
High Prevalence of T354P Sodium\\/Iodide Symporter Gene Mutation in Japanese Patients with Iodide Transport Defect Who Have Heterogeneous Clinical Pictures
A missense and loss of function mutation of the Na1\\/I2 symporter (NIS) gene, T354P (Thr3543 Pro (ACA3 CCA)), was found in the homozygous state in two unrelated Japanese patients with iodide transport defect. In this study we have identified the homozygous T354P NIS germline mutation in seven Japanese patients, including one previously reported, from five unrelated families. No other nu-
Non-invasive imaging of cardiac transgene expression with PET: comparison of the human sodium\\/iodide symporter gene and HSV1-tk as the reporter gene
Purpose: Genes encoding for intracellular en- zymes or transmembrane proteins are suitable as reporters, but may differ in terms of applicability for cardiac im- aging. The aim of this study was to compare the human sodium iodide symporter gene (hNIS) with the herpes simplex virus type 1 thymidine kinase gene (HSV1-tk )a s the reporter gene in non-invasive imaging of
Masao Miyagawa; Martina Anton; Bettina Wagner; Roland Haubner; Michael Souvatzoglou; Bernd Gansbacher; Markus Schwaiger; Frank M. Bengel
The Porcine Sodium\\/Iodide Symporter Gene Exhibits an Uncommon Expression Pattern Related to the Use of Alternative Splice Sites not Present in the Human or Murine Species
The sodium\\/iodide symporter (NIS) is a membrane protein mediating the active transport of iodide into the thyroid gland. NIS, expressed by human, rat, and mouse thyrocytes, is encoded by a single transcript. We identified NIS mRNA spe- cies of 3.5 and 3 kb in porcine thyrocytes. Because porcine thyrocytes in primary culture is a widely used experimental system for thyroid
In vivo Radioiodide Imaging and Treatment of Pancreatic Cancer Xenografts after MUC1 Promoter-Driven Expression of the Human Sodium-Iodide Symporter
Introduction and Aims: Mucin 1 (MUC1) is a transmembrane glycoprotein that is overexpressed in many tumor types, including breast, pancreatic, and ovarian cancer. The aim of this study was to create a construct containing sodium-iodide symporter (NIS) under the control of the 0.8-kb MUC1 promoter to infect pancreatic cancer cells both in vitro and in vivo, to investigate the potential
Ru F. Chen; Zhi H. Li; Qiu H. Pan; Jia J. Zhou; Qi B. Tang; Fen Y. Yu; Quan B. Zhou; Jie Wang; Ji S. Chen
Combination Therapy and Noninvasive Imaging with a Dual Therapeutic Vector Expressing MDR1 Short Hairpin RNA and a Sodium Iodide Symporter
We investigated the feasibility of using combination gene therapy and noninvasive nuclear imaging after expression of the human sodium iodide symporter (hNIS) and inhibition of the multidrug resistance (MDR1) gene in colon cancer cells. Methods: HCT- 15 cells were stably transfected with a dual expression vector, in which the hNIS gene, driven by a constitutive cytomegalovirus promoter, has been coupled
Seung-Yoon Park; Wonjung Kwak; Narendra Tapha; Mi-Yeon Jung; Ju-Ock Nam; In-Seop So; So-Youn Kim; Jeongsoo Yoo; Jaetae Lee; In-San Kim
Effective carbon partitioning driven by exotic phloem-specific regulatory elements fused to the Arabidopsis thaliana AtSUC2 sucrose-proton symporter gene
BACKGROUND: AtSUC2 (At1g22710) from Arabidopsis thaliana encodes a phloem-localized sucrose\\/proton symporter required for efficient photoassimilate transport from source tissues to sink tissues. AtSUC2 plays a key role in coordinating the demands of sink tissues with the output capacity of source leaves, and in maintaining phloem hydrostatic pressure during changes in plant-water balance. Expression and activity are regulated, both positively and
Avinash C Srivastava; Savita Ganesan; Ihab O Ismail; Brian G Ayre
Are proton symports in yeast directly linked to H(+)-ATPase acidification?
Transport of amino acids in Saccharomyces cerevisiae is an H(+)-driven secondary active transport. Inhibitors of the plasma membrane H(+)-ATPase, particularly heavy water, diethylstilbestrol and suloctidil, were shown to affect the H(+)-extruding ATPase activity as well as the ATP-hydrolyzing activity, to a similar degree as they inhibited the transport of amino acids. The inhibitors had virtually no effect on the membrane electric potential or on the delta pH which constitute the thermodynamically relevant source of energy for these transports. Transport of acidic amino acids was affected much more than that of the neutral and especially of the basic ones. The effects were greater with higher amino acid concentrations. All this is taken as evidence that the amino acid carriers respond kinetically to the presence of protons directly at the membrane site where they are extruded by the H(+)-ATPase, rather than to the overall protonmotive force. PMID:1347702
Kotyk, A; Dvor�kov�, M
Asn441 plays a key role in folding and function of the Na+/I- symporter (NIS).
The Na(+)/I(-) symporter (NIS) is a plasma membrane glycoprotein that mediates active I(-) transport in the thyroid, the first step in the biosynthesis of the iodine-containing thyroid hormones T3 and T4. Several NIS mutants have been identified as a cause of congenital I(-) transport defect (ITD), and their investigation has yielded valuable mechanistic information on NIS. Here we report a thorough characterization of the ITD-causing NIS mutation in which the sixth intracellular loop residues 439-443 are missing. This mutant protein was intracellularly retained, incompletely glycosylated, and intrinsically inactive. Engineering 5 Ala at positions 439-443 partially recovered cell surface targeting and activity (?15%). Strikingly, NIS with the sequence 439-AANAA-443, in which Asn was restored at position 441, was targeted to the plasma membrane and exhibited ?95% the transport activity of WT NIS. Based on our NIS homology model, we propose that the side chain of N441, a residue conserved throughout most of the SLC5 family, interacts with the main chain amino group of G444, capping the ?-helix of transmembrane segment XII and thus stabilizing the structure of the molecule. Our data provide insight into a critical interhelical interaction required for NIS folding and activity. PMID:23650190
Li, Wenjing; Nicola, Juan Pablo; Amzel, L Mario; Carrasco, Nancy
The Na(+)/I(-) symporter (NIS) is a transmembrane glycoprotein that mediates iodide uptake into thyroid follicular cells and serves as the molecular basis of radioiodine imaging and therapy for thyroid cancer patients. The finding that NIS protein is present in 80-90% of breast tumors suggests that breast cancer patients may also benefit from NIS-mediated radionuclide imaging and targeted therapy. However, only 17-25% of NIS-positive breast tumors have detectable radionuclide uptake activity. The discrepancy between NIS expression and radionuclide uptake activity is most likely contributed by variable cell surface NIS protein levels. Apart from the prevalent view that NIS cell surface trafficking impairments account for the variability, our current study proposes that differential levels of NIS expression may also account for variable cell surface NIS levels among breast tumors. We address the need to confirm the identity of intracellular NIS staining to reveal the mechanisms underlying variable cell surface NIS levels. In addition, we warrant a quantitative correlation between cell surface NIS levels and radionuclide uptake activity in patients such that the cell surface NIS levels required for radionuclide imaging can be defined and the defects impairing NIS activity can be recognized. PMID:18500672
Beyer, S J; Jimenez, R E; Shapiro, C L; Cho, J Y; Jhiang, S M
Targeting of tumor radioiodine therapy by expression of the sodium iodide symporter under control of the survivin promoter.
To test the feasibility of using the survivin promoter to induce specific expression of sodium/iodide symporter (NIS) in cancer cell lines and tumors for targeted use of radionuclide therapy, a recombinant adenovirus, Ad-SUR-NIS, that expressed the NIS gene under control of the survivin promoter was constructed. Ad-SUR-NIS mediating iodide uptake and cytotoxicity was performed in vitro. Scintigraphic, biodistribution and radioiodine therapy studies were performed in vivo. PC-3 (prostate); HepG2 (hepatoma) and A375 (melanoma) cancer cells all exhibited perchlorate-sensitive iodide uptake after infection with Ad-SUR-NIS, approximately 50 times higher than that of negative control Ad-CMV-GFP-infected cells. No significant iodide uptake was observed in normal human dental pulp fibroblast (DPF) cells after infection with Ad-SUR-NIS. Clonogenic assays demonstrated that Ad-SUR-NIS-infected cancer cells were selectively killed by exposure to (131)I. Ad-SUR-NIS-infected tumors show significant radioiodine accumulation (13.3 � 2.85% ID per g at 2 h post-injection), and the effective half-life was 3.1 h. Moreover, infection with Ad-SUR-NIS in combination with (131)I suppressed tumor growth. These results indicate that expression of NIS under control of the survivin promoter can likely be used to achieve cancer-specific expression of NIS in many types of cancers. In combination with radioiodine therapy, this strategy is a possible method of cancer gene therapy. PMID:21037556
Huang, R; Zhao, Z; Ma, X; Li, S; Gong, R; Kuang, A
AtDUR3 encodes a new type of high-affinity urea/H+ symporter in Arabidopsis.
Urea is the major nitrogen form supplied as fertilizer in agricultural plant production but also an important nitrogen metabolite in plants. We report the cloning and functional characterization of AtDUR3, a high-affinity urea transporter in plants. AtDUR3 contains 14 putative transmembrane-spanning domains and represents an individual member in Arabidopsis that belongs to a superfamily of sodium-solute symporters. Heterologous expression in urea uptake-defective yeast as well as two-electrode voltage clamp and uptake studies using (14)C-labeled urea in AtDUR3-expressing oocytes demonstrated that AtDUR3 mediates urea transport. In both heterologous systems, urea transport was stimulated at low pH. In oocytes, inward currents indicated that urea is cotransported with protons. By contrast, a supply of Na(+) ions could not stimulate urea transport. Transport of (14)C-labeled urea by AtDUR3 in oocytes exhibited saturation kinetics with a K(m) of approximately 3 micro M. AtDUR3 was expressed in shoots and roots and upregulated during early germination and under nitrogen deficiency in roots. We propose a role of AtDUR3 in urea uptake by plant cells at low external urea concentrations. PMID:12615950
Liu, Lai-Hua; Ludewig, Uwe; Frommer, Wolf B; von Wir�n, Nicolaus
HSV-NIS, an oncolytic herpes simplex virus type 1 encoding human sodium iodide symporter for preclinical prostate cancer radiovirotherapy.
Several clinical trials have shown that oncolytic herpes simplex virus type 1 (oHSV-1) can be safely administered to patients. However, virus replication in tumor tissue has generally not been monitored in these oHSV clinical trials, and the data suggest that its oncolytic potency needs to be improved. To facilitate noninvasive monitoring of the in vivo spread of an oHSV and to increase its antitumor efficacy, the gene coding for human sodium iodide symporter (NIS) was incorporated into a recombinant oHSV genome and the corresponding virus (oHSV-NIS) rescued in our laboratory. Our data demonstrate that a human prostate cancer cell line, LNCap, efficiently concentrates radioactive iodine after the cells have been infected in vitro or in vivo. In vivo replication of oHSV-NIS in tumors was noninvasively monitored by computed tomography/single-photon emission computed tomography imaging of the biodistribution of pertechnetate and was confirmed. LNCap xenografts in nude mice were eradicated by intratumoral administration of oHSV-NIS. Systemic administration of oHSV-NIS prolonged the survival of tumor-bearing mice, and the therapeutic effect was further enhanced by administration of (131)I after the intratumoral spread of the virus had peaked. oHSV-NIS has the potential to substantially enhance the outcomes of standard therapy for patients with prostate cancer. PMID:23868101
Li, H; Nakashima, H; Decklever, T D; Nace, R A; Russell, S J
A steep radioiodine dose response scalable to humans in sodium-iodide symporter (NIS)-mediated radiovirotherapy for prostate cancer.
The sodium-iodide symporter (NIS) directs the uptake and concentration of iodide in thyroid cells. We have extended the use of NIS-mediated radioiodine therapy to prostate cancer. We have developed a prostate tumor specific conditionally replicating adenovirus that expresses hNIS (Ad5PB_RSV-NIS). For radiovirotherapy to be effective in humans, the radioiodine dose administered in the pre-clinical animal model should scale to the range of acceptable doses in humans. We performed (131)I dose-response experiments aiming to determine the dose required in mice to achieve efficient radiovirotherapy. Efficacy was determined by measuring tumor growth and survival times. We observed that individual tumors display disparate growth rates that preclude averaging within a treatment modality indicating heterogeneity of growth rate. We further show that a statistic and stochastic approach must be used when comparing the effect of an anti-cancer therapy on a cohort of tumors. Radiovirotherapy improves therapeutic value over virotherapy alone by slowing the rate of tumor growth in a more substantial manner leading to an increase in survival time. We also show that the radioiodine doses needed to achieve this increase scaled well within the current doses used for treatment of thyroid cancer in humans. PMID:23037808
Trujillo, M A; Oneal, M J; McDonough, S; Qin, R; Morris, J C
Effective radiovirotherapy for malignant gliomas by using oncolytic measles virus strains encoding the sodium iodide symporter (MV-NIS).
Engineered measles virus (MV) strains deriving from the vaccine lineage represent a promising oncolytic platform and are currently being tested in phase I trials. In this study, we have demonstrated that MV strains genetically engineered to express the human sodium iodide symporter (NIS) have significant antitumor activity against glioma lines and orthotopic xenografts; this compares favorably with the MV strain expressing the human carcinoembryonic antigen, which is currently in clinical testing. Expression of NIS protein in infected cells results in effective concentration of radioactive iodine, which allows for in vivo monitoring of localization of MV-NIS infection by measuring uptake of (123)I or (99m)Tc. In addition, radiovirotherapy with MV-NIS followed by (131)I administration resulted in significant increase of MV-NIS antitumor activity as compared with virus alone in both subcutaneous (p=0.0003) and orthotopic (p=0.004) glioblastoma models. In conclusion, MV-NIS-based radiovirotherapy has significant antitumor activity against glioblastoma multiforme and represents a promising candidate for clinical translation. PMID:22185260
Opyrchal, Mateusz; Allen, Cory; Iankov, Ianko; Aderca, Ileana; Schroeder, Mark; Sarkaria, Jann; Galanis, Evanthia
The Sodium Iodide Symporter (NIS) as an Imaging Reporter for Gene, Viral, and Cell-based Therapies
Preclinical and clinical tomographic imaging systems increasingly are being utilized for non-invasive imaging of reporter gene products to reveal the distribution of molecular therapeutics within living subjects. Reporter gene and probe combinations can be employed to monitor vectors for gene, viral, and cell-based therapies. There are several reporter systems available; however, those employing radionuclides for positron emission tomography (PET) or singlephoton emission computed tomography (SPECT) offer the highest sensitivity and the greatest promise for deep tissue imaging in humans. Within the category of radionuclide reporters, the thyroidal sodium iodide symporter (NIS) has emerged as one of the most promising for preclinical and translational research. NIS has been incorporated into a remarkable variety of viral and non-viral vectors in which its functionality is conveniently determined by in vitro iodide uptake assays prior to live animal imaging. This review on the NIS reporter will focus on 1) differences between endogenous NIS and heterologously-expressed NIS, 2) qualitative or comparative use of NIS as an imaging reporter in preclinical and translational gene therapy, oncolytic viral therapy, and cell trafficking research, and 3) use of NIS as an absolute quantitative reporter.
Penheiter, Alan R; Russell, Stephen J; Carlson, Stephanie K
Proton/l-Glutamate Symport and the Regulation of Intracellular pH in Isolated Mesophyll Cells 1
Addition of l-[U-14C]glutamate to a suspension of mechanically isolated asparagus (Asparagus sprengeri Regel) mesophyll cells results in (a) alkalinization of the medium, (b) uptake of l-[U-14C]glutamate, and (c) efflux of [14C]4-aminobutyrate, a product of glutamate decarboxylation. All three phenomena were eliminated by treatment with 1 millimolar aminooxyacetate. In vitro glutamate decarboxylase (GAD) assays showed that (a) 2 millimolar aminooxyacetate eliminated enzyme activity, (b) activity was pyridoxal phosphate-dependent, and (c) activity exhibited a sharp pH optimum at 6.0 that decreased to 20% of optimal activity at pH 5.0 and 7.0. Addition of 1.5 millimolar sodium butyrate or sodium acetate to cell suspensions caused immediate alkalinization of the medium followed by a resumption of acidification of the medium at a rate approximately double the initial rate. The data indicate that (a) continued H+/l-glutamate contransport is dependent upon GAD activity, (b) the pH-dependent properties of GAD are consistent with a role in a metabolic pH-stat, and (c) the regulation of intracellular pH during H+/l-Glu symport may involve both H+ consumption during 4-aminobutyrate production and ATP-driven H+ efflux.
Snedden, Wayne A.; Chung, Induk; Pauls, Randy H.; Bown, Alan W.
In rat hepatocytes, the hypertonic activation of Na+ conductance and Na+-K+-2Cl? symport - but not Na+-H+ antiport - is mediated by protein kinase C
The initial event in the regulatory volume increase (RVI) of rat hepatocytes is an import of extracellular Na+ via Na+ conductance, Na+-K+-2Cl? symport, and Na+-H+ antiport. Here, the protein kinase inhibitors staurosporine (100 nmol l?1) and bis-indolyl-maleimide I (400 nmol l?1) were used to test for a possible contribution of protein kinase C (PKC) to the hypertonic activation of these transporters in confluent primary cultures. Stimulation of Na+ conductance was monitored: (i) by use of a differential approach based on Na+ fluxes, (ii) by means of cable analysis, and (iii) in experiments with low Na+ pulses. All three experimental protocols in concert demonstrated a block of the activation of Na+ conductance by staurosporine and bis-indolyl-maleimide I. In addition, both compounds significantly reduced the hypertonic activation of Na+-K+-2Cl? symport (quantified on the basis of furosemide-sensitive 86Rb+ uptake) to approximately 30 %. In contrast, neither staurosporine nor bis-indolyl-maleimide I had any detectable effect on the hypertonicity-induced alkalinization of cell pH via Na+-H+ antiport (determined fluorometrically). Staurosporine and bis-indolyl-maleimide I completely blocked the RVI of rat hepatocytes (quantified by means of confocal laser-scanning microscopy). The high efficiency of the block suggests an additional inhibitory effect of both compounds on the activity of Na+/K+-ATPase (determined as ouabain-sensitive 86Rb+ uptake). It is concluded that the hypertonic activation of rat hepatocyte Na+ conductance and Na+-K+-2Cl? symport - but not Na+-H+ antiport - is probably mediated by PKC.
Heinzinger, Heidrun; van den Boom, Frank; Tinel, Hanna; Wehner, Frank
Dietary iodide controls its own absorption through post-transcriptional regulation of the intestinal Na+/I? symporter
Dietary I? absorption in the gastrointestinal tract is the first step in I? metabolism. Given that I? is an essential constituent of the thyroid hormones, its concentrating mechanism is of significant physiological importance. We recently described the expression of the Na+/I? symporter (NIS) on the apical surface of the intestinal epithelium as a central component of the I? absorption system and reported reduced intestinal NIS expression in response to an I?-rich diet in vivo. Here, we evaluated the mechanism involved in the regulation of NIS expression by I? itself in enterocytes. Excess I? reduced NIS-mediated I? uptake in IEC-6 cells in a dose- and time-dependent fashion, which was correlated with a reduction of NIS expression at the plasma membrane. Perchlorate, a competitive inhibitor of NIS, prevented these effects, indicating that an increase in intracellular I? regulates NIS. Iodide induced rapid intracellular recruitment of plasma membrane NIS molecules and NIS protein degradation. Lower NIS mRNA levels were detected in response to I? treatment, although no transcriptional effect was observed. Interestingly, I? decreased NIS mRNA stability, affecting NIS translation. Heterologous green fluorescent protein-based reporter constructs revealed a significant repressive effect of the I?-targeting NIS mRNA 3? untranslated region. In conclusion, excess I? downregulates NIS expression in enterocytes by virtue of a complex mechanism. Our data suggest that I? regulates intestinal NIS mRNA expression at the post-transcriptional level as part of an autoregulatory effect of I? on its own metabolism.
Nicola, Juan Pablo; Reyna-Neyra, Andrea; Carrasco, Nancy; Masini-Repiso, Ana Maria
Dietary iodide controls its own absorption through post-transcriptional regulation of the intestinal Na+/I- symporter.
Dietary I(-) absorption in the gastrointestinal tract is the first step in I(-) metabolism. Given that I(-) is an essential constituent of the thyroid hormones, its concentrating mechanism is of significant physiological importance. We recently described the expression of the Na(+)/I(-) symporter (NIS) on the apical surface of the intestinal epithelium as a central component of the I(-) absorption system and reported reduced intestinal NIS expression in response to an I(-)-rich diet in vivo. Here, we evaluated the mechanism involved in the regulation of NIS expression by I(-) itself in enterocytes. Excess I(-) reduced NIS-mediated I(-) uptake in IEC-6 cells in a dose- and time-dependent fashion, which was correlated with a reduction of NIS expression at the plasma membrane. Perchlorate, a competitive inhibitor of NIS, prevented these effects, indicating that an increase in intracellular I(-) regulates NIS. Iodide induced rapid intracellular recruitment of plasma membrane NIS molecules and NIS protein degradation. Lower NIS mRNA levels were detected in response to I(-) treatment, although no transcriptional effect was observed. Interestingly, I(-) decreased NIS mRNA stability, affecting NIS translation. Heterologous green fluorescent protein-based reporter constructs revealed a significant repressive effect of the I(-)-targeting NIS mRNA 3 untranslated region. In conclusion, excess I(-) downregulates NIS expression in enterocytes by virtue of a complex mechanism. Our data suggest that I(-) regulates intestinal NIS mRNA expression at the post-transcriptional level as part of an autoregulatory effect of I(-) on its own metabolism. PMID:23006481
Comparison of expressed human and mouse sodium/iodide symporters reveals differences in transport properties and subcellular localization.
The active transport of iodide from the bloodstream into thyroid follicular cells is mediated by the Na+/I- symporter (NIS). We studied mouse NIS (mNIS) and found that it catalyzes iodide transport into transfected cells more efficiently than human NIS (hNIS). To further characterize this difference, we compared (125)I uptake in the transiently transfected human embryonic kidney (HEK) 293 cells. We found that the V(max) for mNIS was four times higher than that for hNIS, and that the iodide transport constant (K(m)) was 2.5-fold lower for hNIS than mNIS. We also performed immunocytolocalization studies and observed that the subcellular distribution of the two orthologs differed. While the mouse protein was predominantly found at the plasma membrane, its human ortholog was intracellular in approximately 40% of the expressing cells. Using cell surface protein-labeling assays, we found that the plasma membrane localization frequency of the mouse protein was only 2.5-fold higher than that of the human protein, and therefore cannot alone account for the difference in the obtained V(max) values. We reasoned that the observed difference could also be caused by a higher turnover number for iodide transport in the mouse protein. We then expressed and analyzed chimeric proteins. The data obtained with these constructs suggest that the iodide recognition site could be located in the region extending from the N-terminus to transmembrane domain 8, and that the region between transmembrane domain 5 and the C-terminus could play a role in the subcellular localization of the protein. PMID:18372236
Dayem, Manal; Basquin, C�cile; Navarro, Val�rie; Carrier, Patricia; Marsault, Robert; Chang, Patrick; Huc, Sylvaine; Darrouzet, Elisabeth; Lindenthal, Sabine; Pourcher, Thierry
Differential expression of the Na+/I? symporter protein in thyroid cancer and adjacent normal and nodular goiter tissues
The ability of differentiated thyroid cancer and adjacent thyroid cells to concentrate iodine is dependent on their expression of a functional NA+/I? symporter (NIS). Thyroid cancer is insensitive to 131I treatment if the thyroid cells lack the ability to concentrate iodide. Thus, in this study, we aimed to determine whether the NIS protein was differentially expressed in thyroid cancer and various surrounding tissues. We recruited 114 cases of papillary thyroid carcinoma (PTC) and divided them into two groups: 60 patients of 9 males and 51 females with a mean age of 49.55 years who had PTC with surrounding nodular goiter tissue (simplified as GNG), and 54 patients of 8 males and 46 females with a mean age of 45.78 years who had PTC with surrounding normal tissue (Gnormal) after total or near total thyroidectomy. Formalin-fixed and paraffin-embedded tissue sections were prepared for immunohistochemical staining of the NIS protein and semi-quantitative analysis. The NIS protein was expressed in the basolateral membrane of the normal epithelium, while PTC and nodular goiter cells expressed NIS in the cytoplasm and basolateral membrane. The expression levels of the NIS protein were higher in the adjacent normal tissues compared with those of the surrounding nodular goiter tissues (P=0.002) and expression levels of the NIS protein were higher in PTC tissues compared with the surrounding nodular goiter tissues (P=0.008). The data from this study indicate that cancer-surrounding tissues may play a significant role in mediating the sensitivity of PTC patients to radioactive iodine treatment.
Optimization of Multimodal Imaging of Mesenchymal Stem Cells Using the Human Sodium Iodide Symporter for PET and Cerenkov Luminescence Imaging
Purpose The use of stably integrated reporter gene imaging provides a manner to monitor the in vivo fate of engrafted cells over time in a non-invasive manner. Here, we optimized multimodal imaging (small-animal PET, Cerenkov luminescence imaging (CLI) and bioluminescence imaging (BLI)) of mesenchymal stem cells (MSCs), by means of the human sodium iodide symporter (hNIS) and firefly luciferase (Fluc) as reporters. Methods First, two multicistronic lentiviral vectors (LV) were generated for multimodal imaging: BLI, 124I PET/SPECT and CLI. Expression of the imaging reporter genes was validated in vitro using 99mTcO4? radioligand uptake experiments and BLI. Uptake kinetics, specificity and tracer elution were determined as well as the effect of the transduction process on the cell's differentiation capacity. MSCs expressing the LV were injected intravenously or subcutaneously and imaged using small-animal PET, CLI and BLI. Results The expression of both imaging reporter genes was functional and specific. An elution of 99mTcO4? from the cells was observed, with 31% retention after 3 h. After labeling cells with 124I in vitro, a significantly higher CLI signal was noted in hNIS expressing murine MSCs. Furthermore, it was possible to visualize cells injected intravenously using BLI or subcutaneously in mice, using 124I small-animal PET, CLI and BLI. Conclusions This study identifies hNIS as a suitable reporter gene for molecular imaging with PET and CLI, as confirmed with BLI through the expression of Fluc. It supports the potential for a wider application of hNIS reporter gene imaging and future clinical applications.
Wolfs, Esther; Holvoet, Bryan; Gijsbers, Rik; Casteels, Cindy; Roberts, Scott J.; Struys, Tom; Maris, Michael; Ibrahimi, Abdelilah; Debyser, Zeger; Van Laere, Koen; Verfaillie, Catherine M.; Deroose, Christophe M.
The sodium/iodide symporter NIS is a transcriptional target of the p53-family members in liver cancer cells.
Thyroid iodide accumulation via the sodium/iodide symporter (NIS; SLC5A5) has been the basis for the longtime use of radio-iodide in the diagnosis and treatment of thyroid cancers. NIS is also expressed, but poorly functional, in some non-thyroid human cancers. In particular, it is much more strongly expressed in cholangiocarcinoma (CCA) and hepatocellular carcinoma (HCC) cell lines than in primary human hepatocytes (PHH). The transcription factors and signaling pathways that control NIS overexpression in these cancers is largely unknown. We identified two putative regulatory clusters of p53-responsive elements (p53REs) in the NIS core promoter, and investigated the regulation of NIS transcription by p53-family members in liver cancer cells. NIS promoter activity and endogenous NIS mRNA expression are stimulated by exogenously expressed p53-family members and significantly reduced by member-specific siRNAs. Chromatin immunoprecipitation analysis shows that the p53-REs clusters in the NIS promoter are differentially occupied by the p53-family members to regulate basal and DNA damage-induced NIS transcription. Doxorubicin strongly induces p53 and p73 binding to the NIS promoter, leading to an increased expression of endogenous NIS mRNA and protein in HCC and CCA cells, but not in PHH. Silencing NIS expression reduced doxorubicin-induced apoptosis in HCC cells, pointing to a possible role of a p53-family-dependent expression of NIS in apoptotic cell death. Altogether, these results indicate that the NIS gene is a direct target of the p53 family and suggests that the modulation of NIS by DNA-damaging agents is potentially exploitable to boost NIS upregulation in vivo. PMID:24052075
Guerrieri, F; Piconese, S; Lacoste, C; Schinzari, V; Testoni, B; Valogne, Y; Gerbal-Chaloin, S; Samuel, D; Br�chot, C; Faivre, J; Levrero, M
The sodium/iodide symporter NIS is a transcriptional target of the p53-family members in liver cancer cells
Thyroid iodide accumulation via the sodium/iodide symporter (NIS; SLC5A5) has been the basis for the longtime use of radio-iodide in the diagnosis and treatment of thyroid cancers. NIS is also expressed, but poorly functional, in some non-thyroid human cancers. In particular, it is much more strongly expressed in cholangiocarcinoma (CCA) and hepatocellular carcinoma (HCC) cell lines than in primary human hepatocytes (PHH). The transcription factors and signaling pathways that control NIS overexpression in these cancers is largely unknown. We identified two putative regulatory clusters of p53-responsive elements (p53REs) in the NIS core promoter, and investigated the regulation of NIS transcription by p53-family members in liver cancer cells. NIS promoter activity and endogenous NIS mRNA expression are stimulated by exogenously expressed p53-family members and significantly reduced by member-specific siRNAs. Chromatin immunoprecipitation analysis shows that the p53�REs clusters in the NIS promoter are differentially occupied by the p53-family members to regulate basal and DNA damage-induced NIS transcription. Doxorubicin strongly induces p53 and p73 binding to the NIS promoter, leading to an increased expression of endogenous NIS mRNA and protein in HCC and CCA cells, but not in PHH. Silencing NIS expression reduced doxorubicin-induced apoptosis in HCC cells, pointing to a possible role of a p53-family-dependent expression of NIS in apoptotic cell death. Altogether, these results indicate that the NIS gene is a direct target of the p53 family and suggests that the modulation of NIS by DNA-damaging agents is potentially exploitable to boost NIS upregulation in vivo.
Guerrieri, F; Piconese, S; Lacoste, C; Schinzari, V; Testoni, B; Valogne, Y; Gerbal-Chaloin, S; Samuel, D; Brechot, C; Faivre, J; Levrero, M
Effects of histone acetylation on sodium iodide symporter promoter and expression of thyroid-specific transcription factors.
Inhibitors of histone deacetylases (HDACs) activate the sodium iodide symporter (NIS) expression in thyroid tumor cells. In this study, mechanisms accounting for these effects were investigated. Various human thyroid tumor cell lines (ARO, BCPAP, FRO, TPC-1) were treated with the HDAC inhibitors Na butyrate (NaB) and tricostatin A (TSA), and the effects on the expression of NIS and several thyroid-specific transcription factors together with the activity of NIS promoter were evaluated. TSA and NaB increased NIS mRNA levels in all cell lines. Among thyroid-specific transcription factors, only expression of PAX8 in ARO cells was increased. Down-regulation of thyroid-specific transcription factor-1 expression was observed in BCPAP and TPC-1 cell lines. Thyroid-specific transcription factor-2 mRNA was reduced in FRO, BCPAP, and TPC-1 cells. Histone acetylation had no significant effects on HEX expression. Altogether, these data indicate that the increase of NIS expression is not mediated by modification of expression of thyroid-specific transcription factors. Accordingly, in transfection experiments performed in the HeLa cell line (which does not express thyroid-specific transcription factors), treatment with TSA and NaB increased NIS promoter activity. Stimulation of NIS promoter activity was also obtained by overexpressing histone acetylating proteins pCAF and p300 in HeLa cells. Conversely, overexpression of the HDAC 1 enzyme inhibited basal activity of the NIS promoter. Effects of TSA and NaB on NIS expression were also evaluated in nonthyroid cell lines MCF-7, Hep-G2, and SAOS-2. In all cell lines TSA and NaB greatly increased NIS mRNA levels. We concluded that control of NIS expression by inhibition of HDAC appears not to be mediated by cell-specific mechanisms, suggesting it as a potential strategy to induce radioiodine sensitivity in different human tumors. PMID:15919754
Puppin, Cinzia; D'Aurizio, Federica; D'Elia, Angela V; Cesaratto, Laura; Tell, Gianluca; Russo, Diego; Filetti, Sebastiano; Ferretti, Elisabetta; Tosi, Emanuele; Mattei, Tiziana; Pianta, Annalisa; Pellizzari, Lucia; Damante, Giuseppe
Cellular Bioenergetics is an Important Determinant of the Molecular Imaging Signal Derived from Luciferase and the Sodium-Iodide Symporter
Rationale Molecular imaging is useful for longitudinal assessment of engraftment. However, it is not known which factors, other than cell number can influence the molecular imaging signal obtained from reporter genes. Objective The effects of cell dissociation/suspension on cellular bioenergetics and the signal obtained by firefly luciferase(fluc) and human Na-I symporter(hNIS) labeling of cardiosphere-derived cells (CDCs) was investigated. Methods and Results 18FDG uptake, ATP levels, 99mTc-pertechnetate uptake and bioluminescence were measured in vitro, in adherent and suspended CDCs. In vivo dual isotope SPECT-CT imaging or bioluminescence imaging (BLI) were performed 1hr and 24hrs following CDC transplantation. SPECT quantification was performed using a phantom for signal calibration. Cell loss between 1hr & 24hrs post-transplantation was quantified by qPCR and ex vivo luciferase assay. Cell dissociation followed by suspension for 1hr resulted in decreased glucose uptake, cellular ATP, 99mTc uptake and BLI signal by 82%, 43%, 42%, and 44% respectively, when compared to adherent cells, in vitro. In vivo 99mTc uptake was significantly lower at 1hr, when compared to 24hrs following cell transplantation in the non-infarct (p<0.001, n=3) and infarct (p<0.001, n =4) model, despite significant cell loss during this period. The in vivo BLI signal was significantly higher at 1hr than at 24hrs (p<0.01), with the BLI signal being higher when CDCs were suspended in glucose-containing medium compared to saline(PBS). Conclusion Adhesion is an important determinant of cellular bioenergetics, 99mTc-pertechnetate uptake and BLI signal. BLI and NIS imaging may be useful for in vivo optimization of bioenergetics in transplanted cells.
Chang, Connie; Chan, Angel; Lin, Xiaoping; Higuchi, Takahiro; Terrovitis, John; Afzal, Junaid M.; Rittenbach, Andrew; Sun, Dongdong; Vakrou, Styliani; Woldemichael, Kirubel; O'Rourke, Brian; Wahl, Richard; Pomper, Martin; Tsui, Benjamin; Abraham, M. Roselle
Treatment of medulloblastoma using an oncolytic measles virus encoding the thyroidal sodium iodide symporter shows enhanced efficacy with radioiodine
Background Medulloblastoma is the most common malignant brain tumor of childhood. Although the clinical outcome for medulloblastoma patients has improved significantly, children afflicted with the disease frequently suffer from debilitating side effects related to the aggressive nature of currently available therapy. Alternative means for treating medulloblastoma are desperately needed. We have previously shown that oncolytic measles virus (MV) can selectively target and destroy medulloblastoma tumor cells in localized and disseminated models of the disease. MV-NIS, an oncolytic measles virus that encodes the human thyroidal sodium iodide symporter (NIS), has the potential to deliver targeted radiotherapy to the tumor site and promote a localized bystander effect above and beyond that achieved by MV alone. Methods We evaluated the efficacy of MV-NIS against medulloblastoma cells in vitro and examined their ability to incorporate radioiodine at various timepoints, finding peak uptake at 48 hours post infection. The effects of MV-NIS were also evaluated in mouse xenograft models of localized and disseminated medulloblastoma. Athymic nude mice were injected with D283med-Luc medulloblastoma cells in the caudate putamen (localized disease) or right lateral ventricle (disseminated disease) and subsequently treated with MV-NIS. Subsets of these mice were given a dose of 131I at 24, 48 or 72 hours later. Results MV-NIS treatment, both by itself and in combination with 131I, elicited tumor stabilization and regression in the treated mice and significantly extended their survival times. Mice given 131I were found to concentrate radioiodine at the site of their tumor implantations. In addition, mice with localized tumors that were given 131I either 24 or 48 hours after MV-NIS treatment exhibited a significant survival advantage over mice given MV-NIS alone. Conclusions These data suggest MV-NIS plus radioiodine may be a potentially useful therapy for the treatment of medulloblastoma.
Propylthiouracil Increases Sodium/Iodide Symporter Gene Expression and Iodide Uptake in Rat Thyroid Cells in the Absence of TSH
Background Propylthiouracil (PTU) and methimazole (MMI) are drugs that are widely used to treat Graves' disease. Although both exert an antithyroid effect primarily by blocking thyroid peroxidase activity, their molecular structure and other actions are different. We hypothesized that PTU and MMI may have differential effects on thyroid-specific gene expression and function. Methods The effects of PTU and MMI on thyroid-specific gene expression and function were examined in rat thyroid FRTL-5 cells using DNA microarray, reverse transcriptase (RT)�polymerase chain reaction (PCR), real-time PCR, Western blot, immunohistochemistry, and radioiodine uptake studies. Results DNA microarray analysis showed a marked increase in sodium/iodide symporter (NIS) gene expression after PTU treatment, whereas MMI had no effect. RT-PCR and real-time PCR analysis revealed that PTU-induced NIS mRNA levels were comparable to those elicited by thyroid-stimulating hormone (TSH). PTU increased 5?-1880-bp and 5?-1052-bp activity of the rat NIS promoter. While PTU treatment also increased NIS protein levels, the size of the induced protein was smaller than that induced by TSH, and the protein localized predominantly in the cytoplasm rather than the plasma membrane. Accumulation of 125I in FRTL-5 cells was increased by PTU stimulation, but this effect was weaker than that produced by TSH. Conclusions We found that PTU induces NIS expression and iodide uptake in rat thyroid FRTL-5 cells in the absence of TSH. Although PTU and MMI share similar antithyroid activity, their effects on other thyroid functions appear to be quite different, which could affect their therapeutic effectiveness.
Sue, Mariko; Akama, Takeshi; Kawashima, Akira; Nakamura, Hannah; Hara, Takeshi; Tanigawa, Kazunari; Wu, Huhehasi; Yoshihara, Aya; Ishido, Yuko; Hiroi, Naoki; Yoshino, Gen; Kohn, Leonard D.; Ishii, Norihisa
Identification of a mammalian H+-myo-inositol symporter expressed predominantly in the brain
Inositol and its phosphorylated derivatives play a major role in brain function, either as osmolytes, second messengers or regulators of vesicle endo- and exocytosis. Here we describe the identification and functional characterization of a novel H+-myo- inositol co-transporter, HMIT, expressed predominantly in the brain. HMIT cDNA encodes a 618 amino acid polypeptide with 12 predicted transmembrane domains. Functional expression of HMIT in Xenopus oocytes showed that transport activity was specific for myo-inositol and related stereoisomers with a Michaelis�Menten constant of ?100��M, and that transport activity was strongly stimulated by decreasing pH. Electrophysiological measurements revealed that transport was electrogenic with a maximal transport activity reached at pH�5.0. In rat brain membrane preparations, HMIT appeared as a 75�90�kDa protein that could be converted to a 67�kDa band upon enzymatic deglycosylation. Immunofluorescence microscopy analysis showed HMIT expression in glial cells and some neurons. These data provide the first characterization of a mammalian H+-coupled myo- inositol transporter. Predominant central expression of HMIT suggests that it has a key role in the control of myo-inositol brain metabolism.
Uldry, Marc; Ibberson, Mark; Horisberger, Jean-Daniel; Chatton, Jean-Yves; Riederer, Beat M.; Thorens, Bernard
Theranostic Studies of Human Sodium Iodide Symporter Imaging and Therapy Using 188Re: A Human Glioma Study in Mice
Objective To investigate the role of 188Re in human sodium iodide symporter (hNIS) theranostic gene-mediated human glioma imaging and therapy in model mice. Methods The human glioma cell line U87 was transfected with recombinant lentivirus encoding the hNIS gene under the control of cytomegalovirus promoter (U87-hNIS). The uptake and efflux of 188Re were determined after incubating the cells with 188Re. 188Re uptake experiments in the presence of various concentrations of sodium perchlorate were carried out. In vitro cell killing tests with 188Re were performed. U87-hNIS mediated 188Re distribution, imaging and therapy in nude mice were also tested. Results U87-hNIS cell line was successfully established. The uptake of 188Re in U87-hNIS cells increased up to 26-fold compared to control cells, but was released rapidly with a half-life of approximately 4 minutes. Sodium perchlorate reduced hNIS-mediated 188Re uptake to levels of control cell lines. U87-hNIS cells were selectively killed following exposure to 188Re, with a survival of 21.4%, while control cells had a survival of 92.1%. Unlike in vitro studies, U87-hNIS tumor showed a markedly increased 188Re retention even 48 hours after 188Re injection. In the therapy study, there was a significant difference in tumor size between U87-hNIS mice (317�67 mm3) and control mice (861�153 mm3) treated with 188Re for 4 weeks (P<0.01). Conclusion The results indicate that inserting the hNIS gene into U87 cells is sufficient to induce specific 188Re uptake, which has a cell killing effect both in vitro and in vivo. Moreover, our study, based on the function of hNIS as a theranostic gene allowing noninvasive imaging of hNIS expression by 188Re scintigraphy, provides detailed characterization of in vivo vector biodistribution and level, localization, essential prerequisites for precise planning and monitoring of clinical gene therapy that aims to individualize gene therapy concept.
Guo, Rui; Zhang, M.; Xi, Yun; Ma, Yufei; Liang, Sheng; Shi, Shuo; Miao, Ying; Li, Biao
Stromal targeting of sodium iodide symporter using mesenchymal stem cells allows enhanced imaging and therapy of hepatocellular carcinoma.
The tumor-homing property of mesenchymal stem cells (MSC) has lead to their use as delivery vehicles for therapeutic genes. The application of the sodium iodide symporter (NIS) as therapy gene allows noninvasive imaging of functional transgene expression by (123)I-scintigraphy or PET-imaging, as well as therapeutic application of (131)I or (188)Re. Based on the critical role of the chemokine RANTES (regulated on activation, normal T-cell expressed and presumably secreted)/CCL5 secreted by MSCs in the course of tumor stroma recruitment, use of the RANTES/CCL5 promoter should allow tumor stroma-targeted expression of NIS after MSC-mediated delivery. Using a human hepatocellular cancer (HCC) xenograft mouse model (Huh7), we investigated distribution and tumor recruitment of RANTES-NIS-engineered MSCs after systemic injection by gamma camera imaging. (123)I-scintigraphy revealed active MSC recruitment and CCL5 promoter activation in the tumor stroma of Huh7 xenografts (6.5% ID/g (123)I, biological half-life: 3.7?hr, tumor-absorbed dose: 44.3?mGy/MBq). In comparison, 7% ID/g (188)Re was accumulated in tumors with a biological half-life of 4.1?hr (tumor-absorbed dose: 128.7?mGy/MBq). Administration of a therapeutic dose of (131)I or (188)Re (55.5?MBq) in RANTES-NIS-MSC-treated mice resulted in a significant delay in tumor growth and improved survival without significant differences between (131)I and (188)Re. These data demonstrate successful stromal targeting of NIS in HCC tumors by selective recruitment of NIS-expressing MSCs and by use of the RANTES/CCL5 promoter. The resulting tumor-selective radionuclide accumulation was high enough for a therapeutic effect of (131)I and (188)Re opening the exciting prospect of NIS-mediated radionuclide therapy of metastatic cancer using genetically engineered MSCs as gene delivery vehicles. PMID:23402366
Knoop, Kerstin; Schwenk, Nathalie; Dolp, Patrick; Willhauck, Michael J; Zischek, Christoph; Zach, Christian; Hacker, Markus; G�ke, Burkhard; Wagner, Ernst; Nelson, Peter J; Spitzweg, Christine
The nucleobase cation symporter 1 of Chlamydomonas reinhardtii and that of the evolutionarily distant Arabidopsis thaliana display parallel function and establish a plant-specific solute transport profile.
The single cell alga Chlamydomonas reinhardtii is capable of importing purines as nitrogen sources. An analysis of the annotated C.�reinhardtii genome reveals at least three distinct gene families encoding for known nucleobase transporters. In this study the solute transport and binding properties for the lone C.�reinhardtii nucleobase cation symporter 1 (CrNCS1) are determined through heterologous expression in Saccharomyces cerevisiae. CrNCS1 acts as a transporter of adenine, guanine, uracil and allantoin, sharing similar - but not identical - solute recognition specificity with the evolutionary distant NCS1 from Arabidopsis thaliana. The results suggest that the solute specificity for plant NCS1 occurred early in plant evolution and are distinct from solute transport specificities of single cell fungal NCS1 proteins. PMID:23770594
Schein, Jessica R; Hunt, Kevin A; Minton, Janet A; Schultes, Neil P; Mourad, George S
Arabidopsis thaliana POLYOL/MONOSACCHARIDE TRANSPORTERS 1 and 2: fructose and xylitol/H+ symporters in pollen and young xylem cells
The genome of Arabidopsis thaliana contains six genes, AtPMT1 to AtPMT6 (Arabidopsis thaliana POLYOL/MONOSACCHARIDE TRANSPORTER 1�6), which form a distinct subfamily within the large family of more than 50 monosaccharide transporter-like (MST-like) genes. So far, only AtPMT5 [formerly named AtPLT5 (At3g18830)] has been characterized and was shown to be a plasma membrane-localized H+-symporter with broad substrate specificity. The characterization of AtPMT1 (At2g16120) and AtPMT2 (At2g16130), two other, almost identical, members of this transporter subfamily, are presented here. Expression of the AtPMT1 and AtPMT2 cDNAs in baker's yeast (Saccharomyces cerevisiae) revealed that these proteins catalyse the energy-dependent, high-capacity transport of fructose and xylitol, and the transport of several other compounds with lower rates. Expression of their cRNAs in Xenopus laevis oocytes showed that both proteins are voltage-dependent and catalyse the symport of their substrates with protons. Fusions of AtPMT1 or AtPMT2 with the green fluorescent protein (GFP) localized to Arabidopsis plasma membranes. Analyses of reporter genes performed with AtPMT1 or AtPMT2 promoter sequences showed expression in mature (AtPMT2) or germinating (AtPMT1) pollen grains, as well as in growing pollen tubes, hydathodes, and young xylem cells (both genes). The expression was confirmed with an anti-AtPMT1/AtPMT2 antiserum (?AtPMT1/2) raised against peptides conserved in AtPMT1 and AtPMT2. The physiological roles of the proteins are discussed and related to plant cell wall modifications.
Klepek, Yvonne-Simone; Konrad, Kai R.; Wippel, Kathrin; Hoth, Stefan; Hedrich, Rainer; Sauer, Norbert
Induction of amino acid transporters expression by endurance exercise in rat skeletal muscle
Highlights: �Regulation of amino acid transporter expression in working muscle remains unclear. �Expression of amino acid transporters for leucine were induced by a bout of exercise. �Requirement of leucine in muscle cells might regulate expression of its transporters. �This information is beneficial for understanding the muscle remodeling by exercise. -- Abstract: We here investigated whether an acute bout of endurance exercise would induce the expression of amino acid transporters that regulate leucine transport across plasma and lysosomal membranes in rat skeletal muscle. Rats ran on a motor-driven treadmill at a speed of 28 m/min for 90 min. Immediately after the exercise, we observed that expression of mRNAs encoding L-type amino acid transporter 1 (LAT1) and CD98 was induced in the gastrocnemius, soleus, and extensor digitorum longus (EDL) muscles. Sodium-coupled neutral amino acid transporter 2 (SNAT2) mRNA was also induced by the exercise in those three muscles. Expression of proton-assisted amino acid transporter 1 (PAT1) mRNA was slightly but not significantly induced by a single bout of exercise in soleus and EDL muscles. Exercise-induced mRNA expression of these amino acid transporters appeared to be attenuated by repeated bouts of the exercise. These results suggested that the expression of amino acid transporters for leucine may be induced in response to an increase in the requirement for this amino acid in the cells of working skeletal muscles.
Murakami, Taro, E-mail: email@example.com; Yoshinaga, Mariko
The Paired-Domain Transcription Factor Pax8 Binds to the Upstream Enhancer of the Rat Sodium\\/Iodide Symporter Gene and Participates in Both Thyroid-Specific and Cyclic-AMP-Dependent Transcription
The gene encoding the Na\\/I symporter (NIS) is expressed at high levels only in thyroid follicular cells, where its expression is regulated by the thyroid-stimulating hormone via the second messenger, cyclic AMP (cAMP). In this study, we demonstrate the presence of an enhancer that is located between nucleotides 22264 and 22495 in the 5*-flanking region of the NIS gene and
The CRE-Like Element Inside the 5'Upstream Region of the Rat Sodium\\/Iodide Symporter Gene Interacts with Diverse Classes of b-Zip Molecules that Regulate Transcriptional Activities through Strong Synergy with Pax8
We previously demonstrated that transcription of the rat sodium\\/iodide symporter (NIS) gene is reg- ulated by NUE, an upstream enhancer located be- tween nucleotides 2264 and 2495 of the 5- flanking region. To elucidate the mechanism of TSH\\/cAMP-mediated regulation of NIS gene ex- pression, we have characterized the putative cAMP response element (CRE)\\/activator protein (AP)-1 site (termed NUC) that is
J. T. Chun; V. DI DATO; B. D'ANDREA; M. ZANNINI; R. DI LAURO
The promoter of the Arabidopsis thaliana SUC2 sucrose-H+ symporter gene directs expression of ?-glucuronidase to the phloem: Evidence for phloem loading and unloading by SUC2
The Arabidopsis thaliana (L.) Heynh. SUC2 gene encodes a plasma-membrane sucrose-H+ symporter. The DNA sequence of the SUC2 promoter has been determined. Using a translational fusion of this promoter to the N-terminus of �-glucuronidase (GUS) and the GUS histochemical assay, the tissue specificity of the SUC2 promoter was studied in Arabidopsis plants transformed with this fusion construct. The SUC2 promoter
Elisabeth Truernit; Norbert Sauer
Functional integration of the HUP1 hexose symporter gene into the genome of C. reinhardtii: Impacts on biological H(2) production.
Phototrophic organisms use photosynthesis to convert solar energy into chemical energy. In nature, the chemical energy is stored in a diverse range of biopolymers. These sunlight-derived, energy-rich biopolymers can be converted into environmentally clean and CO(2) neutral fuels. A select group of photosynthetic microorganisms have developed the ability to extract and divert protons and electrons derived from water to chloroplast hydrogenase(s) to produce molecular H(2) fuel. Here, we describe the development and characterization of C. reinhardtii strains, derived from the high H(2) production mutant Stm6, into which the HUP1 (hexose uptake protein) hexose symporter from Chlorella kessleri was introduced. The isolated cell lines can use externally supplied glucose for heterotrophic growth in the dark. More importantly, external glucose supply (1mM) was shown to increase the H(2) production capacity in strain Stm6Glc4 to approximately 150% of that of the high-H(2) producing strain, Stm6. This establishes the foundations for a new fuel production process in which H(2)O and glucose can simultaneously be used for H(2) production. It also opens new perspectives on future strategies for improving bio-H(2) production efficiency under natural day/night regimes and for using sugar waste material for energy production in green algae as photosynthetic catalysts. PMID:17624461
Doebbe, Anja; Rupprecht, Jens; Beckmann, Julia; Mussgnug, Jan H; Hallmann, Armin; Hankamer, Ben; Kruse, Olaf
Novel, missense and loss-of-function mutations in the sodium/iodide symporter gene causing iodide transport defect in three Japanese patients.
Iodide transport defect is a disorder affecting the active transport of iodide, an essential step in the synthesis of thyroid hormones. We have identified novel germ-line mutations in the Na+/I- symporter (NIS) gene from three Japanese patients with iodide transport defect. One patient had a compound heterozygous mutation of T354P/G93R (Gly93-->Arg [GGC-->CGC]), and two sibling patients had a homozygous mutation of G543E (Gly543-->Glu [GGA-->GGA]). G93R and G543E, two novel mutations, are located in the 3rd and 12th transmembrane domains of NIS which are encoded by exons 1 and 13, respectively. The NIS mutants carrying these mutations had minimal iodide uptake activity when expressed in COS-7 cells, confirming that the identified mutations are the direct cause of the iodide transport defect in these patients. Genotyping of unaffected family members and functional assays of co-transfected COS-7 cells indicate that expression of one normal NIS allele in the heterozygote (T354P, G93R, or G543E) is sufficient to maintain active iodide uptake activity. Thus, none of these NIS mutants acts as a dominant-negative mutant. PMID:9745458
Kosugi, S; Inoue, S; Matsuda, A; Jhiang, S M
AMP-Activated Protein Kinase Activation Leads to Lysome-Mediated NA+/I--Symporter Protein Degradation in Rat Thyroid Cells.
Iodide uptake by thyroid cells is mediated by a transmembrane glycoprotein known as the Na+/I--symporter (NIS). NIS-mediated iodide uptake plays important physiological role in thyroid gland function, as well as in diagnostic and treatment of Graves' disease and thyroid cancer. Although different studies investigated the transcriptional mechanisms of NIS expression, there is no report on the NIS post-translational regulation related to NIS protein degradation in thyroid cells. Recently, our group showed that AMP-activated protein kinase (AMPK) plays a pivotal role in the rat thyroid gland, downregulating iodide uptake, NIS protein, and mRNA content. Since several studies demonstrated that AMPK regulates post-transcriptional mechanisms, such as autophagy-mediated processes in different tissues, we hypothesized that AMPK activation could also regulate NIS protein degradation through the lysosome pathway in thyroid cells. Rat follicular thyroid PCCL3 cells cultivated in Ham's F12 supplemented with 5% calf serum and hormones were exposed to the AMPK pharmacological activator 5-aminoimidazole-4 carboxamide ribonucleoside (AICAR), in the presence or absence of Bafilomycin A1 or MG132 for 24?h. Treatment of PCCL3 cells with Bafilomycin A1 fully prevented the decrease of iodide uptake and NIS protein content mediated by AMPK activation. In contrast, the treatment with MG132 was unable to prevent the effects of AMPK activation on NIS. Our results show that AMPK activation significantly induces NIS protein degradation through a lysosome-mediated mechanism. PMID:24691731
Cazarin, J M; Andrade, B M; Carvalho, D P
Switching from MAPK-dependent to MAPK-independent repression of the sodium-iodide symporter in 2D and 3D cultured normal thyroid cells.
Loss of sodium-iodide symporter (NIS) expression in thyroid tumour cells primarily caused by constitutive MAPK pathway activation is often refractory to small molecule MAPK inhibitors. Suggested mechanisms are rebound MAPK signalling and activation of alternative signalling pathways. Here we provide evidence that failure to recover down-regulated NIS by MEK inhibition is not specific to tumour cells. NIS mRNA levels remained repressed in TSH-stimulated primary thyroid cells co-treated with epidermal growth factor (EGF) and pan-MEK inhibitor U0126 in the presence of 5% fetal bovine serum or, independently of serum, in 3D cultured thyroid follicles. This led to inhibited iodide transport and iodination. In contrast, U0126 restituted thyroglobulin synthesis in EGF-treated follicular cells. Serum potentiated TSH-stimulated NIS expression in 2D culture. U0126 blocked down-regulation of NIS only in serum-starved cells with a diminished TSH response. Together, this suggests that morphogenetic signals modify the expression of NIS and recovery response to MEK inhibition. PMID:23969277
Ingeson-Carlsson, Camilla; Nilsson, Mikael
Saccharomyces cerevisiae glycerol/H+ symporter Stl1p is essential for cold/near-freeze and freeze stress adaptation. A simple recipe with high biotechnological potential is given
Background Freezing is an increasingly important means of preservation and storage of microbial strains used for many types of industrial applications including food processing. However, the yeast mechanisms of tolerance and sensitivity to freeze or near-freeze stress are still poorly understood. More knowledge on this regard would improve their biotechnological potential. Glycerol, in particular intracellular glycerol, has been assigned as a cryoprotectant, also important for cold/near-freeze stress adaptation. The S. cerevisiae glycerol active transporter Stl1p plays an important role on the fast accumulation of glycerol. This gene is expressed under gluconeogenic conditions, under osmotic shock and stress, as well as under high temperatures. Results We found that cells grown on STL1 induction medium (YPGE) and subjected to cold/near-freeze stress, displayed an extremely high expression of this gene, also visible at glycerol/H+ symporter activity level. Under the same conditions, the strains harbouring this transporter accumulated more than 400 mM glycerol, whereas the glycerol/H+ symporter mutant presented less than 1 mM. Consistently, the strains able to accumulate glycerol survive 25-50% more than the stl1? mutant. Conclusions In this work, we report the contribution of the glycerol/H+ symporter Stl1p for the accumulation and maintenance of glycerol intracellular levels, and consequently cell survival at cold/near-freeze and freeze temperatures. These findings have a high biotechnological impact, as they show that any S. cerevisiae strain already in use can become more resistant to cold/freeze-thaw stress just by simply adding glycerol to the broth. The combination of low temperatures with extracellular glycerol will induce the transporter Stl1p. This solution avoids the use of transgenic strains, in particular in food industry.
Role of Na+ conductance, Na+-H+ exchange, and Na+-K+-2Cl? symport in the regulatory volume increase of rat hepatocytes
In rat hepatocytes under hypertonic stress, the entry of Na+ (which is thereafter exchanged for K+ via Na+-K+-ATPase) plays the key role in regulatory volume increase (RVI).In the present study, the contributions of Na+ conductance, Na+-H+ exchange and Na+-K+-2Cl? symport to this process were quantified in confluent primary cultures by means of intracellular microelectrodes and cable analysis, microfluorometric determinations of cell pH and buffer capacity, and measurements of frusemide (furosemide)/bumetanide-sensitive 86Rb+ uptake, respectively. Osmolarity was increased from 300 to 400 mosmol l?1 by addition of sucrose.The experiments indicate a relative contribution of approximately 4:1:1 to hypertonicity-induced Na+ entry for the above-mentioned transporters and the overall Na+ yield equalled 51 mmol l?1 (10 min)?1.This Na+ gain is in good agreement with the stimulation of Na+ extrusion via Na+-K+-ATPase plus the actual increase in cell Na+, namely 55 mmol l?1 (10 min)?1, as was determined on the basis of ouabain-sensitive 86Rb+ uptake and by means of Na+-sensitive microelectrodes, respectively.The overall increase in Na+ and K+ activity plus the expected concomitant increase in cell Cl? equalled 68 mmol l?1, which fits well with the increase in osmotic activity expected to occur from an initial cell shrinkage to 87.5 % and a RVI to 92.6 % of control, namely 53 mosmol l?1.The prominent role of Na+ conductance in the RVI of rat hepatocytes could be confirmed on the basis of the pharmacological profile of this process, which was characterized by means of confocal laser-scanning microscopy.
Wehner, Frank; Tinel, Hanna
Sodium iodide symporter (NIS)-mediated radionuclide ((131)I, (188)Re) therapy of liver cancer after transcriptionally targeted intratumoral in vivo NIS gene delivery.
We reported the therapeutic efficacy of (131)I in hepatocellular carcinoma (HCC) cells stably expressing the sodium iodide symporter (NIS) under the control of the tumor-specific ?-fetoprotein (AFP) promoter. In the current study we investigated the efficacy of adenovirus-mediated in vivo NIS gene transfer followed by (131)I and (188)Re administration for the treatment of HCC xenografts. We used a replication-deficient adenovirus carrying the human NIS gene linked to the mouse AFP promoter (Ad5-AFP-NIS) for in vitro and in vivo NIS gene transfer. Functional NIS expression was confirmed by in vivo ?-camera imaging, followed by analysis of NIS protein and mRNA expression. Human HCC (HepG2) cells infected with Ad5-AFP-NIS concentrated 50% of the applied activity of (125)I, which was sufficiently high for a therapeutic effect in an in vitro clonogenic assay. Four days after intratumoral injection of Ad5-AFP-NIS (3�10(9) plaque-forming units) HepG2 xenografts accumulated 14.5% injected dose (ID)/g (123)I with an effective half-life of 13?hr (tumor-absorbed dose, 318?mGy/MBq (131)I). In comparison, 9.2% ID/g (188)Re was accumulated in tumors with an effective half-life of 12.8?hr (tumor-absorbed dose, 545?mGy/MBq). After adenovirus-mediated NIS gene transfer in HepG2 xenografts administration of a therapeutic dose of (131)I or (188)Re (55.5?MBq) resulted in a significant delay in tumor growth and improved survival without a significant difference between (188)Re and (131)I. In conclusion, a therapeutic effect of (131)I and (188)Re was demonstrated in HepG2 xenografts after tumor-specific adenovirus-mediated in vivo NIS gene transfer. PMID:21488714
Klutz, Kathrin; Willhauck, Michael J; Wunderlich, Nathalie; Zach, Christian; Anton, Martina; Senekowitsch-Schmidtke, Reingard; G�ke, Burkhard; Spitzweg, Christine
Hypermethylation of a New Distal Sodium/Iodide Symporter (NIS) Enhancer (NDE) Is Associated With Reduced NIS Expression in Thyroid Tumors.
Context: In thyroid tumors, reduced radioiodine uptake is associated with worse patient outcome concomitantly with low sodium/iodide symporter (NIS) mRNA expression. Previous studies showed that CpG-island methylation in the NIS proximal promoter does not correlate with mRNA expression. Objectives: The aim of the study was to identify new CpG-islands within the NIS 5'region and investigate the involvement of their methylation in NIS expression. Design: DNA was obtained from 30 pairs of thyroid samples: tumor (T) and surrounding nontumor (NT) samples. All T samples had reduced NIS mRNA expression compared to NT samples. Main Outcome Measures: Methylation degree was quantified by bisulfite sequencing, and NIS expression by real-time PCR and Western blot. Reporter gene assays were performed to determine CpG-island functionality. Tumor cell cultures were treated with 5-Aza demethylating agent to determine NIS expression, methylation status, and (125)I uptake. Results: We identified a new CpG-island2 with 14 CpG sites, located -2152/-1887 relative to ATG site. CpG-island2 was hypermethylated in T compared to NT samples, in both benign and malignant tumor groups. There was a significant inverse correlation between NIS mRNA expression and degree of CpG-island2 methylation in NT and T samples. This sequence increased the expression of a reporter gene; thus, it was considered a new enhancer. Cell culture treatments with 5-Aza reduced CpG-island2 methylation levels concomitantly with restoration of NIS mRNA and protein expression and (125)I uptake. Conclusions: We identified a new distal enhancer, NIS distal enhancer, that regulates gene expression through DNA methylation. This enhancer is hypermethylated in T compared to NT samples and is associated with decreased NIS expression in tumors. This epigenetic deregulation may be an early event in tumorigenesis. PMID:24432988
Galr�o, Ana Luiza; Camargo, Rosalinda Y; Friguglietti, Celso U; Moraes, Lais; Cerutti, Janete Maria; Serrano-Nascimento, Caroline; Suzuki, Miriam F; Medeiros-Neto, Geraldo; Rubio, Ileana G S
We recently demonstrated tumor-selective iodide uptake and therapeutic efficacy of radioiodine in neuroblastoma tumors after systemic nonviral polyplex-mediated sodium iodide symporter (NIS) gene delivery. In the present study, we used novel polyplexes based on linear polyethylenimine (LPEI), polyethylene glycol (PEG), and the synthetic peptide GE11 as an epidermal growth factor receptor (EGFR)-specific ligand to target a NIS-expressing plasmid to hepatocellular carcinoma (HCC) (HuH7). Incubation of HuH7 cells with LPEI-PEG-GE11/NIS polyplexes resulted in a 22-fold increase in iodide uptake, which was confirmed in other cancer cell lines correlating well with EGFR expression levels. Using 123I-scintigraphy and ex vivo ?-counting, HuH7 xenografts accumulated 6.5�9% injected dose per gram (ID/g) 123I, resulting in a tumor-absorbed dose of 47 mGray/Megabecquerel (mGy/MBq) 131Iodide (131I) after intravenous (i.v.) application of LPEI-PEG-GE11/NIS. No iodide uptake was observed in other tissues. After pretreatment with the EGFR-specific antibody cetuximab, tumoral iodide uptake was markedly reduced confirming the specificity of EGFR-targeted polyplexes. After three or four cycles of polyplex/131I application, a significant delay in tumor growth was observed associated with prolonged survival. These results demonstrate that systemic NIS gene transfer using polyplexes coupled with an EGFR-targeting ligand is capable of inducing tumor-specific iodide uptake, which represents a promising innovative strategy for systemic NIS gene therapy in metastatic cancers.
Klutz, Kathrin; Schaffert, David; Willhauck, Michael J; Grunwald, Geoffrey K; Haase, Rudolf; Wunderlich, Nathalie; Zach, Christian; Gildehaus, Franz J; Senekowitsch-Schmidtke, Reingard; Goke, Burkhard; Wagner, Ernst; Ogris, Manfred; Spitzweg, Christine
A novel role for AMP-kinase in the regulation of the Na+/I--symporter and iodide uptake in the rat thyroid gland.
The aim of this study was to investigate the role of AMP-kinase (AMPK) in the regulation of iodide uptake by the thyroid gland. Iodide uptake was assessed in PCCL3 follicular thyroid cells exposed to the AMPK agonist 5-aminoimidazole-4-carboxamide-ribonucleoside (AICAR), and also in rat thyroid glands 24 h after a single intraperitoneal injection of AICAR. In PCCL3 cells, AICAR-induced AMPK and acetyl-CoA carboxylase (ACC) phosphorylation decreased iodide uptake in a concentration-dependent manner, while the AMPK inhibitor compound C prevented this effect. In the thyroid gland of rats injected with AICAR, AMPK and ACC phosphorylation was increased and iodide uptake was reduced by ~35%. Under conditions of increased AMPK phosphorylation/activation such as TSH deprivation or AICAR treatment, significant reductions in cellular Na(+)/I(-)-symporter (NIS) protein (~41%) and mRNA content (~65%) were observed. The transcriptional (actinomycin D) and translational (cycloheximide) inhibitors, as well as the AMPK inhibitor compound C prevented AICAR-induced reduction of NIS protein content in PCCL3 cells. The presence of TSH in the culture medium reduced AMPK phosphorylation in PCCL3 cells, while inhibition of protein kinase A (PKA) with H89 prevented this effect. Conversely, the adenylyl cyclase activator forskolin abolished the AMPK phosphorylation response induced by TSH withdrawal in PCCL3 cells. These findings demonstrate that TSH suppresses AMPK phosphorylation/activation in a cAMP-PKA-dependent manner. In summary, we provide novel evidence that AMPK is involved in the physiological regulation of iodide uptake, which is an essential step for the formation of thyroid hormones as well as for the regulation of thyroid function. PMID:21389275
Andrade, Bruno M; Araujo, Renata L; Perry, Robert L S; Souza, Elaine C L; Cazarin, Juliana M; Carvalho, Denise P; Ceddia, Rolando B
Single-photon emission computed tomography of spontaneous liver metastasis from orthotopically implanted human colon cancer cell line stably expressing human sodium/iodide symporter reporter gene
Background We aimed to develop a mouse spontaneous liver metastasis model from an orthotopically implanted human colon cancer cell line stably expressing a human sodium/iodide symporter (NIS) reporter gene, which can be imaged with single-photon emission computed tomography (SPECT) using 99mTcO4?. Methods A recombinant plasmid containing a constitutively driven NIS gene (pcDNA3-NIS) was transfected into the human colon cancer cell line HCT116, and stable cell lines were established. The stable cells were subcutaneously injected into the nude mice. When the diameter reached 10?mm, the xenografts were excised, cut into small fragments, and orthotopically implanted into the cecal walls of another nude mice. 99mTcO4? SPECT/CT imaging was initiated 8?weeks later and repeated every 1 to 2?weeks. Results The production and function of NIS protein was confirmed in vitro by Western blotting and 99mTcO4? uptake assay. On SPECT/CT imaging, focal 99mTcO4? uptake was detected in the liver. Necropsy revealed local growth of the orthotopic colon xenografts with extensive invasion, microscopic serosal metastasis, and metastatic foci in the corresponding hepatic regions showing focal 99mTcO4? uptake. Immunohistochemistry revealed high levels of NIS expression in cells forming liver tumor, indicating that the liver tumor cells originated from the orthotopic colon xenografts. Conclusions The present proof-of-concept study provided a rationale for employing a radionuclide reporter gene for the specific visualization of spontaneous liver metastasis in living mice. This unique animal model of clinically relevant and externally detectable liver metastasis will be a powerful tool for investigating tumor biology and developing novel therapies for cancer metastasis.
Combined Cerenkov luminescence and nuclear imaging of radioiodine in the thyroid gland and thyroid cancer cells expressing sodium iodide symporter: initial feasibility study.
Radioiodine (RI) such as (131)I or (124)I, can generate luminescent emission and be detected with an optical imaging (OI) device. To evaluate the possibility of a novel Cerenkov luminescence imaging (CLI) for application in thyroid research, we performed feasibility studies of CLI by RI in the thyroid gland and human anaplastic thyroid carcinoma cells expressing sodium iodide symporter gene (ARO-NIS). For in vitro study, FRTL-5 and ARO-NIS were incubated with RI, and the luminometric and CLI intensity was measured with luminometer and OI device. Luminescence intensity was compared with the radioactivity measured with ?-counter. In vivo CLI of the thyroid gland was performed in mice after intravenous injection of RI with and without thyroid blocking. Mice were implanted with ARO-NIS subcutaneously, and CLI was performed with injection of (124)I. Small animal PET or ?-camera imaging was also performed. CLI intensities of thyroid gland and ARO-NIS were quantified, and compared with the radioactivities measured from nuclear images (NI). Luminometric assay and OI confirmed RI uptake in the cells in a dose-dependent manner, and luminescence intensity was well correlated with radioactivity of the cells. CLI clearly demonstrated RI uptake in thyroid gland and xenografted ARO-NIS cells in mice, which was further confirmed by NI. A strong positive correlation was observed between CLI intensity and radioactivity assessed by NI. We successfully demonstrated dual molecular imaging of CLI and NI using RI both in vitro and in vivo. CLI can provide a new OI strategy in preclinical thyroid studies. PMID:21551958
Jeong, Shin Young; Hwang, Mi-Hye; Kim, Jung Eun; Kang, Sungmin; Park, Jeong Chan; Yoo, Jeongsoo; Ha, Jeoung-Hee; Lee, Sang-Woo; Ahn, Byeong-Cheol; Lee, Jaetae
Monitoring the initial delivery of an oncolytic measles virus encoding the human sodium iodide symporter to solid tumors using contrast-enhanced computed tomography
Background We aimed to determine the feasibility of monitoring viral delivery and initial distribution to solid tumors using iodinated contrast agent and micro-computed tomography (CT). Methods Human BxPC-3 pancreatic tumor xenografts were established in nude mice. An oncolytic measles virus with an additional transcriptional unit encoding the sodium iodide symporter (NIS), as a reporter for viral infection, was mixed with a 1:10 dilution of Omnipaque 300 (GE Healthcare, Milwaukee, WI, USA) contrast agent and injected directly into tumors. Mice were imaged with micro-CT immediately before and after injection to determine the location of contrast agent/virus mixture. Mice were imaged again on day 3 after injection with micro-single-photon emission CT/CT to determine the location of NIS-mediated 99mTcO4 transport. Results A 1:10 dilution of Omnipaque had no effect on viral infectivity or cell viability in vitro and was more than adequate for CT imaging of the intratumoral injectate distribution. The volume of tumor coverage with initial CT contrast agent and the 3-day postinfection measurement of virally infected tumor volume were significantly correlated. Additionally, regions of the tumor that did not receive contrast agent from the initial injection were largely devoid of viral infection at early time points. Conclusions Contrast-enhanced viral delivery enables a rapid and accurate prediction of the initial viral distribution within a solid tumor. This technique should enable real-time monitoring of viral propagation from initially infected tumor regions to adjacent tumor regions.
Penheiter, Alan R.; Dingli, David; Bender, Claire E.; Russell, Stephen J.; Carlson, Stephanie K.
An increase in essential amino acid availability upregulates amino acid transporter expression in human skeletal muscle
Essential amino acids (EAA) stimulate skeletal muscle mammalian target of rapamycin complex 1 (mTORC1) signaling and protein synthesis. It has recently been reported that an increase in amino acid (AA) transporter expression during anabolic conditions is rapamycin-sensitive. The purpose of this study was to determine whether an increase in EAA availability increases AA transporter expression in human skeletal muscle. Muscle biopsies were obtained from the vastus lateralis of seven young adult subjects (3 male, 4 female) before and 1�3 h after EAA ingestion (10 g). Blood and muscle samples were analyzed for leucine kinetics using stable isotopic techniques. Quantitative RT-PCR, and immunoblotting were used to determine the mRNA and protein expression, respectively, of AA transporters and members of the general AA control pathway [general control nonrepressed (GCN2), activating transcription factor (ATF4), and eukaryotic initiation factor (eIF2) ?-subunit (Ser52)]. EAA ingestion increased blood leucine concentration, delivery of leucine to muscle, transport of leucine from blood into muscle, intracellular muscle leucine concentration, ribosomal protein S6 (Ser240/244) phosphorylation, and muscle protein synthesis. This was followed with increased L-type AA transporter (LAT1), CD98, sodium-coupled neutral AA transporter (SNAT2), and proton-coupled amino acid transporter (PAT1) mRNA expression at 1 h (P < 0.05) and modest increases in LAT1 protein expression (3 h post-EAA) and SNAT2 protein expression (2 and 3 h post-EAA, P < 0.05). Although there were no changes in GCN2 expression and eIF2? phosphorylation, ATF4 protein expression reached significance by 2 h post-EAA (P < 0.05). We conclude that an increase in EAA availability upregulates human skeletal muscle AA transporter expression, perhaps in an mTORC1-dependent manner, which may be an adaptive response necessary for improved AA intracellular delivery.
Drummond, Micah J.; Glynn, Erin L.; Fry, Christopher S.; Timmerman, Kyle L.; Volpi, Elena
Gamma-aminobutyric acid (GABA) transport across human intestinal epithelial (Caco-2) cell monolayers
Transintestinal absorption of gamma-aminobutyric acid (GABA) via a pH-dependent mechanism is demonstrated in the model human intestinal epithelial cell line Caco-2. Experiments with BCECF [2?,7?,-bis(2-carboxyethyl)-5(6)-carboxyfluorescein]-loaded Caco-2 cells demonstrate that GABA transport across the apical membrane is coupled to proton flow into the cell. Short-circuit current (ISC) measurements using Caco-2 cell monolayers under voltage-clamped conditions demonstrate that pH-dependent GABA transport is a rheogenic process even in the absence of extracellular Na+, consistent with H+/GABA symport. A range of GABA analogues were tested for their abilities to: (a) inhibit pH-dependent [3H]GABA uptake across the apical membrane; (b) stimulate H+ flow across the apical surface of BCECF-loaded Caco-2 cell monolayers; (c) increase inward ISC across voltage-clamped Caco-2 cell monolayers. Nipecotic acid, isonipecotic acid, D,L-?-aminobutyric acid, and 3-amino-1-propanesulphonic acid each caused a marked acidification of intracellular pH and an increase in ISC when superfused at the apical surface of Caco-2 cell monolayers. In contrast L-?-amino-n-butyric acid failed to induce proton flow or ISC. The ability of these compounds to induce proton or current flow across the apical surface of this intestinal epithelium was closely related to the relative inhibitory effects on [3H]GABA uptake. These observations demonstrate H+/GABA symport and suggest that this transport mechanism may be accessible as a route for oral absorption of therapeutically-useful GABA analogues.
Thwaites, David T; Basterfield, Laura; McCleave, Peter M J; Carter, Simon M; Simmons, Nicholas L
GATMD: ?-Aminobutyric Acid Transporter Mutagenesis Database
Since the cloning of the first ?-aminobutyric acid (GABA) transporter (GAT1; SLC6A1) from rat brain in 1990, more than 50 published studies have provided structure�function information on investigator-designed rat and mouse GAT1 mutants. To date, more than 200 of 599 GAT1 residues have been subjected to mutagenesis experiments by substitution with different amino acids, and the resulting transporter functional properties have significantly advanced our understanding of the mechanism of Na+- and Cl�-coupled GABA transport by this important member of the neurotransmitter:sodium symporter family. Moreover, many studies have addressed the functional consequences of amino acid deletion or insertion at various positions along the primary sequence. The enormity of this growing body of structure�function information has prompted us to develop GABA Transporter Mutagenesis Database (GATMD), a web-accessible, relational database of manually annotated biochemical, functional and pharmacological data reported on GAT1�the most intensely studied GABA transporter isoform. As of the last update of GATMD, 52 GAT1 mutagenesis papers have yielded 3360 experimental records, which collectively contain a total of ?100?000 annotated parameters. Database URL: http://physiology.sci.csupomona.edu/GATMD/
Anderson, Cynthia M.; Kidd, Patrick D.; Eskandari, Sepehr
A new gene, encoding an anion transporter, is mutated in sialic acid storage diseases.
Sialic acid storage diseases (SASD, MIM 269920) are autosomal recessive neurodegenerative disorders that may present as a severe infantile form (ISSD) or a slowly progressive adult form, which is prevalent in Finland (Salla disease). The main symptoms are hypotonia, cerebellar ataxia and mental retardation; visceromegaly and coarse features are also present in infantile cases. Progressive cerebellar atrophy and dysmyelination have been documented by magnetic resonance imaging (ref. 4). Enlarged lysosomes are seen on electron microscopic studies and patients excrete large amounts of free sialic acid in urine. A H+/anionic sugar symporter mechanism for sialic acid and glucuronic acid is impaired in lysosomal membranes from Salla and ISSD patients. The locus for Salla disease was assigned to a region of approximately 200 kb on chromosome 6q14-q15 in a linkage study using Finnish families. Salla disease and ISSD were further shown to be allelic disorders. A physical map with P1 and PAC clones was constructed to cover the 200-kb area flanked by the loci D6S280 and D6S1622, providing the basis for precise physical positioning of the gene. Here we describe a new gene, SLC17A5 (also known as AST), encoding a protein (sialin) with a predicted transport function that belongs to a family of anion/cation symporters (ACS). We found a homozygous SLC17A5 mutation (R39C) in five Finnish patients with Salla disease and six different SLC17A5 mutations in six ISSD patients of different ethnic origins. Our observations suggest that mutations in SLC17A5 are the primary cause of lysosomal sialic acid storage diseases. PMID:10581036
Verheijen, F W; Verbeek, E; Aula, N; Beerens, C E; Havelaar, A C; Joosse, M; Peltonen, L; Aula, P; Galjaard, H; van der Spek, P J; Mancini, G M
Image-Guided Tumor-Selective Radioiodine Therapy of Liver Cancer After Systemic Nonviral Delivery of the Sodium Iodide Symporter Gene
Abstract We reported the induction of tumor-selective iodide uptake and therapeutic efficacy of 131I in a hepatocellular carcinoma (HCC) xenograft mouse model, using novel polyplexes based on linear polyethylenimine (LPEI), shielded by polyethylene glycol (PEG), and coupled with the epidermal growth factor receptor-specific peptide GE11 (LPEI-PEG-GE11). The aim of the current study in the same HCC model was to evaluate the potential of biodegradable nanoparticle vectors based on pseudodendritic oligoamines (G2-HD-OEI) for systemic sodium iodide symporter (NIS) gene delivery and to compare efficiency and tumor specificity with LPEI-PEG-GE11. Transfection of HCC cells with NIS cDNA, using G2-HD-OEI, resulted in a 44-fold increase in iodide uptake in vitro as compared with a 22-fold increase using LPEI-PEG-GE11. After intravenous application of G2-HD-OEI/NIS HCC tumors accumulated 6�11% ID/g 123I (percentage of the injected dose per gram tumor tissue) with an effective half-life of 10?hr (tumor-absorbed dose, 281?mGy/MBq) as measured by 123I scintigraphic gamma camera or single-photon emission computed tomography computed tomography (SPECT CT) imaging, as compared with 6.5�9% ID/g with an effective half-life of only 6?hr (tumor-absorbed dose, 47?mGy/MBq) for LPEI-PEG-GE11. After only two cycles of G2-HD-OEI/NIS/131I application, a significant delay in tumor growth was observed with markedly improved survival. A similar degree of therapeutic efficacy had been observed after four cycles of LPEI-PEG-GE11/131I. These results clearly demonstrate that biodegradable nanoparticles based on OEI-grafted oligoamines show increased efficiency for systemic NIS gene transfer in an HCC model with similar tumor selectivity as compared with LPEI-PEG-GE11, and therefore represent a promising strategy for NIS-mediated radioiodine therapy of HCC.
Klutz, Kathrin; Willhauck, Michael J.; Dohmen, Christian; Wunderlich, Nathalie; Knoop, Kerstin; Zach, Christian; Senekowitsch-Schmidtke, Reingard; Gildehaus, Franz-Josef; Ziegler, Sibylle; Furst, Sebastian; Goke, Burkhard; Wagner, Ernst
Image-guided tumor-selective radioiodine therapy of liver cancer after systemic nonviral delivery of the sodium iodide symporter gene.
We reported the induction of tumor-selective iodide uptake and therapeutic efficacy of (131)I in a hepatocellular carcinoma (HCC) xenograft mouse model, using novel polyplexes based on linear polyethylenimine (LPEI), shielded by polyethylene glycol (PEG), and coupled with the epidermal growth factor receptor-specific peptide GE11 (LPEI-PEG-GE11). The aim of the current study in the same HCC model was to evaluate the potential of biodegradable nanoparticle vectors based on pseudodendritic oligoamines (G2-HD-OEI) for systemic sodium iodide symporter (NIS) gene delivery and to compare efficiency and tumor specificity with LPEI-PEG-GE11. Transfection of HCC cells with NIS cDNA, using G2-HD-OEI, resulted in a 44-fold increase in iodide uptake in vitro as compared with a 22-fold increase using LPEI-PEG-GE11. After intravenous application of G2-HD-OEI/NIS HCC tumors accumulated 6-11% ID/g (123)I (percentage of the injected dose per gram tumor tissue) with an effective half-life of 10?hr (tumor-absorbed dose, 281?mGy/MBq) as measured by (123)I scintigraphic gamma camera or single-photon emission computed tomography computed tomography (SPECT CT) imaging, as compared with 6.5-9% ID/g with an effective half-life of only 6?hr (tumor-absorbed dose, 47?mGy/MBq) for LPEI-PEG-GE11. After only two cycles of G2-HD-OEI/NIS/(131)I application, a significant delay in tumor growth was observed with markedly improved survival. A similar degree of therapeutic efficacy had been observed after four cycles of LPEI-PEG-GE11/(131)I. These results clearly demonstrate that biodegradable nanoparticles based on OEI-grafted oligoamines show increased efficiency for systemic NIS gene transfer in an HCC model with similar tumor selectivity as compared with LPEI-PEG-GE11, and therefore represent a promising strategy for NIS-mediated radioiodine therapy of HCC. PMID:21851208
Klutz, Kathrin; Willhauck, Michael J; Dohmen, Christian; Wunderlich, Nathalie; Knoop, Kerstin; Zach, Christian; Senekowitsch-Schmidtke, Reingard; Gildehaus, Franz-Josef; Ziegler, Sibylle; F�rst, Sebastian; G�ke, Burkhard; Wagner, Ernst; Ogris, Manfred; Spitzweg, Christine
Assessing the ability of sequence-based methods to provide functional insight within membrane integral proteins: a case study analyzing the neurotransmitter/Na+ symporter family
Background Efforts to predict functional sites from globular proteins is increasingly common; however, the most successful of these methods generally require structural insight. Unfortunately, despite several recent technological advances, structural coverage of membrane integral proteins continues to be sparse. ConSequently, sequence-based methods represent an important alternative to illuminate functional roles. In this report, we critically examine the ability of several computational methods to provide functional insight within two specific areas. First, can phylogenomic methods accurately describe the functional diversity across a membrane integral protein family? And second, can sequence-based strategies accurately predict key functional sites? Due to the presence of a recently solved structure and a vast amount of experimental mutagenesis data, the neurotransmitter/Na+ symporter (NSS) family is an ideal model system to assess the quality of our predictions. Results The raw NSS sequence dataset contains 181 sequences, which have been aligned by various methods. The resultant phylogenetic trees always contain six major subfamilies are consistent with the functional diversity across the family. Moreover, in well-represented subfamilies, phylogenetic clustering recapitulates several nuanced functional distinctions. Functional sites are predicted using six different methods (phylogenetic motifs, two methods that identify subfamily-specific positions, and three different conservation scores). A canonical set of 34 functional sites identified by Yamashita et al. within the recently solved LeuTAa structure is used to assess the quality of the predictions, most of which are predicted by the bioinformatic methods. Remarkably, the importance of these sites is largely confirmed by experimental mutagenesis. Furthermore, the collective set of functional site predictions qualitatively clusters along the proposed transport pathway, further demonstrating their utility. Interestingly, the various prediction schemes provide results that are predominantly orthogonal to each other. However, when the methods do provide overlapping results, specificity is shown to increase dramatically (e.g., sites predicted by any three methods have both accuracy and coverage greater than 50%). Conclusion The results presented herein clearly establish the viability of sequence-based bioinformatic strategies to provide functional insight within the NSS family. As such, we expect similar bioinformatic investigations will streamline functional investigations within membrane integral families in the absence of structure.
Livesay, Dennis R; Kidd, Patrick D; Eskandari, Sepehr; Roshan, Usman
Molecular dissection of membrane-transport proteins: mass spectrometry and sequence determination of the galactose-H+ symport protein, GalP, of Escherichia coli and quantitative assay of the incorporation of [ring-2-13C]histidine and (15)NH(3).
The molecular mass of the galactose-H(+) symport protein GalP, as its histidine-tagged derivative GalP(His)(6), has been determined by electrospray MS (ESI-MS) with an error of <0.02%. One methionine residue, predicted to be present from the DNA sequence, was deduced to be absent. This is a significant advance on the estimation of the molecular masses of membrane-transport proteins by SDS/PAGE, where there is a consistent under-estimation of the true molecular mass due to anomalous electrophoretic migration. Addition of a size-exclusion chromatography step after Ni(2+)-nitrilotriacetate affinity purification was essential to obtain GalP(His)(6) suitable for ESI-MS. Controlled trypsin, trypsin+chymotrypsin and CNBr digestion of the protein yielded peptide fragments suitable for ESI-MS and tandem MS analysis, and accurate mass determination of the derived fragments resulted in identification of 82% of the GalP(His)(6) protein. Tandem MS analysis of selected peptides then afforded 49% of the actual amino acid sequence of the protein; the absence of the N-terminal methionine was confirmed. Matrix-assisted laser-desorption ionization MS allowed identification of one peptide that was not detected by ESI-MS. All the protein/peptide mass and sequence determinations were in accord with the predictions of amino acid sequence deduced from the DNA sequence of the galP gene. [ring-2-(13)C]Histidine was incorporated into GalP(His)(6) in vivo, and ESI-MS analysis enabled the measurement of a high (80%) and specific incorporation of label into the histidine residues in the protein. MS could also be used to confirm the labelling of the protein by (15)NH(3) (93% enrichment) and [(19)F]tryptophan (83% enrichment). Such MS measurements will serve in the future analysis of the structures of membrane-transport proteins by NMR, and of their topology by indirect techniques. PMID:11931651
Venter, Henrietta; Ashcroft, Alison E; Keen, Jeffrey N; Henderson, Peter J F; Herbert, Richard B
Increased expression of specific intestinal amino acid and peptide transporter mRNA in rats fed by TPN is reversed by GLP-2.
Intestinal function depends on the presence of luminal nutrients and is altered during starvation and refeeding. Amino acids are essential for enterocytes, but the luminal supply is compromised with changes in dietary intake. To test the hypothesis that during periods of restricted luminal nutrient availability mucosal cells undergo adaptations aimed toward preserving amino acid supply, the expression of amino acid and peptide transporter mRNAs was quantified in rats with no oral intake, whose nutritional status was maintained with total parenteral nutrition (TPN). The role of the intestinotrophic hormone glucagon-like peptide-2 (GLP-2) was investigated in the adaptive responses. Rats were administered TPN with or without exogenous GLP-2. Amino acid and peptide transporter mRNAs in small intestine mucosa were measured by semiquantitative RT-PCR. Compared with orally fed rats, removal of luminal nutrition increased the expression of ASCT1, SAT2, and GLYT1 mRNAs in the duodenum and of ASCT2, EAAC1, NBAT, and PepT1 mRNAs in the ileum. CAT1, PAT1, and SN2 mRNA abundances were unaffected. GLP-2 reversed these effects. Three subgroups of transporters were identified by regional differences in response to TPN. This may reflect differing roles for substrates of transporters located apically and basally and along the proximal-distal axis of the intestine. The importance of maintaining amino acid supply for intestinal mucosal cells is illustrated. PMID:15514259
Howard, Alison; Goodlad, Robert A; Walters, Julian R F; Ford, Dianne; Hirst, Barry H
An Acid-activated Nucleobase Transporter from Leishmania major*
Parasitic protozoa are unable to synthesize purines de novo and must import preformed purine nucleobases or nucleosides from their hosts. Leishmania major expresses two purine nucleobase transporters, LmaNT3 and LmaNT4. Previous studies revealed that at neutral pH, LmaNT3 is a broad specificity, high affinity nucleobase transporter, whereas LmaNT4 mediates the uptake of only adenine. Because LmaNT4 is required for optimal viability of the amastigote stage of the parasite that lives within acidified phagolysomal vesicles of mammalian macrophages, the function of this permease was examined under acidic pH conditions. At acidic pH, LmaNT4 acquires the ability to transport adenine, hypoxanthine, guanine, and xanthine with Km values in the micromolar range, indicating that this transporter is activated at low pH. Thus, LmaNT4 is an acid-activated purine nucleobase transporter that functions optimally under the physiological conditions the parasite is exposed to in the macrophage phagolysosome. In contrast, LmaNT3 functions optimally at neutral pH. Two-electrode voltage clamp experiments performed on LmaNT3 and LmaNT4 expressed in Xenopus oocytes revealed substrate-induced inward directed currents at acidic pH, and application of substrates induced acidification of the oocyte cytosol. These observations imply that LmaNT3 and LmaNT4 are nucleobase/proton symporters.
Ortiz, Diana; Sanchez, Marco A.; Koch, Hans P.; Larsson, H. Peter; Landfear, Scott M.
A Thyroid-Specific Far-Upstream Enhancer in the Human Sodium\\/Iodide Symporter Gene Requires Pax8 Binding and Cyclic Adenosine 3',5'Monophosphate Response Element-Like Sequence Binding Proteins for Full Activity and Is Differentially Regulated in Normal and Thyroid Cancer Cells
The sodium\\/iodide symporter (NIS) gene is highly expressed in the thyroid gland and is important for the diagnosis and radioiodide therapy of differen- tiated thyroid cancers. We investigated a human NIS (hNIS) gene 5-far-upstream enhancer (hNUE) (9847 to 8968). The hNUE is TSH responsive in both FRTL-5 cells and primary normal thyroid cells, but not in human papillary thyroid cancer
Molecular basis of essential amino acid transport from studies of insect nutrient amino acid transporters of the SLC6 family (NAT-SLC6)
Two protein families that represent major components of essential amino acid transport in insects have been identified. They are annotated as the SLC6 and SLC7 families of transporters according to phylogenetic proximity to characterized amino acid transporters (HUGO nomenclature). Members of these families have been identified as important apical and basolateral parts of transepithelial essential amino acid absorption in the metazoan alimentary canal. Synergistically, they play critical physiological roles as essential substrate providers to diverse metabolic processes, including generic protein synthesis. This review briefly clarifies the requirements for amino acid transport and a variety of amino acid transport mechanisms, including the aforementioned families. Further it focuses on the large group of Nutrient Amino acid Transporters (NATs), which comprise a recently identified subfamily of the Neurotransmitter Sodium Symporter family (NSS or SLC6). The first insect NAT, cloned from the caterpillar gut, has a broad substrate spectrum similar to mammalian B0 transporters. Several new NAT-SLC6 members have been characterized in an effort to explore mechanisms for the essential amino acid absorption in model dipteran insects. The identification and functional characterization of new B0-like and narrow specificity transporters of essential amino acids in fruit fly and mosquitoes leads to a fundamentally important insight: that NATs evolved and act together as the integrated active core of a transport network that mediates active alimentary absorption and systemic distribution of essential amino acids. This role of NATs is projected from the most primitive prokaryotes to the most complex metazoan organisms, and represents an interesting platform for unraveling the molecular evolution of amino acid transport and modeling amino acid transport disorders. The comparative study of NATs elucidates important adaptive differences between essential amino acid transportomes of invertebrate and vertebrate organisms, outlining a new possibility for selective targeting of essential amino acid absorption mechanisms to control medically and economically important arthropods and other invertebrate organisms.
Boudko, Dmitri Y.
Molecular basis of essential amino acid transport from studies of insect nutrient amino acid transporters of the SLC6 family (NAT-SLC6).
Two protein families that represent major components of essential amino acid transport in insects have been identified. They are annotated as the SLC6 and SLC7 families of transporters according to phylogenetic proximity to characterized amino acid transporters (HUGO nomenclature). Members of these families have been identified as important apical and basolateral parts of transepithelial essential amino acid absorption in the metazoan alimentary canal. Synergistically, they play critical physiological roles as essential substrate providers to diverse metabolic processes, including generic protein synthesis. This review briefly clarifies the requirements for amino acid transport and a variety of amino acid transport mechanisms, including the aforementioned families. Further it focuses on the large group of Nutrient Amino acid Transporters (NATs), which comprise a recently identified subfamily of the Neurotransmitter Sodium Symporter family (NSS or SLC6). The first insect NAT, cloned from the caterpillar gut, has a broad substrate spectrum similar to mammalian B(0) transporters. Several new NAT-SLC6 members have been characterized in an effort to explore mechanisms for the essential amino acid absorption in model dipteran insects. The identification and functional characterization of new B(0)-like and narrow specificity transporters of essential amino acids in fruit fly and mosquitoes leads to a fundamentally important insight: that NATs evolved and act together as the integrated active core of a transport network that mediates active alimentary absorption and systemic distribution of essential amino acids. This role of NATs is projected from the most primitive prokaryotes to the most complex metazoan organisms, and represents an interesting platform for unraveling the molecular evolution of amino acid transport and modeling amino acid transport disorders. The comparative study of NATs elucidates important adaptive differences between essential amino acid transportomes of invertebrate and vertebrate organisms, outlining a new possibility for selective targeting of essential amino acid absorption mechanisms to control medically and economically important arthropods and other invertebrate organisms. PMID:22230793
Boudko, Dmitri Y
Functional Radionuclide Imaging, In-Vitro Radioiodine Uptake Estimation and RT-PCR in the Evaluation of Sodium Iodide Symporter (NIS) Expression and Functionality in Breast Cancer: A Pilot Study.
Breast cancer is a common malignancy in females, which is considered as a systemic disease, whose treatment involves combined modality including systemic as well as local treatment. Recent studies have shown that breast cancer also expresses Sodium Iodide Symporter (NIS) gene, like in the thyroid, which is the factor responsible for the uptake of iodide by the thyroid, enabling radioiodine therapy of thyroid disorders. This study aimed to evaluate various radionuclide imaging characteristics, in vitro radioiodine uptake (RAIU) and evaluation of NIS expression by using Reverse Transcriptase-Polymerase Chain Reaction (RT-PCR) to explore sodium iodide symporter (NIS) expression and iodine uptake in breast cancer and to explor e whether radioiodine can be used for the diagnosis and treatment of breast cancer. Ways of differential regulation of NIS expression in breast cancer has also been explored. Female patients with palpable breast lump and histologically proven infiltrating duct carcinoma were taken up for the study, which included 50 females of mean age 49�years. (range: 23-73�years). The patients were categorized into different groups, depending on the type of the study performed. The uptake patterns in various imaging modalities were analyzed and compared with invitro and RT-PCR studies. 68�% of breast cancer cases showed (99m)Tc-pertechnetate uptake at the initial images. This finding could partly be due to tumor vascularity, which is usually higher compared to the normal tissues. The uptake in the delayed imaging could be related to that due to NIS in the breast. Use of perchlorate or stable iodine did not alter the pertechnetate uptake pattern in breast tumor. Good correlation between (99m)Tc-pertechnetate and (99m)Tc-tetrofosmin uptake in breast cancer was demonstrated. In vitro radioactive iodine uptake in the breast tumor was significantly higher than that in the normal breast tissue. Only 42�% of breast tumor samples studied using RT-PCR showed NIS expression. Correlation between (99m)Tc-pertechnetate uptake and NIS expression could not be well established. Further studies with higher dose of radioiodine and/or mechanisms of differentially blocking the thyroid are required to assess the feasibility of radioiodine therapy for breast cancer. PMID:24426705
Joseph, J K; Patel, R B; Damle, A A; Nair, N; Badwe, R A; Basu, S
Preclinical efficacy of the oncolytic measles virus expressing the sodium iodide symporter in iodine non-avid anaplastic thyroid cancer: a novel therapeutic agent allowing noninvasive imaging and radioiodine therapy.
Anaplastic thyroid cancer is an extremely aggressive disease resistant to radioiodine treatment because of loss of sodium iodide symporter (NIS) expression. To enhance prognosis of this fatal cancer, we validated the preclinical efficacy of measles virus (MV)-NIS, the vaccine strain of the oncolytic MV (MV-Edm), modified to include the NIS gene. Western blotting analysis confirmed that a panel of eight anaplastic thyroid cancer (ATC)-derived cell lines do not express NIS protein, but do express CD46, the MV receptor. In vitro cell death assays and in vivo xenograft studies demonstrate the oncolytic efficacy of MV-NIS in BHT-101 and KTC-3, ATC-derived cell lines. Radioactive iodine uptake along with single-photon emission computed tomography (SPECT)-computed tomography imaging of KTC-3 xenografts after (99)Tc(m) administration confirmed NIS expression in vitro and in vivo, respectively, after virus treatment. Adjuvant administration of RAI, to MV-NIS-treated KTC-3 tumors showed a trend for increased tumor cell killing. As current treatment for ATC is only palliative, and MV-NIS is currently Food and Drug Administration approved for human clinical trials in myeloma, our data indicate that targeting ATC with MV-NIS could prove to be a novel therapeutic strategy for effective treatment of iodine-resistant ATC and will expedite its testing in clinical trials for this aggressive disease. PMID:22790962
Reddi, H V; Madde, P; McDonough, S J; Trujillo, M A; Morris, J C; Myers, R M; Peng, K W; Russell, S J; McIver, B; Eberhardt, N L
Transcriptional repression by glycerol of genes involved in the assimilation of n-alkanes and fatty acids in yeast Yarrowia lipolytica.
The yeast Yarrowia lipolytica assimilates n-alkanes or fatty acids as carbon sources. Transcriptional activation by n-alkanes of ALK1 encoding a cytochrome P450 responsible for the terminal hydroxylation has been well studied so far, but its regulation by other carbon sources is poorly understood. Here, we analyzed the transcriptional regulation of ALK1 by glycerol. Glycerol is a preferable carbon source compared to glucose for Y.�lipolytica. The n-decane-induced transcript levels of ALK1 as well as the reporter gene under the control of ALK1 promoter were significantly decreased in the simultaneous presence of glycerol, but not of glucose. Similarly, the expression of PAT1 encoding acetoacetyl-CoA thiolase involved in ?-oxidation was induced by n-decane or oleic acid, but its transcript level was decreased when glycerol was supplemented. These results indicate that glycerol represses the transcription of the genes involved in the metabolism of hydrophobic carbon sources in Y.�lipolytica. Repression of ALK1 transcription by glycerol was not observed in the deletion mutant of GUT1 encoding glycerol kinase, implying that the phosphorylation of glycerol is required for the glycerol repression. PMID:23241327
Mori, Katsuki; Iwama, Ryo; Kobayashi, Satoshi; Horiuchi, Hiroyuki; Fukuda, Ryouichi; Ohta, Akinori
Human sodium-iodide symporter (hNIS) gene expression is inhibited by a trans-active transcriptional repressor, NIS-repressor, containing PARP-1 in thyroid cancer cells.
Radioiodine remains the only tumoricidal therapy for disseminated thyroid carcinomas; however, dedifferentiated tumors lose the expression of human sodium-iodide symporter (hNIS) gene, and cannot respond to this treatment. Previous studies suggested that a trans-active protein factor (NIS-repressor) represses endogenous hNIS transcription, likely contributing to the loss of radioiodine uptake, and defined the NIS-repressor binding site (NRBS) in the proximal hNIS promoter. Using electrophoretic mobility shift assay (EMSA), we found evidence of NIS-repressor in the nuclear extract from KAK-1 cells, and confirmed this result using nuclear extracts prepared from multiple verified thyroid cell lines. Luciferase reporter assays of hNIS promoter constructs and EMSA were used to define two core sequences, NRBS-P and NRBS-D, in the hNIS promoter as the binding sites for NIS-repressor. Electrophoretic analysis of KAK-1 nuclear extract proteins cross-linked with NRBS-P suggests that NIS-repressor is a protein complex. Analysis of KAK-1 nuclear extract proteins bound to NRBS-P, via liquid chromatography coupled with tandem mass spectroscopy, demonstrated poly(ADP-ribose) polymerase-1 (PARP-1) as a NIS-repressor component. Pharmacological inhibition of PARP-1 enzymatic activity using PJ34 stimulated both the luciferase reporter activity driven by hNIS promoter and the endogenous hNIS mRNA level. Supershift studies suggest that thyroid transcription factor 2 (TTF-2) is also associated with the NIS-repressor complex. NIS-repressor, including its PARP-1 component, presents a potential therapeutic target to restore radioiodine uptake in dedifferentiated thyroid carcinomas. PMID:20228127
Li, Wei; Ain, Kenneth B
Amino acids
Amino acids are organic compounds that combine to form proteins . Amino acids and proteins are the building blocks of life. When proteins are digested or broken down, amino acids are left. The human body uses amino acids ...
Insulin increases mRNA abundance of the amino acid transporter SLC7A5/LAT1 via an mTORC1-dependent mechanism in skeletal muscle cells.
Abstract Amino acid transporters (AATs) provide a link between amino acid availability and mammalian/mechanistic target of rapamycin complex 1 (mTORC1) activation although the direct relationship remains unclear. Previous studies in various cell types have used high insulin concentrations to determine the role of insulin on mTORC1 signaling and AAT mRNA abundance. However, this approach may limit applicability to human physiology. Therefore, we sought to determine the effect of insulin on mTORC1 signaling and whether lower insulin concentrations stimulate AAT mRNA abundance in muscle cells. We hypothesized that lower insulin concentrations would increase mRNA abundance of select AAT via an mTORC1-dependent mechanism in C2C12 myotubes. Insulin (0.5 nmol/L) significantly increased phosphorylation of the mTORC1 downstream effectors p70 ribosomal protein S6 kinase 1 (S6K1) and ribosomal protein S6 (S6). A low rapamycin dose (2.5 nmol/L) significantly reduced the insulin-(0.5 nmol/L) stimulated S6K1 and S6 phosphorylation. A high rapamycin dose (50 nmol/L) further reduced the insulin-(0.5 nmol/L) stimulated phosphorylation of S6K1 and S6. Insulin (0.5 nmol/L) increased mRNA abundance of SLC38A2/SNAT2 (P ? 0.043) and SLC7A5/LAT1 (P ? 0.021) at 240 min and SLC36A1/PAT1 (P = 0.039) at 30 min. High rapamycin prevented an increase in SLC38A2/SNAT2 (P = 0.075) and SLC36A1/PAT1 (P ? 0.06) mRNA abundance whereas both rapamycin doses prevented an increase in SLC7A5/LAT1 (P ? 0.902) mRNA abundance. We conclude that a low insulin concentration increases SLC7A5/LAT1 mRNA abundance in an mTORC1-dependent manner in skeletal muscle cells. PMID:24760501
Walker, Dillon K; Drummond, Micah J; Dickinson, Jared M; Borack, Michael S; Jennings, Kristofer; Volpi, Elena; Rasmussen, Blake B
Insulin increases mRNA abundance of the amino acid transporter SLC7A5/LAT1 via an mTORC1-dependent mechanism in skeletal muscle cells
Abstract Amino acid transporters (AATs) provide a link between amino acid availability and mammalian/mechanistic target of rapamycin complex 1 (mTORC1) activation although the direct relationship remains unclear. Previous studies in various cell types have used high insulin concentrations to determine the role of insulin on mTORC1 signaling and AAT mRNA abundance. However, this approach may limit applicability to human physiology. Therefore, we sought to determine the effect of insulin on mTORC1 signaling and whether lower insulin concentrations stimulate AAT mRNA abundance in muscle cells. We hypothesized that lower insulin concentrations would increase mRNA abundance of select AAT via an mTORC1?dependent mechanism in C2C12 myotubes. Insulin (0.5 nmol/L) significantly increased phosphorylation of the mTORC1 downstream effectors p70 ribosomal protein S6 kinase 1 (S6K1) and ribosomal protein S6 (S6). A low rapamycin dose (2.5 nmol/L) significantly reduced the insulin?(0.5 nmol/L) stimulated S6K1 and S6 phosphorylation. A high rapamycin dose (50 nmol/L) further reduced the insulin?(0.5 nmol/L) stimulated phosphorylation of S6K1 and S6. Insulin (0.5 nmol/L) increased mRNA abundance of SLC38A2/SNAT2 (P ? 0.043) and SLC7A5/LAT1 (P ? 0.021) at 240 min and SLC36A1/PAT1 (P = 0.039) at 30 min. High rapamycin prevented an increase in SLC38A2/SNAT2 (P = 0.075) and SLC36A1/PAT1 (P ? 0.06) mRNA abundance whereas both rapamycin doses prevented an increase in SLC7A5/LAT1 (P ? 0.902) mRNA abundance. We conclude that a low insulin concentration increases SLC7A5/LAT1 mRNA abundance in an mTORC1?dependent manner in skeletal muscle cells.
Walker, Dillon K.; Drummond, Micah J.; Dickinson, Jared M.; Borack, Michael S.; Jennings, Kristofer; Volpi, Elena; Rasmussen, Blake B.
Folic Acid
Folic acid is used to treat or prevent folic acid deficiency. It is a B-complex vitamin needed by ... Folic acid comes in tablets. It usually is taken once a day. Follow the directions on your prescription label ...
Folic acid is a B vitamin. It helps the body make healthy new cells. Everyone needs folic acid. For women who may get pregnant, it is really important. Getting enough folic acid before and during pregnancy can prevent major birth ...
Acid rain
Acid precipitation is a global problem. The effects were first seen in Europe; it affects the Great Lakes and the Midwest because higher-than-normal levels of acidity in rain are found in these areas. Several bays of the Great Lakes are now known to receive substantial runoff from freshwater streams that have been made acidic by acid rains. These areas may
Characterization of a developmentally regulated amino acid transporter (AAT1p) of the rust fungus Uromyces fabae.
summary In the rust fungus Uromyces fabae, invasion of the host plant and haustorium formation are accompanied by the activation of many genes (PIGs =in planta induced genes). In addition to the previously described AAT2 (PIG2), AAT1 (PIG27) was found to encode a protein with a high similarity to fungal amino acid permeases. AAT1 transcripts are present in germinated hyphae and throughout the mycelium later in the infection process, but occur at the highest levels in haustoria. Expression of AAT1p in a histidine uptake-defective yeast mutant revealed energy-dependent transport of (14)C-histidine, with a K(M) value of 25.8 microm. In addition, complementation analysis revealed AAT1-dependent transport for lysine. Using Xenopus oocytes as expression system, AAT1p-dependent symport of protons with a broad spectrum of amino acids was observed, with the highest activities obtained with histidine and lysine. These results confirm that in rust fungi, the expression of amino acid transporters is developmentally regulated and occurs preferentially in the parasitic phase of development. PMID:20569305
Struck, Christine; Ernst, Michael; Hahn, Matthias
Sequencing, bioinformatic characterization and expression pattern of a putative amino acid transporter from the parasitic cestode Echinococcus granulosus.
We have sequenced and partially characterized an Echinococcus granulosus cDNA, termed egat1, from a protoscolex signal sequence trap (SST) cDNA library. The isolated 1627 bp long cDNA contains an ORF of 489 amino acids and shows an amino acid identity of 30% with neutral and excitatory amino acid transporters members of the Dicarboxylate/Amino Acid Na+ and/or H+ Cation Symporter family (DAACS) (TC 2.A.23). Additional bioinformatics analysis of EgAT1, confirmed the results obtained by similarity searches and showed the presence of 9 to 10 transmembrane domains, consensus sequences for N-glycosylation between the third and fourth transmembrane domain, a highly similar hydropathy profile with ASCT1 (a known member of DAACS family), high score with SDF (Sodium Dicarboxilate Family) and similar motifs with EDTRANSPORT, a fingerprint of excitatory amino acid transporters. The localization of the putative amino acid transporter was analyzed by in situ hybridization and immunofluorescence in protoscoleces and associated germinal layer. The in situ hybridization labelling indicates the distribution of egat1 mRNA throughout the tegument. EgAT1 protein, which showed in Western blots a molecular mass of approximately 60 kD, is localized in the subtegumental region of the metacestode, particularly around suckers and rostellum of protoscoleces and layers from brood capsules. The sequence and expression analyses of EgAT1 pave the way for functional analysis of amino acids transporters of E. granulosus and its evaluation as new drug targets against cystic echinococcosis. PMID:18272295
Camicia, Federico; Paredes, Rodolfo; Chalar, Cora; Galanti, Norbel; Kamenetzky, Laura; Gutierrez, Ariana; Rosenzvit, Mara C
Mefenamic Acid
Mefenamic acid is used to relieve mild to moderate pain, including menstrual pain (pain that happens before or during a menstrual period). Mefenamic acid is in a class of medications called NSAIDs. ...
Aminocaproic Acid
Aminocaproic acid is used to control bleeding that occurs when blood clots are broken down too quickly. This type ... the baby is ready to be born). Aminocaproic acid is also used to control bleeding in the ...
Ascorbic Acid
Ascorbic acid is used to prevent and treat scurvy, a disease caused by a lack of vitamin C ... Ascorbic acid comes in extended-release (long-acting) capsules and tablets, lozenges, syrup, chewable tablets, and liquid drops ...
Shikimic Acid
NSDL National Science Digital Library
The molecule for this month comes from the article Isolation of Shikimic Acid from Star Aniseed by Richard Payne and Michael Edmonds. Shikimic acid plays a key role in the biosynthesis of many important natural products including aromatic amino acids, alkaloids, phenolics, and phenylpropanoids. It plays such an important role that one of the key biosynthetic pathways is referred to as the shikimate pathway.
Mechanism of fatty-acid-dependent UCP1 uncoupling in brown fat mitochondria.
Mitochondrial uncoupling protein 1 (UCP1) is responsible for nonshivering thermogenesis in brown adipose tissue (BAT). Upon activation by long-chain fatty acids (LCFAs), UCP1 increases the conductance of the inner mitochondrial membrane (IMM) to make BAT mitochondria generate heat rather than ATP. Despite being a member of the family of mitochondrial anion carriers (SLC25), UCP1 is believed to transport H(+) by an unusual mechanism that has long remained unresolved. Here, we achieved direct patch-clamp measurements of UCP1 currents from the IMM of BAT mitochondria. We show that UCP1 is an LCFA anion/H(+) symporter. However, the LCFA anions cannot dissociate from UCP1 due to hydrophobic interactions established by their hydrophobic tails, and UCP1 effectively operates as an H(+) carrier activated by LCFA. A similar LCFA-dependent mechanism of transmembrane H(+) transport may be employed by other SLC25 members and be responsible for mitochondrial uncoupling and regulation of metabolic efficiency in various tissues. PMID:23063128
Fedorenko, Andriy; Lishko, Polina V; Kirichok, Yuriy
Basically Acids
Students learn the basics of acid/base chemistry in a fun, interactive way by studying instances of acid/base chemistry found in popular films such as Harry Potter and the Prisoner of Azkaban and National Treasure. Students learn what acids, bases and indicators are and how they can be used, including invisible ink. They also learn how engineers use acids and bases every day to better our quality of life. Students' interest is piqued by the use of popular culture in the classroom.
University Of Houston
Sodium ion-dependent amino acid transport in membrane vesicles of Bacillus stearothermophilus.
Amino acid transport in membrane vesicles of Bacillus stearothermophilus was studied. A relatively high concentration of sodium ions is needed for uptake of L-alanine (Kt = 1.0 mM) and L-leucine (Kt = 0.4 mM). In contrast, the Na(+)-H(+)-L-glutamate transport system has a high affinity for sodium ions (Kt less than 5.5 microM). Lithium ions, but no other cations tested, can replace sodium ions in neutral amino acid transport. The stimulatory effect of monensin on the steady-state accumulation level of these amino acids and the absence of transport in the presence of nonactin indicate that these amino acids are translocated by a Na+ symport mechanism. This is confirmed by the observation that an artificial delta psi and delta mu Na+/F but not a delta pH can act as a driving force for uptake. The transport system for L-alanine is rather specific. L-Serine, but not L-glycine or other amino acids tested, was found to be a competitive inhibitor of L-alanine uptake. On the other hand, the transport carrier for L-leucine also translocates the amino acids L-isoleucine and L-valine. The initial rates of L-glutamate and L-alanine uptake are strongly dependent on the medium pH. The uptake rates of both amino acids are highest at low external pH (5.5 to 6.0) and decline with increasing pH. The pH allosterically affects the L-glutamate and L-alanine transport systems. The maximal rate of L-glutamate uptake (Vmax) is independent of the external pH between pH 5.5 and 8.5, whereas the affinity constant (Kt) increases with increasing pH. A specific transport system for the basic amino acids L-lysine and L-arginine in the membrane vesicles has also been observed. Transport of these amino acids occurs most likely by a uniport mechanism.
Heyne, R I; de Vrij, W; Crielaard, W; Konings, W N
Acid clusters
Molecular clusters can be considered to be the smallest size range of an aerosol particle size distribution. Nucleation from the gas phase to particles or droplets involves the formation of clusters in the initial stages. Consequently, knowledge of the properties and formation of clusters containing acids contribute to an understanding of acid rain. This paper presents an overview of results obtained in the laboratory on the formation and stability of both neutral and ionized acid clusters. With free jet expansion techniques, the authors have produced clusters of aqueous nitric acid, aqueous hydrochloric acid, aqueous sulfuric acid, acetic acid and aqueous sulfur dioxide. For analogy to buffering, the formation of clusters containing ammonia have also been examined. These have included ammonia with aqueous nitric acid, hydrogen sulfide and sulfur dioxide. The basic experiment involves expansion of vapor through a nozzle, collimation of the jet with a skimmer to form a well-directed molecular beam, and detection of clusters via electron impact ionization and mass spectrometry. Some variations include the introduction of a reactive gas into vacuum near the expansion as described elsewhere and the implementation of an electrostatic quadrupolar field to examine the polarity of the neutral clusters.
Keesee, R.G.; Castleman, A.W. Jr.
Domoic Acid
This online student report discusses the chemistry of domoic acid, a biotoxin that is produced by the diatom Psuedo-nitzschia and associated with Amnesiac Shellfish Poisoning (ASP). In addition to a descriptive summary and images, the report links to other areas of interest related to domoic acid poisoning including signs and symptoms, modes of action, and treatment.
Bailey, Christina; Kohlen, Corinne
The causes and effects of acid rain are detailed. Sulfur dioxide and nitrogen dioxide emissions from anthropogenic sources are the primary causative agents. These emissions are transported over long distances and transformed into sulfates and nitrates and washed out of the atmosphere. Trends in acidity in precipitation water are reviewed for eastern portions of Canada and the U.S. Adverse effects
R. E. Ghelardi; B. L. Murphy
Acids (GCMP)
Acids: this is a resource in the collection "General Chemistry Multimedia Problems". We will observe the reaction of sodium bicarbonate with three acid solutions. General Chemistry Multimedia Problems ask students questions about experiments they see presented using videos and images. The questions asked apply concepts from different parts of an introductory course, encouraging students to decompartmentalize the material.
This book was written in a concise and readable style for the lay public. It's purpose was to make the public aware of the damage caused by acid rain and to mobilize public opinion to favor the elimination of the causes of acid rain.
Elsworth, S.
Usnic acid.
Since its first isolation in 1844, usnic acid [2,6-diacetyl-7,9-dihydroxy-8,9b-dimethyl-1,3(2H,9bH)-dibenzo-furandione] has become the most extensively studied lichen metabolite and one of the few that is commercially available. Usnic acid is uniquely found in lichens, and is especially abundant in genera such as Alectoria, Cladonia, Usnea, Lecanora, Ramalina and Evernia. Many lichens and extracts containing usnic acid have been utilized for medicinal, perfumery, cosmetic as well as ecological applications. Usnic acid as a pure substance has been formulated in creams, toothpaste, mouthwash, deodorants and sunscreen products, in some cases as an active principle, in others as a preservative. In addition to antimicrobial activity against human and plant pathogens, usnic acid has been shown to exhibit antiviral, antiprotozoal, antiproliferative, anti-inflammatory and analgesic activity. Ecological effects, such as antigrowth, antiherbivore and anti-insect properties, have also been demonstrated. A difference in biological activity has in some cases been observed between the two enantiomeric forms of usnic acid. Recently health food supplements containing usnic acid have been promoted for use in weight reduction, with little scientific support. The emphasis of the current review is on the chemistry and biological activity of usnic acid and its derivatives in addition to rational and ecologically acceptable methods for provision of this natural compound on a large scale. PMID:12453567
Ing�lfsd�ttir, K
Acid test
Baking soda can be used as an indicator of how much acid a substance contains. Lemons and limes have more acid in them than grapefruits and oranges. Indophenol can be used as an indicator of how much vitamin C is in a substance.
Olivia Worland (Purdue University;Biological Sciences)
Acid precipitation includes not only rain but also acidified snow, hail and frost, as well as sulfur and nitrogen dust. The principal source of acid precipitation is pollution emitted by power plants and smelters. Sulfur and nitrogen compounds contained in the emissions combine with moisture to form droplets with a high acid content - sometimes as acidic as vinegar. When sufficiently concentrated, these acids can kill fish and damage material structures. Under certain circumstances they may reduce crop and forest yields and cause or aggravate respiratory diseases in humans. During the summer, especially, pollutants tend to collect over the Great Lakes in high pressure systems. Since winds typically are westerly and rotate clockwise around high pressure systems, the pollutants gradually are dispersed throughout the eastern part of the continent.
Sweet, W.
S. pombe TORC1 activates the ubiquitin-proteasomal degradation of the meiotic regulator Mei2 in cooperation with Pat1 kinase.
Target of rapamycin (TOR) kinase regulates cell metabolism and growth, acting as a subunit of two multi-protein complexes, TORC1 and TORC2. Known TORC substrates are either kinases or general factors involved in growth control. Here, we show that fission yeast TORC1, which promotes vegetative growth and suppresses sexual development, can phosphorylate Mei2 (a specific factor involved in switching the cell fate) in vitro. Alanine substitutions at the nine Mei2 phosphorylation sites stabilize the protein and promote mating and meiosis in vivo. We found that Mei2 is polyubiquitylated in vivo in a TORC1-dependent manner. Based on these data, we propose that TORC1 contributes to the suppression of sexual development by phosphorylating Mei2, in addition to controlling the cellular metabolic status. PMID:24741065
Otsubo, Yoko; Yamashita, Akira; Ohno, Hayao; Yamamoto, Masayuki
Acid Rain
Due to the presence of dissolved gases such as carbon dioxide, rainfall is naturally acidic. The release of other gases and chemicals such as sulfur dioxide during the combustion of coal and oil can cause rainfall to become even more acidic, sometimes to the point of toxicity. In this activity, students will measure the pH of local rainfall to see what effect these gases have in their region. They will also check an online resource to see how the releases of acid rain-causing chemicals have varied over the past 20 years, and answer questions about the information they uncover.
Fox, Chris
Mathematical modelling and assessment of the pH homeostasis mechanisms in Aspergillus niger while in citric acid producing conditions.
In this work we introduce an extended model of the Aspergillus niger metabolism while in citrate production conditions. The model includes many recent findings related to various transport processes. It now considers a new information about the fructose uptake system and the proton and amino acids carriers between cytoplasm and the external medium. It also accounts for recent information about both the malate-citrate antiport between mitochondria and cytoplasm and the dihydrogen citrate ion excretion symport with protons. Finally, the model also accounts for new information about the glycerol-3-phosphate shuttle and pH buffering systems. Provided with this updated representation and after having assessed its quality and dynamic behaviour, we were able to explain the observed pH homoeostasis found in A. niger while in citrate producing conditions. The model also serves to enhance our comprehension of the molecular mechanisms operating in order to keep homoeostasis of pH in A. niger and other fungi, bacteria and yeast of biotechnological relevance. PMID:21549718
Garc�a, Jacqueline; Torres, N�stor
MedlinePLUS Videos and Cool Tools
... register. I'm interested in: Pregnancy Baby growth & care Research Volunteer opportunities Advocacy in government For health ... acid During your pregnancy Your pregnant body Prenatal care Eating and nutrition Physical activity Emotional and life ...
EPA Science Inventory
Acid precipitation has become one of the major environmental problems of this decade. It is a challenge to scientists throughout the world. Researchers from such diverse disciplines as plant pathology, soil science, bacteriology, meteorology and engineering are investigating diff...
NHE(VNAT): an H+ V-ATPase electrically coupled to a Na+:nutrient amino acid transporter (NAT) forms an Na+/H+ exchanger (NHE).
Glycolysis, the citric acid cycle and other metabolic pathways of living organisms generate potentially toxic acids within all cells. One ubiquitous mechanism for ridding cells of the acids is to expel H(+) in exchange for extracellular Na(+), mediated by electroneutral transporters called Na(+)/H(+) exchangers (NHEs) that are driven by Na(+) concentration gradients. The exchange must be important because the human genome contains 10 NHEs along with two Na(+)/H(+) antiporters (NHAs). By contrast, the genomes of two principal disease vector mosquitoes, Anopheles gambiae and Aedes aegypti, contain only three NHEs along with the two NHAs. This shortfall may be explained by the presence of seven nutrient amino acid transporters (NATs) in the mosquito genomes. NATs transport Na(+) stoichiometrically linked to an amino acid into the cells by a process called symport or co-transport. Three of the mosquito NATs and two caterpillar NATs have previously been investigated after heterologous expression in Xenopus laevis oocytes and were found to be voltage driven (electrophoretic). Moreover, the NATs are present in the same membrane as the H(+) V-ATPase, which generates membrane potentials as high as 120 mV. We review evidence that the H(+) V-ATPase moves H(+) out of the cells and the resulting membrane potential (V(m)) drives Na(+) linked to an amino acid into the cells via a NAT. The H(+) efflux by the V-ATPase and Na(+) influx by the NAT comprise the same ion exchange as that mediated by an NHE; so the V and NAT working together constitute an NHE that we call NHE(VNAT). As the H(+) V-ATPase is widely distributed in mosquito epithelial cells and there are seven NATs in the mosquito genomes, there are potentially seven NHE(VNAT)s that could replace the missing NHEs. We review published evidence in support of this hypothesis and speculate about broader functions of NHE(VNAT)s. PMID:19151209
Harvey, William R; Boudko, Dmitri Y; Rheault, Mark R; Okech, Bernard A
NHEVNAT: an H+ V-ATPase electrically coupled to a Na+:nutrient amino acid transporter (NAT) forms an Na+/H+ exchanger (NHE)
Summary Glycolysis, the citric acid cycle and other metabolic pathways of living organisms generate potentially toxic acids within all cells. One ubiquitous mechanism for ridding cells of the acids is to expel H+ in exchange for extracellular Na+, mediated by electroneutral transporters called Na+/H+ exchangers (NHEs) that are driven by Na+ concentration gradients. The exchange must be important because the human genome contains 10 NHEs along with two Na+/H+ antiporters (NHAs). By contrast, the genomes of two principal disease vector mosquitoes, Anopheles gambiae and Aedes aegypti, contain only three NHEs along with the two NHAs. This shortfall may be explained by the presence of seven nutrient amino acid transporters (NATs) in the mosquito genomes. NATs transport Na+ stoichiometrically linked to an amino acid into the cells by a process called symport or co-transport. Three of the mosquito NATs and two caterpillar NATs have previously been investigated after heterologous expression in Xenopus laevis oocytes and were found to be voltage driven (electrophoretic). Moreover, the NATs are present in the same membrane as the H+ V-ATPase, which generates membrane potentials as high as 120 mV. We review evidence that the H+ V-ATPase moves H+ out of the cells and the resulting membrane potential (Vm) drives Na+ linked to an amino acid into the cells via a NAT. The H+ efflux by the V-ATPase and Na+ influx by the NAT comprise the same ion exchange as that mediated by an NHE; so the V and NAT working together constitute an NHE that we call NHEVNAT. As the H+ V-ATPase is widely distributed in mosquito epithelial cells and there are seven NATs in the mosquito genomes, there are potentially seven NHEVNATs that could replace the missing NHEs. We review published evidence in support of this hypothesis and speculate about broader functions of NHEVNATs.
Harvey, William R.; Boudko, Dmitri Y.; Rheault, Mark R.; Okech, Bernard A.
Acid Stomach
This Science NetLinks lesson is intended for a high-school, introductory chemistry class or health class. The lesson begins with an article on the history of the development of aspirin. Students will then complete a lab that compares the reaction of regular aspirin, buffered aspirin, and enteric aspirin in neutral, acidic, and basic solutions. They will then analyze the results of the experiment to gain insight into how this information was used by researchers to solve some of the problems associated with aspirin. To complete the lesson, students must understand acids and bases.
Science Netlinks;
Salicylic acids
Salicylic acid is well known phytohormone, emerging recently as a new paradigm of an array of manifestations of growth regulators. The area unleashed yet encompassed the applied agriculture sector to find the roles to strengthen the crops against plethora of abiotic and biotic stresses. The skipped part of integrated picture, however, was the evolutionary insight of salicylic acid to either allow or discard the microbial invasion depending upon various internal factors of two interactants under the prevailing external conditions. The metabolic status that allows the host invasion either as pathogenesis or symbiosis with possible intermediary stages in close systems has been tried to underpin here.
Hayat, Shamsul; Irfan, Mohd; Wani, Arif; Nasser, Alyemeni; Ahmad, Aqil
Stearic Acid
ERIC Educational Resources Information Center
A chemical laboratory information profile (CLIP) is presented for the chemical, stearic acid. The profile lists the chemical's physical and harmful characteristics, exposure limits, and symptoms of major exposure, for the benefit of teachers and students, who use the chemical in the laboratory.
Young, Jay A.
Amino Acids
The Featured Molecules this month are the 20 standard α-amino acids found in proteins and serve as background to the paper by Barone and Schmidt on the Nonfood Applications of Proteinaceous Renewable Materials. The molecules are presented in two formats, the neutral form and the ionized form found in solution at physiologic pH.
Ethacrynic Acid
... or any of the ingredients in ethacrynic acid tablets. Ask your doctor or pharmacist for a list of the ingredients.tell your doctor and pharmacist what prescription and nonprescription medications, vitamins, nutritional supplements and herbal products you are taking or plan to take. ...
Tranexamic Acid
... or any of the ingredients in tranexamic acid tablets. Ask your pharmacist for a list of the ingredients.tell your doctor and pharmacist what other prescription and nonprescription medications, vitamins, nutritional supplements, and herbal products you are taking or plan to take. ...
Carbonate acidizing
The authors present the first quantitative study and complete model of the wormholing phenomenon, leading to a means of predicting and optimizing carbonate acidizing treatments. Laboratory experiments on a gypsum model system and computer simulations show that for a given geometry, wormholes can be quantified by a unique parameter, their equivalent hydraulic length. The behavior of this quantifying parameter vs. all the system parameters is studied and allows the quantitative prediction of the efficiency of an acidizing treatment. This study highlights the fractal nature of the phenomenon, which is accounted for in the equations, and the strong effect of the sample geometry. Three types of etching can be obtained: compact, wormhole type, or homogeneous. The optimum conditions for achieving the best skin decrease correspond to the creation of wormholes and can then be defined in terms of fluid reactivity and injection rate.
Daccord, G.; Touboul, E.; Lenormand, R.
Levulinic acid
The title compound (systematic name: 4-oxo�penta�noic acid), C5H8O3, is close to planar (r.m.s. deviation = 0.0762?�). In the crystal, the mol�ecules inter�act via O�H?O hydrogen bonds in which the hy�droxy O atoms act as donors and the ketone O atoms in adjacent mol�ecules as acceptors, forming C(7) chains along [20-1].
Hachula, Barbara; Polasz, Anna; Dzida, Marzena; Nowak, Maria; Kusz, Joachim
This highly detailed chemical information page features domoic acid, a toxin associated with Amnesic shellfish poisoning and naturally produced by the red algae Chondria armata and diatoms of the genus Pseudo-nitzschia. Created by the International Programme on Chemical Safety, this web page organizes information under the following sections: Name, Summary, Physio-Chemical Properties, Uses, Routes of Entry, Kinetics, Toxicology, Toxicological and Biomedical Investigations, Clinical Effects, Management, Illustrative Cases, Additional Information, References, and Authors.
Inchem; Safety, International P.
Acid Ocean
The I2I-Acid Ocean virtual lab is an e-learning activity where students become virtual scientists studying the impact of ocean acidification on sea urchin larval growth. Students recreate a real, up-to-date climate change experiment. They also learn important general scientific principles, such as the importance of sample size and numbers of replicates, and discuss what this research into a specific impact of climate change may mean for the future of our oceans. There is a French translation available.
Transport in Halobacterium Halobium: Light-Induced Cation-Gradients, Amino Acid Transport Kinetics, and Properties of Transport Carriers
NASA Technical Reports Server (NTRS)
Cell envelope vesicles prepared from H. halobium contain bacteriorhodopsin and upon illumination protons are ejected. Coupled to the proton motive force is the efflux of Na(+). Measurements of Na-22 flux, exterior pH change, and membrane potential, Delta(psi) (with the dye 3,3'-dipentyloxadicarbocyanine) indicate that the means of Na(+) transport is sodium/proton exchange. The kinetics of the pH changes and other evidence suggests that the antiport is electrogenic (H(+)/Na(++ greater than 1). The resulting large chemical gradient for Na(+) (outside much greater than inside), as well as the membrane potential, will drive the transport of 18 amino acids. The I9th, glutamate, is unique in that its accumulation is indifferent to Delta(psi): this amino acid is transported only when a chemical gradient for Na(+) is present. Thus, when more and more NaCl is included in the vesicles glutamate transport proceeds with longer and longer lags. After illumination the gradient of H+() collapses within 1 min, while the large Na(+) gradient and glutamate transporting activity persists for 10- 15 min, indicating that proton motive force is not necessary for transport. A chemical gradient of Na(+), arranged by suspending vesicles loaded with KCl in NaCl, drives glutamate transport in the dark without other sources of energy, with V(sub max) and K(sub m) comparable to light-induced transport. These and other lines of evidence suggest that the transport of glutamate is facilitated by symport with Na(+), in an electrically neutral fashion, so that only the chemical component of the Na(+) gradient is a driving force.
Lanyi, Janos K.
The SLC36 family of proton-coupled amino acid transporters and their potential role in drug transport
Members of the solute carrier (SLC) 36 family are involved in transmembrane movement of amino acids and derivatives. SLC36 consists of four members. SLC36A1 and SLC36A2 both function as H+-coupled amino acid symporters. SLC36A1 is expressed at the luminal surface of the small intestine but is also commonly found in lysosomes in many cell types (including neurones), suggesting that it is a multipurpose carrier with distinct roles in different cells including absorption in the small intestine and as an efflux pathway following intralysosomal protein breakdown. SLC36A1 has a relatively low affinity (Km 1�10 mM) for its substrates, which include zwitterionic amino and imino acids, heterocyclic amino acids and amino acid-based drugs and derivatives used experimentally and/or clinically to treat epilepsy, schizophrenia, bacterial infections, hyperglycaemia and cancer. SLC36A2 is expressed at the apical surface of the human renal proximal tubule where it functions in the reabsorption of glycine, proline and hydroxyproline. SLC36A2 also transports amino acid derivatives but has a narrower substrate selectivity and higher affinity (Km 0.1�0.7 mM) than SLC36A1. Mutations in SLC36A2 lead to hyperglycinuria and iminoglycinuria. SLC36A3 is expressed only in testes and is an orphan transporter with no known function. SLC36A4 is widely distributed at the mRNA level and is a high-affinity (Km 2�3 �M) transporter for proline and tryptophan. We have much to learn about this family of transporters, but from current knowledge, it seems likely that their function will influence the pharmacokinetic profiles of amino acid-based drugs by mediating transport in both the small intestine and kidney. LINKED ARTICLES This article is part of a themed section on Transporters. To view the other articles in this section visit http://dx.doi.org/10.1111/bph.2011.164.issue-7
Thwaites, David T; Anderson, Catriona MH
Methylmalonic acid blood test
... acid is a substance produced when proteins (called amino acids) in the body break down. A test can ... Cederbaum S, Berry GT. Inborn errors of carbohydrate, ammonia, amino acid, and organic acid metabolism. In: Gleason CA, Devaskar ...
Folic acid - test
Folic acid is a type of B vitamin. This article discusses the test to measure the amount of folic acid in the blood. ... that may interfere with test results, including folic acid supplements. Drugs that can decrease folic acid measurements ...
New method of acidizing or acid fracturing: crosslinked acid gels
Acid polymer gels having pH less than one have been crosslinked for retarding the chemical and physical activity of hydrochloric acid on calcareous formations. Hydrochloric acid concentrations from .0025 to 28% have been successfully crosslinked. This stimulation fluid offers high viscosity with adequate shear stability, perfect support for proppants, and clay stabilization. Additionally, the fluid provides effective fluid loss control and retardation of acid reaction enabling live acid to penetrate deeper into the formation for better formation conductivity and practically a residue-free break for rapid clean-up of the well after the job. Results of lab and field tests show the acid crosslinked system to be an effective stimulation fluid for acidizing and acid fracturing in calcareous and sandstone formations having low formation permeability.
Pabley, A.S.; Holcomb, D.L.
Characterization of functional regions in the Schizosaccharomyces pombe mei3 developmental activator.
The Schizosaccharomyces pombe mei3(+) gene is expressed only in diploid cells undergoing meiosis. Ectopic expression of mei3(+) in haploid cells causes meiotic catastrophe. Mei3 is an inhibitor of Ran1/Pat1 kinase and contains a nine-amino-acid motif, Mei3-RKDIII, that resembles two regions in the Ste11 substrate for Ran1/Pat1. Substitution of serine for Arg-81 within Mei3-RKDIII transforms the inhibitor into a substrate for Ran1/Pat1. Thus, it is likely that Mei3-RKDIII defines a pseudosubstrate sequence. In this study, we constructed a series of mei3 deletion mutations and assayed each for activity. This analysis indicates that the carboxy-terminal domain of Mei3 is sufficient for function in vivo. Alanine-scanning mutagenesis identifies critical residues within the inhibitory domain. Two mutations, SM1 and SM8, fail to cause meiotic catastrophe. The SM1 mutation contains alterations of amino acid residues in Mei3-RKDIII. Recombinant SM1 protein exhibits reduced ability to inhibit Ran1/Pat1 kinase in vitro and interacts inefficiently with the kinase in a two-hybrid assay. The SM8 protein binds to Ran1/Pat1 in a two-hybrid assay but fails to inhibit Ran1/Pat1 substrate phosphorylation in vitro. These findings provide evidence that Mei3-RKDIII defines a Ran1/Pat1-binding site that is necessary but not sufficient for inhibition of the kinase. Using fusions to green fluorescent protein, the cellular localization of Ran1 and Mei3 was examined in living cells. Ran1 is concentrated in the nucleus. Mei3 is also enriched in the nucleus and, consistent with the genetic and biochemical results, the inhibitory domain of Mei3 is sufficient for nuclear localization.
Wang, W; Li, P; Schettino, A; Peng, Z; McLeod, M
Automobiles and acid rain
In his editorial Acid rain Philip H. Abelson writes that everyone who drives an automobile is a contributor to acid rain. Examination of emissions data indicates that controlling automobile emissions will contribute little to solving acid precipitation problems. Of the strong acid anions associated with precipitation acidity, sulfate accounts for about 60% and nitrate for about 40%, on an equivalence
Folic Acid and Pregnancy
... before conception and during early pregnancy . About Folic Acid Folic acid, sometimes called folate, is a B ... well as tissue formation. Continue Getting Enough Folic Acid The Centers for Disease Control and Prevention (CDC) ...
Acid Lipase Disease
NINDS Acid Lipase Disease Information Page Synonym(s): Cholesterol Ester Storage Disease, Wolman�s Disease Table of Contents (click to jump ... research is being done? Clinical Trials What is Acid Lipase Disease ? Acid lipase disease occurs when the ...
Uric acid test (image)
Uric acid urine test is performed to check for the amount of uric acid in urine. Urine is collected over a 24 ... for testing. The most common reason for measuring uric acid levels is in the diagnosis or treatment of ...
Organic acids tunably catalyze carbonic Acid decomposition.
Density functional theory calculations predict that the gas-phase decomposition of carbonic acid, a high-energy, 1,3-hydrogen atom transfer reaction, can be catalyzed by a monocarboxylic acid or a dicarboxylic acid, including carbonic acid itself. Carboxylic acids are found to be more effective catalysts than water. Among the carboxylic acids, the monocarboxylic acids outperform the dicarboxylic ones wherein the presence of an intramolecular hydrogen bond hampers the hydrogen transfer. Further, the calculations reveal a direct correlation between the catalytic activity of a monocarboxylic acid and its pKa, in contrast to prior assumptions about carboxylic-acid-catalyzed hydrogen-transfer reactions. The catalytic efficacy of a dicarboxylic acid, on the other hand, is significantly affected by the strength of an intramolecular hydrogen bond. Transition-state theory estimates indicate that effective rate constants for the acid-catalyzed decomposition are four orders-of-magnitude larger than those for the water-catalyzed reaction. These results offer new insights into the determinants of general acid catalysis with potentially broad implications. PMID:24933150
Kumar, Manoj; Busch, Daryle H; Subramaniam, Bala; Thompson, Ward H
External HCO(3) (-) dehydration maintained by acid zones in the plasma membrane is an important component of the photosynthetic carbon uptake in Ruppia cirrhosa.
Ruppia cirrhosa, a temperate seagrass growing in brackish water, featured a high capacity for HCO(3) (-) utilisation, which could operate over a wide pH range (from 7.5 up to 9.5) with maintained efficiency. Tris buffer inhibited this means of HCO(3) (-) utilisation in a competitive manner, while addition of acetazolamide, an inhibitor of extracellular carbonic anhydrase activity, caused a 40-50% inhibition. A mechanism involving periplasmic carbonic anhydrase-catalysed HCO(3) (-) dehydration in acid zones, followed by a (probably diffusive) transport of the formed CO(2) across the plasma membrane was thus, at least partly, responsible for the HCO(3) (-) utilisation. This mechanism, which comprises a CO(2)-concentrating mechanism (CCM) associated with the plasma membrane, is thus shown for the first time in an aquatic angiosperm. Additional mechanisms involved in the Tris-sensitive HCO(3) (-) utilisation could be direct HCO(3) (-) uptake (e.g., in an H(+)/HCO(3) (-)symport) or (more likely) non-catalysed HCO(3) (-) dehydration in the acid zones. Based on these results, and on earlier investigations on Zostera marina, a general model for analysis of HCO(3) (-) utilisation mechanisms of seagrasses is suggested. In this model, three 'systems' for HCO(3) (-) utilisation are defined which are characterised (and can to some extent be quantified) by their capability to operate at high pH in combination with their response to acetazolamide and Tris. Some consequences of the fact that HCO(3) (-) utilisation and osmoregulation probably depend on the same energy source (ATP via H(+)-ATPase in the plasma membrane) are discussed. PMID:16228374
Hellblom, Frida; Axelsson, Lennart
Acid Rain Study Guide.
Acid rain is a complex, worldwide environmental problem. This study guide is intended to aid teachers of grades 4-12 to help their students understand what acid rain is, why it is a problem, and what possible solutions exist. The document contains specific sections on: (1) the various terms used in conjunction with acid rain (such as acid�
Hunger, Carolyn; And Others
Acid-Base Equilibria
This 9-page PDF document is part of an environmental geochemistry course taught by Dr. David Sherman at the University of Bristol. Topics include acid-base theories, aqueous systems, strong and weak acids and bases, acid-base properties of minerals, the pH of weak acid and buffered systems, and the calculation of titration curves.
Sherman, David W.; Bristol, University O.
Acid tolerance in amphibians
Studies of amphibian acid tolerance provide information about the potential effects of acid deposition on amphibian communities. Amphibians as a group appear to be relatively acid tolerant, with many species suffering increased mortality only below pH 4. However, amphibians exhibit much intraspecific variation in acid tolerance, and some species are sensitive to even low levels of acidity. Furthermore, nonlethal effects, including depression of growth rates and increases in developmental abnormalities, can occur at higher pH.
Pierce, B.A.
Gas-phase Acidities of Aspartic Acid, Glutamic Acid, and their Amino Acid Amides.
Gas-phase acidities (GA or ?Gacid) for the two most acidic common amino acids, aspartic acid and glutamic acid, have been determined for the first time. Because of the amide linkage�s importance in peptides and as an aid in studying side chain versus main chain deprotonation, aspartic acid amide and glutamic acid amide were also studied. Experimental GA values were measured by proton transfer reactions in an electrospray ionization/Fourier transform ion cyclotron resonance mass spectrometer. Calculated GAs were obtained by density functional and molecular orbital theory approaches. The best agreement with experiment was found at the G3MP2 level; the MP2/CBS and B3LYP/aug-cc-pVDZ results are 3�4 kcal/mol more acidic than the G3MP2 results. Experiment shows that aspartic acid is more acidic than glutamic acid by ca. 3 kcal/mol whereas the G3MP2 results show a smaller acidity difference of 0.2 kcal/mol. Similarly, aspartic acid amide is experimentally observed to be ca. 2 kcal/mol more acidic than glutamic acid amide whereas the G3MP2 results show a correspondingly smaller energy difference of 0.7 kcal/mol. The computational results clearly show that the anions are all ring-like structures with strong hydrogen bonds between the OH or NH2 groups and the CO2? group from which the proton is removed. The two amino acids are main-chain deprotonated. In addition, use of the COSMO model for the prediction of the free energy differences in aqueous solution gave values in excellent agreement with the most recent experimental values for pKa. Glutamic acid is predicted to be more acidic than aspartic acid in aqueous solution due to differential solvation effects.
Li, Zhong; Matus, Myrna H.; Velazquez, Hector A.; Dixon, David A.; Cassady, Carolyn J.
Gas-phase acidities of aspartic acid, glutamic acid, and their amino acid amides
NASA Astrophysics Data System (ADS)
Gas-phase acidities (GA or [Delta]Gacid) for the two most acidic common amino acids, aspartic acid and glutamic acid, have been determined for the first time. Because of the amide linkage's importance in peptides and as an aid in studying side chain versus main chain deprotonation, aspartic acid amide and glutamic acid amide were also studied. Experimental GA values were measured by proton transfer reactions in an electrospray ionization/Fourier transform ion cyclotron resonance mass spectrometer. Calculated GAs were obtained by density functional and molecular orbital theory approaches. The best agreement with experiment was found at the G3MP2 level; the MP2/CBS and B3LYP/aug-cc-pVDZ results are 3-4 kcal/mol more acidic than the G3MP2 results. Experiment shows that aspartic acid is more acidic than glutamic acid by ca. 3 kcal/mol whereas the G3MP2 results show a smaller acidity difference of 0.2 kcal/mol. Similarly, aspartic acid amide is experimentally observed to be ca. 2 kcal/mol more acidic than glutamic acid amide whereas the G3MP2 results show a correspondingly smaller energy difference of 0.7 kcal/mol. The computational results clearly show that the anions are all ring-like structures with strong hydrogen bonds between the OH or NH2 groups and the CO2- group from which the proton is removed. The two amino acids are main-chain deprotonated. In addition, use of the COSMO model for the prediction of the free energy differences in aqueous solution gave values in excellent agreement with the most recent experimental values for pKa. Glutamic acid is predicted to be more acidic than aspartic acid in aqueous solution due to differential solvation effects.
Li, Zhong; Matus, Myrna H.; Velazquez, Hector Adam; Dixon, David A.; Cassady, Carolyn J.
Modulating the acidity: highly acidic Br�nsted acids in asymmetric catalysis.
Recently, chiral highly acidic Br�nsted acids have emerged as powerful catalysts for enantioselective C-C and C-X bond-forming reactions. Their strong acidity renders them valuable tools for the activation of imines, carbonyl compounds, and other weakly basic substrates. As a result, new perspectives are opened and highly stereoselective transformations based on the concept of chiral contact-ion-pair catalysis can be realized. This Minireview gives an overview of the design and application of these new organocatalysts and presents recent results in this rapidly growing field. PMID:21678531
Rueping, Magnus; Nachtsheim, Boris J; Ieawsuwan, Winai; Atodiresei, Iuliana
Thin-Layer Separation of Citric Acid Cycle Intermediates, Lactic Acid, and the Amino Acid Taurine.
This paper describes a two-dimensional mixed-layer method for separating citric acid cycle intermediates, lactic acid and the amino acid taurine. The method cleanly separates all citric acid cycle intermediates tested, excepting citric acid and isocitric ...
R. T. Riley M. C. Mix
Succinic Acid in Wines.
Following a brief discussion of methods which are currently employed to determine the succinic acid content of wines, as well as the shortcomings of those methods, procedures used for colorimetric evaluation of this particular acid are explained. The prin...
M. Castino
Omega-3 fatty acids
OBJECTIVE To examine evidence for the role of omega-3 fatty acids in cardiovascular disease. QUALITY OF EVIDENCE PubMed was searched for articles on the role of omega-3 fatty acids in cardiovascular disease. Level I and II evidence indicates that omega-3 fatty acids are beneficial in improving cardiovascular outcomes. MAIN MESSAGE Dietary intake of omega-3 fatty acids has declined by 80% during the last 100 years, while intake of omega-6 fatty acids has greatly increased. Omega-3 fatty acids are cardioprotective mainly due to beneficial effects on arrhythmias, atherosclerosis, inflammation, and thrombosis. There is also evidence that they improve endothelial function, lower blood pressure, and significantly lower triglycerides. CONCLUSION There is good evidence in the literature that increasing intake of omega-3 fatty acids improves cardiac outcomes. Physicians need to integrate dietary recommendations for consumption of omega-3 fatty acids into their usual cardiovascular care.
Schwalfenberg, Gerry
Acid Aerosols Issue Paper.
The report evaluates scientific information on direct health effects associated with exposure to acid aerosols. The present report is not intended as a complete and detailed review of all literature pertaining to acid aerosols. Rather, an attempt has been...
Fatty acid analogs
In one aspect, a radioactively labeled analog of a fatty acid which is capable of being taken up by mammalian tissue and which exhibits an in vivo beta-oxidation rate below that with a corresponding radioactively labeled fatty acid.
Elmaleh, David R. (Newton Center, MA); Livni, Eli (Brookline, MA)
Polymerization of Itaconic Acid.
Itaconic acid has various uses in chain copolymerization reactions, because of its two carboxylic groups and relatively good reactivity with different comonomers. As an unsaturated dicarboxylic acid it also has applications in grafting reactions and polyc...
J. Ahlgren
Sulfuric acid poisoning
Sulfuric acid is a very strong chemical that is corrosive. Corrosive means it can cause severe burns and ... or mucous membranes. This article discusses poisoning from sulfuric acid. This is for information only and not for ...
Folic Acid Quiz
... folic acid supplement d) eat a cup of broccoli 5. Spina bifida and anencephaly: Hint a) are ... acid every day. This question is tricky because broccoli is a natural source of food folate, however ...
Aminocaproic Acid Injection
Aminocaproic acid injection is used to control bleeding that occurs when blood clots are broken down too quickly. This ... the baby is ready to be born). Aminocaproic acid injection is also used to control bleeding in ...
Hydrochloric acid poisoning
Hydrocholoric acid is a clear, poisonous liquid. It is highly corrosive, which means it immediately causes severe damage, such ... poisoning due to swallowing or breathing in hydrochloric acid. This is for information only and not for ...
Facts about Folic Acid
... of the baby's brain and spine ( anencephaly and spina bifida ). How much folic acid a woman needs 400 ... audiocast about folic acid. Related Pages Healthy Pregnancy spina bifida Birth Defects CDC�s National Center on Birth Defects ...
Uric acid - blood
Uric acid is a chemical created when the body breaks down substances called purines. Purines are found in ... mackerel, dried beans and peas, and beer. Most uric acid dissolves in blood and travels to the kidneys. ...
Plasma amino acids
Plasma amino acids is a screening test done on infants that looks at the amounts of amino ... Rheumatoid arthritis High or low concentrations of individual plasma amino acids must be interpreted along with other ...
Refining Lurgi tar acids
There is disclosed a process for removing tar bases and neutral oils from the Lurgi tar acids by treating the tar acids with aqueous sodium bisulfate to change the tar bases to salts and to hydrolyze the neutral oils to hydrolysis products and distilling the tar acids to obtain refined tar acid as the distillate while the tar base salts and neutral oil hydrolysis products remain as residue.
Greco, N.P.
Nucleic acid molecule
US Patent & Trademark Office Database
The invention relates to an isolated nucleic acid molecule encoding a polypeptide capable of producing a triterpenoid hydrocarbon. The invention also relates to the encoded polypeptide, a vector comprising the nucleic acid molecule, a recombinant non-human organism comprising the nucleic acid molecule, and to methods of producing a triterpenoid hydrocarbon or an intermediate of biofuel using the nucleic acid molecule, polypeptide or recombinant organism.
Linolenic acid deficiency
Linolenic acid deficiency has not been demonstrated clearly in warm blooded animals, yet circumstantial evidence suggests\\u000a that n?3 fatty acids may have functions in these animals. The fact that several species of fish definitely require dietary\\u000a n?3 fatty acids indicates that n?3 fatty acids have important and specific functions in these animals and suggests that such\\u000a functions may also be
J. Tinoco; R. Babcock; I. Hincenbergs; B. Medwadowski; P. Miljanich; M. A. Williams
Plant fatty acid hydroxylases
This invention relates to plant fatty acyl hydroxylases. Methods to use conserved amino acid or nucleotide sequences to obtain plant fatty acyl hydroxylases are described. Also described is the use of cDNA clones encoding a plant hydroxylase to produce a family of hydroxylated fatty acids in transgenic plants. In addition, the use of genes encoding fatty acid hydroxylases or desaturases to alter the level of lipid fatty acid unsaturation in transgenic plants is described.
Somerville, Chris (Portola Valley, CA); Broun, Pierre (Burlingame, CA); van de Loo, Frank (Lexington, KY)
Acid-fast stain
... The slide is then washed with an acid solution and a different stain is applied. Bacteria that hold onto the first dye are considered "acid-fast" because they resist the acid wash. This type of bacteria is associated with tuberculosis and other infections.
[alpha]-Oxocarboxylic Acids
Several [alpha]-oxocarboxylic acids play key roles in metabolism in plants and animals. However, there are inconsistencies between the structures as commonly portrayed and the reported acid ionization constants, which result because the acids are predominantly hydrated in aqueous solution; that is, the predominant form is RC(OH)[subscript 2]COOH�
Kerber, Robert C.; Fernando, Marian S.
Energy and acid rain
Acid rain is one of the foremost environmental issues of the 1980s and will be of continuining importance to energy policy for several reasons. First, the pollutants that cause acid rain are projected to increase through the end of the century as the demand for energy grows and as coal replaces oil. Second, many of the effects of acid rain
R R Gould
Acid (and Base) Rainbows
Learners use red cabbage juice and pH indicator paper to test the acidity and basicity of household materials. The activity links this concept of acids and bases to acid rain and other pollutants. Resource contains vocabulary definitions and suggestions for assessment, extensions, and scaling for different levels of learners.
Kolenbrander, Amy; Yowell, Janet; Mach, Natalie; Zarske, Malinda S.; Carlson, Denise; Perez, Sharon
Neutralizing Acids and Bases
Learners use their knowledge of color changes with red cabbage indicator to neutralize an acidic solution with a base and then neutralize a basic solution with an acid. Use this as a follow-up activity to the related activity, "Color Changes with Acids and Bases."
Kessler, James H.; Galvan, Patricia M.
The Acid Rain Reader.
A topic which is often not sufficiently dealt with in elementary school textbooks is acid rain. This student text is designed to supplement classroom materials on the topic. Discussed are: (1) "Rain"; (2) "Water Cycle"; (3) "Fossil Fuels"; (4) "Air Pollution"; (5) "Superstacks"; (6) "Acid/Neutral/Bases"; (7) "pH Scale"; (8) "Acid Rain"; (9)�
Stubbs, Harriett S.; And Others
Crystallization of uric acid
Crystals of uric acid have been grown in tetra methoxy silane and silica gel medium. Small winged, transparent, platy crystals of uric acid of about 0.5x0.5x0.1 mm were grown and were found to be hydrated uric acid.
Kalkura, S. Narayana; Vaidyan, V. K.; Kanakavel, M.; Ramasamy, P.
Transmembrane domain 8 of the {gamma}-aminobutyric acid transporter GAT-1 lines a cytoplasmic accessibility pathway into its binding pocket.
GAT-1 is a sodium- and chloride-coupled gamma-aminobutyric acid (GABA) transporter, which fulfills an essential role in the synaptic transmission by this neurotransmitter. Cysteine-399 is the major site of inhibition of GAT-1 by membrane-permeant sulfhydryl reagents. This cysteine residue was previously thought to reside on a cytoplasmic loop connecting transmembrane domains (TMs) 8 and 9. However, the crystal structure of LeuT, a bacterial homologue of the mammalian neurotransmitter:sodium symporters, revealed that the residue corresponding to Cys-399 is in fact located in the middle of TM 8. This residue is located to the cytoplasmic side of Asp-395 and Ser-396, whose side chains are thought to ligand one of the two cotransported sodium ions. To determine how the sulfhydryl reagents approach cysteine-399, a cysteine scan of all 35 residues of TM 8 was performed. Sulfhydryl reagents inhibited transport when a cysteine residue was present at either of the positions 399, 402, 406, and 410. SKF-89976A and other non-transportable analogues, which are expected to lock the transporter in a conformation facing the extracellular medium, protected against the sulfhydryl modification at positions 399, 402, and 406. Such a protection was not seen by GABA itself, which actually modestly potentiated the modification at positions 399 and 402. Our results point to an alpha-helical stripe on TM8 lining an aqueous access pathway from the cytoplasm into the binding pocket, which gets occluded in the conformation of the transporter where the binding pocket is exposed to the extracellular medium. PMID:19201752
Ben-Yona, Assaf; Kanner, Baruch I
The substrates of the gamma-aminobutyric acid transporter GAT-1 induce structural rearrangements around the interface of transmembrane domains 1 and 6.
The sodium- and chloride-coupled gamma-aminobutyric acid (GABA) transporter GAT-1 is essential for efficient synaptic transmission by this neurotransmitter. GAT-1 is the first cloned member of the neurotransmitter-sodium-symporter family. Here we address the idea that during transport the extracellular halves of transmembrane domains (TM) 1 and 6, TM 1b/TM 6a, move relative to the binding pocket. Therefore, we have probed the aqueous accessibility of TM 6a and its proximity to TM 1b in the presence and absence of its substrates. Cysteines were introduced, one by one, at all TM 6a positions. In several mutants, transport activity was inhibited by the impermeant sulfhydryl reagent (2-trimethylammonium)methanethiosulfonate, whereas wild type GAT-1 was basically insensitive. This inhibition was potentiated by sodium, whereas GABA was protective. Moreover, we used paired cysteine mutagenesis in conjunction with treatments with copper(II)(1,10-phenanthroline)(3) (CuPh). CuPh did not affect the activity of wild type GAT-1 but potently inhibited transport by the TM 6a mutant D287C. Such inhibition was not observed with D287C/C74A, indicating that Asp-287 is close to Cys-74 of TM 1b. Inhibition of transport of D287C by CuPh, but not by (2-trimethylammonium)methanethiosulfonate, was potentiated when sodium and GABA were both removed. Thus, the degree of inhibition by CuPh is not a simple function of the accessibility of the individual cysteines but also involves structural rearrangements around the TM 1b/TM 6a interface. PMID:18381286
Rosenberg, Alex; Kanner, Baruch I
The follicular thyroid cell line PCCL3 responds differently to laminin and to polylaminin, a polymer of laminin assembled in acidic pH.
The extracellular-matrix protein laminin forms polymers both in vivo and in vitro. Acidification of pH leads to the formation of an artificial polymer with biomimetic properties, named polylaminin (polyLM). Follicle cells in the thyroid are in close contact with laminin, but their response to this important extracellular signal is still poorly understood. PCCL3 thyroid follicular cells cultured on glass, on regular laminin (LM) or on laminin previously polymerized in acidic pH (polyLM) showed different cell morphologies and propensities to proliferate, as well as differences in the organization of their actin cytoskeleton. On polyLM, cells displayed a typical epithelial morphology and radially organized actin fibers; whereas on LM, they spread irregularly on the substrate, lost cell contacts, and developed thick actin fibers extending through the entire cytoplasm. Iodide uptake decreased similarly in response to both laminin substrates, in comparison to glass. On both the LM and polyLM substrates, the expression of the sodium iodide symporter (NIS) decreased slightly but not significantly. NIS showed dotted immunostaining at the plasma membrane in the cells cultured on glass; on polyLM, NIS was observed mainly in the perinuclear region, and more diffusely throughout the cytoplasm on the LM substrate. Additionally, polyLM specifically favored the maintenance of cell polarity in culture. These findings indicate that PCCL3 cells can discriminate between LM and polyLM and that they respond to the latter by better preserving the phenotype observed in the thyroid tissue. PMID:23747985
Palmero, C Y; Miranda-Alves, L; Sant'Ana Barroso, M M; Souza, E C L; Machado, D E; Palumbo-Junior, A; Santos, C A N; Portilho, D M; Mermelstein, C S; Takiya, C M; Carvalho, D P; Hochman-Mendez, C; Coelho-Sampaio, T; Nasciutti, L E
Nucleic acid detection compositions
The present invention relates to means for the detection and characterization of nucleic acid sequences, as well as variations in nucleic acid sequences. The present invention also relates to methods for forming a nucleic acid cleavage structure on a target sequence and cleaving the nucleic acid cleavage structure in a site-specific manner. The structure-specific nuclease activity of a variety of enzymes is used to cleave the target-dependent cleavage structure, thereby indicating the presence of specific nucleic acid sequences or specific variations thereof.
Prudent, James R. (Madison, WI) [Madison, WI; Hall, Jeff G. (Madison, WI) [Madison, WI; Lyamichev, Victor I. (Madison, WI) [Madison, WI; Brow, Mary Ann (Madison, WI) [Madison, WI; Dahlberg, James L. (Madison, WI) [Madison, WI
Cleavage of nucleic acids
Prudent, James R. (Madison, WI); Hall, Jeff G. (Madison, WI); Lyamichev, Victor I. (Madison, WI); Brow, Mary Ann D. (Madison, WI); Dahlberg, James E. (Madison, WI)
Prudent, James R. (Madison, WI); Hall, Jeff G. (Waunakee, WI); Lyamichev, Victor I. (Madison, WI); Brow; Mary Ann D. (Madison, WI); Dahlberg, James E. (Madison, WI)
Acid Rain: Students Site
This website introduces students to the concepts and issues surrounding acid rain. They will learn what acid rain is, how it forms, and what its effects are on humans, on the environment (lakes, rivers), and on infrastructure (buildings, monuments). There is also discussion of what is being done to reduce the presence of acid rain, and some suggestions for the students themselves to help reduce acid rain. A set of games, puzzles, and activities provides students with an opportunity to put their new knowledge of acid rain to use, and an animated slide show is provided to visually illustrate its causes and effects. A Spanish translation is available.
Prudent, James R. (Madison, WI); Hall, Jeff G. (Madison, WI); Lyamichev, Victor L. (Madison, WI); Brow, Mary Ann D. (Madison, WI); Dahlberg, James E. (Madison, WI)
Amino acid analysis
The process and apparatus for qualitative and quantitative analysis of the amino acid content of a biological sample are presented. The sample is deposited on a cation exchange resin and then is washed with suitable solvents. The amino acids and various cations and organic material with a basic function remain on the resin. The resin is eluted with an acid eluant, and the eluate containing the amino acids is transferred to a reaction vessel where the eluant is removed. Final analysis of the purified acylated amino acid esters is accomplished by gas-liquid chromatographic techniques.
Winitz, M.; Graff, J. (inventors)
Demospongic Acids Revisited
The well-known fatty acids with a ?5,9 unsaturation system were designated for a long period as demospongic acids, taking into account that they originally occurred in marine Demospongia sponges. However, such acids have also been observed in various marine sources with a large range of chain-lengths (C16�C32) and from some terrestrial plants with short acyl chains (C18�C19). Finally, the ?5,9 fatty acids appear to be a particular type of non-methylene-interrupted fatty acids (NMA FAs). This article reviews the occurrence of these particular fatty acids in marine and terrestrial organisms and shows the biosynthetic connections between ?5,9 fatty acids and other NMI FAs.
Kornprobst, Jean-Michel; Barnathan, Gilles
[Biosynthesis of adipic acid].
Adipic acid is a six-carbon dicarboxylic acid, mainly for the production of polymers such as nylon, chemical fiber and engineering plastics. Its annual demand is close to 3 million tons worldwide. Currently, the industrial production of adipic acid is based on the oxidation of aromatics from non-renewable petroleum resources by chemo-catalytic processes. It is heavily polluted and unsustainable, and the possible alternative method for adipic acid production should be developed. In the past years, with the development of synthetic biology and metabolic engineering, green and clean biotechnological methods for adipic acid production attracted more attention. In this study, the research advances of adipic acid and its precursor production are reviewed, followed by addressing the perspective of the possible new pathways for adipic acid production. PMID:24432653
Han, Li; Chen, Wujiu; Yuan, Fei; Zhang, Yuanyuan; Wang, Qinhong; Ma, Yanhe
Process for the preparation of lactic acid and glyceric acid
Hexose and pentose monosaccharides are degraded to lactic acid and glyceric acid in an aqueous solution in the presence of an excess of a strongly anionic exchange resin, such as AMBERLITE IRN78 and AMBERLITE IRA400. The glyceric acid and lactic acid can be separated from the aqueous solution. Lactic acid and glyceric acid are staple articles of commerce.
Jackson, James E [Haslett, MI; Miller, Dennis J [Okemos, MI; Marincean, Simona [Dewitt, MI
NRPSs and amide ligases producing homopoly(amino acid)s and homooligo(amino acid)s.
Microorganisms are capable of producing a wide variety of biopolymers. Homopoly(amino acid)s and homooligo(amino acid)s, which are made up of only a single type of amino acid, are relatively rare; in fact, only two homopoly(amino acid)s have been known to occur in nature: poly(?-L-lysine) (?-PL) and poly(?-glutamic acid) (?-PGA). Bacterial enzymes that produce homooligo(amino acid)s, such as L-?-lysine-, L-valine-, L-leucine-, L-isoleucine-, L-methionine-, and L-glutamic acid-oligopeptides and poly(?-l-glutamic acid) (?-PGA) have recently been identified, as well as ?-PL synthetase and ?-PGA synthetase. This article reviews the current knowledge about these unique enzymes producing homopoly(amino acid)s and homooligo(amino acid)s. PMID:23817633
Hamano, Yoshimitsu; Arai, Toshinobu; Ashiuchi, Makoto; Kino, Kuniki
Glycolic Acid 15% Plus Salicylic Acid 2%
Background: Facial flat warts are a contagious viral disease that can cause disturbing cosmetic problems. Topical glycolic acid has been reported to be effective in dermatological treatment depending on the exfoliant capacity, but has not often been reported to be effective in the treatment of facial flat warts. Objective: The aim of this paper was to evaluate the efficacy and safety of glycolic acid 15% topical gel plus salicylic acid 2% in the treatment of recalcitrant facial flat warts. Methods: A total of 20 consecutive patients 7 to 16 years of age with recalcitrant facial flat warts were enrolled in this study. Patients having warts by the eye and lip regions were excluded from the study. A fine layer of face gel was applied to the treatment area once daily. Most of the participants had tried different treatments with no success. Assessments for the response and the occurrence of side effects were performed every two weeks at Weeks 2, 4, 6, and 8. Results: All the patients were clinically cured within eight weeks. Seven patients cleared in four weeks, and 13 patients cleared in eight weeks. No noticeable adverse events were related to the skin. Conclusion: Topical gel of glycolic acid 15% plus salicylic acid 2% is safe and effective when applied to facial flat warts once daily until clearance and may be considered as first-line treatment.
Sanchez-Blanco, Elena
21 CFR 172.860 - Fatty acids.
Code of Federal Regulations, 2010 CFR
...and oils derived from edible sources: Capric acid, caprylic acid, lauric acid, myristic acid, oleic acid, palmitic acid, and stearic acid. (b) The food additive meets the following specifications: (1) Unsaponifiable matter does...
Code of Federal Regulations, 2013 CFR
Kinetic acidity of cubane
The authors have determined the kinetic acidity of cubane by the application of a {sup 3}H NMR spectroscopic approach. An earlier measurement of the acidity of cubane has been subject to some controversy. Kinetic acidities are a useful measure of the acidity of weak carbon acids and are obtained by measuring rates of base-catalyzed proton-exchange reactions. It has been found that one-bond {sup 13}C-{sup 1}H NMR coupling constants ({sup 1}J{sub CH}) correlate closely with kinetic acidities for cyclic aliphatic hydrocarbons. This correlation holds even for strongly strained systems such as cyclopropane. Cubane, a strained polycycloalkane, would be anticipated to also fit this correlation.
Dixon, R.E.; Streitwieser, A. (Univ. of California, Berkeley (USA)); Williams, P.G. (Lawrence Berkeley Lab., CA (USA)); Eaton, P.E. (Univ. of Chicago, IL (USA))
Acid Rain Learning Activities
These suggestions for activities allow students to learn about acid deposition in new and interactive ways, both in and out of the classroom. The suggestions are for individuals and small groups, the class as a whole, or for field trips. Students may contact local experts about acid rain issues, investigate the energy sources used to generate electricity by their local power companies, collect cartoons about acid rain and air pollution, or play the roles of scientists or interested parties involved in investigations of acid rain issues. Field trip ideas include visiting a local museum or science center to see exhibits or resources on acid rain, and visiting a local cemetary to examine the effects of acid rain on the headstones.
of natural amino acids
A new method for the synthesis of protected N a -(�-Y-alkyl) amino acids (Y is a thio, amino or carboxy group) and related compounds by reductive alkylation of natural amino acids is reported. These new amino acids serve as building units for the synthesis of backbone-cyclic peptides. They are orthogonally protected at the �-amino position by butoxycarbonyl (Boc) or 9-fluorenylmethoxycarbonyl
Gal Bitan; Dan Muller; Ron Kasher; Evgenia V. Gluhov; Chaim Gilon
Acid-Base Solutions
How do strong and weak acids differ? Use lab tools on your computer to find out! Dip the paper or the probe into solution to measure the pH, or put in the electrodes to measure the conductivity. Then see how concentration and strength affect pH. Can a weak acid solution have the same pH as a strong acid solution?
Simulations, Phet I.; Lancaster, Kelly; Malley, Chris; Loeblein, Patricia; Parson, Robert; Perkins, Kathy
Characterization of acid tars
Acid tars from the processing of petroleum and petrochemicals using sulfuric acid were characterized by gas chromatography\\/mass spectrometry (GC\\/MS), inductively coupled plasma\\/optical emission spectrometry (ICP\\/OES), differential scanning calorimetry (DSC), Fourier transform infrared (FTIR) spectrometry, and scanning electron microscopy\\/energy dispersive X-ray (SEM\\/EDX) micro-analysis. Leaching of contaminants from the acid tars in 48h batch tests with distilled water at a liquid-to-solid ratio
Sunday A. Leonard; Julia A. Stegemann; Amitava Roy
This paper describes a two-dimensional mixed-layer method for separating citric acid cycle intermediates, lactic acid and the amino acid taurine. The method cleanly separates all citric acid cycle intermediates tested, excepting citric acid and isocitric acid. The solvents are in...
USGS Tracks Acid Rain
USGS Publications Warehouse
The U.S. Geological Survey (USGS) has been actively studying acid rain for the past 15 years. When scientists learned that acid rain could harm fish, fear of damage to our natural environment from acid rain concerned the American public. Research by USGS scientists and other groups began to show that the processes resulting in acid rain are very complex. Scientists were puzzled by the fact that in some cases it was difficult to demonstrate that the pollution from automobiles and factories was causing streams or lakes to become more acidic. Further experiments showed how the natural ability of many soils to neutralize acids would reduce the effects of acid rain in some locations--at least as long as the neutralizing ability lasted (Young, 1991). The USGS has played a key role in establishing and maintaining the only nationwide network of acid rain monitoring stations. This program is called the National Atmospheric Deposition Program/National Trends Network (NADP/NTN). Each week, at approximately 220 NADP/NTN sites across the country, rain and snow samples are collected for analysis. NADP/NTN site in Montana. The USGS supports about 72 of these sites. The information gained from monitoring the chemistry of our nation's rain and snow is important for testing the results of pollution control laws on acid rain.
Gordon, John D.; Nilles, Mark A.; Schroder, LeRoy J.
In his editorial Acid rain Philip H. Abelson writes that everyone who drives an automobile is a contributor to acid rain. Examination of emissions data indicates that controlling automobile emissions will contribute little to solving acid precipitation problems. Of the strong acid anions associated with precipitation acidity, sulfate accounts for about 60% and nitrate for about 40%, on an equivalence basis. The contribution to national SO/sub 2/ emissions by all forms of transportation is about 3%. The corresponding value for national NO/sub x/ emissions is about 44%. If ground-level emissions from highway vehicles contribute to long-range transport of atmospheric pollutants and to precipitation acidity on an equal basis with other sources, for example, power plant smokestacks, then the respective contributions to precipitation acidity can be approximated. The atmospheric chemistry of NO/sub x/ and its interaction with SO/sub 2/ is poorly known. However, it is likely that automobiles account for less than 14% of total equivalents of strong acid anions in either wet or dry deposition in the eastern US. The implication of these data for regulatory policies aimed at controlling acid precipitation by reducing SO/sub 2/ and NO/sub x/ emissions is obvious.
Hendrey, G.R.
Molecular Structure of Fumaric acid
Fumaric acid is odorless and colorless or white crystalline powder with a fruit acid taste. Fumaric acid is used as a substitute of tartaric acid in beverages and baking powders and as a replacement for citric acid in fruits drinks. It is also used as antioxidant to prevent rancidity in butter, cheese, powdered milk, and other foodstuff. In addition, fumaric acid is a chemical intermediate in the manufacture of pharmaceuticals, synthetic resins and plastics. Fumaric acid can be prepared by catalytic oxidation of benzene or by bacterial action on glucose and it is involved in the production of energy from food. Fumaric acid (known as trans-butanedioic acid) is the trans isomer of maleic acid (also called cis-butanedioic acid). Fumaric acid is more stable than maleic acid and can be prepared by heating maleic acid.
Structure of Acid phosphatases.
Acid phosphatases are enzymes that have been studied extensively due to the fact that their dysregulation is associated with pathophysiological conditions. This characteristic has been exploited for the development of diagnostic and therapeutic methods. As an example, prostatic acid phosphatase was the first marker for metastatic prostate cancer diagnosis and the dysregulation of tartrate resistant acid phosphatase is associated with abnormal bone resorption linked to osteoporosis. The pioneering crystallization studies on prostatic acid phosphatase and mammalian tartrate-resistant acid phosphatase conformed significant milestones towards the elucidation of the mechanisms followed by these enzymes (Schneider et al., EMBO J 12:2609-2615, 1993). Acid phosphatases are also found in nonmammalian species such as bacteria, fungi, parasites, and plants, and most of them share structural similarities with mammalian acid phosphatase enzymes. Acid phosphatase (EC 22.214.171.124) enzymes catalyze the hydrolysis of phosphate monoesters following the general equation. Phosphate monoester + H2O -->/<-- alcohol + phosphate. The general classification "acid phosphatase" relies only on the optimum acidic pH for the enzymatic activity in assay conditions using non-physiological substrates. These enzymes accept a wide range of substrates in vitro, ranging from small organic molecules to phosphoproteins, constituting a heterogeneous group of enzymes from the structural point of view. These structural differences account for the divergence in cofactor dependences and behavior against substrates, inhibitors, and activators. In this group only the tartrate-resistant acid phosphatase is a metallo-enzyme whereas the other members do not require metal-ion binding for their catalytic activity. In addition, tartrate-resistant acid phosphatase and erythrocytic acid phosphatase are not inhibited by L-(+)-tartrate ion while the prostatic acid phosphatase is tartrate-sensitive. This is an important difference that can be exploited in in vitro assays to differentiate between different kinds of phosphatase activity. The search for more sensitive and specific methods of detection in clinical laboratory applications led to the development of radioimmunoassays (RIA) for determination of prostatic acid phosphatase in serum. These methods permit the direct quantification of the enzyme regardless of its activity status. Therefore, an independent structural classification exists that helps to group these enzymes according to their structural features and mechanisms. Based on this we can distinguish the histidine acid phosphatases (Van Etten, Ann N Y Acad Sci 390:27-51, 1982), the low molecular weight protein tyrosine acid phosphatases and the metal-ion dependent phosphatases. A note of caution is worthwhile mentioning here. The nomenclature of acid phosphatases has not been particularly easy for those new to the subject. Unfortunately, the acronym PAP is very common in the literature about purple acid phosphatases and prostatic acid phosphatase. In addition, LPAP is the acronym chosen to refer to the lysophosphatidic acid phosphatase which is a different enzyme. It is important to bear in mind this distinction while reviewing the literature to avoid confusion. PMID:23860654
Araujo, C�sar L; Vihko, Pirkko T
Exocarpic acid inhibits mycolic acid biosynthesis in Mycobacterium tuberculosis.
Exocarpic acid (13 E-octadecene-9,11-diynoic acid) from Exocarpos latifolius R.Br. (Santalaceae) was previously shown to have specific antimycobacterial activity. Microarray data suggested inhibition of fatty acid metabolism as a potential mode of action. Experiments designed to elucidate the mechanism of action showed that exocarpic acid was effective at inhibition of mycolic acid biosynthesis and did not act by dissipating the proton gradient in treated M. tuberculosis. Amide derivatives of exocarpic acid displayed similar properties to exocarpic acid, while other polyacetylenic fatty acids varied in their effects on mycolic acid biosynthesis. PMID:20506078
Koch, Michael; Bugni, Tim S; Sondossi, Mohammad; Ireland, Chris M; Barrows, Louis R
What Causes Acid Rain?
The phenomenon is the formation of acid rain. The resource explains the chemical reaction that begins when compounds like sulfur dioxide and nitrogen oxides are released into the air, mix and react with water and other chemicals to form more acidic pollutants that dissolve very easily in water and can be carried long distances where they become part of rain, sleet, snow, and fog.
Iodinated humic acids
Humic acids are iodinated by elemental iodine and, if the iodine is present as iodide, by peroxidase-mediated reactions. It is demonstrated that iodination of humic acids leads to a product with a uniform distribution of iodine. It could not be unambiguously verified whether the enzymatically mediated iodination is a direct reaction between a peroxidase-iodine complex and the humic acid molecule or a two-step reaction in which the enzyme creates elemental iodine, which consecutively reacts with the humic acid. Based on a simple model of a reaction between sites in the humic acids available for iodination and the electrophilic iodinating species, it was concluded that the reaction should be described as an equilibrium with a logarithmic equilibrium constant of approximately 4. The number of sites available for iodination was, in the humic acids studied, determined to be approximately 4�10-4 per gram humic acid. The different parameters influencing the enzymatically controlled iodination of humic acids are discussed.
Christiansen, Jesper V.; Carlsen, Lars
Characterization of acid tars.
Acid tars from the processing of petroleum and petrochemicals using sulfuric acid were characterized by gas chromatography/mass spectrometry (GC/MS), inductively coupled plasma/optical emission spectrometry (ICP/OES), differential scanning calorimetry (DSC), Fourier transform infrared (FTIR) spectrometry, and scanning electron microscopy/energy dispersive X-ray (SEM/EDX) micro-analysis. Leaching of contaminants from the acid tars in 48 h batch tests with distilled water at a liquid-to-solid ratio 10:1 was also studied. GC/MS results show that the samples contained aliphatic hydrocarbons, cyclic hydrocarbons, up to 12 of the 16 USEPA priority polycyclic aromatic hydrocarbons (PAHs), and numerous other organic groups, including organic acids (sulfonic acids, carboxylic acids and aromatic acids), phenyl, nitrile, amide, furans, thiophenes, pyrroles, and phthalates, many of which are toxic. Metals analysis shows that Pb was present in significant concentration. DSC results show different transition peaks in the studied samples, demonstrating their complexity and variability. FTIR analysis further confirmed the presence of the organic groups detected by GC/MS. The SEM/EDX micro-analysis results provided insight on the surface characteristics of the samples and show that contaminants distribution was heterogeneous. The results provide useful data on the composition, complexity, and variability of acid tars; information which hitherto have been scarce in public domain. PMID:19857924
Leonard, Sunday A; Stegemann, Julia A; Roy, Amitava
Recent reviews of available data indicate that precipitation in a large region of North America is highly acidic when its pH is compared with the expected pH value of 5.65 for pure rain water in equilibrium with CO2. A growing body of evidence suggests that acid rain is responsib...
Acid rain options
A number of contributions made to a series of seminars on acid rain options held by the Air Pollution Control Association are presented. They include statements on US and Canadian policy and legislation, and views from coal producers, electric utilities and motor vehicle manufacturers on measures to counteract or prevent the effects of acid rain.
Perhac, R.M.
Analysis of Organic Acids.
Presented are the procedures and a discussion of the results for an experiment in which students select unknown carboxylic acids, determine their melting points, and investigate their solubility behavior in water and ethanol. A table of selected carboxylic acids is included. (CW)
Griswold, John R.; Rauner, Richard A.
Altered retmoic acid receptors
Structurally and functionally altered retinoic acid receptors have been associated with rare human neoplasms: acute promyelocytic leuke- mia and hepatoceilular carcinoma. Whereas the ret- inoic acid receptor 13 (RAR13) rearrangement in hepatocellular carcinoma is unique, in acute promyelocytic leukemia (APL), RARU fusion to the promyelocytic leukemia (PML) gene by the t(15;17) translocation is a general feature of the disease. APL
CNRS UPR; Service de Biochimie
Humus Acids of Soils.
Humus acids are known to occur widely in soils, peats, and natural waters. Soil fertility, migration and accumulation of minerals in natural landscapes, and mineral nutrition of plants are all associated with these acids. In recent times they have been us...
D. S. Orlov
Lead-acid cell
A lead-acid storage battery is described that has a lead negative electrode, a lead dioxide positive electrode and a sulfuric acid electrolyte having an organic catalyst dissolved therein which prevents dissolution of the electrodes into lead sulfate whereby in the course of discharge, the lead dioxide is reduced to lead oxide and the lead is oxidized.
Hradcovsky, R.J.; Kozak, O.R.
Students are introduced to the differences between acids and bases and how to use indicators, such as pH paper and red cabbage juice, to distinguish between them. They learn why it is important for engineers to understand acids and bases.
Integrated Teaching And Learning Program
Acid in water
Plants and animals that live in water create some amount of acid in the water. The carbon dioxide that plants and animals release into the water makes the water acidic and unsafe for living organisms. This is why the water of captive aquatic animals and plants must be changed often.
Laszlo Ilyes (None;)
Acids in Proteins
This lesson describes how amino acids build proteins in a person's body. Amino acids are the chemical building blocks for the structure of an organism. A link to a quiz is provided at the end of the lesson to check comprehension.
Strong Acids (GCMP)
Strong Acids: this is a resource in the collection "General Chemistry Multimedia Problems". This problem will explore the properties of common strong acids. General Chemistry Multimedia Problems ask students questions about experiments they see presented using videos and images. The questions asked apply concepts from different parts of an introductory course, encouraging students to decompartmentalize the material.
Acids and Salts (GCMP)
Acids and Salts: this is a resource in the collection "General Chemistry Multimedia Problems". This problem will explore a few properties of common acids and their salts. General Chemistry Multimedia Problems ask students questions about experiments they see presented using videos and images. The questions asked apply concepts from different parts of an introductory course, encouraging students to decompartmentalize the material.
Lead-acid batteries
A method of producing a lead-acid battery capable of activation by the addition of electrolyte thereto, comprises the steps of: starting with a battery container accommodating at least one pack of battery plate grids having insulating separators interposed between adjacent grids, each of said grids carrying the lead-acid battery paste required to produce a positive or a negative battery plate
J. A. Bant; V. J. Raban
The report evaluates scientific information on direct health effects associated with exposure to acid aerosols. The present report is not intended as a complete and detailed review of all literature pertaining to acid aerosols. Rather, an attempt has been made to focus on the eva...
Acid Rain Revisited
The results of a long term study of the effects of acidic deposition in the Northeast were published in Bioscience this week, and they suggest that forests, lakes, and streams of the Northeastern US are not recovering from the toxic effects of acid rain despite significant cuts in the power plant emissions of sulfur dioxide and nitrogen oxide -- two major contributors to the problem. "Acid rain," more accurately called acidic deposition, causes toxic forms of aluminum to concentrate in soil and water, vital calcium and magnesium to be leached from trees, and surface waters to become inhospitable to aquatic biota. The study showed that, after 30 years of federally mandated air emission reductions, sulfur dioxide emissions have decreased while nitrogen oxide emissions have remained the same and that acidic deposition-related problems continue to plague New York and New England.
Sanders, Hilary C.
Acid rain is one of the foremost environmental issues of the 1980s and will be of continuining importance to energy policy for several reasons. First, the pollutants that cause acid rain are projected to increase through the end of the century as the demand for energy grows and as coal replaces oil. Second, many of the effects of acid rain are cumulative, so that even at current levels of emissions the problem is expected to intensify and to spread geographically. Third, the transport of air pollutants across state and national boundaries has raised fundamental issues of equity that are likely to be disputed for some time. Finally, any serious program for reducing acid rain will exert a profound influence on the future development of energy supplies. This review summarizes the causes, effects, and transport of acid rain, and discusses possible strategies for mitigating the problem.
Gould, R.R.
Airborne haloacetic acids.
Haloacetic acid (HAA) concentrations were measured in air samples from a semi-rural and a highly urbanized site in southern Ontario throughout 2000 to investigate their sources and gas-particle partitioning behavior. Denuders were efficient for collection of gaseous HAAs, and the particle phase was collected on a downstream quartz filter with negligible breakthrough. Total HAA concentrations (i.e., gas + particles) ranged between <0.025 and 19 ng m(-3) for individual HAAs at both sites. The dominant airborne HAA was monochloroacetic acid (MCA), followed in decreasing order by dichloroacetic acid (DCA), trifluoroacetic acid (TFA), and trichloroacetic acid (TCA). Difluoroacetic acid (DFA), monofluoroacetic acid (MFA), and chlorodifluoroacetic acid (CDFA) were also frequently detected at lower concentrations. Between sites, TFA, DFA, MFA, and TCA concentrations were significantly higher in Toronto, while CDFA concentrations were higher in Guelph. HAAs were primarily in the gas phase all year; however, during colder months, particle-phase HAA concentrations increased relative to the gas phase. Trichloroacetic acid had the highest particle fraction (phi) for all detected HAAs, with a mean phi of 0.51 and 0.56 for Guelph and Toronto, respectively, and both vapor pressure and acid strength appeared to influence gas-particle partitioning. Temporal trends at both sites were partially explained by temperature, short-wave radiation, and particle mass (PM10), leading to indications of the respective sources. A simple deposition model indicated that dry deposition of TFA and TCA should not be neglected in temperate mid-latitude environments and that precipitation concentrations can be successfully predicted by the Henry's law constant. PMID:12875391
Martin, Jonathan W; Mabury, Scott A; Wong, Charles S; Noventa, Francis; Solomon, Keith R; Alaee, Mehran; Muir, Derek C G
Contribution of organic acids to the acidity of Finnish lakes.
This study provides a comprehensive assessment of the contribution of organic carbon and organic acids to the acid-base status of Finnish lakes, summarizing empirical organic acidity measurements (from 16 lakes) combined with the Finnish Lake Survey data ...
P. Kortelainen
Acid recovery from waste sulfuric acid by diffusion dialysis
In the process of sulfuric acid production from pyrite, there is a lot of waste acid produced in fume washing with dilute\\u000a acid. Acid recovery from this sort of waste sulfuric acid by diffusion dialysis is studied in the paper. The mass transfer\\u000a dialysis coefficient of sulfuric acid of the membrane AFX is measured, the effect of the flowrate of
Guiqing Zhang; Qixiu Zhang; Kanggen Zhou
Trans Fatty Acids
Fats and their various fatty acid components seem to be a perennial concern of nutritionists and persons concerned with healthful diets. Advice on the consumption of saturated, polyunsaturated, monounsaturated, and total fat bombards us from magazines and newspapers. One of the newer players in this field is the group of trans fatty acids found predominantly in partially hydrogenated fats such as margarines and cooking fats. The controversy concerning dietary trans fatty acids was recently addressed in an American Heart Association (AHA) science advisory (1) and in a position paper from the American Society of Clinical Nutrition/American Institute of Nutrition (ASCN/AIN) (2). Both reports emphasize that the best preventive strategy for reducing risk for cardiovascular disease and some types of cancer is a reduction in total and saturated fats in the diet, but a reduction in the intake of trans fatty acids was also recommended. Although the actual health effects of trans fatty acids remain uncertain, experimental evidence indicates that consumption of trans fatty acids adversely affects serum lipid levels. Since elevated levels of serum cholesterol and triacylglycerols are associated with increased risk of cardiovascular disease, it follows that intake of trans fatty acids should be minimized.
Doyle, Ellin
Strongly Acidic Auxin Indole-3-Methanesulfonic Acid
A radiochemical synthesis is described for [14C]indole-3-methanesulfonic acid (IMS), a strongly acidic auxin analog. Techniques were developed for fractionation and purification of IMS using normal and reverse phase chromatography. In addition, the utility of both Fourier transform infrared spectrometry and fast atom bombardment mass spectrometry for analysis of IMS has been demonstrated. IMS was shown to be an active auxin, stimulating soybean hypocotyl elongation, bean first internode curvature, and ethylene production. IMS uptake by thin sections of soybean hypocotyl was essentially independent of solution pH and, when applied at a 100 micromolar concentration, IMS exhibited a basipetal polarity in its transport in both corn coleoptile and soybean hypocotyl sections. [14C]IMS should, therefore, be a useful compound to study fundamental processes related to the movement of auxins in plant tissues and organelles.
Cohen, Jerry D.; Baldi, Bruce G.; Bialek, Krystyna
Understanding acid rain
The complexities of the phenomenon of acid rain are described. Many factors, including meteorology, geology, chemistry, and biology, all play parts. Varying weather, varying soils, the presence of other pollutants and species differences all act to blur the connections between industrial emissions, acid rain, and environmental damage. Some experts believe that the greatest pH shock to lakes occurs during snow melt and runoff in the spring; others believe that much of the plant damage ascribed to acid rain is actually due to the effects of ozone. Much work needs to be done in the area of sampling. Historical data are lacking and sampling methods are not sufficiently accurate. (JMT)
Budiansky, S.
Nucleic acid-based matrixes
Various nucleic acid-based matrixes are provided, comprising nucleic acid monomers as building blocks, as well as nucleic acids encoding proteins, so as to produce novel biomaterials. Methods of utilizing such biomaterials include cell-free protein synthesis.
Waste Acid Detoxification and Reclamation.
Economically feasible processes that reduce the volume, quantity, and toxicity of metal-bearing waste acids by reclaiming, reusing, and recycling spent acids and metal salts are being developed and demonstrated. The acids used in the demonstrations are ge...
T. M. Brouns T. L. Stewart
This Environmental Security Technology Certification Program (ESTCP) project demonstrated the Waste Acid Detoxification and Reclamation (WADR) systems ability to recover waste electropolish acid solutions generated during the manufacturing of gun-tubes, and reuse the clean acid. ...
Hydroxy-Conjugated Fatty Acids.
This invention relates to a method of producing hydroxy-conjugated octadecadienoic acid from linoleic acid contained in vegetable oils and soap stocks from alkali-refined vegetable oils. Linoleic acid soaps are dispersed in an aqueous medium containing di...
E. A. Emkeu
Nucleic Acid Cloning.
The present invention provides an improved system for linking nucleic acids to one another. In particular, the present invention provides techniques for producing DNA product molecules that may be easily and directly ligated to recipient molecules. The pr...
K. A. Jarrell V. W. Coljee W. Donahue S. Mikheeva
(Acid rain workshop)
The traveler presented a paper entitled Susceptibility of Asian Ecosystems to Soil-Mediated Acid Rain Damage'' at the Second Workshop on Acid Rain in Asia. The workshop was organized by the Asian Institute of Technology (Bangkok, Thailand), Argonne National Laboratory (Argonne, Illinois), and Resource Management Associates (Madison, Wisconsin) and was sponsored by the US Department of Energy, the United Nations Environment Program, the United Nations Economic and Social Commission for Asia and the Pacific, and the World Bank. Papers presented on the first day discussed how the experience gained with acid rain in North America and Europe might be applied to the Asian situation. Papers describing energy use projections, sulfur emissions, and effects of acid rain in several Asian countries were presented on the second day. The remaining time was allotted to discussion, planning, and writing plans for a future research program.
Turner, R.S.
Aminolevulinic Acid Topical
... under the skin that result from exposure to sunlight and can develop into skin cancer) of the ... acid will make your skin very sensitive to sunlight (likely to get sunburn). Avoid exposure of treated ...
Acid-Base Tutorial
Website for anyone wanting to become more familiar with the physiology of acid-base balance in clinical medicine. Several pages are interactive. Numerical results are accompanied by text interpretations to facilitate recognition and understanding.
MD Alan W. Grogono (Tulane University School of Medicine Dept. of Anesthesiology)
Stomach acid test
Gastric acid secretion test ... The test is done after a period of not eating so that fluid is all that remains in the ... the stomach through the esophagus (food pipe). To test the ability of the cells in the stomach ...
Amino Acids and Chirality
Amino acids are among the most heavily studied organic compound class in carbonaceous chondrites. The abundance, distributions, enantiomeric compositions, and stable isotopic ratios of amino acids have been determined in carbonaceous chondrites fi'om a range of classes and petrographic types, with interesting correlations observed between these properties and the class and typc of the chondritcs. In particular, isomeric distributions appear to correlate with parent bodies (chondrite class). In addition, certain chiral amino acids are found in enantiomeric excess in some chondrites. The delivery of these enantiomeric excesses to the early Earth may have contributed to the origin of the homochirality that is central to life on Earth today. This talk will explore the amino acids in carbonaceous chondritcs and their relevance to the origin of life.
Cook, Jamie E.
Molecular Structure of Gallic acid
Gallic acid is found in its free state and combined with the tannin molecule, from which it can be extracted by the hydrolysis of tannic acid with sulfuric acid. Since one molecule of gallic acid has a carboxylic acid group and hydroxyl groups, it can react with another molecule of gallic acid to form an ester, digallic acid. When heated above 200 degrees C, gallic acid loses carbon dioxide to form pyrogallol (1,2,3-trihydroxybenzene, C6H3(OH)3), which is used in the production of azo dyes, photographic developers, and in laboratories for absorbing oxygen.
Acid Rain Lesson Plan
Five articulated lessons focus on air quality using classroom and field data collection activities. Case study in Great Smoky Mountains has broader application. Background and data for lessons on: the pH scale, understanding acid vs. base, collecting data, mapping relationship of weather events to acid rain. Links to NPS data on air quality, current values, atlas and reports, packaged datasets on ozone, meteorological conditions and other parameters. Also available: teacher resources; educator workshops.
Inflammation, acid and ulcers.
Chronic active type B gastritis is invariably the result of Helicobacter pylori infection and is an important factor in duodenal ulcer disease. The actions of mediators produced (a protein factor, a lipid soluble "pore-forming factor" and urease) or induced (immune/inflammatory cell mediators) by this bacterium on the control of gastric acid secretion are currently being investigated. These studies are reviewed in light of our current knowledge of the physiological control of gastric acid secretion.
Muller, M. J.; Hunt, R. H.
Domoic Acid Fact Sheet
This online fact sheet illustrates the transfer of domoic acid through the food web. Domoic acid is a nerve toxin produced by a naturally occurring Harmful Algal Bloom (HAB) usually (but not always) of the genus Pseudonitzchia. The sheet explains what to do if you find a sick or dead animal and includes contact information for injured/sick/entangled animal rescue networks in California.
Sanctuary, Channel I.
Effects of acid precipitation
Acid precipitation, including dry deposition, is recognized as coming from pollution-caused strong acid precursors that result from the burning of fossil fuels. Recent studies suggest that ecosystems susceptible to acidification occur over the entire length of the Appalachian Mountains and certain other regions of the eastern U.S. This paper reviews a short-term assessment by the EPA to survey the extent
Norman R. Glass; Dean E. Arnold; James N. Galloway; George R. Hendrey; Jeffrey J. Lee; W. W. McFee; Stephen A. Norton; Charles F. Powers; Danny L. Rambo; Carl L. Schofield
Acid-base chemistry
The book is not a research compendium and there are no references to the literature. It is a teaching text covering the entire range of undergraduate subject matter dealing with acid-base chemistry (some of it remotely) as taught in inorganic, analytical, and organic chemistry courses. The excellent chapters VII through IX deal in detail with the quantitative aspects of aqueous acid-base equilibria (salt hydrolysis and buffer, titrations, polyprotic and amphoteric substances).
Hand, C.W.; Blewit, H.L.
X-ray structures of LeuT in substrate-free outward-open and apo inward-open states
Neurotransmitter sodium symporters are integral membrane proteins that remove chemical transmitters from the synapse and terminate neurotransmission mediated by serotonin, dopamine, noradrenaline, glycine and GABA (?-aminobutyric acid). Crystal structures of the bacterial homologue, LeuT, in substrate-bound outward-occluded and competitive inhibitor-bound outward-facing states have advanced our mechanistic understanding of neurotransmitter sodium symporters but have left fundamental questions unanswered. Here we report
Harini Krishnamurthy; Eric Gouaux
Gas-phase acidities of the 20 protein amino acids
The gas-phase acidities of the 20 protein amino acids (PAAs) have been determined using an electrospray ionization-quadrupole ion trap instrument. Three different methods were used to determine both the absolute acidities and the relative acidity ordering of the PAAs. The extended kinetic method was used to determine absolute acidities for all 20 PAAs with substituted carboxylic acids and substituted phenols as reference acids. Acidities were obtained with an average uncertainty of �10 kJ/mol, which is large compared to some of the differences between amino acids with similar acidities. To determine the relative acidity ordering, single-reference kinetic method experiments were performed using both the reference acids from the absolute acidity studies and tryptophan and threonine as reference acids. Additional ordering information was obtained from kinetic method experiments in which proton-bound dimer ions comprising pairs of amino acids were generated and dissociated in the ion trap. The recommended acidity ordering is Gly < Ala < Pro < Val < Leu < Ile < Lys < Trp < Phe < Tyr < Met < Ser < Thr < Cys < Gln < Gln < Arg < Asn < His < Glu < Asp. Isodesmic acidity values were also obtained at the B3LYP/6-311++G**//B3LYP/6-31+G* level of theory with acetic acid as the reference acid. The theoretical acidities are in excellent agreement with the absolute acidities obtained from the extended kinetic method studies. The calculations predict that the preferred isomer for protonated cysteine and tyrosine is not a carboxylate anion, but rather a thiolate anion and a phenoxide anion, respectively.
Jones, Christopher M.; Bernier, Matthew; Carson, Erin; Colyer, Kathryn Ee; Metz, Rachel; Pawlow, Anna; Wischow, Emily D.; Webb, Ian; Andriole, Erica J.; Poutsma, John C.
Optimizing acid suppression for treatment of acid-related diseases
Gastric acid is of central importance in the pathogenesis of duodenal ulcer, gastric ulcer, and gastroesophageal reflux disease. Pharmacological reduction of acid secretion is, therefore, the mainstay of current treatment, but the optimal degree of acid suppression remains incompletely understood. This paper considers the ideal ways of assessing and reporting the pharmacological effectiveness of acid-inhibiting drugs and relating such data
Richard H. Hunt; Christer Cederberg; John Dent; Fred Halter; Colin Howden; I. N. Solly Marks; Simon Rune; Robert P. Walt
Thin-layer chromatography of gallic acid, methyl gallate, pyrogallol, phloroglucinol, catechol, resorcinol, hydroquinone, catechin, epicatechin, cinnamic acid, p-coumaric acid, ferulic acid and tannic acid
Six solvent systems of varying suitability are reported for the thin-layer chromatographic separation of simple phenolics and related compounds such as gallic acid, methyl gallate, pyrogallol, phloroglucinol, catechol, resorcinol, hydroquinone, catechin, epicatechin, cinnamic acid, p-coumaric acid, ferulic acid and tannic acid. The solvent system chloroform-ethyl acetate-acetic acid (50:50:1) facilitated the separation of all the compounds except pyrogallol and ferulic acid;
Om Prakash Sharma; Tej Krishan Bhat; Bhupinder Singh
Levulinic acid in organic synthesis
Data concerning the methods of synthesis, chemical transformations and application of levulinic acid are analysed and generalised. The wide synthetic potential of levulinic acid, particularly as a key compound in the synthesis of various heterocyclic systems, saturated and unsaturated ketones and diketones, difficultly accessible acids and other compounds is demonstrated. The accessibility of levulinic acid from hexose-containing wood-processing and agricultural
Boris V Timokhin; V A Baransky; G D Eliseeva
New look at sandstone acidizing
The acid mutual solvent (AMS) technique is a 3-step process which involves a preflush, a mixed HF-HCl stage, and an afterflush employing the mutual solvent. The preflush is normally regular hydrochloric acid (15% HCl). This step is designed to serve as a buffer between formation water and hydrofluoric acid. Normally an adequate preflush is 50 gal of regular acid per
Fungal production of citric acid
Citric acid is the principal organic acid found in citrus fruits. To meet increasing demands it is produced from carbohydrate feedstock by fermentation with the fungus Aspergillus niger and the yeasts of Candida spp. Effect of various fermentation conditions and the biochemistry of citric acid formation by A. niger have been discussed. Commercially citric acid is produced by surface, submerged
H. S. Grewal; K. L. Kalra
Acid rain: a background report
This Staff Brief was prepared for the Wisconsin Legislative Council's Special Committee on Acid Rain to provide an introduction to the issue of acid rain. It is divided into four parts. Part I provides an overview on the controversies surrounding the measurement, formation and effects of acid rain. As described in Part I, the term acid rain is used to
L. Glustrom; J. Stolzenberg
Molecular Structure of Malonic acid
Propanedioic acid is a dibasic carboxylic acid that was first synthesized by oxidizing malic acid in 1858 by a scientist named Dessaigne. Naturally, propandioic acid is found in apples. This chemical is relatively unstable and has few uses, but its ester derivative, diethyl malonate, is used to synthesize useful compounds such as barbiturates, flavors, fragrances, and vitamins (B1 and B6).
Optical high acidity sensor
An apparatus and method for determining acid concentrations in solutions having acid concentrations of from about 0.1 Molar to about 16 Molar is disclosed. The apparatus includes a chamber for interrogation of the sample solution, a fiber optic light source for passing light transversely through the chamber, a fiber optic collector for receiving the collimated light after transmission through the chamber, a coating of an acid resistant polymeric composition upon at least one fiber end or lens, the polymeric composition in contact with the sample solution within the chamber and having a detectable response to acid concentrations within the range of from about 0.1 Molar to about 16 Molar, a measurer for the response of the polymeric composition in contact with the sample solution, and a comparer of the measured response to predetermined standards whereby the acid molarity of the sample solution within the chamber can be determined. Preferably, a first lens is attached to the end of the fiber optic light source, the first lens adapted to collimate light from the fiber optic light source, and a second lens is attached to the end of the fiber optic collector for focusing the collimated light after transmission through the chamber. 10 figs.
Jorgensen, B.S.; Nekimken, H.L.; Carey, W.P.; O`Rourke, P.E.
Molecular Structure of Succinic acid
Succinic acid is an odorless and colorless crystal, triclinic or monoclinic prism with a very acid taste. Succinic acid is one of the natural acids found in broccoli, rhubarb, beets, asparagus, fresh meat extracts, sauerkraut and cheese. It is also a constituent of almost all plant and animal tissues and plays an important role in intermediary metabolism. Succinic acid is produced commercially by catalytic hydrogenation of maleic or fumaric acid or by acid hydrolysis of succinonitrile. Succinic acid is used in flavoring for food and beverages, and in the manufacture of lacquers, dyes, esters for perfumes, succinates, in photography and in foods as a sequestrant, buffer and neutralizing agent. Succinic acid has uses in certain drug compounds and in agricultural production. An interesting fact, succcinic acid has also been found in meteorites.
Exposures to acidic aerosols.
Ambient monitoring of acid aerosols in four U.S. cities and in a rural region of southern Ontario clearly show distinct periods of strong acidity. Measurements made in Kingston, TN, and Steubenville, OH, resulted in 24-hr H+ ion concentrations exceeding 100 nmole/m3 more than 10 times during summer months. Periods of elevated acidic aerosols occur less frequently in winter months. The H+ determined during episodic conditions in southern Ontario indicates that respiratory tract deposition can exceed the effects level reported in clinical studies. Observed 12-hr H+ concentrations exceeded 550 nmole/m3 (approximately 27 micrograms/m3 H2SO4). The maximum estimated 1-hr concentration exceeded 1500 nmole/m3 for H+ ions. At these concentrations, an active child might receive more than 2000 nmole of H+ ion in 12 hr and in excess of 900 nmole during the hour when H2SO4 exceeded 50 micrograms/m3.
Spengler, J D; Keeler, G J; Koutrakis, P; Ryan, P B; Raizenne, M; Franklin, C A
Amino acid analysis.
Amino acid analysis (AAA) is one of the best methods to quantify peptides and proteins. Two general approaches to quantitative AAA exist, namely, classical postcolumn derivatization following ion-exchange chromatography and precolumn derivatization followed by reversed-phase HPLC (RP-HPLC). Excellent instrumentation and several specific methodologies are available for both approaches, and both have advantages and disadvantages. This unit focuses on picomole-level AAA of peptides and proteins using the most popular precolumn-derivatization method, namely, phenylthiocarbamyl amino acid analysis (PTC-AAA). It is directed primarily toward those interested in establishing the technology with a modest budget. PTC derivatization and analysis conditions are described, and support and alternate protocols describe additional techniques necessary or useful for most any AAA method--e.g., sample preparation, hydrolysis, instrument calibration, data interpretation, and analysis of difficult or unusual residues such as cysteine, tryptophan, phosphoamino acids, and hydroxyproline. PMID:18429107
Crabb, J W; West, K A; Dodson, W S; Hulmes, J D
Reducing acid rain
The effects of acidic rainfall are most evident and highly publicized in Europe and the northeastern US. Greatest damage is done to lakes that are poorly buffered. The various strategies for reducing acid rain involve possible investments of billions of dollars annually. To minimize acid rain in a cost-effective manner, the authors must develop a better understanding of the chemistry of the oxides of nitrogen and sulfur as well as hydrogen peroxide, ozone, formaldehyde, and other species in cloud droplets and in the vapor state. The roles of reactive species, such as the radicals of OH, OOH, and NO/sub 3/, in oxidation reactions leading to scavenging of pollutants from the atmosphere need to be better understood in the heterogeneous atmosphere.
Not Available
Biodegradation of Cyanuric Acid
Cyanuric acid biodegrades readily under a wide variety of natural conditions, and particularly well in systems of either low or zero dissolved-oxygen level, such as anaerobic activated sludge and sewage, soils, muds, and muddy streams and river waters, as well as ordinary aerated activated sludge systems with typically low (1 to 3 ppm) dissolved-oxygen levels. Degradation also proceeds in 3.5% sodium chloride solution. Consequently, there are degradation pathways widely available for breaking down cyanuric acid discharged in domestic effluents. The overall degradation reaction is merely a hydrolysis; CO2 and ammonia are the initial hydrolytic breakdown products. Since no net oxidation occurs during this breakdown, biodegradation of cyanuric acid exerts no primary biological oxygen demand. However, eventual nitrification of the ammonia released will exert its usual biological oxygen demand.
Saldick, Jerome
Coronary vasodilatation by fatty acids
Summary Fatty acids increase the coronary flow rate of rat hearts, perfused according to theLangendorff technique. Long-chain and medium-chain fatty acids are more effective vasodilators than short-chain fatty acids. The vasodilatation by fatty acids does not proceed through the intermediate formation of the vasodilator adenosine, nor by stimulation of adenylcyclase activity. Since at low Ca2+ concentrations fatty acids not only
W. C. H�lsmann
Molecular Structure of Citric Acid
Citric Acid was first isolated in 1734 by Carl Wilhelm Scheele. Citric acid is found in many fruits, in particular lemons, grapefruit, and oranges. Several types of bacteria and fungi are also known to produce citric acid. In fact, the fungus Aspergillus niger produces the vast majority of citric acid, which is used in almost all carbonated sodas. Additionally, citric acid is also used to clean stainless steel.
Acid Deposition Lab
In this activity students will design an apparatus and carry out tests of fossil fuels to determine their impact on acid deposition by placing a small amount of a fossil fuel on a cotton puff and burning it to measure if acidic compounds are given off. Students will provide a diagram of their collection device and describe how it should function. Students will then draw a map showing the location of their precipitation collector and develop a graph or chart based on the results they have collected.
Liposomal spherical nucleic acids.
A novel class of metal-free spherical nucleic acid nanostructures was synthesized from readily available starting components. These particles consist of 30 nm liposomal cores, composed of an FDA-approved 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) lipid monomer. The surface of the liposomes was functionalized with DNA strands modified with a tocopherol tail that intercalates into the phospholipid layer of the liposomal core via hydrophobic interactions. The spherical nucleic acid architecture not only stabilizes these constructs but also facilitates cellular internalization and gene regulation in SKOV-3 cells. PMID:24983505
Banga, Resham J; Chernyak, Natalia; Narayan, Suguna P; Nguyen, SonBinh T; Mirkin, Chad A
Polyvalent Nucleic Acid Nanostructures
Polyvalent oligonucleotide-nanoparticle conjugates possess several unique emergent properties including enhanced cellular uptake, high antisense bioactivity, and nuclease resistance, which hypothetically originate from the dense packing and orientation of oligonucleotides on the surface of the nanoparticle. In this communication, we describe a new class of polyvalent nucleic acid nanostructures (PNANs), which comprise only crosslinked and oriented nucleic acids. We demonstrate that these particles are capable of effecting high cellular uptake and gene regulation without the need of a cationic polymer co-carrier. The PNANs also exhibit cooperative binding behavior and nuclease resistance properties.
Cutler, Joshua I.; Zhang, Ke; Zheng, Dan; Auyeung, Evelyn; Prigodich, Andrew E.
Acid rain revisited
This article reviews calculations of the estimates reported in this Newletter in 1983 on the contributions of nitric and sulfuric acids to deposition that may originate with motor vehicles. Previous estimates can now be updated, based on the recently released 1985 emissions inventory that was compiled by the National Acid Precipitation Assessment Program (NAPAP). This inventory relied in turn on the US Environmental Protection Agency's National emissions Data System (NEDS), the 1985 version of which is the latest containing both point source and area source information in sufficient detail to be useful for this purpose.
Ethylenediaminetetraacetic acid in endodontics
Ethylenediaminetetraacetic acid (EDTA) is a chelating agent can bind to metals via four carboxylate and two amine groups. It is a polyamino carboxylic acid and a colorless, water-soluble solid, which is widely used to dissolve lime scale. It is produced as several salts, notably disodium EDTA and calcium disodium EDTA. EDTA reacts with the calcium ions in dentine and forms soluble calcium chelates. A review of the literature and a discussion of the different indications and considerations for its usage are presented.
Mohammadi, Zahed; Shalavi, Sousan; Jafarzadeh, Hamid
Bacterial Degradation of 4-Hydroxyphenylacetic Acid and Homoprotocatechuic Acid
A species of Acinetobacter and two strains of Pseudomonas putida when grown with 4-hydroxyphenylacetic acid gave cell extracts that converted 3,4-dihydroxyphenylacetic acid (homoprotocatechuic acid) into carbon dioxide, pyruvate, and succinate. The sequence of enzyme-catalyzed steps was as follows: ring-fission by a 2,3-dioxygenase, nicotinamide adenine dinucleotide-dependent dehydrogenation, decarboxylation, hydration, aldol fission, and oxidation of succinic semialdehyde. Two new metabolites, 5-carboxymethyl-2-hydroxymuconic acid and 2-hydroxyhepta-2,4-diene-1,7-dioic acid, were isolated from reaction mixtures and a third, 4-hydroxy-2-ketopimelic acid, was shown to be cleaved by extracts to give pyruvate and succinic semialdehyde. Enzymes of this metabolic pathway were present in Acinetobacter grown with 4-hydroxyphenylacetic acid but were effectively absent when 3-hydroxyphenylacetic acid or phenylacetic acid served as sources of carbon.
Sparnins, Velta L.; Chapman, Peter J.; Dagley, Stanley
The Acid-Base Titration of a Very Weak Acid: Boric Acid
A laboratory experiment based on the titration of boric acid with strong base in the presence of d-mannitol is described. Boric acid is a very weak acid and direct titration with NaOH is not possible. An auxiliary reagent that contributes to the release of protons in a known stoichiometry facilitates the acid-base titration. Students obtain the�
Celeste, M.; Azevedo, C.; Cavaleiro, Ana M. V.
The second acidic constant of salicylic acid.
The second dissociation constant of salicylic acid (H2L) has been determined, at 25 degrees C, in NaCl ionic media by UV spectrophotometric measurements. The investigated ionic strength values were 0.16, 0.25, 0.50, 1.0, 2.0 and 3.0 M. The protolysis constants calculated at the different ionic strengths yielded, with the Specific Interaction Theory, the infinite dilution constant, log beta1(0) = 13.62 +/- 0.03, for the equilibrium L2- + H+ <==> HL-. The interaction coefficient between Na+ and L2-, b(Na+, L2-) = 0.02 +/- 0.07, has been also calculated. PMID:16235788
Porto, Raffaella; De Tommaso, Gaetano; Furia, Emilia
A Direct, Biomass-Based Synthesis of Benzoic Acid: Formic Acid-Mediated Deoxygenation of the Glucose-Derived Materials Quinic Acid and Shikimic Acid
An alternative biomass-based route to benzoic acid from the renewable starting materials quinic acid and shikimic acid is described. Benzoic acid is obtained selectively using a highly efficient, one-step formic acid-mediated deoxygenation method.
Arceo, Elena; Ellman, Jonathan; Bergman, Robert
Alkyl phosphonic acids and sulfonic acids in the Murchison meteorite
Homologous series of alkyl phosphonic acids and alkyl sulfonic acids, along with inorganic orthophosphate and sulfate, are identified in water extracts of the Murchison meteorite after conversion to their t-butyl dimethylsilyl derivatives. The methyl, ethyl, propyl, and butyl compounds are observed in both series. Five of the eight possible alkyl phosphonic acids and seven of the eight possible alkyl sulfonic acids through C4 are identified. Abundances decrease with increasing carbon number as observed of other homologous series indigenous to Murchison. Concentrations range downward from approximately 380 nmol/gram in the alkyl sulfonic acid series, and from 9 nmol/gram in the alkyl phosphonic acid series.
Cooper, George W.; Onwo, Wilfred M.; Cronin, John R.
Production of conjugated fatty acids by lactic acid bacteria.
Conjugated fatty acids have attracted much attention as a novel type of biologically beneficial functional lipid. Some isomers of conjugated linoleic acid (CLA) reduce carcinogenesis, atherosclerosis, and body fat. Considering the use of CLA for medicinal and nutraceutical purposes, a safe isomer-selective process is required. The introduction of biological reactions for CLA production could be an answer. We screened microbial reactions useful for CLA production, and found several unique reactions in lactic acid bacteria. Lactic acid bacteria produced CLA from linoleic acid. The produced CLA comprised a mixture of cis-9,trans-11-octadecadienoic acid (18:2) and trans-9,trans-11-18:2. Lactobacillus plantarum AKU 1009a was selected as a potential CLA producer. Using washed cells of L. plantarum AKU 1009a as a catalyst, CLA production from linoleic acid reached 40 mg/ml under the optimized conditions. The CLA-producing reaction was found to consist of two successive reactions, i.e., hydration of linoleic acid to 10-hydroxy-12-octadecenoic acid and dehydrating isomerization of the hydroxy fatty acid to CLA. On the basis of these results, the transformation of hydroxy fatty acids by lactic acid bacteria was investigated. Lactic acid bacteria transformed ricinoleic acid (12-hydroxy-cis-9-octadecenoic acid) to CLA (a mixture of cis-9,trans-11-18:2 and trans-9,trans-11-18:2). Castor oil, which is rich in the triacylglycerol form of ricinoleic acid, was also found to act as a substrate for CLA production by lactic acid bacteria with the aid of lipase-catalyzed triacylglycerol hydrolysis. L. plantarum AKU 1009a produced conjugated trienoic fatty acids from alpha- and gamma-linolenic acid. The trienoic fatty acids produced from alpha-linolenic acid were identified as cis-9,trans-11,cis-15-octadecatrienoic acid (18:3) and trans-9,trans-11,cis-15-18:3. Those produced from gamma-linolenic were cis-6,cis-9,trans-11-18:3 and cis-6,trans-9,trans-11-18:3. The conjugated trienoic fatty acids produced from alpha- and gamma-linolenic acid were further saturated by L. plantarum AKU 1009a to trans-10,cis-15-18:2 and cis-6,trans-10-18:2, respectively. PMID:16310724
Ogawa, Jun; Kishino, Shigenobu; Ando, Akinori; Sugimoto, Satoshi; Mihara, Kousuke; Shimizu, Sakayu
Synthesis of acid addition salt of delta-aminolevulinic acid from 5-bromo levulinic acid esters
A process of preparing an acid addition salt of delta-aminolevulinc acid comprising: a) dissolving a lower alkyl 5-bromolevulinate and hexamethylenetetramine in a solvent selected from the group consisting of water, ethyl acetate, chloroform, acetone, ethanol, tetrahydrofuran and acetonitrile, to form a quaternary ammonium salt of the lower alkyl 5-bromolevulinate; and b) hydrolyzing the quaternary ammonium salt with an inorganic acid to form an acid addition salt of delta-aminolevulinic acid.
Moens, Luc (Lakewood, CO)
Gamma-hydroxybutyric acid
Gamma-hydroxybutyric acid (GHB; liquid ecstasy, sodium oxybate) is a drug of abuse which is increasingly used recreationally and has been implicated in cases of �drug rape�. GHB-intoxicated patients may appear sedated or may show paradoxical agitation. After ingestion of a toxic amount of GHB, rapid onset of respiratory depression and deep coma may occur, resolving spontaneously within 24 hours with
Ruben Thanacoody
Plant fatty acid hydroxylase
The present invention relates to the identification of nucleic acid sequences and constructs, and methods related to the use of these sequences and constructs to produce genetically modified plants for the purpose of altering the composition of plant oils, waxes and related compounds.
Somerville, C.; Loo, F. van de
Acid rain game II.
In this paper we have developed a conceptual model of the issues connected to cooperative efforts in Europe to solve acid rains problems. The basic model is a simple game theoretic model with a unique equilibrium in dominant strategies. In order to illust...
K. G. Maeler
Acid rain in Asia
Acid rain has been an issue of great concern in North America and Europe during the past several decades. However, due to the passage of a number of recent regulations, most notably the Clean Air Act in the United States in 1990, there is an emerging perception that the problem in these Western nations is nearing solution. The situation in
N. Bhatti; D. G. Streets; W. K. Foell
Neeloo Bhatti; David G. Streets; Wesley K. Foell
Antimicrobial Fatty Acid Derivatives.
Acetylthio amides and esters were prepared by the free radical addition of thiolacetic acid to the terminal and nonterminal double bonds of N-substituted fatty amides and fatty esters. These new sulfur or halogen-containing compounds were found to have an...
R. R. Mod J. A. Harris F. C. Magne G. Sumrell A. F. Novak
Targeting tumor acidity
One of the main features of solid tumors is extracellular acidity, which correlates with tumor aggressiveness and metastatic potential. We introduced novel approach in targeting of acidic tumors, and translocation of cell-impermeable cargo molecules across cellular membrane. Our approach is based on main principle of insertion and folding of a polypeptide in lipid bilayer of membrane. We have identified family of pH Low Insertion Peptides (pHLIPs), which are capable spontaneous insertion and folding in membrane at mild acidic conditions. The affinity of peptides of pHLIP family to membrane at low pH is several times higher than at neutral pH. The process of peptides folding occurs within milliseconds. The energy released in a result of folding (about 2 kcal/mol) could be used to move polar cargo across a membrane, which is a novel concept in drug delivery. pHLIP peptides could be considered as a pH-sensitive single peptide molecular transporters and conjugated with imaging probes for fluorescence, MR, PET and SPECT imaging, they represent a novel in vivo marker of acidity. The work is supported by NIH grants CA133890 and GM073857 to OAA, DME, YRK.
Reshetnyak, Yana K.; Engelman, Donald M.; Andreev, Oleg A.
A growing body of evidence suggests that acid rain is responsible for substantial adverse effects on the public welfare. Such effects include: the acidification of lakes and rivers, with resultant damage to fish and other components of aquatic ecosystems; acidification and demineralization of soils; and possible reductions in crop and forest productivity. Affected areas include Canada and the northeastern US.
Norman R. Glass; Gary E. Glass; Peter J. Rennie
Ascorbic Acid in Oranges
IN 1934 Bacharach, Cook and Smith1 reported that the concentration of ascorbic acid in the peel of oranges was greater than that in the juice. This was the result of tests carried out on five bitter oranges and one sweet orange, and was afterwards confirmed by various workers2-6. In connexion with an investigation on methods of preparing orange juices for
L. H. Lampitt; L. C. Baker
[Acid hydrolysis of mandioca].
The influence of time of hydrolysis, pression of the process, ratio of mass of flour and volume and concentration of the acid solution was studied in the hydrolytic processes for Cassava flour. The aim was to obtain fermentable sugars, and the results were submitted to variance analysis. PMID:1228838
Colombo, A J; Schneiderman, B; Baruffaldi, R; Nacco, R
The present invention relates to the identification of nucleic acid sequences and constructs, and methods related thereto, and the use of these sequences and constructs to produce genetically modified plants for the purpose of altering the composition of plant oils, waxes and related compounds.
Somerville, Chris (Portola Valley, CA); van de Loo, Frank (Lexington, KY)
Citric acid urine test
Urine - citric acid test ... to discontinue drugs that may interfere with the test. On day 1, urinate into the toilet when ... No special preparation is necessary for this test. However, the ... performed while you are eating regularly. Ask your health care ...
40 CFR 721.3620 - Fatty acid amine condensate, polycarboxylic acid salts.
...acid amine condensate, polycarboxylic acid salts. 721.3620 Section 721.3620 ...acid amine condensate, polycarboxylic acid salts. (a) Chemical substance and significant...acid amine condensate, polycarboxylic acid salts. (PMN P-92-445) is subject...
Code of Federal Regulations, 2012 CFR
Code of Federal Regulations, 2011 CFR
Molecular Structure of Aspartic Acid
Aspartate was first isolated in 1868 from legumin in plant seeds. Aspartic acid forms colorless crystals that are soluble in water and insoluble in alcohols and ethers. This is a naturally occurring nonessential amino acid that is produced in the liver from oxaloacetic acid, but is plentiful in meats and sprouting seeds. The amino acid is important in the Krebs cycle as well as the urea cycle, where it is vital in the elimination of dietary waste products. Aspartic acid is required for stamina, brain and neural health. This acid has been found to be important in the functioning of ribonucleic acid (RNA), deoxyribonucleic acid (DNA), and in the production of immunoglobin and antibody synthesis. A deficiency of aspartate will lead to fatigue and depression. Aspartic acid has many uses that include biological and clinical studies, preparation of culture media, and it also functions as a detergent, fungicide, germicide, and metal complexation.
Specific bile acids inhibit hepatic fatty acid uptake
Bile acids are known to play important roles as detergents in the absorption of hydrophobic nutrients and as signaling molecules in the regulation of metabolism. Here we tested the novel hypothesis that naturally occurring bile acids interfere with protein-mediated hepatic long chain free fatty acid (LCFA) uptake. To this end stable cell lines expressing fatty acid transporters as well as primary hepatocytes from mouse and human livers were incubated with primary and secondary bile acids to determine their effects on LCFA uptake rates. We identified ursodeoxycholic acid (UDCA) and deoxycholic acid (DCA) as the two most potent inhibitors of the liver-specific fatty acid transport protein 5 (FATP5). Both UDCA and DCA were able to inhibit LCFA uptake by primary hepatocytes in a FATP5-dependent manner. Subsequently, mice were treated with these secondary bile acids in vivo to assess their ability to inhibit diet-induced hepatic triglyceride accumulation. Administration of DCA in vivo via injection or as part of a high-fat diet significantly inhibited hepatic fatty acid uptake and reduced liver triglycerides by more than 50%. In summary, the data demonstrate a novel role for specific bile acids, and the secondary bile acid DCA in particular, in the regulation of hepatic LCFA uptake. The results illuminate a previously unappreciated means by which specific bile acids, such as UDCA and DCA, can impact hepatic triglyceride metabolism and may lead to novel approaches to combat obesity-associated fatty liver disease.
Nie, Biao; Park, Hyo Min; Kazantzis, Melissa; Lin, Min; Henkin, Amy; Ng, Stephanie; Song, Sujin; Chen, Yuli; Tran, Heather; Lai, Robin; Her, Chris; Maher, Jacquelyn J.; Forman, Barry M.; Stahl, Andreas
Effects of acidic precipitation and acidity on soil microbial processes
Effects of oil acidity on microbial decomposition of organic matter and transformation of nitrogen in an acid forest soil were investigated. In the oak-leaf-amended pH-adjusted acid soils, CO/sub 2/ production in 14- and 150-day preincubated samples decreased by about 6 and 37%, respectively. In the control (unamended) acidified soils, reductions in CO/sub 2/ production of 14% in 14-day preincubated samples and 52% in 150-day samples were observed. Ammonia formation in the pH-adjusted acid soil was about 50% less than in the naturally acid soil. Increased rates of ammonification and nitrification were observed in the pH-adjusted neutral soil. Little autotrophic and heterotrophic nitrifying activity was detected in naturally acid and acidified forest soils. The rate of denitrification was rather slow in acid soils, and at greater acidities N/sub 2/O was the predominant end product. The abundance of nitrogen-fixing free-living bacteria was very low in acidic and acidified forest soils, and nitrogen gains by asymbiotic bacterial fixation in an acid forest ecosystem may be insignificant. These results suggest that further acidification of acid forest soils by addition of sulfuric acid or by acid precipitation may lead to significant reductions in the leaf litter decomposition, ammonification, nitrification, and denitrification and thus reduce nutrient recycling in the forest ecosystem.
Francis, A.J.
Fatty acid-producing hosts
Described are hosts for overproducing a fatty acid product such as a fatty acid. The hosts include an exogenous nucleic acid encoding a thioesterase and, optionally, an exogenous nucleic acid encoding an acetyl-CoA carboxylase, wherein an acyl-CoA synthetase in the hosts are functionally delected. The hosts prefereably include the nucleic acid encoding the thioesterase at an intermediate copy number. The hosts are preferably recominantly stable and growth-competent at 37.degree. C. Methods of producing a fatty acid product comprising culturing such hosts at 37.degree. C. are also described.
Pfleger, Brian F; Lennen, Rebecca M
[Clinical significance of bile acids].
During the last years bile acids have gained more and more clinical importance. They play a decisive part in intestinal fat resorption. Increased bile acid content in the colon will result in diarrhea. By determination of serum bile acids the liver function can be judged exactly. It seems probable that bile acids take part in the pathogenesis of gastritis gastric ulcer and colonic cancer. By administration of chenodeoxycholic acid and ursodeoxycholic acid dissolution of cholesterol stones within the gall bladder is possible. PMID:456969
Schmack, B
Fatty Acids of Mycobacterium kansasii
The cellular fatty acids of 35 strains of Mycobacterium kansasii isolated from clinical material were analyzed to establish properties by which we could identify and characterize these acid-fast microorganisms. The fatty acids were extracted from cells grown in liquid synthetic media, and they were analyzed as methyl esters by gas-liquid chromatography. The fatty acid profiles of all strains were similar. They differed from fatty acid profiles of other mycobacteria by their content of a saturated fatty acid with a methyl group at C2. Images
Thoen, Charles O.; Karlson, Alfred G.; Ellefson, Ralph D.
Process for Synthesis of Phosphonic Acids and Phosphinic Acids.
A process is claimed for the synthesis of phosphonic and phosphinic acids by hydrolysis of the corresponding esters. The phosphonic and phosphinic acids produced act as catalysts in the process, which is characterized by the fact that the hydrolysis proce...
H. D. Block H. Kohnen
Boswellic acid inhibits expression of acid sphingomyelinase in intestinal cells
Background Boswellic acid is a type of triterpenoids with antiinflammatory and antiproliferative properties. Sphingomyelin metabolism generates multiple lipid signals affecting cell proliferation, inflammation, and apoptosis. Upregulation of acid sphingomyelinase (SMase) has been found in several inflammation-related diseases such as inflammatory bowel diseases, atherosclerosis, and diabetes. Methods The present study is to examine the effect of 3-acetyl-11-keto-?-boswellic acids (AKBA), a potent boswellic acid, on acid SMase activity and expression in intestinal cells. Both transformed Caco-2 cells and non-transformed Int407 cells were incubated with AKBA. After incubation, the change of acid SMase activity was assayed biochemically, the enzyme protein was examined by Western blot, and acid SMase mRNA was quantified by qPCR. Results We found that AKBA decreased acid SMase activity in both intestinal cell lines in dose and time dependent manners without affecting the secretion of the enzyme to the cell culture medium. The effect of AKBA was more effective in the fetal bovine serum-free culture medium. Among different types of boswellic acid, AKBA was the most potent one. The inhibitory effect on acid SMase activity occurred only in the intact cells but not in cell-free extract in the test tubes. At low concentration, AKBA only decreased the acid SMase activity but not the quantity of the enzyme protein. However, at high concentration, AKBA decreased both the mass of acid SMase protein and the mRNA levels of acid SMase in the cells, as demonstrated by Western blot and qPCR, respectively. Under the concentrations decreasing acid SMase activity, AKBA significantly inhibited cell proliferation. Conclusion We identified a novel inhibitory effect of boswellic acids on acid SMase expression, which may have implications in human diseases and health.
Do trans Fatty Acids Impair Linoleic Acid Metabolism in Children?
Trans isomeric fatty acids disturb themetabolism of essential polyunsaturated fatty acids in animals and in premature infants. We assessed whether similar effects may also occur in healthy children. Plasma phospholipid fatty acid composition was analysed in 53 apparently healthy children aged 1-15 years (mean 7.5 years). Trans fatty acids were found in all samples and contributed 1.78 � 0.10% (w\\/w,
Tam�s Decsi; Berthold Koletzko
Prophylactic Tranexamic Acid and ?-Aminocaproic Acid for Primary Myocardial Revascularization
Background. The efficacy of prophylactic ?-aminocaproic acid and tranexamic acid to reduce transfusions after primary myocardial revascularization was evaluated in a teaching hospital context.Methods. Patients (n = 134) received either ?-aminocaproic acid (15-g bolus + infusion of 1 g\\/h), high-dose tranexamic acid (10-g bolus + placebo infusion), or normal saline solution in a double-blind fashion. Anticoagulation and conduct of cardiopulmonary
Jean-Fran�ois Hardy; Sylvain B�lisle; Charles Dupont; Fran�ois Harel; Danielle Robitaille; Micheline Roy; Lyne Gagnon
Effect of phenolic acids on glucose and organic acid metabolism by lactic acid bacteria from wine.
The influence of phenolic (p-coumaric, caffeic, ferulic, gallic and protocatechuic) acids on glucose and organic acid metabolism by two strains of wine lactic acid bacteria (Oenococcus oeni VF and Lactobacillus hilgardii 5) was investigated. Cultures were grown in modified MRS medium supplemented with different phenolic acids. Cellular growth was monitored and metabolite concentrations were determined by HPLC-RI. Despite the strong inhibitory effect of most tested phenolic acids on the growth of O. oeni VF, the malolactic activity of this strain was not considerably affected by these compounds. While less affected in its growth, the capacity of L. hilgardii 5 to degrade malic acid was clearly diminished. Except for gallic acid, the addition of phenolic acids delayed the metabolism of glucose and citric acid in both strains tested. It was also found that the presence of hydroxycinnamic acids (p-coumaric, caffeic and ferulic) increased the yield of lactic and acetic acid production from glucose by O. oeni VF and not by L. hilgardii 5. The results show that important oenological characteristics of wine lactic acid bacteria, such as the malolactic activity and the production of volatile organic acids, may be differently affected by the presence of phenolic acids, depending on the bacterial species or strain. PMID:19376463
Campos, Francisco M; Figueiredo, Ana R; Hogg, Tim A; Couto, Jos� A
An amino acid transporter involved in gastric acid secretion
Gastric acid secretion is regulated by a variety of stimuli, in particular histamine and acetyl choline. In addition, dietary factors such as the acute intake of a protein-rich diet and the subsequent increase in serum amino acids can stimulate gastric acid secretion only through partially characterized pathways. Recently, we described in mouse stomach parietal cells the expression of the system
Philipp Kirchhoff; Mital H. Dave; Christine Remy; Ortrud Kosiek; Stephanie M. Busque; Matthias Dufner; John P. Geibel; Francois Verrey; Carsten A. Wagner
College Chemistry Students' Mental Models of Acids and Acid Strength
The central goal of this study was to characterize the mental models of acids and acid strength expressed by advanced college chemistry students when engaged in prediction, explanation, and justification tasks that asked them to rank chemical compounds based on their relative acid strength. For that purpose we completed a qualitative research�
McClary, LaKeisha; Talanquer, Vicente
Acid Earth--The Global Threat of Acid Pollution.
Acid pollution is a major international problem, but the debate it has elicited has often clouded the distinction between myth and facts. This publication attempts to concerning the acid pollution situation. This publication attempts to identify available facts. It is the first global review of the problem of acid pollution and the first to�
McCormick, John
Oxalic acid excretion after intravenous ascorbic acid administration
Ascorbic acid is frequently administered intravenously by alternative health practitioners and, occasionally, by mainstream physicians. Intravenous administration can greatly increase the amount of ascorbic acid that reaches the circulation, potentially increasing the risk of oxalate crystallization in the urinary space. To investigate this possibility, we developed gas chromatography mass spectrometry methodology and sampling and storage procedures for oxalic acid analysis
Line Robitaille; Orval A. Mamer; Wilson H. Miller Jr.; Mark Levine; Sarit Assouline; David Melnychuk; Caroline Rousseau; L. John Hoffer
Molecular Structure of Sulfuric Acid
H2SO4 was discovered by alchemists and made from heating a compound of iron sulfate. In 1740, sulfuric acid was produced for commercial sale. Sulfuric acid is a very strong acid which is used in car batteries. The acid disassociates in water to give two protons and sulfate. This acid can destroy flesh and cause blindness. It was discovered in the 19th century that adding sulfuric acid to soil produces phosphorus, which is beneficial to plants; hence, sulfuric acid is used as a fertilizer in the form of super phosphate and ammonium sulfate. Sulfuric acid is also used to refine petroleum and process metals, and is found in paints and car batteries.
Crosslinked acid gels offer advantages
Acid polymer gels having a pH less than one have been crosslinked for retarding the chemical and physical activity of hydrochloric acid on calcareous formations. Hydrochloric acid concentrations from /one quarter/% to 28% have been successfully crosslinked. This unique stimulation fluid offers high viscosity with adequate shear stability, perfect support for propants, and clay stabilization. Additionally, the fluid provided effective fluid loss control and retardation of acid reaction enabling live acid to penetrate deeper into the formation for better conductivity; furthermore, there is practically a residue free break for rapid cleanup of the well after the job. Results of lab and field tests show this new acid crosslinked system to be an effective stimulation fluid for acidizing and acid fracturing in calcareous and sandstone formations having low permeability. 5 refs.
Molecular Structure of Glutaric acid
Glutaric acid is a colorless liquid and white crystals as a solid occurring in plants and animal tissues. It is used in organic synthesis and as an intermediate for the manufacture of polymers such as polyamides and polyesters, ester plasticizers and corrosion inhibitors. It is also useful in the application of decreasing polymer elasticity and in a variety of industrial applications. In addition glutaric acid plays an important role as an intermediary in the Krebs cycle and is used in medication against a large number of viruses and in animal diabetes. Glutaric acid can be prepared from cyclopentanone by oxidative ring fission with nitric acid and in the presence of a catalyst. Glutaric acid has the lowest melting point among dicarboxylic acids (98 C); it is very soluble in water and the solution in water is a medium strong acid. Short-term exposure to glutaric acid may cause irritation to the eyes, skin and the respiratory tract.
Ethylene Production from Linolenic Acid.
The possibility that the ethylene that is evolved from plants arises from linolenic acid was investigated by determining the relationship between ethylene evolution and linolenic acid levels from a number of plants. The evidence indicates that linolenic a...
F. B. Abeles
Omega-3-acid Ethyl Esters
Omega-3-acid ethyl esters are used together with lifestyle changes (diet, weight-loss, exercise) to reduce the amount ... a fat-like substance) in your blood. Omega-3-acid ethyl esters are in a class of ...
Omega-3 fatty acids (image)
Omega-3 fatty acids are a form of polyunsaturated fat that the body derives from food. Omega-3s (and omega-6s) are known as essential fatty acids (EFAs) because they are important for good health. ...
Ideas about Acids and Alkalis.
Investigates students' ideas, conceptions, and misconceptions about acids and alkalis before and after a teaching sequence in a small-scale research project. Concludes that student understanding of acids and alkalis is lacking. (DDR)
Toplis, Rob
Antilisterial activity of selected phenolic acids
Phenolic acids are known to exhibit antimicrobial activity against a variety of micro-organisms. Their influence on the behavior of Listeria monocytogenes, particularly in foods derived from plants, is not well understood. Several phenolic acids including chlorogenic acid and the hydroxycinnamic acids, caffeic acid, p-coumaric acid and ferulic acid, were screened for activity against five strains of L. monocytogenes using a
Aimin Wen; Pascal Delaquis; Kareen Stanich; Peter Toivonen
(Radioiodinated free fatty acids)
The traveler participated in the Second International Workshop on Radioiodinated Free Fatty Acids in Amsterdam, The Netherlands where he presented an invited paper describing the pioneering work at the Oak Ridge National Laboratory (ORNL) involving the design, development and testing of new radioiodinated methyl-branched fatty acids for evaluation of heart disease. He also chaired a technical session on the testing of new agents in various in vitro and in vivo systems. He also visited the Institute for Clinical and Experimental Nuclear Medicine in Bonn, West Germany, to review, discuss, plan and coordinate collaborative investigations with that institution. In addition, he visited the Cyclotron Research Center in Liege, Belgium, to discuss continuing collaborative studies with the Osmium-191/Iridium-191m radionuclide generator system, and to complete manuscripts and plan future studies.
Knapp, Jr., F. F.
Acid rain abatement
This patent describes a method of obtaining acid rain abatement from a flue gas containing nitrogen oxides (NOX) and sulfur oxides (SOX). It comprises the steps of treating the flue gas with a reducing agent to remove the remaining oxygen and produce an effluent, the reducing agent being selected from group consisting of natural gas, methane, a mixture of CO and hydrogen derived from steam, hydrocarbon, and hydrogen, passing effluent over a catalyst to simultaneously reduce the NOX to water and elemental nitrogen and the SOX to H{sub 2}S or elemental sulfur, the catalyst being selected from the group consisting of heteropoly acids and their salts, the reduction of the NOX and SOX taking place in a temperature range of 200{degrees} - 900{degrees} C., and removing the sulfur or sulfur compounds from the reduced flue gas to thereby remove essentially all of the NOX and SOX.
Stiles, A.B.
Industrial ecotoxicology "acid rain".
The acid rain phenomenon was studied in the province of Cordoba, Argentina. This study, based on a previously outlined framework, determined the anthropogenic origin of the low pH due to the presence of industrial hydrochloric acid wastage. This industrial ecotoxicological phenomenon seriously affected the forest wealth, causing a great defoliation of trees and shrubs, with a lower effect on crops. A survey on its effects on human beings has not been carried out, but considering the corrosion caused to different metals and its denouncing biocide effect on plants and animals, we should expect to find some kind of harm to the health of the workers involved or others engaged in farming, and even to those who are far away from the polluting agent. PMID:3758667
Astolfi, E; Gotelli, C; Higa, J
Weak Acid Equilibrium
Students are asked to calculate the pH of a weak acid aqueous solution. The problems involve a series of generic acids with assigned equilibrium constants (Ka) and total concentrations (Ct). Initially, students are required to hand calculate all problems by algebraic manipulation of the mathematical relationships of the system. The solution is a cubic equation. Through a series of assumptions, the solution is simplified. The assumptions are based on the chemistry of the system given the Ka and Ct for the problem. The problems are then graphically solved. Ultimately, the students develop an Excel worksheet to solve the problems and a Bjerrum plot to display the speciation as a function of pH.
Stapleton, Michael
An amino acid transporter involved in gastric acid secretion.
Gastric acid secretion is regulated by a variety of stimuli, in particular histamine and acetyl choline. In addition, dietary factors such as the acute intake of a protein-rich diet and the subsequent increase in serum amino acids can stimulate gastric acid secretion only through partially characterized pathways. Recently, we described in mouse stomach parietal cells the expression of the system L heteromeric amino acid transporter comprised of the LAT2-4F2hc dimer. Here we address the potential role of the system L amino acid transporter in gastric acid secretion by parietal cells in freshly isolated rat gastric glands. RT-PCR, western blotting and immunohistochemistry confirmed the expression of 4F2-LAT2 amino acid transporters in rat parietal cells. In addition, mRNA was detected for the B(0)AT1, ASCT2, and ATB(0+) amino acid transporters. Intracellular pH measurements in parietal cells showed histamine-induced and omeprazole-sensitive H+-extrusion which was enhanced by about 50% in the presence of glutamine or cysteine (1 mM), two substrates of system L amino acid transporters. BCH, a non-metabolizable substrate and a competitive inhibitor of system L amino acid transport, abolished the stimulation of acid secretion by glutamine or cysteine suggesting that this stimulation required the uptake of amino acids by system L. In the absence of histamine glutamine also stimulated H+-extrusion, whereas glutamate did not. Also, phenylalanine was effective in stimulating H+/K+-ATPase activity. Glutamine did not increase intracellular Ca2+ levels indicating that it did not act via the recently described amino acid modulated Ca2+-sensing receptor. These data suggest a novel role for heterodimeric amino acid transporters and may elucidate a pathway by which protein-rich diets stimulate gastric acid secretion. PMID:16308696
Kirchhoff, Philipp; Dave, Mital H; Remy, Christine; Kosiek, Ortrud; Busque, Stephanie M; Dufner, Matthias; Geibel, John P; Verrey, Francois; Wagner, Carsten A
Acid rain has been an issue of great concern in North America and Europe during the past several decades. However, due to\\u000a the passage of a number of recent regulations, most notably the Clean Air Act in the United States in 1990, there is an emerging\\u000a perception that the problem in these Western nations is nearing solution. The situation in
Acid Deposition Sampling
This learning activity from the Advanced Technology Environmental and Energy Center (ATEEC) provides students with the opportunity to use analytical meters and instruments and perform acid deposition sampling. Students will collect samples from various sources over a period of time, then measure pH and develop graphs or charts. Users must download this resource for viewing, which requires a free log-in. There is no cost to download the item.
Willey, Babe
Spermatotoxicity of dichloroacetic acid
The testicular toxicity of dichloroacetic acid (DCA), a disinfection byproduct of drinking water, was evaluated in adult male rats given both single and multiple (up to 14 d) oral doses. Delayed spermiation and altered resorption of residual bodies were observed in rats given single doses of 1500 and 3000 mg\\/kg; these effects persisted to varying degrees on post-treatment days 2,14,
Ralph E. Linder; Gary R. Klinefelter; Lillian F. Strader; Juan D. Suarez; Naomi L. Roberts
Lipoic acid biosynthesis defects.
Lipoate is a covalently bound cofactor essential for five redox reactions in humans: in four 2-oxoacid dehydrogenases and the glycine cleavage system (GCS). Two enzymes are from the energy metabolism, ?-ketoglutarate dehydrogenase and pyruvate dehydrogenase; and three are from the amino acid metabolism, branched-chain ketoacid dehydrogenase, 2-oxoadipate dehydrogenase, and the GCS. All these enzymes consist of multiple subunits and share a similar architecture. Lipoate synthesis in mitochondria involves mitochondrial fatty acid synthesis up to octanoyl-acyl-carrier protein; and three lipoate-specific steps, including octanoic acid transfer to glycine cleavage H protein by lipoyl(octanoyl) transferase 2 (putative) (LIPT2), lipoate synthesis by lipoic acid synthetase (LIAS), and lipoate transfer by lipoyltransferase 1 (LIPT1), which is necessary to lipoylate the E2 subunits of the 2-oxoacid dehydrogenases. The reduced form dihydrolipoate is reactivated by dihydrolipoyl dehydrogenase (DLD). Mutations in LIAS have been identified that result in a variant form of nonketotic hyperglycinemia with early-onset convulsions combined with a defect in mitochondrial energy metabolism with encephalopathy and cardiomyopathy. LIPT1 deficiency spares the GCS, and resulted in a combined 2-oxoacid dehydrogenase deficiency and early death in one patient and in a less severely affected individual with a Leigh-like phenotype. As LIAS is an iron-sulphur-cluster-dependent enzyme, a number of recently identified defects in mitochondrial iron-sulphur cluster synthesis, including NFU1, BOLA3, IBA57, GLRX5 presented with deficiency of LIAS and a LIAS-like phenotype. As in DLD deficiency, a broader clinical spectrum can be anticipated for lipoate synthesis defects depending on which of the affected enzymes is most rate limiting. PMID:24777537
Mayr, Johannes A; Feichtinger, Ren� G; Tort, Frederic; Ribes, Antonia; Sperl, Wolfgang
Acid Rain Effects
Learners conduct a simple experiment to model and explore the harmful effects of acid rain (vinegar) on living (green leaf and eggshell) and non-living (paper clip) objects. Learners observe the effects over a period of days. This activity has links to other activities which can be combined to make a larger lesson. Resource contains vocabulary definitions and suggestions for assessment, extensions, and scaling for different levels of learners.
Kolenbrander, Amy; Yowell, Janet; Mach, Natalie; Zarske, Malinda S.; Carlson, Denise
Acid hydrolysis of chitosans
The hydrolysis of the O-glycosidic linkages (depolymerization) and the N-acetyl linkage (de-N-acetylation) of partially N-acetylated chitosans were studied in dilute and concentrated HCl. The rate of hydrolysis of the glycosidic linkages was found to be equal to the rate of de-N-acetylation in dilute acid, while the glycosidic linkages was hydrolysed more than 10 times faster than the N-acetyl linkage in
K. M. V�rum; M. H. Ott�y; O. Smidsr�d
Enviropedia: Introduction to Acid Rain
This resource provides information about acid rain, a widespread term used to describe all forms of acid precipitation. The sources, nature, and chemistry of acid rain are discussed, along with its impact on buildings, soils, freshwater lakes, trees, and wildlife. Other topics include measuring, modeling, and monitoring acid rain; and vehicle and industrial emission controls. The problem of airborne pollutants migrating across international borders is also discussed.
Molecular Structure of Octanoic acid
Caprylic acid is a colorless oil manufactured from 1-heptene or 1-octanol. Octanoic acid has an unpleasant rancid taste. When converted from the carboxlic acid to an ester, it has a pleasant taste. In addition, esters of caprylic acid are used in the preparation of dyes, perfumes, and food preservatives. This compound has also been found to have antifungal activity and is used to treat yeast infections.
Fatty acid signaling in Arabidopsis
.�� Many organisms use fatty acid derivatives as biological regulators. In plants, for example, fatty acid-derived signals have\\u000a established roles in the regulation of developmental and defense gene expression. Growing numbers of these compounds, mostly\\u000a derived from fatty acid hydroperoxides, are being characterized. The model plant Arabidopsis thaliana is serving a vital role in the discovery of fatty acid-derived signal
Edward E. Farmer; Hans Weber; Sabine Vollenweider
Exposures to acidic aerosols
Ambient monitoring of acid aerosols in four U.S. cities and in a rural region of southern Ontario clearly show distinct periods of strong acidity. Measurements made in Kingston, TN, and Steubenville, OH, resulted in 24-hr H+ ion concentrations exceeding 100 nmole/m/sup 3/ more than 10 times during summer months. Periods of elevated acidic aerosols occur less frequently in winter months. The H+ determined during episodic conditions in southern Ontario indicates that respiratory tract deposition can exceed the effects level reported in clinical studies. Observed 12-hr H+ concentrations exceeded 550 nmole/m/sup 3/ (approximately 27 micrograms/m/sup 3/ H/sub 2/SO/sub 4/). The maximum estimated 1-hr concentration exceeded 1500 nmole/m/sup 3/ for H+ ions. At these concentrations, an active child might receive more than 2000 nmole of H+ ion in 12 hr and in excess of 900 nmole during the hour when H/sub 2/SO/sub 4/ exceeded 50 micrograms/m/sup 3/.
Spengler, J.D.; Keeler, G.J.; Koutrakis, P.; Ryan, P.B.; Raizenne, M.; Franklin, C.A.
Ambient monitoring of acid aerosols in four U.S. cities and in a rural region of southern Ontario clearly show distinct periods of strong acidity. Measurements made in Kingston, TN, and Steubenville, OH, resulted in 24-hr H(+) ion concentrations exceeding 100 nmole/cu m more than 10 times during summer months. Periods of elevated acidic aerosols occur less frequently in winter months. The H(+) determined during episodic conditions in southern Ontario indicates that respiratory tract deposition can exceed the effects level reported in clinical studies. Observed 12-hr (H+) concentrations exceeded 550 nmole/cu m (approximately 27 microgram/cu m H{sub 2}SO{sub 4}). The maximum estimated 1-hr concentration exceeded 1500 nmole/cu m for H(+) ions. At these concentrations, an active child might receive more than 2000 nmole of H(+) ion in 12 hr and in excess of 900 nmole during the hour when H{sub 2}SO{sub 4} exceeded 50 microgram/cu m.
Spengler, J.D.; Keeler, G.J.; Koutrakis, P.; Ryan, P.B.; Raizenne, M.
Acid rain has been an issue of widespread concern in North America and Europe for more than fifteen years. However, there is an emerging feeling that the problem in Europe and North America is nearing solution, largely as a result of existing and newly enacted legislation, decreased energy use due to conservation and efficiency improvements, and/or trends in energy policy away from fossil fuels. The situation in Asia appears much bleaker. Fossil fuels are already used in large quantities, such that local air pollution is becoming a serious problem and high deposition levels are being measured. Emission regulations in most countries (with the notable exception of Japan) are not very stringent. Energy plans in many countries (particularly PRC, India, Thailand, and South Korea) call for very large increases in coal combustion in the future. Finally, there is not presently a strong scientific or public constituency for action to mitigate the potential effects of acid deposition. These factors imply potentially serious problems in the future for long-range transport and deposition of sulfur and nitrogen species and consequent damage to ecosystems and materials. The political ramifications of transboundary environmental pollution in this region are also potentially serious. The purpose of this paper is to provide background information on the acid deposition situation in Asia, with the intention of laying the foundation for the development of a possible research program for this region. 36 refs., 8 figs., 8 tabs.
Bhatti, N.; Streets, D.G. (Argonne National Lab., IL (USA)); Foell, W.K. (Resource Management Associates, Madison, WI (USA))
In Asia, fossil fuels are used in large quantities, such that local air pollution is becoming a serious problem and high deposition levels are being measured. Emission regulations in most countries (with the exception of Japan) are not very stringent. Energy plans in many countries call for very large increases in coal combustion in the future. Finally, there is not presently a strong constituency for action to mitigate the potential effects of acid deposition. These factors imply potentially serious problems in the future for long-range transport and deposition of sulfur and nitrogen species and consequent damage to ecosystems and materials. The political ramifications of transboundary environmental pollution in this region are also potentially serious. A predictive tool could be built to help decision makers project future trends in emissions, estimate the regional consequences for acid deposition levels, evaluate the vulnerability of natural and man-made systems, and determine the costs and effectiveness of alternative mitigative actions that might be taken. Such a policy analysis exercise can start to raise environmental awareness in the region and begin a dialogue that could help ameliorate an environmental problem in its early stages. The purpose of this paper is to provide background information on the acid deposition situation in Asia, with the intention of laying the foundation for the design of a possible research program for this region. 41 refs., 10 figs., 9 tabs.
Bhatti, N.; Streets, D.G. (Argonne National Lab., IL (USA)); Foell, W.K. (Resource Management Associates, Madison, WI (USA) Wisconsin Univ., Madison, WI (USA))
Cannabinoid acids analysis.
The cannabinoid pattern of vegetable preparations from Cannabis sativa (hashish, marijuana) allows to recognize the phenotype of the plants, to be used as drug or for fiber. Cannabinoid determination by analytical point of view has represented some problems caused by the complex composition of the hexane extract. Capillary gas chromatography of the hexane extracts of vegetable samples, shows the presence of rather polar constituents that eluted, with noticeable interactions, only on polar phase. The compounds can be methylated by diazomethane and silanized (TMS) by silylating reagents. The methyl and methyl-TMS derivatives are analyzed by high resolution gas chromatography (HRGC) and by gas chromatography-mass spectrometry (GC-MS). The identification of the compounds shows their nature of cannabinoid acids, which the main by quantitative point of view results the cannabidiolic acid (CBDA). It is known that the cannabinoid acids are thermally unstable and are transformed in the corresponding cannabinoids by decarboxilation. This is of interest in forensic analysis with the aim to establish the total amount of THC in the Cannabis preparations, as the active component. PMID:1503600
Lercker, G; Bocci, F; Frega, N; Bortolomeazzi, R
Docosahexaenoic acid: A valuable nutraceutical?
A high content of docosahexaenoic acid (DHA) in the brain and retina is important for proper nervous system and visual functions. Although it is difficult to deplete the mammalian nervous system of its essential fatty acids, particularly in adults, a well-balanced intake of both w-3 and ?-6 polyunsaturated fatty acids (PUFAs) appears to be necessary for the satisfactory development of
Y.-Y. Linko; K. Hayakawa
Molecular Structure of Carbonic acid
The hypothetical acid formed with carbon dioxide and water; it is only in the H2CO3 form when in solution. This acid is found in everyday products, the most prominent of which include carbonated beverages. The conversion of carbonic acid into water and carbon dioxide in sodas is the reason the beverage looses the bubbling.
Factors Controlling Naphthenic Acid Corrosion
A laboratory study was conducted to elucidate the influence of chemical and physical parameters on corrosion of type 1018 carbon steel (CS, UNS G10180) and 5% Cr-0.5% Mo steel in oils containing naphthenic acids (NAs) for application to crude oil refinery systems. Effects of test duration, temperature, and acid concentration were assessed for a range of single acids of varying
Alan Turnbull; Evelina Slavcheva; Bryan Shone
Uranium Extraction from Phosphoric Acid.
A study has been carried out for the extraction of uranium from phosphoric acid produced in Algeria. First of all, the Algerian phosphoric acid produced by SONATRACH has been characterised. This study helped the authors to synthesize a phosphoric acid tha...
A. Lounis
New politics of acid rain
The acid rain problem is not nationwide across the USA but the politicians want to spread the cost of emission reductions. An overview of acid rain and its environmental impacts is given, and a cost-benefit analysis of acid rain control is outlined. USA policies are discussed. 6 references.
Trisko, E.M.
Acid rain: the international response
Acid rain or technically �acid deposition� has far reaching environmental, economic, political and international implications. It has been blamed for large?scale damage to aquatic ecosystems and forests in Scandinavia, southeastern Canada and the northeastern United States. A number of other countries have expressed concern about the possible affects of acid rain on water bodies, forests, agricultural crops and material structures.Never
Gordon L. Brady; Joseph C. Selle
Carboxylic acid sorption regeneration process
Carboxylic acids are sorbed from aqueous feedstocks into an organic liquid phase or onto a solid adsorbent. The acids are freed from the sorbent phase by treating it with aqueous alkylamine thus forming an alkylammonium carboxylate which is dewatered and decomposed to the desired carboxylic acid and the alkylamine.
King, C. Judson (Kensington, CA); Poole, Loree J. (Baton Rouge, LA)
Acid rain. what is it
Acid rain and its effects are difficult to measure. Even harder to determine is whether reducing sulfur and nitrogen oxide emissions will reduce acid rain and, if so, where the effect will take place. A better understanding of atmospheric chemistry and the role of oxidizing agents and clouds in forming acids is needed. Several bills before Congress call for reductions
The possibility of linolenic acid serving as the precursor of ethylene in vivo was investigated. A comparison of the amount of linolenic acid present in plant tissues with the rates of ethylene production indicated that linolenic acid does not serve as th...
Acid rain: Reign of controversy
Acid Rain is a primer on the science and politics of acid rain. Several introductory chapters describe in simple terms the relevant principles of water chemistry, soil chemistry, and plant physiology and discuss the demonstrated or postulated effects of acid rain on fresh waters and forests as well as on statuary and other exposed objects. There follow discussions on the
Determination of Mine Waste Acidity.
Pollution from acid mine drainage is a chronic problem. The current vigorous interest has led to a re-examination of the methods of measuring total acidity, which are clearly based on the limited laboratory capabilities of prior years. Defined acid-metal ...
B. V. Salotto E. F. Barth M. B. Ettinger W. E. Tolliver
Scientists Puzzle Over Acid Rain
Reports on a growing concern over increased acidity in atmospheric percipitation. Explores possible causes of the increased acidity, identifies chemical components of precipitation in various parts of the world, and presents environmental changes that might be attributed to the acidity. (GS)
Chemical and Engineering News, 1975
Experimental study of the hydrothermal reactivity of organic acids and acid anions: II. Acetic acid, acetate, and valeric acid
Organic acids and acid anions occur in substantial concentrations in many aqueous geologic fluids and are thought to take part in a variety of geochemical processes ranging from the transport of metals in ore-forming fluids to the formation of natural gas to serving as a metabolic energy source for microbes in subsurface habitats. The widespread occurrence of organic acids and their potential role in diverse geologic processes has led to numerous experimental studies of their thermal stability, yet there remain substantial gaps in our knowledge of the factors that control the rates and reaction pathways for the decomposition of these compounds under geologic conditions. In order to address some of these uncertainties, a series of laboratory experiments were conducted to examine the behavior of organic acids and acid anions under hydrothermal conditions in the presence of minerals. Reported here are results of experiments where aqueous solutions of acetic acid, sodium acetate, or valeric acid ( n-pentanoic acid) were heated at 325�C, 350 bars in the presence of the mineral assemblages hematite + magnetite + pyrite, pyrite + pyrrhotite + magnetite, and hematite + magnetite. The results indicate that aqueous acetic acid and acetate decompose by a combination of two reaction pathways: decarboxylation and oxidation. Both reactions are promoted by minerals, with hematite catalyzing the oxidation reaction while magnetite catalyzes decarboxylation. The oxidation reaction is much faster, so that oxidation dominates the decomposition of acetic acid and acetate when hematite is present. In contrast to previous reports that acetate decomposed more slowly than acetic acid, we found that acetate decomposed at slightly faster rates than the acid in the presence of minerals. Although longer-chain monocarboxylic acids are generally thought to decompose by decarboxylation, valeric acid appeared to decompose primarily by "deformylation" to 1-butene plus formic acid. Subsequent decomposition of 1-butene and formic acid generated a variety of short-chain (?C 4) hydrocarbons and moncarboxylic acids as well as CO 2. Valeric acid decomposition proceeded more rapidly (by a factor of 2) in the presence of hematite-magnetite-pyrite than with the other mineral assemblages, with the greater reaction rate apparently attributable to the effects of fluid chemistry. Valeric acid was observed to decompose at a substantially faster rate than acetic acid under similar conditions. The results suggest that decomposition of aqueous monocarboxylic acids may make a significant contribution to the conversion of petroleum to light hydrocarbons in natural gas and thermal fluids.
McCollom, Thomas M.; Seewald, Jeffrey S.
Acid-hydrolysis of fish wastes for lactic acid fermentation.
In this study, two acid-hydrolysis processes, process A and process B, were proposed to produce low-cost nutrients for the production of lactic acid. Process A was a direct way to hydrolyze protein with diluted acid while process B was process A plus fish wastes pretreatment (an extraction by water). The two methods could both treat fish wastes to be suitable nutrient sources for promoting lactic acid production. As the pretreatment indicated some favorable effect on fish waste hydrolyzate (FWH), process B increased lactic acid productivity by 22%. Compared with 20 g/L yeast extract (YE), 6.8% FWH hydrolyzed by process B had more efficiency in lactic acid production, indicating that process B was suitable to produce high performance nutrients for lactic acid production and FWH hydrolyzed by process B would be an substitute for YE. PMID:16293413
Gao, Min-Tian; Hirata, Makoto; Toorisaka, Eiichi; Hano, Tadashi
Composition for nucleic acid sequencing
The present invention is directed to a method of sequencing a target nucleic acid molecule having a plurality of bases. In its principle, the temporal order of base additions during the polymerization reaction is measured on a molecule of nucleic acid, i.e. the activity of a nucleic acid polymerizing enzyme on the template nucleic acid molecule to be sequenced is followed in real time. The sequence is deduced by identifying which base is being incorporated into the growing complementary strand of the target nucleic acid by the catalytic activity of the nucleic acid polymerizing enzyme at each step in the sequence of base additions. A polymerase on the target nucleic acid molecule complex is provided in a position suitable to move along the target nucleic acid molecule and extend the oligonucleotide primer at an active site. A plurality of labelled types of nucleotide analogs are provided proximate to the active site, with each distinguishable type of nucleotide analog being complementary to a different nucleotide in the target nucleic acid sequence. The growing nucleic acid strand is extended by using the polymerase to add a nucleotide analog to the nucleic acid strand at the active site, where the nucleotide analog being added is complementary to the nucleotide of the target nucleic acid at the active site. The nucleotide analog added to the oligonucleotide primer as a result of the polymerizing step is identified. The steps of providing labelled nucleotide analogs, polymerizing the growing nucleic acid strand, and identifying the added nucleotide analog are repeated so that the nucleic acid strand is further extended and the sequence of the target nucleic acid is determined.
Korlach, Jonas (Ithaca, NY) [Ithaca, NY; Webb, Watt W. (Ithaca, NY) [Ithaca, NY; Levene, Michael (Ithaca, NY) [Ithaca, NY; Turner, Stephen (Ithaca, NY) [Ithaca, NY; Craighead, Harold G. (Ithaca, NY) [Ithaca, NY; Foquet, Mathieu (Ithaca, NY) [Ithaca, NY
Evolution of rosmarinic acid biosynthesis.
Rosmarinic acid and chlorogenic acid are caffeic acid esters widely found in the plant kingdom and presumably accumulated as defense compounds. In a survey, more than 240 plant species have been screened for the presence of rosmarinic and chlorogenic acids. Several rosmarinic acid-containing species have been detected. The rosmarinic acid accumulation in species of the Marantaceae has not been known before. Rosmarinic acid is found in hornworts, in the fern family Blechnaceae and in species of several orders of mono- and dicotyledonous angiosperms. The biosyntheses of caffeoylshikimate, chlorogenic acid and rosmarinic acid use 4-coumaroyl-CoA from the general phenylpropanoid pathway as hydroxycinnamoyl donor. The hydroxycinnamoyl acceptor substrate comes from the shikimate pathway: shikimic acid, quinic acid and hydroxyphenyllactic acid derived from l-tyrosine. Similar steps are involved in the biosyntheses of rosmarinic, chlorogenic and caffeoylshikimic acids: the transfer of the 4-coumaroyl moiety to an acceptor molecule by a hydroxycinnamoyltransferase from the BAHD acyltransferase family and the meta-hydroxylation of the 4-coumaroyl moiety in the ester by a cytochrome P450 monooxygenase from the CYP98A family. The hydroxycinnamoyltransferases as well as the meta-hydroxylases show high sequence similarities and thus seem to be closely related. The hydroxycinnamoyltransferase and CYP98A14 from Coleus blumei (Lamiaceae) are nevertheless specific for substrates involved in RA biosynthesis showing an evolutionary diversification in phenolic ester metabolism. Our current view is that only a few enzymes had to be "invented" for rosmarinic acid biosynthesis probably on the basis of genes needed for the formation of chlorogenic and caffeoylshikimic acid while further biosynthetic steps might have been recruited from phenylpropanoid metabolism, tocopherol/plastoquinone biosynthesis and photorespiration. PMID:19560175
Petersen, Maike; Abdullah, Yana; Benner, Johannes; Eberle, David; Gehlen, Katja; H�cherig, Stephanie; Janiak, Verena; Kim, Kyung Hee; Sander, Marion; Weitzel, Corinna; Wolters, Stefan