Structure and reaction geometry of geranylgeranyl diphosphate synthase from Sinapis alba.
The crystal structure of the geranylgeranyl diphosphate synthase from Sinapis alba (mustard) has been solved in two crystal forms at 1.8 and 2.0 A resolutions. In one of these forms, the dimeric enzyme binds one molecule of the final product geranylgeranyl diphosphate in one subunit. The chainfold of the enzyme corresponds to that of other members of the farnesyl diphosphate synthase family. Whereas the binding modes of the two substrates dimethylallyl diphosphate and isopentenyl diphosphate at the allyl and isopentenyl sites, respectively, have been established with other members of the family, the complex structure presented reveals for the first time the binding mode of a reaction product at the isopentenyl site. The binding geometry of substrates and product in conjunction with the protein environment and the established chemistry of the reaction provide a clear picture of the reaction steps and atom displacements. Moreover, a comparison with a ligated homologous structure outlined an appreciable induced fit: helix alpha8 and its environment undergo a large conformational change when either the substrate dimethylallyl diphosphate or an analogue is bound to the allyl site; only a minor conformational change occurs when the other substrate isopentenyl diphosphate or the product is bound to the isopentenyl site.


Fibroblasts of two patients with trisomy 18 show 1.5-fold increase in peptidase A activity over normal human diploid fibroblasts.
Peptidase A (Pep A) activity assigned to chromosome 18q23 was biochemically examined in fibroblasts cultured from two patients with trisomy 18 and in fibroblasts derived from normal individuals. The trisomy 18 fibroblasts showed approximately a 1.5-fold increase in Pep A activity over that of the control fibroblasts. Agar gel electrophoretic analysis revealed no detectable differences in the electrophoretic mobility and isoenzyme patterns of Pep A between the trisomy 18 fibroblasts and normal ones. The present results show the trisomy 18 fibroblasts to be suitable for study of the gene dosage effect of Pep A.


Formation of vesicular stomatitis virus pseudotypes bearing surface proteins of hepatitis B virus.
It has been difficult to propagate and titrate hepatitis B virus (HBV) in tissue culture. We examined whether vesicular stomatitis virus (VSV) pseudotypes bearing HBV surface (HBs) proteins infectious for human cell lines could be prepared. For this, expression plasmids for three surface proteins, L, M, and S, of HBV were made. 293T cells were then transfected with these plasmids either individually or in different combinations. 293T cells expressing HBs proteins were infected with VSVdeltaG*-G, a recombinant VSV expressing green fluorescent protein (GFP), to make VSV pseudotypes. Culture supernatants together with cells were harvested and sonicated for a short time. The infectivities of freshly harvested supernatants were determined by quantifying the number of cells expressing GFP after neutralization with anti-VSV serum and mouse monoclonal antibodies (MAbs) against HBs protein. Among 14 cell lines tested for susceptibility to HBV pseudotype samples, HepG2, JHH-7, and 293T cells were judged to be the most susceptible. Namely, the infectious units (IU) of the culture supernatant samples neutralized with anti-VSV in the absence and presence of anti-HBs S MAbs and titrated on HepG2 cells ranged from 1,000 to 4,000 IU/ml and 200 to 400 IU/ml, respectively, suggesting the presence of VSVdeltaG*(HBV) pseudotypes. This infectivity was inhibited by treatment with lactoferrin or dextran sulfate. Pretreatment of the cells with trypsin or tunicamycin inhibited plating of the pseudotype samples. The HBV pseudotypes can be used to analyze early steps of HBV infection, including the entry mechanism of HBV.


Home Polysomnography Reveals a First-Night Effect in Patients With Low Sleep Bruxism Activity.
To examine the presence of a first-night effect (FNE) and the level of internight variability in sleep bruxism (SB) activity when a self-applicable electrode set is used in home polysomnography (PSG) in a sample of subjects with possible SB. Fourteen females and two males aged 38.3 ± 9.1 years (mean ± standard deviation) with self-reported SB underwent home-PSG on three consecutive nights. The subjects applied PSG sensors themselves, including self-applicable electrode sets used to record sleep and masseter muscle activity. Repeated-measures analysis of variance was used to compare SB and sleep variables between the nights. Surprisingly, there were statistically significant elevations in the rhythmic masticatory muscle activity (RMMA) episode index (P = .009), burst index (P = .016), and bruxism time index (P = .049) throughout the course of 3 nights. More bruxers were diagnosed on the second (6 bruxers, ≥ 2 episodes/h) and third night (7 bruxers) compared to the first night (2 bruxers). Most subjects (14/16) had their highest RMMA index on the second or third night. The mean coefficient of variation for RMMA episode index was 50.7%. No statistically significant differences were detected in other sleep variables. The results indicate that a FNE may be present in SB activity, possibly lasting several nights in some subjects. Furthermore, FNE appears to be combined with high internight variability of SB activity without indications of internight changes in sleep macrostructure. To confirm the level of ongoing SB activity, several nights of PSG may be required, especially in subjects with low first-night SB activity. A commentary on this article appears in this issue on page 1281.


Characterization of polyacrylate membrane-coated fibers used in chemical absorption studies with programmed thermal treatment and FT-IR microscopy.
A polyacrylate (PA) film was coated onto a fused-silica fiber as a permeation membrane in a membrane-coated fiber (MCF) technique and a solid-phase microextraction technique. The molecular changes of the PA membrane after different temperature treatments were studied with FT-IR microscopy. The absorption bands of the PA aliphatic backbone at 2902, 2795, and 2740 cm(-)(1) remained unchanged over the elevated thermal treatments, indicating that the polymer backbone was stable over these conditions. The spectra of the PA membrane remained unchanged when the thermal treatment temperature was under 150 degrees C. When the temperature was 250 degrees C, the O-H stretching band in the -COOH groups of the poly(acrylic acid) at 3315 cm(-)(1) was significantly reduced. When the temperature was higher than 280 degrees C, this O-H band disappeared. These evidences suggested that the PA membrane underwent dehydroxyl reaction to form an anhydride when the thermal treatments were higher than 250 degrees C. Thermal treatments of a deuterated PA MCF confirmed the anhydride formation mechanism. The anhydride formation explained the absorption property of PA MCFs in GC applications where they must be preconditioned at 300 degrees C. The absorption data suggest that a PA fiber does not preferably absorb polar compounds (with permanent dipole moment); instead, it absorbs preferably aromatic compounds.


Influence of cataract extraction on cornea and conjunctiva, assessed by vital staining.
A study of the post-operative course in 100 patients subjected to cataract extraction revealed fairly pronounced dehydration of the corneal epithelium and the generally non-exposed segment of the conjunctiva, estimated by Rose Bengal stainability. This dehydration was to become appreciably reduced from the first to the third post-operative day, reaching normal levels of Rose Bengal stainability. The exposed conjunctival segment showed a surprisingly low Rose Bengal stainability on the first post-operative day, in 10% declining until the third day. Disinfection with polyvinylpyrrolidine iodide was found not to alter the stainability of cornea and conjunctiva, compared with the commonly employed method of disinfection with tincture of iodine. In 34% 'bulla'-like formations localized in the superior half of the cornea were noticed on the first post-operative day. These were supposed to represent oedema. The percentage had declined to 20 on the third day.


Methionine requirement of developing turkeys from 8-12 weeks of age.
Two experiments were conducted with Large White male turkeys from 8-12 weeks of age to estimate the methionine requirement for optimum growth rate and feed efficiency. The first experiment was conducted with five pens of 22 turkeys per treatment when the mean minimum temperature was 17.2 C and the maximum 32.2 C. The second experiment was conducted with four groups of 14 turkeys per pen when the mean minimum temperature was 1.4 C and the maximum 12.4 C. Initial body weight and rate of gain in the first experiment was lower than that in Experiment 2. Maximum growth and feed efficiency in Experiment 1 were obtained with a methionine level between 1.32 and 1.41 g./mcal. of M.E. and in Experiment 2 between 1.27 and 1.43 g. Adding 0.1% l-cystine to the basal diet in Experiment 2 failed to improve growth or feed efficiency significantly. The estimated requirement for methionine in these experiments agrees with the present estimate of the National Research Council.


Field-dependence factors in American Indian and Caucasian obesity.
Compared urban obese and non-obese American Indians and Caucasians on the Group Embedded Figures Test (N = 160). Results indicated an apparent relationship between obesity and field-dependence for Caucasians, but not for American Indians. American Indians were more field-dependent than Caucasians overall, which implies a social conformity factor. These results suggest that psychological factors related to obesity vary across ethnic groups because of apparent cultural differences.


Reduced cell surface expression of HLA-C molecules correlates with restricted peptide binding and stable TAP interaction.
HLA-C molecules are poorly expressed at the cell surface compared with HLA-A and HLA-B locus products. The reason for the low surface expression and the underlying mechanism is unclear. We show that the HLA-C4 allele is expressed intracellularly in amounts similar to HLA-A and HLA-B alleles. However, the majority of the HLA-C4 molecules is not transported, but is retained in the endoplasmic reticulum by stable interaction with TAP. This pool does not appear to participate in the formation of HLA-C4/peptide complexes, but is degraded in the endoplasmic reticulum. HLA-C4 molecules can dissociate from TAP upon binding of specific peptide. However, they require a 10-fold higher concentration of a completely degenerated 9-mer peptide mixture for release from TAP than the HLA-A and HLA-B alleles. Our data show that the HLA-C molecules tested are more selective in their peptide binding than HLA-A and HLA-B molecules, resulting in prolonged association with TAP and a reduced formation of intracellular HLA-C/peptide complexes. The restricted peptide binding of certain HLA-C alleles provides one explanation for the reduced expression of HLA-C molecules at the cell surface. Other mechanisms will be discussed.


Effects of lower body positive pressure on cardiovascular responses during walking in elderly women.
This study was undertaken to investigate the effects of lower body positive pressure (LBPP) on cardiovascular responses during a 15-min walking trial in young (22.1+/-0.4 years) and elderly women (67.8+/-1.1 years). The application of 20 mm Hg LBPP reduced ground reaction forces by 31.2+/-0.5 kgw in both groups. We hypothesized that cardiovascular responses to LBPP during walking were different between the young and elderly subjects. Applying 20 mm Hg of LBPP increased diastolic and mean blood pressure but not systolic blood pressure in both groups. LBPP-induced reduction in heart rate (HR) occurred more quickly in the young group compared to the elderly group (p<0.05). Applying LBPP also decreased double product (systolic blood pressure x HR) in both groups, suggesting that LBPP reduces myocardial oxygen consumption during exercise. These results suggest that heart rate responses to LBPP during exercise vary with increasing age.


Ethical issues in cross-cultural neuropsychology.
Clinical neuropsychologists who assess patients from diverse cultural and linguistic backgrounds face unique ethical challenges. In this article, we address 4 critical questions relevant to ethics of cross-cultural neuropsychology: (a) Should culture or race be considered in neuropsychological testing? (b) Should race- and ethnicity-specific normative data be used in the clinical neuropsychological evaluation? (c) Who is competent to design and translate tests for ethnic minority groups and non-English speakers and who is competent to administer and interpret them? and (d) Are neuropsychology training programs adequately preparing clinicians to be competent in the assessment of cross-cultural groups? The overall aims of the article are to highlight the complexity of these clinical and ethical issues, to provide comprehensive and balanced information to help guide clinician choices, and to stimulate future research in this area.


Global warming and allergy in Asia Minor.
The earth is warming, and it is warming quickly. Epidemiological studies have demonstrated that global warming is correlated with the frequency of pollen-induced respiratory allergy and allergic diseases. There is a body of evidence suggesting that the prevalence of allergic diseases induced by pollens is increasing in developed countries, a trend that is also evident in the Mediterranean area. Because of its mild winters and sunny days with dry summers, the Mediterranean area is different from the areas of central and northern Europe. Classical examples of allergenic pollen-producing plants of the Mediterranean climate include Parietaria, Olea and Cupressaceae. Asia Minor is a Mediterranean region that connects Asia and Europe, and it includes considerable coastal areas. Gramineae pollens are the major cause of seasonal allergic rhinitis in Asia Minor, affecting 1.3-6.4 % of the population, in accordance with other European regions. This article emphasizes the importance of global climate change and anticipated increases in the prevalence and severity of allergic disease in Asia Minor, mediated through worsening air pollution and altered local and regional pollen production, from an otolaryngologic perspective.


N-cadherin mediates axon-aligned process growth and cell-cell interaction in rat Schwann cells.
The molecular mechanisms underlying the contact behavior of Schwann cells (SCs) and SC-axon association are poorly understood. SC-SC and SC-axon interactions were studied using purified adult rat SCs and cocultures of SCs with embryonic dorsal root ganglion neurons. After contact of SCs with axons, SCs start to extend processes in alignment with axons. This unique alignment was quantitated using a new assay. SC-axon alignment and SC-SC band formation were disrupted in medium containing low extracellular calcium, indicating the involvement of calcium-dependent adhesion molecules. N-cadherin expression was strong in developing rat sciatic nerves but weak in adult sciatic nerves. In purified adult-derived rat SCs, N-cadherin expression was increased by mitogens (neuregulins) and decreased by high cell density. High-resolution confocal images show intense N-cadherin signals in SC process tips. Subcellular N-cadherin was accumulated in bands at intercellular junctions between SCs and was clustered at axon-SC contact sites. Blocking antibodies (rabbit and guinea pig IgG directed against the first extracellular domain of N-cadherin) and cyclic pentapeptides (including the HAV motif) were used to perturb N-cadherin function. All blocking agents reduced the number of N-cadherin-positive SC-SC junctions and perturbed axon-aligned growth of SC processes. Averaging over all N-cadherin-perturbation experiments, in controls 67-86% of SCs exhibited axon-aligned process growth, whereas in treated cultures only 41% of the SCs aligned with axons. These results are evidence that in mammals N-cadherin is important for formation of SC-SC junctions and SC process growth in alignment with axons.


Cardioneuroablation for Treating Vasovagal Syncope: Current Status and Future Directions.
Syncope is defined by transient and spontaneous loss of consciousness with rapid recovery. Vasovagal syncope (VVS) is the most common form of syncope and is strongly associated with hypervagotonia. There is, however, a lack of effective therapies for VVS. Cardioneuroablation (CNA) is an emerging and promising intervention for VVS with favourable outcomes. CNA has been shown to suppress excessive excitation of vagal activity through ablating the cardiac ganglionated plexi. CNA in the management of VVS requires more structured and comprehensive studies and several issues concerning patient selection, selection of ablation targets, ablation endpoints and the long-term effect of CNA are yet to be determined. This review describes its clinical applications and future directions based on current research data and the authors' own experiences.


Topology of megagauss magnetic fields and of heat-carrying electrons produced in a high-power laser-solid interaction.
The intricate spatial and energy distribution of magnetic fields, self-generated during high power laser irradiation (at Iλ^{2}∼10^{13}-10^{14} W.cm^{-2}.μm^{2}) of a solid target, and of the heat-carrying electron currents, is studied in inertial confinement fusion (ICF) relevant conditions. This is done by comparing proton radiography measurements of the fields to an improved magnetohydrodynamic description that fully takes into account the nonlocality of the heat transport. We show that, in these conditions, magnetic fields are rapidly advected radially along the target surface and compressed over long time scales into the dense parts of the target. As a consequence, the electrons are weakly magnetized in most parts of the plasma flow, and we observe a reemergence of nonlocality which is a crucial effect for a correct description of the energetics of ICF experiments.


Quantitative fluorescence spectroscopy in turbid media: a practical solution to the problem of scattering and absorption.
The presence of practically unavoidable scatterers and background absorbers in turbid media such as biological tissue or cell suspensions can significantly distort the shape and intensity of fluorescence spectra of fluorophores and, hence, greatly hinder the in situ quantitative determination of fluorophores in turbid media. In this contribution, a quantitative fluorescence model (QFM) was proposed to explicitly model the effects of the scattering and absorption on fluorescence measurements. On the basis of the proposed model, a calibration strategy was developed to remove the detrimental effects of scattering and absorption and, hence, realize accurate quantitative analysis of fluorophores in turbid media. A proof-of-concept model system, the determination of free Ca(2+) in turbid media using Fura-2, was utilized to evaluate the performance of the proposed method. Experimental results showed that QFM can provide quite precise concentration predictions for free Ca(2+) in turbid media with an average relative error of about 7%, probably the best results ever achieved for turbid media without the use of advanced optical technologies. QFM has not only good performance but also simplicity of implementation. It does not require characterization of the light scattering properties of turbid media, provided that the light scattering and absorption properties of the test samples are reasonably close to those of the calibration samples. QFM can be developed and extended in many application areas such as ratiometric fluorescent sensors for quantitative live cell imaging.


Carbohydrates in plant immunity and plant protection: roles and potential application as foliar sprays.
Increasing interest is devoted to carbohydrates for their roles in plant immunity. Some of them are elicitors of plant defenses whereas other ones act as signaling molecules in a manner similar to phytohormones. This review first describes the main classes of carbohydrates associated to plant immunity, their role and mode of action. More precisely, the state of the art about perception of "PAMP, MAMP, and DAMP (Pathogen-, Microbe-, Damage-Associated Molecular Patterns) type" oligosaccharides is presented and examples of induced defense events are provided. A particular attention is paid to the structure/activity relationships of these compounds. The role of sugars as signaling molecules, especially in plant microbe interactions, is also presented. Secondly, the potentialities and limits of foliar sprays of carbohydrates to stimulate plant immunity for crop protection against diseases are discussed, with focus on the roles of the leaf cuticle and phyllosphere microflora.


Ecology and Function of the Transmissible Locus of Stress Tolerance in Escherichia coli and Plant-Associated Enterobacteriaceae.
The transmissible locus of stress tolerance (tLST) is a genomic island which confers resistance to heat and chlorine. In this study, we determined that the tLST is frequent in genomes of those Enterobacteriaceae that occur in association with plants as well as the intestines of humans and animals and are relevant as nosocomial pathogens, e.g., Klebsiella and Cronobacter species. The tLST is more frequent in environmental and clinical isolates of Klebsiella pneumoniae than in animal isolates, and heat and chlorine resistance of tLST-positive strains of K. pneumoniae matched the resistance of tLST-positive strains of Escherichia coli. The function of 13 tLST genes was determined by assessing the heat and chlorine resistance of E. coli MG1655 mutants. The deletion of sHsp20, clpKGI, sHspGI, pscA, pscB, and hdeDGI reduced both heat and chlorine resistance; deletion of kefB reduced only chlorine resistance. Genes coding for heat shock proteins sHsp20, clpKGI, and sHspGI decreased the oxidation of cytoplasmic proteins, while kefB decreased the oxidation of membrane lipids. The fitness cost of the tLST for E. coli MG1655 was assessed by pairwise competition experiments with isogenic tLST-positive or tLST-negative strains. The tLST imposes a fitness cost that is compensated for by frequent and lethal challenges with chlorine. All core genes need to be present to maintain the ecological advantage relative to the fitness cost. Taken together, core tLST genes are necessary to provide protection for E. coli against heat and chlorine stress, and the selective pressure for the tLST maintains core genes. IMPORTANCE The transmissible locus of stress tolerance (tLST) is a genomic island comprising 10 core genes that occurs in diverse Enterobacteriaceae and confers resistance to heat and chlorine. Experimentation described in the manuscript describes the physiological function of the core genes by characterization of the resistance of 13 single-knockout (KO) mutants and by characterization of protein and membrane oxidation in these strains after chlorine challenge. Results identify tLST resistance as a genomic island that is specific for those Enterobacteriaceae that occur in plant-associated habitats as well in the intestines of vertebrates. In addition, the ecological function of the genomic island was characterized by large-scale genomic analysis and competition experiments of wild-type and mutant strains. Results suggest that tLST-mediated resistance to chlorine may contribute to the persistence of nosocomial pathogens in hospitals.


Reproducibility of the DrGoniometer app for field-based assessment of the break-point angle in Nordic Hamstring exercise.
Smartphone-based measurement could represent a portable and cheap solution to assess the Nordic Hamstring break-point angle (BPA). We investigated the following: (1) the reproducibility of the DrGoniometer app, (2) its agreement with a computerized video analysis software used as a reference method, and (3) the effects of raters' experience. Fifty recordings were processed by four independent raters. Intraclass correlation coefficient (ICC), SEM, and minimum detectable change (MDC) were calculated. Bland-Altman plots verified the limits of agreement (LoA) with the reference method. The effect of raters' clinical experience was investigated by unpaired t-tests. The app showed excellent intra-rater (ICC = 0.89, 95% CI, 0.84-0.92) and inter-rater reliability (ICC = 0.82, 95% CI, 0.74-0.89), low error (SEM = 2°; MDC = 5.5°), and substantial agreement with video analysis [95% LoA = ±6.7°; mean difference = 1.0°]. Rater experience did not affect results (P > 0.05). The smartphone-based app offers a reliable, valid, quick, and portable method for measuring the BPA in Nordic Hamstring exercise.


GTPase Sar1 regulates the trafficking and secretion of the virulence factor gp63 in Leishmania.
Metalloprotease gp63 (Leishmania donovani gp63 (Ldgp63)) is a critical virulence factor secreted by Leishmania However, how newly synthesized Ldgp63 exits the endoplasmic reticulum (ER) and is secreted by this parasite is unknown. Here, we cloned, expressed, and characterized the GTPase LdSar1 and other COPII components like LdSec23, LdSec24, LdSec13, and LdSec31 from Leishmania to understand their role in ER exit of Ldgp63. Using dominant-positive (LdSar1:H74L) and dominant-negative (LdSar1:T34N) mutants of LdSar1, we found that GTP-bound LdSar1 specifically binds to LdSec23, which binds, in turn, with LdSec24(1-702) to form a prebudding complex. Moreover, LdSec13 specifically interacted with His6-LdSec31(1-603), and LdSec31 bound the prebudding complex via LdSec23. Interestingly, dileucine 594/595 and valine 597 residues present in the Ldgp63 C-terminal domain were critical for binding with LdSec24(703-966), and GFP-Ldgp63L594A/L595A or GFP-Ldgp63V597S mutants failed to exit from the ER. Moreover, Ldgp63-containing COPII vesicle budding from the ER was inhibited by LdSar1:T34N in an in vitro budding assay, indicating that GTP-bound LdSar1 is required for budding of Ldgp63-containing COPII vesicles. To directly demonstrate the function of LdSar1 in Ldgp63 trafficking, we coexpressed RFP-Ldgp63 along with LdSar1:WT-GFP or LdSar1:T34N-GFP and found that LdSar1:T34N overexpression blocks Ldgp63 trafficking and secretion in Leishmania Finally, we noted significantly compromised survival of LdSar1:T34N-GFP-overexpressing transgenic parasites in macrophages. Taken together, these results indicated that Ldgp63 interacts with the COPII complex via LdSec24 for Ldgp63 ER exit and subsequent secretion.


[Case report--polydactyly in a German holstein calf].
A female German Holstein twin calf showed polydactyly in both front limbs and in one of the hind limbs. In three limbs an additional medial toe was seen. No malformations of the limbs were present in the male twin partner and four maternal half sibs. The analysis of the pedigree revealed that the affected calf had common ancestors. The coefficient of inbreeding of the affected calf was 1.312%. As a monogenic or digenic inheritance was suspected in similar cases, it may be probable that the polydactyly observed here has a genetic background.


Roles of the narJ and narI gene products in the expression of nitrate reductase in Escherichia coli.
Nitrate reductase, released and purified from membrane fractions of Escherichia coli, is composed of three subunits. Formation of the enzyme depends on induction of the nar operon, narGHJI, which is composed of four open reading frames (ORF). Previous studies established that the first two genes in the operon narG and narH encode the alpha and beta subunits, respectively, while formation of the gamma subunit, cytochrome bNR, depends on expression of the promoter distal genes. The narJ and narI genes were subcloned separately into plasmids where each was under the control of the nar promoter. Expression of these plasmids in a mutant which forms only alpha and beta subunits revealed that expression of the narI gene is sufficient to restore normal levels of cytochrome bNR, but expression of both genes is required for assembly of fully active, membrane-bound nitrate reductase. The amino acid composition, the N-terminal sequence, and the sequence of cyanogen bromide fragments derived from the isolated gamma subunit corresponds to that expected for a protein produced by the narI ORF. A protein corresponding to the narJ ORF did not appear to be associated with the purified nitrate reductase complex or with the complex immunoprecipitated from Triton X-100-solubilized membrane preparations. We conclude that narI encodes the gamma subunit (cytochrome bNR) and that while the product of the narJ gene is required for assembly of fully active membrane-bound enzyme it is not tightly associated with the active enzyme.


MYC oncogene involved in a t(8;22) chromosome translocation is not altered in its putative regulatory regions.
We have cloned the translocation-associated MYC gene from the Burkitt lymphoma cell line (BL2) with a t(8;22) chromosomal translocation and have determined the nucleotide sequence of the first exon and of the 3' and 5' flanking regions, where sequences with putative regulatory functions have been identified. The nucleotide sequence of the 5' flanking region, which contains regions of DNase hypersensitivity and binding sites for putative regulatory proteins, is the same as that of the normal MYC. Accordingly, mutations in these regulatory regions are not required for the transcriptional deregulation of MYC in the BL2 cell line. The nucleotide sequence of the first exon is similar to that of the normal MYC [Gazin, C., Dupont de Direchin, S., Hampe, A., Masson, J. M., Martin, P., Stehelin, D. & Galibert, F. (1984) EMBO J. 3, 383-387] and has the coding capacity for a 188-residue polypeptide. However, six nucleotide changes that occur in the middle of this reading frame could result in amino acid substitutions. We also have cloned and sequenced the t(8;22) chromosomal breakpoint that is located 10 kilobases 3' of the MYC exon 3 and near the C lambda 3 gene on chromosome 22. Sequences with homology to immunoglobulin joining signals occur close to the breakpoint both on chromosome 8 and 22, providing further evidence that the immunoglobulin joining enzymes may be involved in the recombinations associated with a variety of chromosomal translocations in B and T cells.


Organizational Culture and Climate and Mental Health Provider Attitudes Toward Evidence-Based Practice.
Mental health provider attitudes toward adopting evidence-based practice (EBP) are associated with organizational context and provider individual differences. Organizational culture and climate are contextual factors that can affect staff acceptance of innovation. This study examined the association of organizational culture and climate with attitudes toward adopting EBP. Participants were 301 public sector mental health service providers from 49 programs providing mental health services for youths and families. Correlation analyses and multilevel hierarchical regressions, controlling for effects of provider characteristics, showed that constructive culture was associated with more positive attitudes toward adoption of EBP and poor organizational climates with perceived divergence of usual practice and EBP. Behavioral health organizations may benefit from consideration of how culture and climate affect staff attitudes toward change in practice.


Nucleoid sedimentation analysis of DNA superstructure, gamma-radiation-induced damage and repair in human and chacma baboon (Papia ursinus) peripheral lymphocytes.
The use of the chacma baboon (Papio ursinus) in radiobiological investigations justifies special attention because it answers many of the criteria of parallelism to the human. The present study was undertaken to establish whether in vitro gamma-radiation effects in chacma baboon and human lymphocytes are comparable. The sensitive and rapid nucleoid sedimentation technique was employed to evaluate in vitro DNA superstructure, damage and repair in readily obtainable radiosensitive peripheral lymphocytes. Dose-response curves after 60Co gamma-irradiation were obtained, and by applying single-hit kinetics of the target theory, an estimation of molecular masses of the supercoiled domains was made. The baboon and human lymphocytes produced analogous results, while an ethidium bromide intercalation study also revealed a similarity in average DNA superhelical density. Lymphocyte DNA repair after 0.5-4.0 Gy gamma-irradiation and repair times from 0.5 to 5.0 h were evaluated. The repair data obtained from baboon and human cells after 2.0 Gy irradiation compared favourably in extent of DNA repair as well as the profiles of the kinetic curves. These findings indicate that the chacma baboon would be a useful and relevant model for further in vivo radiobiological studies on lymphocytes. The effects of sedimentation conditions and advantages of using vertical-tube rotors in the nucleoid sedimentation technique are also discussed.


Increasing whole blood oxygen affinity during rapid exchange transfusion: a potential hazard.
Whole blood cell separators are being used to achieve partial exchange transfusions in patients with hemoglobin S. The rate and extent of the alteration of whole blood oxygen affinity, incurred by this procedure, has been determined in a patient with Hb S/C disease. The implications in terms of oxygen delivery to the tissues are described for this patient and also for patients with the more severe disease associated with Hb S/S. It is concluded that whole blood cell separators should be used with an awareness of the possible detrimental effects of acute increases of whole blood oxygen affinity in these patients.


[Influencing factors of the occurrence of propagated sensation effects at Zusanli (ST 36) based on artificial intelligence technology].
To explore the influencing factors of the occurrence of propagated sensation effects at Zusanli (ST 36) based on artificial intelligence technology. A total of 272 undergraduates and postgraduates of Guangzhou University of TCM were recruited. The basic information, including TCM constitution, body weight index (BMI), pulse, tongue and coating, face complexion, as well as the occurrence of propagated sensation effects at Zusanli (ST 36) with different acupuncture needles and techniques, were recorded. The repeated incremental pruning to produce error reduction (RIPPER) algorithm was applied to predict the results and summarize the rules of relevant factors. ① The propagated sensation effects were often observed in the subject with yin-deficiency constitution (55 cases in total, of them 1 case was misclassified). ② The propagated sensation effects were observed in the subject with balanced yin-yang constitution (39 cases in total, of them 0 case was misclassified). ③ The propagated sensation effects were observed in the subject with phlegm-damp constitution combined with thin tongue and white coating (31 cases in total, of them 8 cases were misclassified). ④ The propagated sensation effects were observed in the subject with phlegm-damp constitution combined with light tongue coating (7 cases in total, of them 2 cases were misclassified). ⑤ The propagated sensation effects were observed in all the subject with special constitution (6 cases in total, of them 0 cases were misclassified). ⑥ The propagated sensation effects were not observed in the subject with other constitution or the combination of constitution and symptoms (134 cases in total, of them 4 cases were misclassified). The accuracy rate of classification was 94.49%. The difference of acupuncture needles and techniques was not significant in propagated sensation effects (P>0.05). The TCM constitution may be the most essential factor in the occurrence of propagated sensation effects.


Optical imaging of the heart.
Optical techniques have revolutionized the investigation of cardiac cellular physiology and advanced our understanding of basic mechanisms of electrical activity, calcium homeostasis, and metabolism. Although optical methods are widely accepted and have been at the forefront of scientific discoveries, they have been primarily applied at cellular and subcellular levels and considerably less to whole heart organ physiology. Numerous technical difficulties had to be overcome to dynamically map physiological processes in intact hearts by optical methods. Problems of contraction artifacts, cellular heterogeneities, spatial and temporal resolution, limitations of surface images, depth-of-field, and need for large fields of view (ranging from 2x2 mm2 to 3x3 cm2) have all led to the development of new devices and optical probes to monitor physiological parameters in intact hearts. This review aims to provide a critical overview of current approaches, their contributions to the field of cardiac electrophysiology, and future directions of various optical imaging modalities as applied to cardiac physiology at organ and tissue levels.


Prefabrication of vascularized bone graft using an interconnected porous calcium hydroxyapatite ceramic in presence of vascular endothelial growth factor and bone marrow mesenchymal stem cells: Experimental study in rats.
The purpose of this experimental pilot study was to create a prefabricated vascularized bone graft using interconnected porous calcium hydroxyapatite ceramic (PCHC) block by combining vascular bundle implantation, rat bone marrow mesenchymal stem cells and administration of vascular endothelial growth factor (VEGF) in a rat model. Sixty male Sprague-Dawley rats were used. Experimental animals were divided into six groups, each of which comprised 10 rats. The PCHC blocks were implanted in the medial thigh region in groups I, III, and V without vascular bundle implantation. The PCHC blocks were vascularized by the superficial inferior epigastric artery and vein in groups II, IV and VI. These vessels were passed through the hole of the PCHC blocks. Mesenchymal stem cells were administered into the PCHC in groups III, IV, V and VI. In addition, both mesenchymal stem cells and VEGF were administered in group V and VI. The presence and density of any new bone formation and neovascularization from the vascular bundle was evaluated by X-ray, microangiography, scintigraphy, biochemical analysis and histomorphometry. The newly formed vessels and bone formations were significantly greater in group VI, in which both mesenchymal stem cells and VEGF were applied. THIS PRELIMINARY STUDY SUGGESTS THAT: Both mesenchymal stem cells and VEGF provide vascularized bone prefabrication by enhancing neovascularization and osteogenesis in a shorter time compared to only VEGF application.


Clinical tolerance of olsalazine.
Experience of 804 patients treated with olsalazine has shown that this new 5-ASA based drug has a safe clinical profile and good tolerance. The only common side-effect was loose stools or overt diarrhoea, which necessitated withdrawal of the drug in 6.3% of the patients.


Lesions of the posterior paraventricular thalamus block habituation of hypothalamic-pituitary-adrenal responses to repeated restraint.
We examined the role of the posterior division of the paraventricular nucleus of the thalamus (pPVTh) in habituation of hypothalamic-pituitary-adrenal (HPA) responses to repeated restraint. Habituation refers to the decrement in HPA activity that occurs with repeated exposure to the same or homotypic stressor. To date, the pPVTh has been shown to inhibit the enhanced or facilitated HPA responses to novel, heterotypic restraint in previously chronically cold stressed rats. We hypothesized that the pPVTh also inhibits HPA activity under conditions of habituation. In the first experiment, we lesioned the pPVTh and examined adrenocorticotropic hormone (ACTH) and corticosterone responses to the first or eighth restraint exposure. In sham-lesioned rats, we found lower ACTH and corticosterone responses to the eighth period of 30 min restraint compared to the first exposure, evidence for habituation. In pPVTh-lesioned rats, there was no difference in ACTH and corticosterone responses to the eighth compared to the first restraint exposure. Therefore, pPVTh lesions prevented the habituation of HPA responses to repeated restraint. In the second experiment, we examined whether habituation to restraint is observable in response to an acute, single restraint on day 28 in sham and pPVTh lesioned rats that were exposed to restraint only on days 1 through 8. In this experiment, we replicated the results from the first experiment, and found evidence that habituation to restraint can be observed weeks after chronic stress has been terminated. Furthermore, pPVTh lesions had no additional effects on HPA responses to acute stress on day 28. In summary, pPVTh lesions inhibit habituation of HPA activity to a homotypic stressor, without altering HPA responses to the first restraint. Thus, the intact pPVTh inhibits HPA activity under conditions of habituation, as well as facilitation, and represents an important regulator of HPA activity under conditions of chronic stress.


Satisfaction, quality of life and perception of patients regarding burdens and benefits of vitamin K antagonists compared with direct oral anticoagulants in patients with nonvalvular atrial fibrillation.
To compare the satisfaction of patients treated with vitamin K antagonists (VKA) with that of patients treated with direct oral anticoagulants (DOACs) and to determine the impact on quality of life of both treatments in patients with nonvalvular atrial fibrillation (NVAF). Cross-sectional multicenter study in which outpatients with NVAF completed the ACTS (Anti-Clot Treatment Scale), SAT-Q (Satisfaction Questionnaire) and EQ-5D-3L (EuroQol 5 dimensions questionnaire, 3 level version) questionnaires. The study population comprised 1337 patients, of whom 587 were taking DOACs and 750 VKAs. Compared with VKAs, DOACs were more commonly prescribed in patients with a history of stroke and in patients with a higher thromboembolic risk. The study scores were as follows: SAT-Q: 63.8 ± 17.8; EQ-5D-3L total score: 75.6 ± 20.9; visual analog scale: 63.1 ± 20.6; ACTS Burdens: 51.8 ± 8.4 and ACTS Benefits: 11.9 ± 2.4. The ACTS Burdens score and ACTS Benefits score were higher with DOACs than with VKAs (54.83 ± 6.11 vs 49.50 ± 9.15; p < 0.001 and 12.36 ± 2.34 vs 11.48 ± 2.46; p < 0.001 respectively). NVAF patients treated with oral anticoagulants had many comorbidities and a high thromboembolic risk. Satisfaction and quality of life with oral anticoagulants were high, although they were both better with DOACs than with VKAs.


Ultrastructural alterations produced in mammalian myocardium by variation in perfusate ionic composition.
This study describes the changes produced in the subcellular morphology of mammalian myocardium when perfusate sodium, calcium, and chloride concentrations are varied. By means of a recently developed perfusion technique, functioning dog papillary muscles were perfused with isotonic solutions of varying ionic compositions. Examination of the tissue in the electron microscope revealed that control muscles showed satisfactory preservation of ultrastructure, demonstrating that the protocol itself did not create significant morphological artefact. Low sodium chloride perfusion produced dilatation of both transverse tubules and longitudinal sarcoplasmic reticulum elements. Low sodium or high calcium concentrations produced dilation of tubular elements of the longitudinal sarcoplasmic reticulum while leaving transverse tubules intact. High calcium perfusion produced mitochondrial swelling and vacuolization. Mitochondrial precipitate, both crystalline and amorphous in form, was observed and presumed to be calcium phosphate, either alone or mixed with calcium carbonate. The possibility that the morphological changes observed might indicate subcellular loci of specific ion permeability is discussed. A correlation of the known kinetic behavior of sodium and calcium ions in mammalian myocardium with the ultrastructural alterations produced is suggested.


Synthesis and antibacterial activity of a new derivative of the Meldrun acid: 2,2-dimethyl-5-(4H-1,2,4-triazol-4-ylaminomethylene)-1,3-dioxane-4,6-dione (C9H10N4O4).
The discovery of new substances with proven antimicrobial activity is the current study goal of various researchers. Usage of synthetic products has grown considerably in the past few years due to processing agility, and capability of going through previous chemical modifications in order to enhance its biological activity. Widespread careless use of antimicrobials has made the number of resistant microorganisms rise significantly, thus demanding more efficient drugs to fight them. One of these synthetic candidates for this purpose is the substance 2,2-Dimethyl-5-(4H-1,2,4-triazol-4-ylaminomethylene)-1,3-dioxane-4,6-dione (C9H10N4O4), aminomethylene derivative from Meldrum's acid. This substance, alone and in association with common antibiotics, were evaluated in vitro for antimicrobial activity, and had their minimum inhibitory concentration (MIC) towards Staphylococcus aureus ATCC 25923, Escherichia coli ATCC 10536 and Pseudomonas aeruginosa ATCC 15442, as well as multiresistant strains Escherichia coli 27, Staphylococcus aureus 358 and Pseudomonas aeruginosa 03 determined. The antimicrobial modulation action tests of the aminoglycosides with C9H10N4O4 were performed according to the microdilution method, and resulted in observation of a positive synergic effect.


Physiologically Based Pharmacokinetic Modeling of Tamoxifen and its Metabolites in Women of Different CYP2D6 Phenotypes Provides New Insight into the Tamoxifen Mass Balance.
Tamoxifen is a first-line endocrine agent in the mechanism-based treatment of estrogen receptor positive (ER(+)) mammary carcinoma and applied to breast cancer patients all over the world. Endoxifen is a secondary and highly active metabolite of tamoxifen that is formed among others by the polymorphic cytochrome P450 2D6 (CYP2D6). It is widely accepted that CYP2D6 poor metabolizers exert a pronounced decrease in endoxifen steady-state plasma concentrations compared to CYP2D6 extensive metabolizers. Nevertheless, an in-depth understanding of the chain of cause and effect between CYP2D6 genotype, endoxifen steady-state plasma concentration, and subsequent tamoxifen treatment benefit still remains to be evolved. In this study, physiologically based pharmacokinetic (PBPK)-modeling was applied to mechanistically investigate the impact of CYP2D6 phenotype on endoxifen formation in female breast cancer patients undergoing tamoxifen therapy. A PBPK-model of tamoxifen and its pharmacologically important metabolites N-desmethyltamoxifen (NDM-TAM), 4-hydroxytamoxifen (4-OH-TAM), and endoxifen was developed and validated. This model is able to simulate the pharmacokinetics (PK) after single and repeated oral tamoxifen doses in female breast cancer patients in dependence of the CYP2D6 phenotype. A detailed model-based analysis of the mass balance offered support for a recent hypothesis stating a more prominent role for endoxifen formation from 4-OH-TAM. In the future this model provides a good basis to further investigate the linkage of PK, mode of action, and treatment outcome in dependence of factors such as phenotype, ethnicity, or co-treatment with CYP2D6 inhibitors.


Rosafuningensis (Rosaceae), a new species from Yunnan, China.
A new species Rosafuningensis and its variant R.funingensisf.rosea, both collected from Yunnan Province, China, are, for the first time, documented and illustrated in this study. Morphological analysis in comparison with two related species in the wild, R.gigantea and R.rubus, presents distinguishable features through leaf surfaces, inflorescences and the shape of styles. R.funingensis leaf surfaces are abaxially villous, purple-red, pale green when mature, adaxially glabrous, dark green; inflorescences solitary or 2-5(7) in corymbose cyme; and styles connate into a column or not, exserted.


In vivo and in vitro antiarrhythmic and arrhythmogenic effects of N-acetyl procainamide.
We studied the effects of N-acetyl procainamide (NAPA) on isolated canine cardiac tissues by using standard microelectrode techniques. NAPA (10-40 mg/l) does not suppress the rate of phase 4 depolarization of Purkinje fibers and does not change resting membrane potential, action potential amplitude or maximum upstroke velocity of phase O of the action potentials of Purkinje fibers or ventricular muscle cells. In contrast, action potential duration of both types of cells is significantly prolonged by NAPA in a dose-dependent manner. In toxic concentrations (80-240 mg/l), NAPA can produce a "secondary plateau" at about -55 mV during phase 3 of the action potential of Purkinje fibers driven at cycle lengths of 2000 to 4000 msec. Early afterdepolarizations and single or multiple spontaneous action potentials were often triggered during the secondary plateau. When NAPA is given to conscious or anesthetized dogs (50-100 mg/kg i.v.), ventricular extrasystoles occur at constant coupling intervals if the basic rate of ventricular activation is slow (60 per minute or slower). Single extrasystoles, or the first extrasystole in a salvo, appear at coupling intervals of 400 to 600 msec. The single and multiple extrasystoles induced by NAPA may be caused by action potentials triggered during the secondary plateau and may degenerate into ventricular fibrillation. In doses of up to 100 mg/kg i.v., NAPA exerts slight antiarrhythmic effects in dogs with 24-hr myocardial infarcts.


Thymus-derived (T) cell immunoglobulins. Presence of a receptor site for IgG and absence of large amounts of "buried" Ig determinants on T cells.
Quantitation of surface and total cell Ig obtained after lysis by detergent, urea-acid treatment, and freeze-thawing were determined on spleen cells, thymus cells, and spleen cells specifically depleted of B cells. A two- to four-fold increase in measurable Ig was found after cell lysis. All cell populations showed a similar increase in measurable Ig indicating that no discordantly large amounts of buried Ig determinants were associated with the surface of T cells. The lack of appreciable amounts of T cell Ig was confirmed by immunoprecipitation of radioiodinated cells. A theta-positive lymphoma was described which, when grown in culture, lacked detectable surface Ig but contained a receptor site for IgG. This resulted in appreciable amounts of surface IgG being associated with the tumor line when isolated from ascitic fluid of tumor-bearing mice or after preincubation of cultured cells with either heat-aggregated IgG or normal mouse serum.


Insights into an autonomously formed oxygen-evacuated Cu2O electrode for the selective production of C2H4 from CO2.
Electrochemical conversion of carbon dioxide (CO2) to small organic fuels (e.g. formate, methanol, ethylene, ethanol) is touted as one of the most promising approaches for solving the problems of climate change and energy security. In this study, we report the highly efficient electrochemical reduction of CO2 using cuprous oxide (Cu2O) electrodes to produce ethylene (C2H4) primarily. During CO2 electrolysis using electrodeposited Cu2O on a carbon electrode, we observe the transformation of a compact metal oxide layer to a metal oxide structure with oxygen vacant sites at the bulk region. In contrast to previous studies, our results clearly indicate that Cu2O remains at the surface of the catalyst and it efficiently catalyzes the conversion process of CO2 at low overpotential, exhibiting a high selective faradaic efficiency of over 20% towards C2H4 formation even in long-term electrolysis.


Does c-erbB-2 expression have a role in medulloblastoma prognosis?
The prognosis of patients with medulloblastoma has remained same for the last two decades. This study evaluated the role of c-erbB-2 expression in medulloblastoma as a prognostic marker. Fifty cases of medulloblastomas were investigated for the expression of c-erbB-2 protein using immunohistochemistry. The expression of c-erbB-2 was correlated with age, histology and disease-free survival (DFS). Thirty-five (70%) tumors were c-erbB-2 positive. Immunoreactivity for c-erbB-2 receptor was observed as mixed cytoplasmic and membrane positivity of tumour cells. The mean DFS in c-erbB-2 positive cases was 19.81 months compared to 48.33 months in c-erbB-2 negative cases. c-erbB-2 positivity was found to be an independent predictor of poor outcome in medulloblastoma (p value < 0.05). No correlation of c-erbB-2 expression was observed with the age of patient and the histological type of tumour. Additionally, c-erbB-4 expression was also evaluated in these tumours. Thirty-three cases showed co-expression of c-erbB-2 and c-erbB-4 proteins. However, c-erbB-4 expression alone was not associated with poor outcome, whereas its co-expression with c-erbB-2 was associated with shorter DFS (p < 0.05).


High baseline C-reactive protein levels predict partial type 2 diabetes mellitus remission after biliopancreatic diversion.
Several studies demonstrated that surgery can improve inflammation parameters, such as C-reactive protein (CRP). Few biomarkers have been investigated to potentially predict type 2 diabetes mellitus (T2DM) remission. We aimed at determining whether pre-surgery serum CRP levels could predict T2DM remission after 3 years in patients undergoing bariatric surgery, especially biliopancreatic diversion (BPD). This study was conducted from 2007 to 2009 at the Surgical Department of the University of Genoa, Italy. Forty-four patients with T2DM undergoing BPD (n = 38) or Roux-en-Y gastric bypass (n = 6) were enrolled. The primary endpoint was to evaluate whether pre-surgery CRP levels could predict T2DM partial remission at 3-year follow-up. Secondary endpoints were to assess whether glycaemic, lipid, and inflammatory parameters modified during the follow-up. At baseline, patients with T2DM ranged from overweight to morbid obesity, had mild dyslipidaemia, and a low-grade inflammation. Bariatric surgery improved body weight, lipid and glycaemic profile both at 1- and 3-year follow-up. Pre-surgery CRP levels progressively decreased at 1- and 3-year follow-up. Among inflammatory pre-surgery parameters, only high CRP levels were shown to predict T2DM partial remission after 3 years. Multivariate analysis confirmed the predictive value of pre-surgery CRP levels independently of age, gender, type of surgery, and body mass index. Bariatric surgery, in particular BPD, improved both metabolic and inflammatory biomarkers at 1- and 3-year follow-up. Pre-surgery high CRP levels predicted 3-year T2DM partial remission, indicating a promising target population to be especially treated with BPD.


RAS GTPases and Interleaflet Coupling in the Plasma Membrane.
RAS genes are frequently mutated in cancer. The primary signaling compartment of wild-type and constitutively active oncogenic mutant RAS proteins is the inner leaflet of the plasma membrane (PM). Thus, a better understanding of the unique environment of the PM inner leaflet is important to shed further light on RAS function. Over the past few decades, an integrated approach of superresolution imaging, molecular dynamic simulations, and biophysical assays has yielded new insights into the capacity of RAS proteins to sort lipids with specific headgroups and acyl chains, to assemble signaling nanoclusters on the inner PM. RAS proteins also sense and respond to changes in components of the outer PM leaflet, including glycophosphatidylinositol-anchored proteins, sphingophospholipids, glycosphingolipids, and galectins, as well as cholesterol that translocates between the two leaflets. Such communication between the inner and outer leaflets of the PM, called interleaflet coupling, allows RAS to potentially integrate extracellular mechanical and electrostatic information with intracellular biochemical signaling events, and reciprocally allows mutant RAS-transformed tumor cells to modify tumor microenvironments. Here, we review RAS-lipid interactions and speculate on potential mechanisms that allow communication between the opposing leaflets of the PM.


The Role of Exposomes in the Pathophysiology of Autoimmune Diseases I: Toxic Chemicals and Food.
Autoimmune diseases affect 5-9% of the world's population. It is now known that genetics play a relatively small part in the pathophysiology of autoimmune disorders in general, and that environmental factors have a greater role. In this review, we examine the role of the exposome, an individual's lifetime exposure to external and internal factors, in the pathophysiology of autoimmune diseases. The most common of these environmental factors are toxic chemicals, food/diet, and infections. Toxic chemicals are in our food, drink, common products, the air, and even the land we walk on. Toxic chemicals can directly damage self-tissue and cause the release of autoantigens, or can bind to human tissue antigens and form neoantigens, which can provoke autoimmune response leading to autoimmunity. Other types of autoimmune responses can also be induced by toxic chemicals through various effects at the cellular and biochemical levels. The food we eat every day commonly has colorants, preservatives, or packaging-related chemical contamination. The food itself may be antigenic for susceptible individuals. The most common mechanism for food-related autoimmunity is molecular mimicry, in which the food's molecular structure bears a similarity with the structure of one or more self-tissues. The solution is to detect the trigger, remove it from the environment or diet, then repair the damage to the individual's body and health.


biochem4j: Integrated and extensible biochemical knowledge through graph databases.
Biologists and biochemists have at their disposal a number of excellent, publicly available data resources such as UniProt, KEGG, and NCBI Taxonomy, which catalogue biological entities. Despite the usefulness of these resources, they remain fundamentally unconnected. While links may appear between entries across these databases, users are typically only able to follow such links by manual browsing or through specialised workflows. Although many of the resources provide web-service interfaces for computational access, performing federated queries across databases remains a non-trivial but essential activity in interdisciplinary systems and synthetic biology programmes. What is needed are integrated repositories to catalogue both biological entities and-crucially-the relationships between them. Such a resource should be extensible, such that newly discovered relationships-for example, those between novel, synthetic enzymes and non-natural products-can be added over time. With the introduction of graph databases, the barrier to the rapid generation, extension and querying of such a resource has been lowered considerably. With a particular focus on metabolic engineering as an illustrative application domain, biochem4j, freely available at http://biochem4j.org, is introduced to provide an integrated, queryable database that warehouses chemical, reaction, enzyme and taxonomic data from a range of reliable resources. The biochem4j framework establishes a starting point for the flexible integration and exploitation of an ever-wider range of biological data sources, from public databases to laboratory-specific experimental datasets, for the benefit of systems biologists, biosystems engineers and the wider community of molecular biologists and biological chemists.


Poly(glycoamidoamine) vehicles promote pDNA uptake through multiple routes and efficient gene expression via caveolae-mediated endocytosis.
The use of synthetic polymers for the delivery of nucleic acids holds considerable promise for understanding and treating disease at the molecular level. This work aims to decipher the cellular internalization mechanisms for a series of synthetic glycopolymer DNA delivery vehicles we have termed poly(glycoamidoamine)s (PGAAs). To this end, we have performed cellular delivery experiments in the presence of pharmacological endocytosis inhibitors. Confocal microscopy analysis showed colocalization of labeled pDNA in polyplexes with immunolabeled endocytic molecules to identify the cellular internalization pathways in HeLa cells. Direct membrane penetration was also investigated through various methods, including cellular energy depletion and leakage of a cytosolic enzyme from the cell. The data suggests that the cellular internalization of PGAA polyplexes occurs through a multifaceted internalization mechanism primarily involving caveolae, yet clathrin-coated vesicles and macropinosomes were also involved to a lesser degree. The primary mechanism that leads to efficient nuclear delivery and transgene expression appears to be caveolae/raft-mediated endocytosis. The cellular internalization pathways for PGAAs were not identical to those for polyethylenimine, illustrating that differences in the chemical structure of materials directly impacts the cellular internalization mechanisms.


Ketamine-induced changes in metabolic and endocrine parameters of normal and 2-kidney 1-clip rats.
The aim of this study was to investigate the effect of ketamine on the endocrine and lipid metabolic status of the renal-banded animals. Forty male rats were randomly divided into four groups. Group A served as control, Group B animals received ketamine intraperitoneally at a dose of 100 mg kg(-1), Group C was submitted to 2-kidney 1-clip experimental hypertension and Group D received ketamine as above, as well as being submitted to renal artery clipping. Atrial natriuretic peptide, angiotensin II and free fatty acid concentrations were measured in serum. In addition, adipose tissue lipoprotein lipase activity and angiotensin II content were determined, while the left ventricular weight relative to body weight was used as a cardiac hypertrophy index. In renal-banded rats (Groups C and D) serum atrial natriuretic peptide, free fatty acid and angiotensin II concentrations as well as ventricular weight were increased, while adipose tissue lipoprotein lipase activity was lower than in control animals (Groups A and B). Ketamine administration did not influence angiotensin II concentrations either in normal (Group B) or banded rats (Group D). Ketamine increased serum atrial natriuretic peptide and free fatty acid concentrations only in normal animals (Group B). It had no influence on adipose tissue lipoprotein lipase activity either in normal (Group B) or banded animals (Group D). Adipose angiotensin II content did not differ between the four groups. Ketamine increased the atrial natriuretic peptide and free fatty acid concentration in normal rats. In 2-kidney 1-clip animals, ketamine did not elicit an additional response of serum atrial natriuretic peptide or free fatty acids levels. Its contribution to these factors was not significant.


Innate heart regeneration: endogenous cellular sources and exogenous therapeutic amplification.
The -once viewed as heretical- concept of the adult mammalian heart as a dynamic organ capable of endogenous regeneration has recently gained traction. However, estimated rates of myocyte turnover vary wildly and the underlying mechanisms of cardiac plasticity remain controversial. It is still unclear whether the adult mammalian heart gives birth to new myocytes through proliferation of resident myocytes, through cardiomyogenic differentiation of endogenous progenitors or through both mechanisms. In this review, the authors discuss the cellular origins of postnatal mammalian cardiomyogenesis and touch upon therapeutic strategies that could potentially amplify innate cardiac regeneration. The adult mammalian heart harbors a limited but detectable capacity for spontaneous endogenous regeneration. During normal aging, proliferation of pre-existing cardiomyocytes is the dominant mechanism for generation of new cardiomyocytes. Following myocardial injury, myocyte proliferation increases modestly, but differentiation of endogenous progenitor cells appears to also contribute to cardiomyogenesis (although agreement on the latter point is not universal). Since cardiomyocyte deficiency underlies almost all types of heart disease, development of therapeutic strategies that amplify endogenous regeneration to a clinically-meaningful degree is of utmost importance.


[Contention on the theory of processing techniques of Chinese materia medica in the Ming-Qing period].
On the basis of the golden stage of development of processing techniques of medicinals in the Song dynasty, the theory and techniques of processing in the Ming-Qing dynasties developed and accomplished further. The knowledge of some physicians on the processing of common medicinal, such as Radix rehmannia and Radixophiopogonis, was questioned, with new idea of processing methods put forward and argued against those insisting traditional ones, marking the progress of the art of processing. By reviewing the contention of technical theory of medicinal processing in the Ming-Qing period, useful references can be provided for the inheritance and development of the traditional art of processing medicinals.


[Interaction between microorganisms and heavy metals and its application].
Interactions between microorganisms and heavy metals were expatriated in many ways. Because of their resistance and detoxification to heavy metals, microorganisms are able to leach, absorb and transform heavy metals, which has been actively applied to extract heavy metals from low-rate ore in mine exploitation. Although heavy metals are toxic to microorganisms population and harmful to its processes, some special microorganisms have been applied to the treatment of industrial waste materials and the remediation of soils polluted by heavy metals. The application of microbiological biomass and activity to evaluating the pollution situation and ecological risk of heavy metals in environment media is also significant.


How does the use of antiplatelet and anticoagulants affect the success of mechanical thrombectomy in acute ischemic stroke cases?
In this study, we wanted to investigate the effect of antiplatelet and anticoagulant use on the success of mechanical thrombectomy in acute ischemic stroke cases. 174 patients who were brought to the Stroke Center of Gaziantep University Şahinbey Research and Practice Hospital between January 2018 and February 2019 due to acute ischemic stroke and who underwent mechanical thrombectomy were retrospectively analyzed. The demographic characteristics, antiplatelet/anticoagulant use before the stroke and mTICI (modified-Throm-bolysis-In-Cerebral-Infarction) scores used for reperfusion in mechanical thrombectomy were evaluated. The findings were analyzed statistically (p<0.05). The mean age was 63.3 ± 13.5 in 174 patients who underwent mechanical thrombectomy. 23/174 (13.2%) patients were using anticoagulant therapy (warfarin-OAC or new generation oral anticoagulant-NOAC) and 28/174 (16.1%) were using antiplatelet therapy. A history of atrial fibrillation (AF) was significantly higher in patients receiving anticoagulant therapy before acute ischemic stroke (p=0.001). Patients with a history of hyper tension (HT), diabetes mellitus (DM) and coronary artery disease (CAD) before acute ischemic stroke were receiving antiplatelet therapy in higher rates (respectively; p=0.003, p=0.037, p=0.005). Successful recanalization (mTICI ≥ 2b) was higher in patients with a history of anticoagulant use and who underwent mechanical thrombectomy (p=0.025). Our study showed that the use of anti-platelet or anticoagulants before mechanical thrombec-tomy may have an indirect positive effect on the success of the procedure.


Effect of oocyte dysmorphisms on intracytoplasmic sperm injection cycle outcomes in normal ovarian responders.
The aim of this study was to identify the influence of oocyte dysmorphisms on clinical outcomes after intracytoplasmic sperm injection cycle in normal responders. In the prospective study, morphology of 1999 metaphase II oocytes retrieved from 316 intracytoplasmic sperm injection cycles was evaluated from March 2011 to March 2013 at Royan Institute. Controlled ovarian stimulation was performed by long standard agonist protocol. Oocyte morphology was assessed before sperm injection by one embryologist. The associations between fertilization rate, embryo quality and the independent variables were analyzed using odds ratio (OR) calculated with unconditional logistic regression test. From all retrieved oocytes, 1543 (77.1%) showed at least one morphologic aberration. Presence of cytoplasmic vacuoles and high cytoplasmic viscosity were associated with a significant decrease in the fertilization rate (OR: 0.5; P = 0.004 and OR: 0.6; P = 0.03, respectively). The results showed that oocyte morphology did not affect embryo quality. The number of gonadotrophin injections used showed a direct relation with presence of large perivitelline space. No significant difference was observed among four groups (women with total normal morphologic oocytes [group 1], women with total extracytoplasmic dysmorphic oocytes [group 2], women with total cytoplasmic dysmorphic oocytes [group 3] and women with total oocytes containing multiple dysmorphic features [group 4]) in terms of implantation and clinical pregnancy rates. Metaphase II oocyte morphology had minor impacts on fertilization rate, pronuclear morphology and embryo quality in women with normal ovarian response.


Comparison of the Effect of Nd:YAG and Diode Lasers and Photodynamic Therapy on Microleakage of Class V Composite Resin Restorations.
Considering the importance of disinfecting dentin after cavity preparation and the possible effect of disinfection methods on induction of various reactions between the tooth structure and the adhesive restorative material, the aim of the present study was to evaluate microleakage of composite resin restorations after disinfecting the prepared dentin surface with Nd:YAG and Diode lasers and photodynamic therapy. Standard Class V cavities were prepared on buccal surfaces of 96 sound bovine teeth. The samples were randomly divided into 4 groups based on the disinfection method: Group 1: Nd:YAG laser; Group 2: Diode laser; Group 3: photodynamic therapy; and Group 4: the control. Self-etch bonding agent (Clearfil SE Bond) was applied and all the cavities were restored with composite resin (Z100). After thermocycling and immersing in 0.5% basic fuchsin, the samples were prepared for microleakage evaluation under a stereomicroscope. Data was analyzed with Kruskal-Wallis and Wilcoxon signed-rank tests at P<0.05. There were no significant differences in the microleakage of occlusal and gingival margins between the study groups (P>0.05). There were no significant differences in microleakage between the occlusal and gingival margins in the Nd:YAG laser group (P>0.05). In the other groups, microleakage at gingival margins was significantly higher than that at the occlusal margins (P<0.05). Nd:YAG and Diode lasers and photodynamic therapy can be used to disinfect cavity preparations before composite resin restorations.


Prophylactic minitracheostomy after thoracotomy: a prospective, random control, clinical trial.
The effect of prophylactic minitracheostomy on the incidence of postoperative atelectasis and pulmonary gas exchange was evaluated in a prospective, random control study of 29 patients undergoing thoracotomy. Fourteen were randomly allocated to have a minitracheostomy, which was done in the recovery room immediately after operation, and the remainder acted as controls. Increased radiodensity and atelectasis were seen more often on the chest radiographs of the control patients on the first (p less than 0.05) and on the seventh postoperative days (p less than 0.01), irrespective of the extent of the pulmonary operation. Bronchial lavage under general anaesthesia or sedation was required by 6 of the 15 control patients, but by none of those who had had a minitracheostomy (p less than 0.05). These results support the hypothesis that minitracheostomy is a useful adjunct to postoperative care after thoracotomy.


Hybrid Methods in Iron-Sulfur Cluster Biogenesis.
Hybrid methods, which combine and integrate several biochemical and biophysical techniques, have rapidly caught up in the last twenty years to provide a way to obtain a fuller description of proteins and molecular complexes with sizes and complexity otherwise not easily affordable. Here, we review the use of a robust hybrid methodology based on a mixture of NMR, SAXS, site directed mutagenesis and molecular docking which we have developed to determine the structure of weakly interacting molecular complexes. We applied this technique to gain insights into the structure of complexes formed amongst proteins involved in the molecular machine, which produces the essential iron-sulfur cluster prosthetic groups. Our results were validated both by X-ray structures and by other groups who adopted the same approach. We discuss the advantages and the limitations of our methodology and propose new avenues, which could improve it.


Macroscopic Polarization Enhancement Promoting Photo- and Piezoelectric-Induced Charge Separation and Molecular Oxygen Activation.
Efficient photo- and piezoelectric-induced molecular oxygen activation are both achieved by macroscopic polarization enhancement on a noncentrosymmetric piezoelectric semiconductor BiOIO3 . The replacement of V5+ ions for I5+ in IO3 polyhedra gives rise to strengthened macroscopic polarization of BiOIO3 , which facilitates the charge separation in the photocatalytic and piezoelectric catalytic process, and renders largely promoted photo- and piezoelectric induced reactive oxygen species (ROS) evolution, such as superoxide radicals (. O2- ) and hydroxyl radicals (. OH). This work advances piezoelectricity as a new route to efficient ROS generation, and also discloses macroscopic polarization engineering on improvement of multi-responsive catalysis.


Three-year Outcomes of a Randomized Controlled Trial Comparing a 4.5-mm-Wide to a 3.75-mm-Wide Titanium Implant for Bone Conduction Hearing.
To compare 3-year implant stability, survival, and tolerability of a 4.5-mm-wide (test) and a 3.75-mm-wide (control) percutaneous titanium implant for bone-conduction hearing, loaded with the sound processor after 3 weeks. Sixty implants were allocated in a 2:1 ratio (test-control) in 57 adult patients included in this prospective randomized controlled clinical trial. Follow-up visits were performed at 7, 14, 21, and 28 days; 6 and 12 weeks; 6 months; and at 1, 2, and 3 years after implantation. During these visits, the implant stability quotient (ISQ) was measured by means of resonance frequency analysis (RFA). The peri-abutment soft tissue status was assessed according to the Holgers classification. Skin height around the abutment was evaluated. The mean area-under-the-curve (AUC) of ISQ-low was statistically significantly higher for the test implant (65.7 versus 61.4, p = 0.0002). Both implants showed high survival rates (97.4% versus 95.0%, p = 0.6374). Adverse soft tissue reactions were observed sporadically, with no significant inter-group differences. Skin thickening was seen in the majority of the patients, but no correlation with adverse soft tissue reactions or implant type was observed. The 4.5-mm-wide implant provides significantly higher ISQ values during the first 3 years after surgery compared with the previous generation 3.75-mm-wide implant. Both implants showed high survival rates and good tolerability. These long-term results indicate that the wider implant, loaded with a sound processor at 3 weeks, is a safe and well-performing option for hearing rehabilitation in specific types of hearing loss.


Promising Lipophilic PyTri Extractant for Selective Trivalent Actinide Separation from High Active Raffinate.
Within a spent nuclear fuel recycling strategy, in the past few years, the pyridine-bis-triazole unit was found to be rather effective and selective in minor actinide (MA) separation from synthetic high active raffinate (HAR). In this research work, the main features of the recently studied PTEH ligand were investigated in order to evaluate its potentialities in SANEX-like processes. Its applicability in advanced separation processes was demonstrated, even at process temperatures. It manifested satisfactory extraction properties for a successful selective An separation from Ln, easy cation release, and adequate extraction kinetics as well as outstanding hydrolytical and radiolytical stability. All the results collected in this work allowed the scientists on the committee of the H2020 GENIORS project to promote PTEH as a concrete alternative to the reference CyMe4-BTBP ligand.


Functional neurogenomics of the courtship song of male Drosophila melanogaster.
The genetically programmed courtship song of male Drosophila melanogaster has a rhythmic repetitive structure of sound pulses that is a critical component of mate recognition. This study investigates the neuronal basis of courtship song production by using the GAL4 enhancer trap paradigm to drive the expression of UASG-tetanus toxin (TNT) and UASG-transformer (tra) in subsets of cells. Of the 727 enhancer trap lines screened for courtship song defects in this way, 38 produced a phenotype, and these were used to drive the expression of Green Fluorescent Protein (GFP), thereby implicating the corresponding neuronal structures. Twelve of 13 lines which gave vigorous courtship but did not produce song, showed GFP expression in the mushroom bodies (MBs) and the giant descending neuronal tract and/or the larval interhemispheric junction (IJ). Furthermore, the relevant DNA in which the enhancer trap was inserted was isolated and sequenced in 27 of the lines, and a number of candidate genes that have been implicated in Ras and hedgehog signalling were identified. Other candidate genes that could plausibly affect courtship song were involved in mesothoracic or axonal development.


Integrated Analysis Identifies Upregulated SAMD9L as a Potential Biomarker Correlating with the Severity of Primary Sjögren's Syndrome.
Primary Sjögren's syndrome (pSS) is an autoimmune disease with lymphocytic infiltration of the salivary and lachrymal glands, whose present disease-specific objective indicators are few and have shortcomings that should be addressed. An integrated analysis of sequencing data from different cohorts has the potential to unveil novel biomarkers in pSS. We identified 3 GEO datasets, including gene expression data from minor salivary gland (MSG) biopsy samples of 49 patients with pSS and 31 non-pSS and whole blood cells of 30 pSS patients and 30 healthy controls (HCs). Differentially expressed genes (DEGs) involved in pSS were identified from these datasets. Function Enrichment Analyses of common upregulated DEGs and PPI (protein-protein interaction) networks were performed. Furthermore, we have carried out further analysis of these DEGs to explore their potential clinical significance and diagnostic efficacy as a biomarker for pSS. Sterile Alpha Motif Domain Containing 9 Like (SAMD9L), one of the DEGs, has been identified as a promising candidate biomarker that correlates with the severity of pSS. This has been validated by analyzing local clinical samples from 30 pSS and non-pSS patients' MSG biopsies, as well as serum samples of 18 pSS and HC individuals. Finally, we performed correlation analysis to understand the relationship between SAMD9L and infiltrated immune cells. We identified 10 common highly expressed DEGs in pSS of different tissues. These genes were mainly involved in virus infection-related pathways and inferno-related pathways. GEO data and our clinical data showed that SAMD9L increases with disease severity. Public and local cohorts showed that SAMD9L has high diagnostic performance (AUC=0.845-0.867) as a biomarker, and its AUC was comparable to the Focus score when combined with RF or SSA. Up-regulated SAMD9L may serve as a promising novel pSS diagnostic biomarker and have potential value for evaluating the severity of pSS.


Exome sequencing identifies MXRA5 as a novel cancer gene frequently mutated in non-small cell lung carcinoma from Chinese patients.
Lung cancer has become the top killer among malignant tumors in China and is significantly associated with somatic genetic alterations. We performed exome sequencing of 14 non-small cell lung carcinomas (NSCLCs) with matched adjacent normal lung tissues extracted from Chinese patients. In addition to the lung cancer-related genes (TP53, EGFR, KRAS, PIK3CA, and ROS1), this study revealed "novel" genes not previously implicated in NSCLC. Especially, matrix-remodeling associated 5 was the second most frequently mutated gene in NSCLC (first is TP53). Subsequent Sanger sequencing of matrix-remodeling associated 5 in an additional sample set consisting of 52 paired tumor-normal DNA samples revealed that 15% of Chinese NSCLCs contained somatic mutations in matrix-remodeling associated 5. These findings, together with the results from pathway analysis, strongly indicate that altered extracellular matrix-remodeling may be involved in the etiology of NSCLC.


Aromatase inhibitors--mechanisms of steroidal inhibitors.
Inhibition of aromatase has been an attractive approach for examining the roles of estrogen biosynthesis in various physiological or pathological processes. Effective aromatase inhibitors can serve as potential therapeutic agents for controlling estrogen-dependent diseases such as hormone-dependent breast cancer. Investigations on the development of aromatase inhibitors have therefore expanded greatly in the past two decades. Numerous steroidal agents have been developed that have high affinities for the aromatase enzyme complex and exhibit either competitive inhibition, irreversible inhibition, or mechanism-based (enzyme-activated) inhibition. Mechanism-based inhibitors have distinct advantages in drug design, since these inhibitors are highly enzyme specific, produce prolonged inhibition, and exhibit minimal toxicities. Examination of the structure-activity relationships of the numerous steroidal aromatase inhibitors suggest that the spacial requirements for interaction of agents with the active site of aromatase are very restrictive, permitting only small structural changes to be made on the A-ring and at C-19. Incorporation of small polar substituents at the C-4 position, such as a hydroxyl group, or addition of aryl functionalities at the 7 alpha-position of the steroid, are the exceptions, and inhibitors with such modifications exhibit enhanced affinity for the enzyme. Future investigations of steroidal aromatase inhibitors as probes of the active site of purified aromatase will provide valuable information on enzyme structure at the molecular level, will permit a more detailed examination of the mechanisms of inhibition, and will enhance the development of more specific and effective inhibitors for the treatment of estrogen-dependent breast cancer.


The workbook approach to making estimates and projecting future scenarios of HIV/AIDS in countries with low level and concentrated epidemics.
This paper describes an approach to making estimates and short term projections of future scenarios of HIV/AIDS in countries with low level and concentrated epidemics. This approach focuses on identifying populations which through their behaviour are at higher risk of infection with HIV or who are exposed through the risk behaviour of their sexual partners. Estimates of the size and HIV prevalence of these populations allow the total number of HIV infected people in a country or region to be estimated. Subsequently, assumptions about the possible level and timing of saturation of HIV prevalence among each population can be used to explore future scenarios of HIV prevalence. The basic structure of the software used to make estimates and projections is described. This software includes a set of consistency and audit checks to help exclude unrealistic projections. The paper also discusses the strengths and weakness to this approach to making estimates and projections of HIV/AIDS in countries with low level and concentrated epidemics.


Cryptic DNA-binding domain in the C terminus of RNA polymerase II general transcription factor RAP30.
The C terminus of mammalian transcription factor RAP30 has been found to be a cryptic DNA-binding domain strikingly similar to the C-terminal DNA-binding domain present in conserved region 4 of members of the sigma 70 family of bacterial sigma factors. This RAP30 domain shares strongest sequence similarity with the DNA-binding domain present in region 4 of Bacillus subtilis sporulation-specific sigma K. Like the region 4 DNA-binding activity of Escherichia coli sigma 70, the RAP30 C-terminal DNA binding activity is masked in intact RAP30 but is readily detectable when the RAP30 C terminus is expressed as a fusion protein. Consistent with a role for RAP30 DNA-binding activity in transcription, mutations that abolish DNA binding also abolish transcription. Therefore, RAP30 may function at least in part through the action of an evolutionarily ancient DNA-binding domain that first appeared prior to the divergence of bacteria and eukaryotes.


Factors that affect stabilisation times of canine spontaneous hypoadrenocorticism.
The objective of this retrospective study was to examine factors that may have affected the stabilisation times of 50 dogs with spontaneous hypoadrenocorticism that were being treated with fludrocortisone acetate, with particular emphasis on dosing frequency and the concurrent use of prednisolone. Stabilisation was defined as an absence of clinical signs with a sodium:potassium ratio >27:1 and both electrolyte concentrations within a laboratory reference range. It was found that the median time till stabilisation was three months. The frequency of fludrocortisone treatment (once, twice or changed from once to twice a day) had no effect on the stabilisation time. The two groups of dogs that were started and stabilised on once a day or twice a day dosing had a median stabilisation time of two months. However, dogs that failed to stabilise on once a day dosing of fludrocortisone and were then changed onto twice a day dosing then stabilised a median of one month later. Concurrent use of prednisolone resulted in significantly faster stabilisation times. It was concluded that dogs with hypoadrenocorticism should be continued on prednisolone therapy until they are stabilised. If a dog is failing to stabilise on once a day fludrocortisone acetate, a change to twice a day administration could be considered.


Fluorescent Carbon Dots Derived from Maillard Reaction Products: Their Properties, Biodistribution, Cytotoxicity, and Antioxidant Activity.
Food-borne nanoparticles have received great attention because of their unique physicochemical properties and potential health risk. In this study, carbon dots (CDs) formed during one of the most important chemical reactions in the food processing field, the Maillard reaction from the model system including glucose and lysine, were investigated. The CDs purified from Maillard reaction products emitted a strong blue fluorescence under ultraviolet light with a fluorescent quantum yield of 16.30%. In addition, they were roughly spherical, with sizes of around 4.3 nm, and mainly composed of carbon, oxygen, hydrogen, and nitrogen. Their surface groups such as hydroxyl, amino, and carboxyl groups were found to possibly enable CDs to scavenge DPPH and hydroxyl radicals. Furthermore, the cytotoxicity assessment of CDs showed that they could readily enter HepG2 cells while causing negligible cell death at low concentration. However, high CDs concentrations were highly cytotoxic and led to cell death via interference of the glycolytic pathway.


Nitrate regulates rhizobial and mycorrhizal symbiosis in common bean (Phaseolus vulgaris L.).
Nitrogen-limited conditions are considered to be a prerequisite for legume-rhizobial symbiosis, but the effects of nitrate-rich conditions on symbiotic status remain poorly understood. We addressed this issue by examining rhizobial (Rhizobim tropici) and arbusclar mycorrhizal (Glomus intraradices) symbiosis in Phaseolus vulgaris L. cv. Negro Jamapa under nitrate pre-incubation and continuous nitrate conditions. Our results indicate that nitrate pre-incubation, independent of the concentration, did not affect nodule development. However, the continuous supply of nitrate at high concentrations impaired nodule maturation and nodule numbers. Low nitrate conditions, in addition to positively regulating nodule number, biomass, and nitrogenase activity, also extended the span of nitrogen-fixing activity. By contrast, for arbuscular mycorrhizae, continuous 10 and 50 mmol/L nitrate increased the percent root length colonization, concomitantly reduced arbuscule size, and enhanced ammonia transport without affecting phosphate transport. Therefore, in this manuscript, we have proposed the importance of nitrate as a positive regulator in promoting both rhizobial and mycorrhizal symbiosis in the common bean.


Executive summary from the Intraoperative Advisory Council on Donation After Cardiac Death of the United Network for Organ Sharing: practice guidelines.
The number of organs recovered from asystolic donors is less than anticipated and is explained partially by negative attitudes held by health care providers. To understand the reasons for these beliefs and find solutions, the United Network for Organ Sharing under contract with the Department of Health and Human Services convened the Intraoperative Advisory Council on Donation After Cardiac Death in September, 2001. The Council found that, unlike other medical specialties, operating room health care providers were uncertain of their roles and duties in the care of donors declared dead with cardiopulmonary criteria, known as donation after cardiac death. They were reluctant to care for terminally ill patients in whom death was an expected outcome. Council members deliberated these issues, seeking solutions to repatriate operating room health care providers with the national effort to provide reliable and compassionate care to organ donors and their families. The Council requested the construction of practice guidelines, believing that the structure provided by guidelines will improve health provider confidence in donation after cardiac death and thus improve the quality of care. Physician and nonphysician health care providers from the operating room met to create the Guidelines for the United Network for Organ Sharing, which they believe will improve the quality of care of asystolic organ donors.


The dynamics of an anaerobic digestion of crop substrates with an unfavourable carbon to nitrogen ratio.
The purpose of this study was to investigate the effects of the characteristics of basic crop substrates, such as the carbon, nitrogen, ash and volatile fatty acids contents, on the dynamics of the anaerobic digestion process. For this purpose, the stepwise anaerobic digestion of silage from six different plant species was carried out. Scaled probability distributions (log-normal, log-logistic, logistic, Weibull and Gompertz) were used to approximate the cumulative methane production curves obtained. The results indicated that the Gompertz distribution best fit the process. The hazard function of the Gompertz distribution was used to describe the process change dynamics. Ridge regression models were made and tested to clarify the impact of the crop properties on the distribution parameters. The analysis results indicated that the initial rate of the process depended on the reactor acidity and that the nitrogen content of the substrate was a key factor that affected the process dynamics.


A novel aco-type cytochrome-c oxidase from a facultative alkalophilic Bacillus: purification, and some molecular and enzymatic features.
A novel aco-type cytochrome-c oxidase was highly purified from the facultative alkalophilic bacterium, Bacillus YN-2000, grown at pH 10. The enzyme contained 9.0 nmol heme a/mg protein. It contained 1.23 mol of protoheme, 1.06 mol of heme c, 2.0 g atoms of copper, 2.5 g atoms of iron, and 1.8 g atoms of magnesium per mol of heme a. The enzyme molecule seemed to be composed of two subunits with Mrs of 52,000 and 41,600. On the basis of these results, the enzyme seemed to contain one molecule each of heme a, protoheme, and heme c per minimal structural unit (Mr, 93,600). Only protoheme among the three kinds of hemes in the enzyme reacted with CO and CN-. Heme a did not react with CO; cytochrome a3 did not seem to be present in the enzyme. The enzyme oxidized 314 mol of horse ferrocytochrome c per heme a per sec at pH 6.5 and the catalytic activity was 50% inhibited by 7.65 microM KCN. The enzymatic activity was found to be optimal at pH 6.0.


Three-dimensional solution structure of the cytoplasmic B domain of the mannitol transporter IImannitol of the Escherichia coli phosphotransferase system.
The solution structure of the cytoplasmic B domain of the mannitol (Mtl) transporter (II(Mtl)) from the mannitol branch of the Escherichia coli phosphotransferase system has been solved by multidimensional NMR spectroscopy with extensive use of residual dipolar couplings. The ordered IIB(Mtl) domain (residues 375-471 of II(Mtl)) consists of a four-stranded parallel beta-sheet flanked by two helices (alpha(1) and alpha(3)) on one face and helix alpha(2) on the opposite face with a characteristic Rossmann fold comprising two right-handed beta(1)alpha(1)beta(2) and beta(3)alpha(2)beta(4) motifs. The active site loop is structurally very similar to that of the eukaryotic protein tyrosine phosphatases, with the active site cysteine (Cys-384) primed in the thiolate state (pK(a) < 5.6) for nucleophilic attack at the phosphorylated histidine (His-554) of the IIA(Mtl) domain through stabilization by hydrogen bonding interactions with neighboring backbone amide groups at positions i + 2/3/4 from Cys-384 and with the hydroxyl group of Ser-391 at position i + 7. Modeling of the phosphorylated state of IIB(Mtl) suggests that the phosphoryl group can be readily stabilized by hydrogen bonding interactions with backbone amides in the i + 2/4/5/6/7 positions as well as with the hydroxyl group of Ser390 at position i + 6. Despite the absence of any significant sequence identity, the structure of IIB(Mtl) is remarkably similar to the structures of bovine protein tyrosine phosphatase (which contains two long insertions relative to IIB(Mtl)) and the cytoplasmic B component of enzyme II(Chb), which fulfills an analogous role to IIB(Mtl) in the N,N'-diacetylchitobiose branch of the phosphotransferase system. All three proteins utilize a cysteine residue in the nucleophilic attack of a phosphoryl group covalently bound to another protein.


Isolation and Flow Cytometric Analysis of Human Endocervical Gamma Delta T Cells.
The female reproductive tract (FRT) mucosal immune system serves as the first line of defense. Better knowledge of the genital mucosa is therefore essential for understanding pathogenicity of different pathogens including HIV. Gamma delta (GD) T cells are the prototype of 'unconventional' T cells and represent a relatively small subset of T cells defined by their expression of heterodimeric T-cell receptors (TCRs) composed of gamma and delta chains. This sets them apart from the classical and much better known CD4+ helper T cells and CD8+ cytotoxic T cells that are defined by alpha-beta TCRs. GD T cells often show tissue-specific localization and are enriched in epithelium. GD T cells orchestrate immune responses in inflammation, tumor surveillance, infectious disease, and autoimmunity. Here, we present a method to reproducibly isolate and analyze human endocervical intraepithelial GD T lymphocytes. We have used endocervical cytobrush samples from women participating in the Women's Interagency HIV Infection Study (WIHS). Knowledge about GD T cells interactions during conditions in which there is an insult to the vaginal mucosal could be applied to any clinical study in which mucosal vulnerability is addressed, including the development of vaginal microbicides.In addition, knowledge about mucosal GD T cell responses has potential for application of GD T cell-based immune therapy in treating infectious diseases.


[Masked and labelled food allergens].
Several food proteins are used as food additives or may be contained in foods as contaminants. These masked allergens are at high risk of inducing severe clinical reactions: acute asthma, laryngeal angioedema, anaphylactic shock or even death. The present regulation requires labelling only if the level of food ingredients is superior to 25% of the final product. Several immunologic methods can detect low levels of proteins, equal or inferior to 2%. However a level of 1% or less may be noxious, suggesting a need for a specific line of food products which could be guaranteed free of ... and which might be recommended to patients with food allergy.


Thermal death of Bacillus subtilis spores in oil-water systems.
The thermal death of the spores of Bacillus subtilis 168 in oil-water systems including emulsions and separated layers consisting of phosphate buffer and soybean oil or n-hexadecane was investigated. The resultant survivor curve consisted of two phases, an initial rapid reduction followed by a slow reduction, possibly as reflected by the death in the water phase and the oil phase, respectively. The concentration of oil in the system strikingly affected the pattern of thermal death. These results suggest that the spore location in the oil-water system may be a critical factor in determining the heat resistance.


Iodine-Mediated Oxidative Annulation of β,γ-Unsaturated Hydrazones in Dimethyl Sulfoxide: A Strategy to Build 1,6-Dihydropyridazines and Pyrroles.
Simple, commercially available iodine was successfully employed as a highly efficient and chemoselective catalyst for the oxidative annulation of β,γ-unsaturated hydrazones to produce 1,6-dihydropyridazines under mild conditions for the first time. Interestingly, when active β,γ-unsaturated hydrazone compounds containing electron-donating groups, such as furyl, thienyl, and cycloalkyl, were used, pyrroles were obtained. A gram-scale preparation experiment and further derivatization of pyridazines demonstrated the potential applicability of our synthesis method. Experimental studies and density functional theory calculations unveiled the origin of the chemoselectivity determining the formation of different products.