Excretion of 2-methyl-3-oxovaleric acid in propionic acidemia.
A new case of propionic acidemia is presented, paying special attention to the early symptoms of this disease, such as increased drowsiness, muscular hypotonia, poor feeding, hypothermia, metabolic acidosis, ketonuria and vomiting. Investigation by gas chromatography (GC) and gas chromatography-mass spectrometry (GC-MS) revealed the excretion of fairly high amounts of 2-methyl-3-oxovaleric acid, a condensation product of two molecules of propionyl-CoA, as well as the known pathological metabolites such as propionic, 3-hydroxypropionic and methylcitric acids. Among the post mortem findings the histological studies of the liver were the most remarkable.


Acute Respiratory Distress Syndrome, Mechanical Ventilation, and Inhalation Injury in Burn Patients.
Respiratory failure occurs with some frequency in seriously burned patients, driven by a combination of inflammatory and infection factors. Inhalation injury contributes to respiratory failure in some burn patients via direct mucosal injury and indirect inflammation. In burn patients, respiratory failure leading to acute respiratory distress syndrome, with or without inhalation injury, is effectively managed using principles evolved for non-burn critically ill patients.


A comprehensive continuous-time model for the appearance of CGH signal due to chromosomal missegregations during mitosis.
Aneuploidy, the gain or loss of large regions of the genome, is a common feature in cancer cells. Irregularities in chromosomal copy number caused by missegregations of chromosomes during mitosis can be visualized by cytogenetic techniques including fluorescence in situ hybridization (FISH), spectral karyotyping (SKY) and comparative genomic hybridization (CGH). In the current work, we consider the propagation of irregular copy numbers throughout a cell population as the individual cells progress through ordinary mitotic cell cycles. We use an algebraic model to track the different copy numbers as states in a stochastic process, based on the model of chromosome instability of Gusev, Kagansky, and Dooley, and consider the average copy number of a particular chromosome within a cell population as a function of the cell division rate. We review a number of mathematical models for determining the length of the cell cycle, including the Smith-Martin transition probability model and the 'sloppy size' model of Wheals, Tyson and Diekmann. The program MITOSIM simulates the growth of a population of cells using the aforementioned models of the cell cycle. MITOSIM allows the cell population to grow, with occasional resampling, until the average copy number of a given chromosome in the population reaches a preset threshold signifying a positive copy number alteration in this region. MITOSIM calculates the relationship between the missegregation rate and the growth rate of the cell population. This allows the user to test hypotheses regarding the effect chromosomal aberrations have upon the cell cycle, cell growth rates, and time to population dominance.


Inhibition of cell proliferation and migration by chondroitin sulfate-g-polyethylenimine-mediated miR-34a delivery.
Chondroitin sulfate was chemically conjugated to PEI25K through Michael addition to construct a non-viral gene carrier CS-PEI, and then the carrier was employed in miR-34a delivery to achieve the inhibition of cell proliferation and migration, using prostate tumor cell PC-3 as a model. The nanoparticle from CS-PEI and miR-34a at a mass ratio of 10 was prepared with particle size and zeta potential of 170.7 nm and +42.2 mV, respectively. Flow cytometry and fluorescence microscopy revealed that CS-PEI could efficiently induce the cellular uptake of miR-34a in a CD44-mediated endocytosis manner. Through CS-PEI-mediated miR-34a transfection, obvious cell apoptosis was observed with early apoptotic cells of 47.49%, and meanwhile the activation of caspase-3, -8 and -9, and decreased expression level of Bcl-2 were detected. Moreover, wound healing assay showed that CS-PEI/miR-34a transfection could inhibit the cell migration. Overall, CS-PEI is potentially employed as a promising tumor-targeting system for miR-34a delivery in tumor gene therapy.


Monoamines and monoamine metabolites in brains from demented schizophrenics.
The brains from 12 schizophrenic patients were investigated post-mortem for their content of noradrenaline (NA), dopamine (DA), 5-hydroxytryptamine (5-HT), homovanillic acid (HVA), and 5-hydroxyindolacetic acid (5-HIAA). Six of the schizophrenics had been lobotomized 25--30 years prior to death. A control group matched for age was collected in the autopsy room. The concentrations of NA, DA, and HVA in different parts of the brain from the schizophrenic group did not differ from those of the controls. 5-HT was determined in 11 nuclei or areas of the brain. The schizophrenic group had lower mean values compared with the controls, and in the hypothalamus, medulla oblongata, and hippocampus the difference was at a significant level. 5-HIAA was determined in six areas of the brain but only in a few cases. There was a trend towards lower means of 5-HIAA in the schizophrenics. Cause of death, medication, food intake, age, time between death and autopsy, time the corpses have lain in room temperature, and dissection technique are discussed in relation to these findings. These variables have to be kept under careful control before changes can be claimed as having pathogenetic importance for schizophrenia or for the progressing dementia in this disease.


Measurement of placental volume with real-time ultrasound in mid-pregnancy.
A new method to measure placental volume in mid-pregnancy using real-time ultrasound is described. The method is an adaptation of the B-mode parallel planimetric technique. The mean error of in vitro measurements on 45 balloons (mean volume 186 mL range: 88-485 mL) was -2.24 mL, standard deviation 6.76 mL. The error standard deviation of in vivo placental volume measurements was 16.4 mL (volume range: 173-396.5 mL). Placental measurements were made on 175 women between 14.3 weeks and 21.4 weeks, menstrual age (MA) and a smooth centile curve derived. Median volume increased linearly from 144 to 329 mL. The size of the ultrasound probe limits the technique to use before 20 weeks, MA.


Wettability modification of human tooth surface by water and UV and electron-beam radiation.
The wettability of the human tooth enamel and dentin was analyzed by measuring the contact angles of a drop of distilled water deposited on the surface. The samples were cut along the transverse and longitudinal directions, and their surfaces were subjected to metallographic mirror-finish polishing. Some samples were also acid etched until their microstructure became exposed. Wettability measurements of the samples were done in dry and wet conditions and after ultraviolet (UV) and electron beam (EB) irradiations. The results indicate that water by itself was able to increase the hydrophobicity of these materials. The UV irradiation momentarily reduced the contact angle values, but they recovered after a short time. EB irradiation raised the contact angle and maintained it for a long time. Both enamel and dentin surfaces showed a wide range of contact angles, from approximately 10° (hydrophilic) to 90° (hydrophobic), although the contact angle showed more variability on enamel than on dentin surfaces. Whether the sample's surface had been polished or etched did not influence the contact angle value in wet conditions.


Phototransformation of 3-alkoxychromenones: regioselective photocyclisation and dealkoxylation.
The phototransformation of some 2-(3-methoxyphenyl)-4H-chromen-4-ones bearing a propynyloxy moiety at the 3-position has been described. On photolysis with pyrex-filtered UV light from a Hg lamp (125 W), these chromenones produced a major amount of 5-ethynyl-2-methoxy-6-oxa-benzo[5,6-c]xanthen-7-ones consisting of an exotic tetracyclic scaffold. These photoproducts have been envisioned to be produced through regioselective ring closure at the 6'-position of the 2-(3'-methoxy)phenyl moiety of the initially formed 1,4-biradical via a γ-H abstraction mechanism. No product whatsoever was observed through ring closure at the 2'-position. This behaviour has been found to be in accordance with the directive influence observed in free radical aromatic substitutions. This regioselective photocyclisation is further supported by calculations made from 3D structures (MM2 program). In addition, during the irradiation of these substrates, 2-(3-methoxyphenyl)-4H-chromen-4-ones were also realised through dealkoxylation. The structures of the substrates and photoproduct(s) have been determined by their spectroscopic (IR, NMR, mass spectrometry) studies.


Preliminary observations on the co-existence of regulatory peptides in cells of the baboon endocrine pancreas.
Immunocytochemical procedures at ultrastructural and light microscopy level revealed, in the Chacma baboon endocrine pancreas, cells which were immunoreactive for glucagon and pancreatic polypeptide (PP). Some D cells were observed to contain secretory granules with both the appearance and immunoreactivity of A cell secretory granules.


On-demand therapeutic delivery of hydrogen sulfide aided by biomolecules.
Hydrogen sulfide (H2S), known as the third gasotransmitter, exerts various physiological functions including cardiac protection, angiogenesis, anti-inflammatory, and anti-cancer capability. Given its promising therapeutic potential as well as severe perniciousness if improper use, the sustained and tunable H2S delivery systems are highly required for H2S-based gas therapy with enhanced bioactivity and reduced side effects. To this end, a series of stimuli-responsive compounds capable of releasing H2S (termed H2S donors) have been designed over the past two decades to mimic the endogenous generation of H2S and elucidate the biological functions. Further to improve the stability of H2S donors and achieve the targeted delivery, various delivery systems have been constructed. In this review, we focus on the recent advances of an emerging subset, biomolecular-based H2S delivery systems, which combine H2S donors with biomolecular vectors including polysaccharide, peptide, and protein. We demonstrated their basic structures, building strategies, and therapeutic applications respectively to unfold their inherent merits endued by biomolecules including biocompatibility, biodegradability as well as expansibility. The varied development potentials of biomolecular-based H2S delivery systems based on their specific properties are also discussed. At the end, brief future outlooks and upcoming challenges are presented as well.


Assessment of activated gut-homing CD8+ T cells in blood by flow cytometry during a 3-day gluten challenge.
Accurate celiac disease (CD) diagnosis must be performed in individuals following a gluten containing diet. Diagnostic procedures for individuals already on a gluten-free diet (GFD) avoiding long gluten reintroductions are still challenging. To deal with this issue, we developed an accurate but simple method that requires only a 3-day gluten challenge and circumvents the main limitations of previously suggested proposals such as requirement of specific peptides and unusual specialized lab facilities or high cost. In an attempt to standardize this methodology to be used in daily clinical practice, we describe here an optimized protocol for assessing activated gut-homing CD8+ T cells in blood combined with a short gluten challenge. Details about the amount and type of gluten antigen and the starting material are included, as well as the strategy to easily characterize and identify the cells of interest using flow cytometry. This methodology constitutes a diagnostic tool for CD diagnosis of high specificity and sensitivity for seropositive disease (>95%) as an alternative to long-term gluten challenge and open new possibilities to test the response to gluten in research and clinical trials.


Chalcone-based Selective Inhibitors of a C4 Plant Key Enzyme as Novel Potential Herbicides.
Weeds are a challenge for global food production due to their rapidly evolving resistance against herbicides. We have identified chalcones as selective inhibitors of phosphoenolpyruvate carboxylase (PEPC), a key enzyme for carbon fixation and biomass increase in the C4 photosynthetic pathway of many of the world's most damaging weeds. In contrast, many of the most important crop plants use C3 photosynthesis. Here, we show that 2',3',4',3,4-Pentahydroxychalcone (IC50 = 600 nM) and 2',3',4'-Trihydroxychalcone (IC50 = 4.2 μM) are potent inhibitors of C4 PEPC but do not affect C3 PEPC at a same concentration range (selectivity factor: 15-45). Binding and modeling studies indicate that the active compounds bind at the same site as malate/aspartate, the natural feedback inhibitors of the C4 pathway. At the whole plant level, both substances showed pronounced growth-inhibitory effects on the C4 weed Amaranthus retroflexus, while there were no measurable effects on oilseed rape, a C3 plant. Growth of selected soil bacteria was not affected by these substances. Our chalcone compounds are the most potent and selective C4 PEPC inhibitors known to date. They offer a novel approach to combat C4 weeds based on a hitherto unexplored mode of allosteric inhibition of a C4 plant key enzyme.


ALEX1, a novel tumor suppressor gene, inhibits gastric cancer metastasis via the PAR-1/Rho GTPase signaling pathway.
The ALEX is a novel member of the armadillo family and ALEX1 was reported to be reduced or even lost in multiple solid tumors. However, its expression profile and oncogenic role in gastric cancer (GC) remains largely unknown. ALEX1 expression was detected in 161 GC samples by immunohistochemistry staining. NCI-N87 cells transfected by ALEX1 lentivirus vectors and MKN28 cells transfected by ALEX1 shRNA were used for biological function investigation. Western blot was applied to explore the molecular mechanism and pull-down assays were applied to measure the activity of Rho GTPases. In vivo tumorigenicity, peritoneal and lung metastasis experiments were performed by tumor cell engraftment into nude mice. Bisulfite genomic sequencing and methylation-specific PCR were applied to check the methylation status of the ALEX1 gene. The expression rate of ALEX1 was significantly reduced in gastric tumor samples compared to non-tumor samples (43.5 vs. 90.2%), and its expression was closely related to the tumor differentiation, TNM staging, and lymph nodes metastasis. ALEX1 overexpression in NCI-N87 cells significantly inhibited cell proliferation, migration, and invasion in vitro, and disrupted the structure of the cytoskeleton. ALEX1 overexpression attenuated xenografts growth, peritoneal, and lung metastasis in nude mice. Mechanistically, the overexpression of ALEX1 inhibits thrombin-induced metastasis and Rho GTPases activation. Bisulfite genomic sequencing and methylation-specific PCR revealed that the promoter of ALEX1 is highly methylated in GC cells and tissues. ALEX1 expression is reduced in GC and is involved in diverse cellular functions. ALEX1 inhibits metastasis through the PAR-1/Rho GTPase signaling pathway.


The investigation of lipid peroxidation, anti-oxidant levels and some hematological parameters in sheep naturally infested with Wohlfahrtia magnifica larvae.
Wohlfahrtia magnifica is the main agent of traumatic myiasis in many central and eastern European countries as well as in the Mediterranean basin and Asian countries. The present study was aimed to investigate the effect of myiasis infestation on lipid peroxidation, antioxidant levels and several hematological parameters in the sheep naturally infested with W. magnifica larvae. A total of 38 sheep, including 19 awassi sheep naturally infested with W. magnifica larvae and 19 clinically healthy awassi sheep, were studied. The infected animals were divided into three groups depending on the number of larvae (Group 1: <50 larvae; Group 2: 50-100 larvae; Group 3: >100 larvae). In blood samples, red blood cell counts, hemoglobin, hematocrit concentration and white blood cell, neutrophil, lymphocyte, eosinophil, basophil counts, plasma malondialdehyde, erythrocyte glutathione levels and erythrocyte glutathione peroxidase activity were studied. The results revealed a marked decrease in red blood cell counts and hemoglobin concentrations along with a significant increase in white blood cell and neutrophil counts. The elevation in plasma malondialdehyde levels, a function of lipid peroxidation, established a significant difference between the control group and groups 2, 3. Decreased activity of erythrocyte glutathione peroxidase was found significant in the control group and all parasitemia. We conclude that in natural infestations with W. magnifica, as in infestation with Lucilia cuprina, there is a notable increase in inflammatory activities resulting from the movements, secretions, and toxins of the larvae and form the toxins secreted by the bacteria - which leads to an impression of anemia - and that the tissue injury results in an increase in level of free radicals in the organism.


Frameshift lesions induced by oxazolopyridocarbazoles are recognized by the mismatch repair system in Escherichia coli.
The simple reversible intercalating agent isopropyl-OPC (iPr-OPC) induces frameshift-1 mutations in Salmonella typhimurium and Escherichia coli. The mutagenic responses of S. typhimurium and E. coli wild-type strains are not proportional to the amount of drug intercalated into double-stranded nucleic acids in living bacteria; it occurs only above a minimum level of binding. The fact that mismatch-repair-deficient (mutS) as well as adenine-methylation-deficient (dam) E. coli mutants are hypermutable at low concentrations of iPr-OPC suggests that the majority of mutants induced by this intercalating drug occur as mismatch-repairable mutations (or lesions) in the newly synthesized DNA strand close to the replication fork.


The glutathione antioxidant system as a biomarker suite for the assessment of heavy metal exposure and effect in the grey mangrove, Avicennia marina (Forsk.) Vierh.
Alterations in the glutathione antioxidant system and lipid peroxidation in Avicennia marina were studied under laboratory and field conditions. The activity of glutathione peroxidase (GPx) was found to respond to Zn exposure, and a significant positive relationship between leaf Zn concentration and GPx activity was observed after 96 h and 8 weeks. Lipid hydroperoxides increased proportionally with increasing leaf Zn concentration after 2 and 8 weeks, while no changes in total glutathione were observed. Induction of GPx at 96 h predicted effects at the individual level at a later time interval (reduced biomass at 8 weeks). Results from the field revealed that increasing leaf metal concentration (Zn, Cu or Pb) produced a proportional increase in GPx activity whereas lipid hydroperoxides and total glutathione were not affected. The utility of GPx as an early warning biomarker is suggested, since GPx activity increases in a dose-dependant fashion in response to accumulated leaf metals, and is predictive of later effects on growth.


Chemiluminescent bead-based hybridization assay for the detection of genomic DNA from E. coli in purified plasmid samples.
A bead-based hybridization assay was developed for detection of traces of E. coli genomic DNA (gDNA) present in purified plasmid DNA (pDNA) samples. Standards of gDNA and pDNA samples were sheared by sonication and adsorbed onto aminopropyl controlled pore glass (CPG) particles (130 microm). A preliminary study was conducted to optimize the amount of DNA adsorbed on the particles. Results indicated that maximum attachment efficiency was obtained by adsorbing DNA for 2 h in 0.2 x SSC, pH 5.7. The DNA-bound particles were hybridized overnight with a 181-bp digoxigenin-labeled probe, specific for gDNA. Following a chemiluminescent detection protocol, signal intensities of the standards were plotted as a function of initial gDNA concentration. The calculated detection limit (LOD) was 1.4 pM of gDNA. The assay was able to detect gDNA in pure plasmid preparations at the 1% level even in the presence of 1,000-fold excess of noncomplementary target. Hybridization results were compared with a quantitative real-time PCR assay. Both methods afforded similar accurate results at the 95% confidence level.


Triple gene expressions in yeast, Escherichia coli, and mammalian cells by transferring DNA fragments amplified from a mother yeast expression plasmid.
Escherichia coli, Saccharomyces cerevisiae, and mammalian culture cells are standard host organisms for genetic engineering and research, thus various plasmid vectors have been developed. However, the vectors are designed only for a single host owing to their host-specific genetic elements such as promoters and selection markers. In this study, we developed a yeast expression plasmid that enables the expression of the same gene in E. coli and mammalian cells via the transfer of PCR products amplified from the plasmid as a template. The yeast plasmid YHp26352 was constructed to contain the following regions sequentially: yeast TDH3 promoter (TDH3p), red fluorescent protein (eEmRFP), SV40 terminator (SVpA), E. coli origin (ori), ampicillin resistant gene (AmpR), mammalian cytomegalovirus promoter (CMVp), E. coli srlA promoter (srlAp), and yeast selection marker URA3, which expressed eEmRFP in yeast. To express eEmRFP in mammalian cells, an end-promoter DNA fragment encompassing the eEmRFP-SVpA-ori-AmpR-CMVp region was amplified by PCR and directly used for transfection to mammalian culture cells, resulting in gene expression in mammalian cells through non-homologous end joining. Homologous recombination-mediated circularization was carried out for E. coli cloning and expression by attaching a short overlapping sequence to the 5'-end of a PCR primer, which was used to amplify the eEmRFP-SVpA-ori-AmpR-CMVp-srlAp fragment, after which E. coli transformation was performed. Proof-of-concept experiments were performed by expressing GFP-fused human synaptobrevin VAMP1, and wild-type and codon-changed CLuc luciferase genes in yeast, E. coli, and HEK293 cells. This is the first all-in-one plasmid applicable for expression in three host organisms.


Modular total syntheses of trans-clerodanes and sesquiterpene (hydro)quinones via tail-to-head cyclization and reductive coupling strategies.
The trans-clerodanes and sesquiterpene (hydro)quinones are a growing class of natural products that exhibit a wide range of biological activities. Although they are different types of natural products, some of them feature the same trans-decalin core structure. Here, we report the total syntheses of two members of trans-clerodanes, five members of sesquiterpene (hydro)quinones as well as the proposed structure of dysidavarone D via a modular synthetic route. A bioinspired tail-to-head cyclization strategy was developed to syntheses of the trans-decalin architectures by using two diastereochemically complementary radical polyene cyclization reactions catalyzed by Ti(III) and mediated by Mn(III), respectively. The different types of side chains were introduced by challenging nickel catalyzed reductive couplings of sterically hindered alkyl halides. The synthesis of the proposed dysidavarone D proved a wrong structural assignment of the natural product.


Expression and efficient secretion of a functional chitinase from Chromobacterium violaceum in Escherichia coli.
Chromobacterium violaceum is a free-living β-proteobacterium found in tropical and subtropical regions. The genomic sequencing of C. violaceum ATCC 12472 has revealed many genes that underpin its adaptability to diverse ecosystems. Moreover, C. violaceum genes with potential applications in industry, medicine and agriculture have also been identified, such as those encoding chitinases. However, none of the chitinase genes of the ATCC 12472 strain have been subjected to experimental validation. Chitinases (EC 126.96.36.199) hydrolyze the β-(1,4) linkages in chitin, an abundant biopolymer found in arthropods, mollusks and fungi. These enzymes are of great biotechnological interest as potential biocontrol agents against pests and pathogens. This work aimed to experimentally validate one of the chitinases from C. violaceum. The open reading frame (ORF) CV2935 of C. violaceum ATCC 12472 encodes a protein (439 residues) that is composed of a signal peptide, a chitin-binding domain, a linker region, and a C-terminal catalytic domain belonging to family 18 of the glycoside hydrolases. The ORF was amplified by PCR and cloned into the expression vector pET303/CT-His. High levels of chitinolytic activity were detected in the cell-free culture supernatant of E. coli BL21(DE3) cells harboring the recombinant plasmid and induced with IPTG. The secreted recombinant protein was purified by affinity chromatography on a chitin matrix and showed an apparent molecular mass of 43.8 kDa, as estimated by denaturing polyacrylamide gel electrophoresis. N-terminal sequencing confirmed the proper removal of the native signal peptide during the secretion of the recombinant product. The enzyme was able to hydrolyze colloidal chitin and the synthetic substrates p-nitrophenyl-β-D-N,N'-diacetylchitobiose and p-nitrophenyl-β-D-N,N',N"-triacetylchitotriose. The optimum pH for its activity was 5.0, and the enzyme retained ~32% of its activity when heated to 60°C for 30 min. A C. violaceum chitinase was expressed in E. coli and purified by affinity chromatography on a chitin matrix. The secretion of the recombinant protein into the culture medium was directed by its native signal peptide. The mature enzyme was able to hydrolyze colloidal chitin and synthetic substrates. This newly identified signal peptide is a promising secretion factor that should be further investigated in future studies, aiming to demonstrate its usefulness as an alternative tool for the extracellular production of recombinant proteins in E. coli.


Evaluating the specificity of a new type of urine collection bag for infants.
For diagnosing urinary tract infection (UTI) in infants, the urine collection bag is a common sampling method. It has several advantages versus other methods but a high risk of contamination makes culture results difficult to evaluate. Previous studies report a specificity ranging from 14% to 84%. The objective of this study was to evaluate the specificity of a new type of urine collection bag. Urine samples were collected from healthy infants with a new type of collection bag. As the urine is expected to be sterile, any bacterial growth would be considered a contamination. Forty-four samples were included: 40 samples showed <10,000 cfu/ml of mixed growth or no growth at all, three samples showed <10,000 cfu/ml of single-strain growth and one sample showed >10,000<100,000 cfu/ml of single-strain growth. No samples showed any growth >100,000 cfu/ml. According to Kass criteria, 97.7% of the samples would exclude a UTI and 2.3% would be considered inconclusive. None of the samples had a contamination level that, falsely, would be interpreted as positive. Further studies will be valuable as a specificity of 97.7% suggests that this collection device could give the clinician a non-invasive option for diagnosing UTI in infants.


Antimicrobial effect of essential oils on the seafood spoilage micro-organism Photobacterium phosphoreum in liquid media and fish products.
To evaluate the antimicrobial effect of nine essential oils (EO) on P. phosphoreum and determine the effect of oregano oil on the shelf-life of modified atmosphere-packed (MAP) cod fillets. The antimicrobial effect of EO was studied in a liquid medium and in product storage trials. Oils of oregano and cinnamon had strongest antimicrobial activity, followed by lemongrass, thyme, clove, bay, marjoram, sage and basil oils. Oregano oil (0.05%, v/w) reduced growth of P. phosphoreum in naturally contaminated MAP cod fillets and extended shelf-life from 11-12 d to 21-26 d at 2 degrees C. Oregano oil reduced the growth of P. phosphoreum and extended the shelf-life of MAP cod fillets. Mild and natural preservation using EO can extend the shelf-life of MAP seafood through inhibiting the specific spoilage organism P. phosphoreum.


Müllerian agenesis and ovarian torsion. A case report and review of literature.
A case report of a premenarcheal patient with an ovarian torsion and müllerian agenesis is presented. An 11-year-old prepubertal girl presented with severe left lower quadrant abdominal pain and mild rebound. A computed tomography showed a normal appendix; an ultrasound showed a left ovary measuring 3 x 2 cm with multiple 0.5-mm simple cysts. A diagnostic laparoscopy showed the left ovary tube to be twisted, but the uterus was absent, and the right ovary and tube were not visualized in the appropriate location; instead, a right adnexal structure was buried in the right sidewall. The ovary was untwisted and fixed to the pelvic sidewall. Chromosomes were 46,XX, and her hormonal evaluation was normal. Ovarian torsion and müllerian agenesis are rarely reported. An association between the lax attachment of the adnexa and torsion may be a contributing factor in this condition.


Clinicopathologic correlation of vitamin D receptor expression with retinoid X receptor and MIB-1 expression in primary and metastatic osteosarcoma.
Vitamin D, in addition to its effects on bone, is important in cell cycle regulation. Vitamin D receptor (VDR) has been identified in breast, prostate, and colon cancers, as well as in canine and human osteosarcoma (OS) cell lines; however, it has not been well investigated in human OS-archived specimens. We correlated VDR, retinoid X receptor (RXR), and MIB-1 (Ki-67) expression in 110 archived OS cases with several clinicopathologic parameters including patient's age, sex, tumor location, tumor grade, and type and metastatic status. The expression of VDR and RXR was identified in human OS tissue obtained from primary and metastatic OS archival tissue. No statistically significant difference was found in VDR expression in relation with tumor grade, type, age, sex, or location. The expression of RXR was highest in higher-grade (P = .0006) and metastatic tumors but remained unchanged when correlated with tumor type, age, sex, or location. The expression of MIB-1 was statistically elevated in higher-grade tumors (P = .001), patients 25 years or younger (P = .04), tumors located in extremities (P = .005), and metastatic lesions, but was not impacted by tumor type or patient's sex. Proliferative activity was significantly reduced after treatment, as the mean MIB-1 expression dropped from 11% in primary biopsy samples to 6% in resection specimens. There appears to be a relationship between proliferative tumor activity and tumor grade, location, and metastasis. Additional studies on the analysis of the effects of vitamin D and RXR on OS proliferation, apoptosis, and differentiation are critical to further evaluate their potential role in OS treatment.


Purification and properties of the fatty acid synthetase complex from the marine dinoflagellate, Crypthecodinium cohnii.
De novo biosynthesis of fatty acids in the heterotrophic marine dinoflagellate, Crypthecodinium cohnii, has been studied in vitro. Fatty acid synthetase was located in the cytosol and its activity was dependent on acetyl-CoA, malonyl-CoA, NADPH2 and NADH2. The enzyme was purified 100-fold using ion-exchange chromatography on DEAE-Sephadex A-25, adsorption to hydroxyapatite and gel filtration on Sepharose 4B columns. Very active endogenous proteases were separated from the fatty acid synthetase at the first step of purification. The purified enzyme had a molecular weight of about 400000, as judged from gel filtration, sucrose density gradient centrifugation and polyacrylamide gel electrophoresis under non-denaturing conditions. Polyacrylamide gel electrophoresis under denaturing conditions in the presence of SDS and urea revealed one major protein band of Mr 180000, suggesting that the enzyme is composed of two multifunctional subunits of apparently identical molecular weight. Reaction products of the C. cohnii fatty acid synthetase are free fatty acids due to the presence of a thioesterase activity in the purified enzyme complex. The main product is palmitate. Docosahexaenoic acid (C22:6, n-3), the major fatty acid component of C. cohnii lipids, is not directly synthesized by the enzyme.


Prevalence of mood disorders and utility of the PRIME-MD in patients undergoing radiation therapy.
To validate a short, structured interview procedure that allows practicing oncologists to quickly and reliably identify mood disorders in their patients, and to estimate the prevalence and types of mood disorders in a radiation therapy patient setting, noting relationships between mood disorders and patient characteristics. Consecutive, eligible adult patients from the practices of two radiation oncologists were administered the Primary Care Evaluation of Mental Disorders (PRIME-MD) by the treating physician. A subset of these patients was also evaluated with the SCID, administered by trained mental health care personnel. Agreement between the two instruments was examined using the kappa statistic. Prevalence of mood disorders was determined from the PRIME-MD. The significance of relationships between patient characteristics and mood disorders was examined by chi-square and ANOVA analysis, and subsequently by multivariate logistic regression analysis. One hundred twenty-two patients were studied. Fifty-three of these were administered the SCID. Agreement between the two instruments was very good (kappa = 0.70). A diagnosis of a depressive or anxiety disorder by the PRIME-MD was made in 59 of the 122 patients (48%, 95% confidence interval = 39%, 58%). Multivariate analysis showed that a diagnosis of a depressive mood disorder was significantly related to pain intensity and prior history of depression. We have demonstrated the validity and feasibility of the PRIME-MD administered by oncologists in making diagnoses of mood disorders. The prevalence of mood disorders in our set of patients undergoing a course of RT was nearly 50%. Future studies should describe the natural history of these disorders, and determine optimal intervention strategies.


Perspective: Early direct repair of recurrent postoperative cerebrospinal (CSF) fluid leaks: No good evidence epidural blood patches (EBP) work.
"Targeted" epidural blood patches (EBP)" successfully treat "focal dural tears (DT)" diagnosed on thin-cut MR or Myelo-CT studies. These DT are largely attributed to; epidural steroid injections (ESI), lumbar punctures (LP), spinal anesthesia (SA), or spontaneous intracranial hypotension (SICH). Here we asked whether "targeted EBP" could similarly treat MR/Myelo-CT documented recurrent post-surgical CSF leaks/DT that have classically been effectively managed with direct surgical repair. Utilizing ultrasound, fluoroscopy, or O-arm guidance, "targeted EBP" effectively manage "focal DT" attributed to ESI, LP, SA, or SICH. Here we reviewed the literature to determine whether similar "targeted EBP" could effectively manage recurrent postoperative CSF leaks/DT. We were only able to identify 3 studies involving just 20 patients that attempted to utilize EBP to control postoperative CSF fistulas/DT. EBP controlled CSF fistulas/DT in 6 patients in the first study, and 9 of 10 patients (i.e. 90%: 2/2 cervical; 7/8 lumbar) in the second study. However, in the third study, 3 (60%) of 5 EBP failed to avert recurrent CSF leaks/DT in 4 patients (i.e. 1 cervical patient (2 EBP failed attempts), 3 lumbar patients (1 failed EBP)). Early direct surgical repair of recurrent postoperative spinal CSF leaks/DT remains the treatment of choice. Our literature review revealed 3 underpowered studies including just 20 patients where 20% of EBP failed to control recurrent postoperative fistulas (range of failure from 0-60% per study). Although there are likely other studies we failed to identify in this review, they too are likely insufficiently powered to document significant efficacy for performing EBP over direct surgical repair for recurrent postoperative CSF leaks/DT.


Effect of progesterone and/or estradiol treatments prior to induction of ovulation on subsequent luteal lifespan in anestrous Nelore cows.
Three experiments evaluated effects of estradiol (E(2)) and/or progesterone (P(4)) prior to induction of ovulation with GnRH on subsequent luteal lifespan in anestrous Nelore cows. In Experiment 1, cows (25-65 days post-partum [DPP]; n=114) were assigned randomly to receive a 6-day treatment with an intravaginal P(4) device (CIDR) and/or 1mg i.m. injection of 17beta-E(2) (4 groups; 2x2 factorial design) prior to ovulation. Blood samples were collected on days 0, 5, 7, 9, 12, 15 and 19 for evaluation of luteal function. Pre-treatment with P(4) reduced occurrence of premature luteolysis (PL; 79.2% in non-treated vs. 13.5% in treated cows; P<0.01), but there was no effect of treatment with 17beta-E(2) on percentage of PL. In Experiment 2, cows (30-40 DPP; n=35) were assigned randomly to receive either 0.5mL i.m. injection of cottonseed oil (placebo) or 1mg i.m. injection of E(2) cypionate. Blood samples were collected on days 0, 5, 9 and 15 for evaluation of luteal function. Incidence of PL (83.0% in Control Group vs. 75.0% in ECP Group; P>0.1) and mean serum P(4) did not differ between treatment groups. In Experiment 3, cows (30-60 DPP; n=109) were randomly assigned to receive either a 6-day (6-d Group) or a 3-day (3-d Group) treatment with CIDR. Blood samples were collected on days 0, 5, 7 and 9 for luteal function evaluation. Incidence of PL (5.5% in 6-day vs. 5.5% in 3-day groups; P>0.1) and mean serum P(4) did not differ between treatment groups. In conclusion, both 3- and 6-day treatments with P(4) prior to induction of ovulation in anestrous Nelore cows increased percentage of normal luteal lifespan, while administration of 1mg of 17beta-E(2) or E(2) cypionate failed to prevent occurrence of PL.


The prognosis and clinicopathological significance of histone deacetylase in hepatocellular carcinoma: a meta-analysis.
The value of the different types of HDACs (histone deacetylases) for HCC (hepatocellular carcinoma) prognosis and clinicopathological features is still controversial. Here, we performed a meta-analysis to investigate the possible role of different types of HDACs in HCC. Until October 28, 2021, we have searched the Embase, Cochrane, PubMed, Scopus, Web of Science (WOS), SinoMed, Chinese China National Knowledge Infrastructure (CNKI), Chinese WanFang, and Chinese Weipu databases and evaluated eligible studies according to the criteria. We used hazard ratio (HR) and 95% confidence interval (95% CI) to evaluate the prognostic effects of different types of HDACs on overall survival (OS), disease-free survival (DFS)/recurrence-free survival (RFS) and used odds ratio (OR) and corresponding 95% CI to evaluate the significance of HDACs on clinicopathological characteristics. The I2 statistic and chi-square-based Q test were used to assess the heterogeneity. When the heterogeneity was significant, we conducted a subgroup analysis. In addition, Egger's test and funnel chart were used to assess publication bias. The high expression of class I HDACs was associated with poorer OS, DFS/RFS and differentiation, intrahepatic metastasis, tumor-node-metastasis (TNM), tumor number, tumor size, vascular invasion, and other poor clinicopathological characteristics. The high expression of class II HDACs was related to poor OS and multiple and larger tumors. After subgroup analysis, class II HDACs may also be related to worse TNM and Edmondson grading. The high expression of class III HDACs was related to poor OS, hepatitis B, liver cirrhosis, serum AFP, and vascular invasion. But it was more common in women and was related to single, smaller tumors. Type I, II, and III HDACs are associated with poor prognosis, and there are also correlations with some clinicopathological features, suggesting that different types of HDACs may be valuable biomarkers for HCC.


Prevention of hepatotoxic responses to chemicals by glycyrrhizin in rats.
To clarify whether glycyrrhizin, the aqueous extract of licorice root and a drug for treatment of chronic active hepatitis, prevents the development of hepatic injury induced by carbon tetrachloride, allyl formate, and endotoxin, the present study was undertaken in rats. The treatment with glycyrrhizin 20 hr before carbon tetrachloride administration protected the development of the pericentral hepatocellular necrosis. Glycyrrhizin treatment 2 hr prior to the administration of allyl formate also inhibited the development of the periportal hepatocellular necrosis. However, glycyrrhizin did not protect the development of endotoxin-induced focal and random hepatocellular necrosis. These experimental results suggest that glycyrrhizin has no protective effect on hepatic injury following sinusoidal circulatory disturbance as seen in the case of endotoxin and that glycyrrhizin can protect against hepatotoxicity induced by the direct action on the hepatocytes due to hepatotoxins, such as carbon tetrachloride and allyl formate.


A novel thermometric biosensor for fast surveillance of β-lactamase activity in milk.
Regulatory restrictions on antibiotic residues in dairy products have resulted in the illegal addition of β-lactamase to lower antibiotic levels in milk in China. Here we demonstrate a fast, sensitive and convenient method based on enzyme thermistor (ET) for the surveillance of β-lactamase in milk. A fixed amount of penicillin G, which is a specific substrate of β-lactamase, was incubated with the milk sample, and an aliquot of the mixture was directly injected into the ET system to give a temperature change corresponding to the remained penicillin G. The amount of β-lactamase present in sample was deduced by the penicillin G consumed during incubation. This method was successfully applied to quantify β-lactamase in milk with the linear range of 1.1-20 UmL(-1) and the detection limit of 1.1 UmL(-1). The recoveries ranged from 93% to 105%, with relative standard deviations (RSDs) below 8%. The stability of the column equipped in ET was also studied, and only 5% decrease of activity was observed after 60 days of use. Compared with the conventional culture-based assay, the advantages of high throughput, timesaving and accurate quantification have made this method an ideal alternative for routine use.


[Recurrent signet-ring cell gastric carcinoma evidenced by FDG-PET].
To present the case report of a patient with undifferentiated and diffuse signet-ring cell gastric carcinoma in which FDG-PET evidenced recurrent disease. The patient was diagnosed of a stage III gastric carcinoma in 1994 and was treated with a subtotal gastrectomy. In February 2003, recurrent disease was detected in mediastinal and left supraclavicular lymph nodes. The patient was treated with chemotherapy and radiotherapy, reaching a complete response. After 6 months free of disease, he presented an elevation of the tumor markers with negative results in conventional imaging methods (upper digestive endoscopy, bone scintigraphy, and CT). An FDG-PET scan was performed to rule out recurrent disease. FDG-PET detected pathologic findings suggestive of malignant disease in right supraclavicular and mediastinal lymph nodes. These findings were confirmed by clinical follow-up and with another CT scan performed 4 months later. In this case report we stress the importance of early recurrence by FDG-PET in a non-intestinal gastric carcinoma. This is of interest given the greater difficulty to detect mucous secreting and/or producing carcinomas with the PET-FDG.


Event-related potential concomitants of information processing dysfunction in schizophrenic children.
ERPs were recorded from 10 schizophrenic and 13 normal children during the performance of the Span of Apprehension task (Span). This task involves the discrimination of a randomly placed target letter among distractors, and it has been shown to discriminate between normal and schizophrenic individuals. The EEG was recorded at 7 scalp loci, and ERPs were averaged over a 1500 msec interval initiated by a warning tone which preceded the visual Span stimuli by 500 msec. Stimulus arrays were grouped into 4 levels of difficulty. The data from both subject groups were combined in a single principal components analysis (separate PCAs exhibited few differences between groups) generating 8 rotated factors which were readily interpreted in terms of conventional ERP components. Factor scores for the two groups were examined using Analysis of Variance. The schizophrenic children produced a small CNV which was slow to develop and resolve as well as diminished amplitudes for the N1, P3 and slow wave components. This suggests that these children are impaired in their ability to regulate processes involved in the mobilization and direction of attention and the discrimination of target stimuli. Significantly, the schizophrenic children did not show progressive increases in N1 and SW amplitudes in response to increases in information processing demand (array difficulty) as was the case in the normal children. ERP components of the schizophrenic children were most aberrant at frontal leads, but midline and lateralized deficits were also seen at vertex and posterior recording sites.


Radiotherapy of extranodal low-grade follicular and marginal zone lymphomas: long-term follow-up of 159 patients.
To evaluate clinical, histopathologic, and radiation (RT) dose parameters in patients with extranodal low-grade (ENLG) non-Hodgkin lymphoma (NHL) and their possible impact on local control (LC) and survival. The medical records of 159 patients with 181 histologically confirmed ENLG-NHL lesions treated at our institution were reviewed retrospectively. The predominant histological subtype (73%) was marginal zone lymphoma (MZL). Common lesion sites were the gastrointestinal tract (GIT; 33%), skin (26%), and orbit (21%). The majority of patients (88%) presented with stage I/II disease. Thirty-three (20%) lesions were treated with reduced-dose RT (≤30.6 Gy) and 148 lesions (80%) with conventional-dose RT (>30.6 Gy), with an overall median dose of 39.6 Gy (range 4-63). The median follow-up period was 72 months. The 10-year local control (LC), Progression-free survival (PFS), and overall survival (OS) rates were 96, 65, and 82%, respectively. Higher overall response rate (ORR; 98% vs. 94%, p = 0.001) and complete response rate (CRR; 95% vs. 73%, p = 0.001) were observed in patients treated with conventional-dose regimens than in those treated with reduced-dose regimens. Ten-year PFS (p = 0.90) and OS (p = 0.40) was similar between the two dose groups. RT was well tolerated in both dose groups, with no grade 4/5 toxicities. In the multivariate analysis, RT dose and timing (upfront or salvage) were related to LC, whereas age, histology, and complete response (CR) to RT were associated with PFS. Patient age and radiation field size impacted OS. RT is an effective and curative local treatment for early-stage FL and MZL at conventional and reduced radiation doses. Conventional-doses seems to be associated with local response improvement, without significant differences in PFS rates. Age, histology, and response to RT may influence the PFS.


Comparison of resection margin status after single or double radiopaque marker insertion for tumor localization in breast cancer patients receiving neoadjuvant chemotherapy.
Insertion of radiopaque markers is helpful for tumor localization in patients receiving neoadjuvant chemotherapy (NAC) followed by breast-conserving surgery (BCS). The aim of this retrospective study was to investigate the pathologic margin status in patients with single or double marker insertion. We reviewed the records of 130 patients with marker insertion prior to NAC followed by BCS from January 2016 to September 2019. Under ultrasonography guidance, single or double markers were inserted to localize a tumor in the breast. The incidence of additional resection after frozen biopsy and re-excision after permanent pathologic diagnosis was analyzed. In a total of 130 patients, 104 had a single marker in the center of the tumor and 26 had double markers at the periphery of the tumor before NAC. Among 69 patients with residual invasive tumors after NAC, there was no difference in the additional resection rate after frozen biopsy (single vs. double markers; 14.3% vs. 38.5%, P = .059) or the re-excision rate after final pathologic diagnosis (0% vs. 7.7%, P = .188). After propensity score matching for tumor size and subtypes, the two groups showed no differences in the additional resection rate after frozen biopsy (7.7% vs. 19.2%, P = .139) or the re-excision rate (0% vs. 3.8%, P = .308). After a median follow-up of 19 months (range 8-48 months), local recurrence-free survival did not differ between the two groups (log-rank P = .456). Number of inserted markers for tumor localization did not affect the pathologic margin status after BCS.


Papillary thyroid carcinoma: prognostic factors and the role of radioiodine and external radiotherapy.
To evaluate the role of radioiodine and external radiotherapy treatment in papillary thyroid carcinoma (PTC). This is a retrospective study of 842 patients with the diagnosis of PTC registered from 1960 to 1997 at the Department of Clinical Oncology, Queen Elizabeth Hospital, Hong Kong. The mean follow-up was 9.2 years. The stage distribution according to UICC/AJCC TNM staging was as follows: 58.6%, Stage I; 9.6%, Stage II; 26.1%, Stage III; 2.3%, Stage IV; and 3.4%, not stated. The 10-year cause-specific survival (CSS) rates were as follows: Stage I, 99.8%; Stage II, 91.8%; Stage III, 77.4%; and Stage IV, 37.1%. Multivariate analysis showed that the statistically significant poor prognostic factors for CSS were as follows: age older than 45, postoperative gross locoregional (LR) residual disease, distant metastasis (DM) at presentation, and lack of radioactive iodine (RAI) treatment. In patients with no DM and no postoperative LR disease, adjuvant RAI ablation reduced both LR failure (RR [relative risk] = 0.29) and DM (RR = 0.2), although the CSS was not affected. In the subgroup of T1N0 M0 disease, no patient with RAI treatment had a relapse. External radiotherapy reduced the risk of LR failure to 0.35. Subgroup analysis revealed that external radiotherapy was particularly effective in increasing the probability of LR control of disease in patients with gross postoperative LR disease (RR = 0.36). Both RAI and external radiotherapy were effective treatment in PTC. Total or near-total thyroidectomy followed by RAI treatment appears to result in the best outcome. External radiotherapy to improve LR control is indicated in patients with gross postoperative residual disease. Treatment should be individualized for patients with T1N0 M0 disease.


Sudden cardiac death: ethical considerations in the return to play.
The team physician-athlete relationship prompts many basic questions in medical ethics. Return-to-play decisions form many of the core responsibilities facing team physicians, and occasionally these decisions can have overriding ethical dilemmas. Therefore, a structured ethical decision-making process is a valuable skill for every successful sports medicine physician. An ethical question is confronted here in a case presentation that weighs the risk of repeat sudden cardiac death and the potential for failed cardiac resuscitation against the athlete's interest to play competitive basketball. The article applies a four-step framework for ethical decision making in sports medicine. The important first step includes gathering medical information and understanding the preferences of the athlete. Step 2 brings together the decision-making stakeholders, the team physician as a member, to define ethical issues and apply ethical principles: beneficence, non-maleficence, and patient autonomy. Step 3 selects a course of action with unbiased analysis and arrives at a good choice that merits an action plan in step 4. This decision need not be perfect, but should reinforce the team physician's responsibilities to the athlete and center on the athlete's welfare.


Pre-operative haematological assessment in patients scheduled for major surgery.
Peri-operative anaemia, blood loss and allogeneic blood transfusion are associated with increased postoperative morbidity and mortality, and prolonged hospital stay. A multidisciplinary, multimodal, individualised strategy, collectively termed 'patient blood management', may reduce or eliminate allogeneic blood transfusion and improve outcomes. This approach has three objectives: the detection and treatment of peri-operative anaemia; the reduction of peri-operative bleeding and coagulopathy; and harnessing and optimising the physiological tolerance of anaemia. This review focuses on the pre-operative evaluation of erythropoiesis, coagulation status and platelet function. Where possible, evidence is graded systematically and recommended therapies follow recently published consensus guidance.


The effect of anticonvulsant drugs on thymidine and deoxyribosenucleic acid synthesis by human marrow cells.
An abnormal deoxyuridine suppression test was found in treated epileptic patients that did not correlate with other evidence of abnormal folate metabolism. Diphenylhydantoin at a level of 100 microgram/ml interfered with the incorporation of [3H]thymidine into DNA. It is suggested that anticonvulsants may exert their effect at a stage of DNA synthesis beyond that at which folate coenzymes operate. Folate deficiency may then arise as a result of death of cells in vivo causing an increased folate requirement.


Large energy storage density performance of epitaxial BCT/BZT heterostructures via interface engineering.
We grew lead-free BaZr0.2Ti0.8O3 (BZT)/Ba0.7Ca0.3TiO3 (BCT) epitaxial heterostructures and studied their structural, dielectric, ferroelectric and energy density characteristics. The BZT/BCT epitaxial heterostructures were grown on SrRuO3 (SRO) buffered SrTiO3 (STO) single crystal substrate by optimized pulsed laser deposition (PLD) technique. These high-quality nanostructures exhibit high dielectric permittivity (∼1300), slim electric field-dependent polarization (P-E) curve with high saturation polarization (∼100 µC/cm2) and low remnant polarization (∼20 µC/cm2) through interface engineering to develop new lead-free ferroelectric system for energy storage devices. We observe an ultrahigh discharge and charge energy densities of 42.10 and 97.13 J/cm3, respectively, with high efficiency, which might be highly promising for both high power and energy storage electrical devices.


Presence of Toxoplasma gondii tissue cysts in human semen: Toxoplasmosis as a potential sexually transmissible infection.
Toxoplasma gondii is a widely prevalent protozoan parasite in human populations. This parasite is thought to be primarily transmitted through undercooked meat and contamination by cat feces. Here, we seek to determine if Toxoplasma gondii cysts can be found within human semen. We used a mixture of histological and immunofluorescence stains to visualize Toxoplasma gondii cysts in thin smears of human semen. Further, we probed for presence of bradyzoite-specific mRNA transcription using in-situ hybridization. We visualized Toxoplasma gondii cysts in ejaculates of immune-competent and latently infected human volunteers. We confirmed the encystment by probing transcription of a bradyzoite-specific gene in these structures. These observations extend previous observations of the parasite in semen of several non-human host species, including rats, dogs, and sheep. Toxoplasma gondii infection is a clinically significant infection, in view of its high prevalence, its purported role in neuropsychiatric disorders such as schizophrenia, as well as in the more serious form of congenital toxoplasmosis. Our demonstration of intact Toxoplasma gondii cysts in the ejaculate supports the possibility of sexual transmission of the parasite and provides an impetus for further investigations.


Lipoxin recognition sites. Specific binding of labeled lipoxin A4 with human neutrophils.
Lipoxin A4 stimulates rapid lipid remodeling and a pertussis toxin-sensitive release of arachidonic acid in polymorphonuclear leukocytes (PMN) (Nigam, S., Fiore, S., Luscinskas, F.W., and Serhan, C.N. (1990) J. Cell. Physiol. 143, 512-523) and has been shown to inhibit leukocyte responses in several systems. To examine the basis underlying these actions, we have prepared [11,12-3H]lipoxin A4 (LXA4) and characterized its interactions with human PMN. Time course studies (0-90 min) with intact PMN demonstrated cell association of 3H label which was specific and reversible. PMN bound [3H]LXA4 with a Kd of 0.5 +/- 0.3 nM, representing approximately 1,830 sites/PMN, and the Hill plot value of 1.9 suggests cooperative binding. [3H]LXA4 binding was stereoselective since neither leukotriene B4 (LTB4), lipoxin B4 (LXB4), (6S)-LXA4, 11-trans-LXA4, nor SKF 104353 competed for [3H]LXA4-specific binding while LTD4 and LTC4 partially competed. Subcellular fractionation revealed that specific binding with [3H]LXA4 was associated with membrane (42.1%)-, granule (34.5%)-, and nuclear (23.3%)-enriched fractions, a distribution distinct from that of [14,15-3H] LTB4 binding. [11,12-3H]LXA4-specific binding was modulated by guanosine analogs, suggesting the involvement of G proteins. A fluorescent LXA4 derivative (methyl-7-methoxycoumarin-LXA4) competed with [3H]LXA4 binding to intact PMN and showed specific and reversible binding as monitored by flow cytometric analysis. These results indicate that PMN possess specific recognition sites for LXA4 which may mediate its actions.


BinSPreader: Refine binning results for fuller MAG reconstruction.
Despite the recent advances in high-throughput sequencing, metagenome analysis of microbial populations still remains a challenge. In particular, the metagenome-assembled genomes (MAGs) are often fragmented due to interspecies repeats, uneven coverage, and varying strain abundance. MAGs are constructed via a binning process that uses features of input data in order to cluster long contigs presumably belonging to the same species. In this work, we present BinSPreader-a binning refiner tool that exploits the assembly graph topology and other connectivity information to refine binning, correct binning errors, and propagate binning to shorter contigs. We show that BinSPreader could increase the completeness of the bins without sacrificing the purity and could predict contigs belonging to several MAGs. BinSPreader is effective in binning shorter contigs that often contain important conservative sequences that might be of great interest to researchers.


Osteotesticular protein tyrosine phosphatase expression in rodent testis.
In the last decade the novel receptor-like protein tyrosine phosphatase was identified and termed osteotesticular tyrosine phosphatase (OST-PTP) due to its restricted expression in bone and testis. OST-PTP expression is regulated during osteoblast differentiation and it shows stage specific expression in the testis. Confined OST-PTP expression in the basal compartment of the seminiferous tubules suggests that this protein may be a good candidate for a rodent germ stem cell marker. To test this hypothesis we determined the exact pattern of OST-PTP expression in the rodent testis. Adult mouse and rat paraffinized testis sections were subjected to in situ hybridization using riboprobes against the receptor and catalytic domains of the protein OST-PTP. OST-PTP testicular expression in rodents is not confined to the spermatogonia, as previously suggested, but is also present in Sertoli cells in a stage independent pattern. This finding excludes the hypothesis that OST-PTP is a germ stem cell identification marker in rodents. In addition, we report the identification of a testicular OST-PTP isoform lacking part of a catalytic domain that is widely expressed during spermatogenesis in all cell types. This finding implies tight control over OST-PTP expression in the testis, which in turn suggests an important role for OST-PTP and its isoform in male germ cell differentiation.


Immobilization of fractures of the zygomatic bone with an antral pack.
Reduction and fixation of malar complex fractures has been emphasized and re-emphasized. Simple methods of reduction for complicated fractures are still advocated with clinically poor results. The necessity for a three point fixation and overcorrection for the complex fracture is emphasized. While it is true that simple fractures may be reduced by simple techniques, these simple methods cannot be applied to more comminuted fractures.


Occurrence of intestinal adenocarcinomas in sheep in the southwestern part of Norway.
From 1256 adult sheep received for clinical/post-mortem examination over an 8 1/2-year period, intestinal adenocarcinomas were diagnosed in 37 (2.9%). Mean age of the affected animals was 6.1 years. Clinical pathology and post-mortem findings are summarized.


Plasma miR-940 may serve as a novel biomarker for gastric cancer.
It was reported that circulating microRNAs could be applied as non-invasive biomarkers for cancer monitoring. The purpose of this study was to identify plasma miRNA that may serve as a novel biomarker for gastric cancer and to evaluate its clinical application. MicroRNA profiles were generated from plasma samples of 5 patients with gastric cancer (GC) versus 5 healthy controls (HC). MicroRNA-940 (miR-940) was one of the most downregulated miRNAs with fold change of 0.164. It was revealed that the expression of miR-940 was downregulated in both the initial set (N = 30, P < 0.0001) and the validation set (N = 80, P < 0.0001) of plasma samples of patients with gastric cancer. The sensitivity was obviously higher than the current biomarkers CEA and CA19-9 (81.25 % vs. 22.54 % and 15.71 %). MiR-940 was also significantly downregulated in gastric cancer tissue samples (N = 34, P = 0.0015), as well as in cancer cell lines (N = 7). Importantly, miR-940 was significantly highly expressed in stomach tissue samples than in other types of tissue samples including the liver, breast, thyroid, and lung. Moreover, there was a trend of lower expression of miR-940 from early to advanced stage of gastric cancer. Target prediction suggested that miR-940 regulated cell signaling including NF-κB and Wnt/β-catenin, as well as pathways of cell communication and adhesion. These pathways play critical roles in gastric cancer initiation and progression. It is the first report that miR-940 may mainly express in the stomach. Downregulation of plasma miR-940 may serve as a novel biomarker for detection of gastric cancer.


Modulation of subfamily B/R4 RGS protein function by 14-3-3 proteins.
Regulator of G protein signalling (RGS) proteins are primarily known for their ability to act as GTPase activating proteins (GAPs) and thus attenuate G protein function within G protein-coupled receptor (GPCR) signalling pathways. However, RGS proteins have been found to interact with additional binding partners, and this has introduced more complexity to our understanding of their potential role in vivo. Here, we identify a novel interaction between RGS proteins (RGS4, RGS5, RGS16) and the multifunctional protein 14-3-3. Two isoforms, 14-3-3beta and 14-3-3epsilon, directly interact with all three purified RGS proteins and data from in vitro steady state GTP hydrolysis assays show that 14-3-3 inhibits the GTPase activity of RGS4 and RGS16, but has limited effects on RGS5 under comparable conditions. Moreover in a competitive pull-down experiment, 14-3-3epsilon competes with Galphao for RGS4, but not for RGS5. This mechanism is further reinforced in living cells, where 14-3-3epsilon sequesters RGS4 in the cytoplasm and impedes its recruitment to the plasma membrane by Galpha protein. Thus, 14-3-3 might act as a molecular chelator, preventing RGS proteins from interacting with Galpha, and ultimately prolonging the signal transduction pathway. In conclusion, our findings suggest that 14-3-3 proteins may indirectly promote GPCR signalling via their inhibitory effects on RGS GAP function.


The structure and function of glutamate receptor ion channels.
As in the case of many ligand-gated ion channels, the biochemical and electrophysiological properties of the ionotropic glutamate receptors have been studied extensively. Nevertheless, we still do not understand the molecular mechanisms that harness the free energy of agonist binding, first to drive channel opening, and then to allow the channel to close (desensitize) even though agonist remains bound. Recent crystallographic analyses of the ligand-binding domains of these receptors have identified conformational changes associated with agonist binding, yielding a working hypothesis of channel function. This opens the way to determining how the domains and subunits are assembled into an oligomeric channel, how the domains are connected, how the channel is formed, and where it is located relative to the ligand-binding domains, all of which govern the processes of channel activation and desensitization.


Cause, seasonality of death and co-morbidities in progressive supranuclear palsy (PSP).
The aim of this study was to investigate the causes and seasonality of death and co-morbid conditions among progressive supranuclear palsy (PSP) patients. 22 consecutive clinicopathologically confirmed PSP patients were compared with 22 gender and age-matched Parkinson's disease (PD) patients and 26 non-neurological controls. All subjects consented during life to donate their brains to the University of Miami Brain Endowment Bank. Respiratory-related deaths were significantly more frequent in PSP when compared to PD patients. Recurrent respiratory infections were also very frequent in PSP patients and were commonly associated with respiratory-related deaths. Deaths that occurred during winter and spring months accounted for about 70% of deaths among PSP patients. The most common co-morbid condition in PSP was hypertension, present in 50% of cases.


Antibody levels remain high to one-year's follow-up after moderate and severe COVID-19, but not after mild cases.
Understanding the longevity of antibodies against SARS-CoV-2 infection is of utmost importance in predicting the further course of the pandemic and to plan vaccination strategies. Here we report a cohort of COVID-19 patients with different disease severities whose antibody dynamics we evaluated during one-year of follow-up. We conducted a longitudinal study of 123 COVID-19 patients and 45 SARS CoV-2 negative outpatients with upper respiratory tract infection. We analyzed the demographic and clinical features of the patients with COVID-19 in relation to different disease severities according to the WHO classification. The antibody response was evaluated by a Luciferase Immunoprecipitation System (LIPS) assay at 3, 6, and 12 months after the acute infection. Amongst the enrolled COVID-19 patients, 15 (12%) had mild, 42 (34%) had moderate, 39 (32%) had severe and 27 (22%) had critical disease courses; 79% of the patients were hospitalized. During follow-up, all patients had anti-SARS RBD-IgG levels above the cut-off value on all visits, but the antibody levels varied significantly between the different disease severity groups. Between the six- and 12-month follow-up visits, 41% of patients were vaccinated, which enhanced their antibody levels significantly. Our data demonstrate sustained antibody levels at one-year after moderate and severe COVID-19 infection. Vaccination of patients with the mild disease is important to raise the antibody levels to a protective level.


Eutrophication and salinization elevate the dissolved organic matter content in arid lakes.
Dissolved organic matter (DOM) plays an essential role in the global lake carbon cycle. Understanding DOM composition and monitoring its spatiotemporal dynamics are of great significance for understanding the lake carbon cycle, controlling water pollution, and protecting water resources. However, previous studies have focused mainly on eutrophic freshwater lakes, with limited attention given to saline lakes. Based on in situ data collected in ten lakes in northwestern China, this study reported the changes in DOM components in different lake types. Parallel factor analysis (PARAFAC) was used to analyze the three-dimensional excitation emission matrix (EEMs) to obtain the DOM fluorescence components. The contributions of different environmental factors to the changes in DOM components were quantified by the generalized linear model (GLM). The results showed that the eutrophication index was significantly positively related to dissolved organic carbon (DOC) (R2 = 0.95, p < 0.01) and colored DOM (CDOM) (R2 = 0.96, p < 0.01) concentrations. Terrestrial humic-like and tryptophan-like components, which are highly correlated with human activities, explained 62% and 64% of the variations in DOC and CDOM, respectively. In sum, the contributions of human activities to the DOC and CDOM variations were 61% and 57%, respectively. Salinity also showed significant positive correlations with both DOC (R2 = 0.88, p < 0.01) and CDOM (R2 = 0.87, p < 0.01). Lake salinization led to increases in DOM concentration, and salinity contributed 20% and 16% to the DOC and CDOM variations, respectively. Therefore, human activities and salinity codetermined the DOM concentration and its composition in the western arid lakes. Based on these findings, this study proposed a feasible flowchart for remotely estimating DOM in saline lakes using satellite data. This study is significant for the long-term monitoring of the carbon cycle and the effective protection of lake water resources in saline lakes.


North Carolina Youth Connected: Lessons Learned in Engaging Community Leaders in Preventing Teen Pregnancy.
The North Carolina Youth Connected initiative aimed to reduce teen pregnancy rates in two large North Carolina counties by mobilizing local leaders to raise awareness of teen pregnancy and guide solutions. We describe the strategies used to mobilize community leaders, the nature of engagement, and the challenges and lessons learned with engaging community leaders around teen pregnancy prevention. A 22-member Community Advisory Council carried out community mobilization activities. Meeting minutes and internal logs tracked activities. Surveys and group discussions assessed members' challenges and lessons learned. Results were analyzed for salient themes. Lessons learned center on the importance of building meaningful relationships with local leaders, ensuring representation from all sectors, and key considerations for engaging leaders around a common goal when facing local pushback. Cross-sector collaborations, supported by a backbone organization, take time and commitment but can be effective in tackling teen pregnancy prevention.


Infection ability of mycelium and spores ofMicrodochium nivale (Fr) samuels hallett toLolium Perenne L.
Mycelium and spores ofMicrodochium nivale /Syn.Fusarium nivale/ were compared according to their ability to infect plants ofLolium perenne. The experiments were carried out according to the "cold chamber" method (Cormack, Lebeau 1956 modified by Pronczuk 1987). Between these two types of inoculum significant differences were found. The spore inoculum did not give any symptoms while the mycelial inoculum incited a severe disease in plants ofLolium perenne during one month of incubation.Under laboratory conditions it was found that the spore cultures ofMicrodochium nivale grew very slowly at 0 - 1°C, whereas their growth at 18 - 20°C was very fast. Growth of the mycelial cultures was not as profoundly affected by temperatures studied as the spore ones.It was concluded that to incite a disease the spore inoculum require longer incubation time than mycelial ones. The mycelial inoculum is more useful for screening of plants for resistsance.


Memory T cell responses targeting the SARS coronavirus persist up to 11 years post-infection.
Severe acute respiratory syndrome (SARS) is a highly contagious infectious disease which first emerged in late 2002, caused by a then novel human coronavirus, SARS coronavirus (SARS-CoV). The virus is believed to have originated from bats and transmitted to human through intermediate animals such as civet cats. The re-emergence of SARS-CoV remains a valid concern due to the continual persistence of zoonotic SARS-CoVs and SARS-like CoVs (SL-CoVs) in bat reservoirs. In this study, the screening for the presence of SARS-specific T cells in a cohort of three SARS-recovered individuals at 9 and 11 years post-infection was carried out, and all memory T cell responses detected target the SARS-CoV structural proteins. Two CD8(+) T cell responses targeting the SARS-CoV membrane (M) and nucleocapsid (N) proteins were characterized by determining their HLA restriction and minimal T cell epitope regions. Furthermore, these responses were found to persist up to 11 years post-infection. An absence of cross-reactivity of these CD8(+) T cell responses against the newly-emerged Middle East respiratory syndrome coronavirus (MERS-CoV) was also demonstrated. The knowledge of the persistence of SARS-specific celullar immunity targeting the viral structural proteins in SARS-recovered individuals is important in the design and development of SARS vaccines, which are currently unavailable.


Interaction of immunosuppressants with HCV antivirals daclatasvir and asunaprevir: combined effects with mycophenolic acid.
To investigate the specific effects of immunosuppressants on the antiviral action of daclatasvir and asunaprevir. The antiviral activity of daclatasvir (DCV) and asunaprevir (ASV) combined with immunosuppressants was tested using two in vitro models for hepatitis C virus (HCV) infection. Tacrolimus, rapamycin and cyclosporine did not negatively affect the antiviral action of DCV or ASV. Mycophenolic acid (MPA) showed additive antiviral effects combined with these direct acting antivirals (DAAs). MPA induces interferon-stimulated genes (ISGs) and is a potent GTP synthesis inhibitor. DCV or ASV did not induce ISGs expression nor affected ISG induction by MPA. Rather, the combined antiviral effect of MPA with DCV and ASV was partly mediated via inhibition of GTP synthesis. Immunosuppressants do not negatively affect the antiviral activity of DAAs. MPA has additive effect on the antiviral action of DCV and ASV. This combined benefit needs to be confirmed in prospective clinical trials.


Anti-Müllerian hormone is a predictor of medium-term cumulative live birth following in vitro fertilization/intracytoplasmic sperm injection: A retrospective study.
This study aims to examine the capacity of anti-Müllerian hormone (AMH) to predict cumulative live birth rate (CLBR) following IVF/ICSI within 36 months since start of treatment. This is a cohort study of women seeking IVF/ICSI fertility treatment in a private Australian IVF clinic in a single calendar year. Live births were monitored over three years following start date of IVF/ICSI. The impact of serum AMH level on the CLBR was assessed using Cox's proportional hazard models, and its incremental values in the prediction of CLBR were evaluated. The CLBRs were significantly higher in women with AMH levels in the highest (>44.5 pmol/L; 87.0%, 95% CI 79.2% - 95.1%) and in the middle two quartiles (between 11.5 and 44.5 pmol/L; 81.0%, 95% CI 74.2% - 87.6%), compared with AMH levels below the 25th percentile (≤11.5 pmol/L; 63.2%, 95% CI 53.2% - 74.5%). Approximately half of the women with AMH in the lowest quartile conceived a live birth within 12 months of starting IVF compared with two-thirds of the women in the upper three quartiles. After adjusting for confounders, AMH remained a significant, albeit slight predictor of CLBR with a fall of 3 pmol/L equating to an 1% decrease in CLBR. The AMH's added values into the prediction of live birth were slight, indicated by a net reclassification improvement of 13.8%. The value is lower than that of maternal age (35.1%). Serum AMH level was a significant slight predictor of CLBR following IVF/ICSI. AMH should not be used to exclude women from IVF/ICSI however, women with low AMH should be counselled on the likelihood of taking longer to achieve a live birth than individuals with normal AMH levels.


A splicing variant of TFEB negatively regulates the TFEB-autophagy pathway.
Transcription factor EB (TFEB) is a master regulator of the autophagy-lysosomal pathway (ALP). Here, we cloned a novel splicing variant of TFEB, comprising 281 amino acids (hereafter referred to as small TFEB), and lacking the helix-loop-helix (HLH) and leucine zipper (LZ) motifs present in the full-length TFEB (TFEB-L). The TFEB variant is widely expressed in several tissues, including the brain, although its expression level is considerably lower than that of TFEB-L. Intriguingly, in cells stably expressing small TFEB, the expression profile of genes was inverted compared to that in cells ectopically expressing TFEB-L. In addition, fisetin-induced luciferase activity of promoter containing either coordinated lysosomal expression and regulation (CLEAR) element or antioxidant response element (ARE) was significantly repressed by co-transfection with small TFEB. Moreover, fisetin-mediated clearance of phosphorylated tau or α-synuclein was attenuated in the presence of small TFEB. Taken together, the results suggest that small TFEB is a novel splicing variant of TFEB that might act as a negative regulator of TFEB-L, thus fine tuning the activity of ALP during cellular stress.


Exclusive radiotherapy for primary squamous cell carcinoma of the vagina.
To retrospectively analyze results of external beam therapy (EBT) with brachytherapy (BT) for primary vaginal squamous cell carcinoma (PVSCC). From 1970 to 2001, 91 patients were included. FIGO stages were: I (29%), II (38%), III (29%) and IVa (4%). EBT delivered a median total dose of 50 Gy to the pelvis. BT was performed with a customized intra-vaginal applicator and in 36% of applications combined endocavitary and interstitial BT. ICRU Report 38 parameters were reported. The 5-year cause specific survival (CSS) rates were: 83% for stage I, 76% for stage II, 52% for stage III, and 2 of the 4 stage IVa patients died 9 and 36 months after treatment. The 5-year pelvis control rates were: 79% for stage I and II and 62% for stage III. Recurrences as a first event were local only in 68% of cases, nodal only in 10%, metastatic only in 13% and combined in 9%. In multivariate analysis: stage (I and II versus II and IV), response to EBT (evaluated at BT), and the number of BT applications were statistically significant for CSS. Grade 2-3 toxicities were as follows (Franco-Italian Glossary): rectum (n=3), sigmoid colon and small bowel (n=8), bladder (n=5), ureter (n=4) and vagina (n=13). Anterior location of the tumor increased bladder toxicity (p=0.01) and total reference air kerma was higher in patients who experienced grade 2-3 urinary or digestive toxicity (p=0.03). EBT with BT is an effective treatment for patients with stage I-II PVSCC. The incidence and severity of late toxicity were relatively low. Recent advances in the treatment of cervix carcinoma emphasize the need for concomitant radio-chemotherapy in stages III-IV and the use of MRI for treatment planning.


Fiber Ball White Matter Modeling Reveals Microstructural Alterations in Healthy Brain Aging.
Age-related white matter degeneration is characterized by myelin breakdown and neuronal fiber loss that preferentially occur in regions that myelinate later in development. Conventional diffusion MRI (dMRI) has demonstrated age-related increases in diffusivity but provide limited information regarding the tissue-specific changes driving these effects. A recently developed dMRI biophysical modeling technique, Fiber Ball White Matter (FBWM) modeling, offers enhanced biological interpretability by estimating microstructural properties specific to the intra-axonal and extra-axonal spaces. We used FBWM to illustrate the biological mechanisms underlying changes throughout white matter in healthy aging using data from 63 cognitively unimpaired adults ages 45-85 with no radiological evidence of neurodegeneration or incipient Alzheimer's disease. Conventional dMRI and FBWM metrics were computed for two late-myelinating (genu of the corpus callosum and association tracts) and two early-myelinating regions (splenium of the corpus callosum and projection tracts). We examined the associations between age and these metrics in each region and tested whether age was differentially associated with these metrics in late- vs. early-myelinating regions. We found that conventional metrics replicated patterns of age-related increases in diffusivity in late-myelinating regions. FBWM additionally revealed specific intra- and extra-axonal changes suggestive of myelin breakdown and preferential loss of smaller-diameter axons, yielding in vivo corroboration of findings from histopathological studies of aged brains. These results demonstrate that advanced biophysical modeling approaches, such as FBWM, offer novel information about the microstructure-specific alterations contributing to white matter changes in healthy aging. These tools hold promise as sensitive indicators of early pathological changes related to neurodegenerative disease.


Effects of autotransfusion of mediastinal shed blood on biochemical markers of myocardial damage in coronary surgery.
Previous studies have shown conflicting results regarding the effect of autotransfusion of mediastinal shed blood after coronary artery bypass grafting (CABG) on the serum levels of myocardial band (MB) isoenzymes of creatine kinase (CK-MB) and cardiac troponins. The effect of autotransfusion on serum levels of human heart fatty acid binding protein (H-FABP), another marker of myocardial necrosis, has not been studied. The aim of the present study was to investigate the effects of autotransfusion of mediastinal shed blood on the serum levels of CK-MB, cardiac troponin T (cTnT), and H-FABP after uncomplicated primary CABG. Fifty patients were randomized to post-operative autotransfusion of mediastinal shed blood or no autotransfusion. Blood samples for the analysis of the biochemical markers of myocardial damage were drawn pre-operatively and 1, 4, 12, 24, 48, and 72 h after the termination of cardiopulmonary bypass. Samples from the mediastinal shed blood were collected after 1 and 4 h. The levels of the biochemical markers of myocardial injury were all markedly elevated in mediastinal shed blood. Autotransfusion did not significantly affect the serum levels of cTnT or H-FABP. However, during the early post-operative hours, there was a trend towards a higher level of cTnT and H-FABP in the autotransfusion group. During the first 24 h after surgery, the autotransfusion group had a significantly higher serum level of CK-MB. Post-operative autotransfusion of mediastinal shed blood may contribute to elevated serum levels of biochemical markers of myocardial injury.


[Knee orthosis. Evaluation of clinical efficiency with a review of the literature].
The aim of this work was to collect literature data to assess the knee orthosis use with controlled clinical trials. The international literature relating to the years 1980-2000 was carried out with the Medline data bank. Five hundred and twelve articles were collected and 35 were retained, 32 related to clinical studies and three related to toxicity. The evaluations were performed with either functional tests or clinical studies. The open-non controlled clinical trials are far higher than prospective comparative clinical trials that were developed only during last years. Main criteria of judgement are articular stability, functional pain and clinical index. Most of studies focused the anterior cruciate ligament lesions, patellar pains and osteoarthritis. This review of the literature may help to determine the interest and the limits of orthosis use as well as side effect occurrence. Both these clinical and experimental data point out the methodological difficulties facing the investigatory clinician in knee orthosis evaluation. Many studies contributed to the clinical evaluation of the knee orthosis. This work highlights various clinical indications for the same orthosis and variable results for the same indication. These methodological difficulties may not prevent a further evaluation of these devices to improve their use in current practice.


[Four-stage gradual expanding approach of problem based learning in otorhinolaryngology].
The aim of the study is to cover the shortages of PBL, such as time-consuming, abstract, lacking of the course of clinic practice, and to introduce PBL to the teaching of otorhinolaryngology. By the improvement of the international classic teaching model of PBL, we put forward "four-stage gradual expanding approach of PBL" and establish "four-stage gradual expanding approach of PBL in otorhinolaryngology". Through the four stages of watching PBL, simulation PBL, internship PBL, practice PBL, we have accomplished the organic integration of theory teaching and clinical practice. This teaching method is more adaptive to the teaching of otorhinolaryngology, and it can help the medicine students to establish the whole concept of medicine and can stimulate them to form the good habits of self-regulated learning and life-long learning efficiently.


Modeling effects of cadmium on population growth of Palaemonetes pugio: results of a full life cycle exposure.
In an 8-month laboratory experiment, Palaemonetes pugio (grass shrimp) were exposed to aqueous cadmium (free cadmium ion concentrations of 1.51 or 2.51 microg Cd(2+)/L) for an entire life cycle, from larva to reproductive adult and through to production of second-generation larva. Individual-level effects on survival, life stage duration, and reproduction were measured, and population growth was projected using two models: a stage-based matrix model and a z-transformed life cycle graph analysis. Adult survival was significantly reduced at 2.51 microg Cd(2+)/L, but cadmium exposure had no effects on survival or stage duration of embryos, larvae, or juveniles. Survival of second-generation larvae was unaffected by maternal exposure. Brood size was reduced by 27% at 1.51 microg Cd(2+)/L and by 36% at 2.51 microg Cd(2+)/L. The percent of females in the population that was gravid was approximately 50% lower at 2.51 microg Cd(2+)/L, compared to controls. Both population models projected a dose-dependent decrease in population growth rate (lambda), up to a 12% reduction at 2.51 microg Cd(2+)/L, which can be attributed mainly to contributions from reproductive effects. Elasticity analysis revealed that population growth rate was most sensitive to changes in survival of juveniles and adults. However, lethal effects of cadmium made only a small contribution to the effect on population growth rate. Even though both models project positive growth (lambda>1) of grass shrimp populations exposed to low concentrations of cadmium, the ability of populations to withstand predation pressure would be compromised.


Correlation between five minerals and the healing effect of Brazilian medicinal plants.
A great many Brazilian medicinal plants are used in wound healing. They are usually applied directly to wounds, some in natura as poultices, some as dried powders, and others as water extracts (teas) for bathing. Sixteen plants from the Atlantic Rain Forest of the State of Rio de Janeiro in Brazil were analyzed to determine the concentration of five minerals: silicon, manganese, iron, copper, and zinc to study their possible role in the healing processes. The analysis was performed in plant samples and in teas prepared from them. There appears to be a correlation between the healing effect of the plants and their content of these element.


Development and distribution of serotonin in the central nervous system of Manduca sexta during embryogenesis. I. The brain and frontal ganglion.
Development of the serotonergic system in the brain and frontal ganglion of the Manduca embryo between 35 and 100% of development was studied immunocytochemically with an antiserum to serotonin (5-HT). Serotonin immunoreactivity was initially detectable at 40-45% development in short fibers in the head region, prior to differentiation of the brain. Immunoreactive cell bodies were first seen in the brain at 60% development, located in the protocerebrum and tritocerebrum. Thick fiber tracts crossing the midline (commissures) could also be observed at this early stage. As development of the embryo progressed, eight immunoreactive cell groups, containing a total of about 38-40 cells, and four commissures with terminal arborizations appeared successively in the brain. From 75 to 100% development, no obvious changes occurred in the number or distribution of cells, and the brain exhibited the same pattern of 5-HT immunoreactive cells, fiber tracts and arborizations as in last instar larvae of Manduca. However, an increase in the size of the cells in both the brain and frontal ganglion was noted between 75 and 80% development, followed by a decrease by 100% development. The frontal ganglion was found to contain three 5-HT immunoreactive cells, which appeared to send bilateral projections into the frontal connectives and the recurrent nerve. During embryonic development, the dendritic arborizations of these frontal ganglion cells increased, while the amount of 5-HT immunoreactivity in the cell bodies decreased. Thus, the serotonergic system first appears in the Manduca embryo at an early stage of development, similar to the situation in other insects as well as vertebrates. By the end of the embryonic period, the same number of serotonergic neurons are present in the brain as in larval and adult Manduca, suggesting that once formed, these cells persist through postembryonic development and metamorphosis.


Dynamic Changes in High-Sensitivity Cardiac Troponin I Are Associated with Dynamic Changes in Sum Absolute QRST Integral on Surface Electrocardiogram in Acute Decompensated Heart Failure.
A three-dimensional electrocardiographic (ECG) metric, the sum absolute QRST integral (SAI QRST), predicts ventricular arrhythmias in heart failure (HF) patients with implantable cardioverter defibrillator and mechanical response to cardiac resynchronization therapy. We hypothesized that there is an association between patient-specific changes in SAI QRST and myocardial injury as measured by high-sensitivity troponin I (hsTnI). Sum absolute integral QRST on resting 12-lead ECG and hsTnI were measured simultaneously, every 3 hours, and during 12-hour observation period in a prospective cohort of emergency department patients (n = 398; mean age 57.8 ± 13.2 years; 54% female, 64% black), diagnosed with acute coronary syndrome (ACS, n = 28), acutely decompensated HF (acute decompensated heart failure, n = 35), cardiac non-ACS (n = 19), or noncardiac condition (n = 316). Random-effects linear regression analysis assessed the association of SAI QRST and myocardial injury, with adjustment for demographics (age, sex, race), prevalent cardiovascular disease (myocardial infarction, history of revascularization, stroke, and HF), risk factors (diabetes, smoking, hypercholesterolemia, hypertension, and cocaine use), and left bundle branch block. Within the entire cohort, SAI QRST decreased by 3 (95%CI -5 to -1) mV*ms every 3 hours. A 10-fold increase in hsTnI was associated with a 7.7 (0.6-14.9) mV*ms increase in SAI QRST. In the subgroup of acutely decompensated HF patients (n = 35), a 10-fold increase in hsTnI was associated with a 61.0 (5.9-116.1) mV*ms increase in SAI QRST. Patient-specific time-varying changes in the surface ECG scalar measure of global electrical heterogeneity, as measured by SAI QRST, and in myocardial injury as measured by hsTnI, are independently and directly associated with each other, likely reflecting a common underlying mechanism.


Effects of a nitrate reductase inactivating enzyme and NAD(P)H on the nitrate reductase from higher plants and Neurospora.
Evidence is presented which suggests that the NAD(P)H-cytochrome c reductase component of nitrate reductase is the main site of action of the inactivating enzyme. When tested on the nitrate reductase (NADH) from the maize root and scutella, the NADH-cytochrome c reductase was inactivated at a greater rate than was the FADH2-nitrate reductase component. With the Neurospora nitrate reductase (NADPH) only the NADPH-cytochrome c reductase was inactivated. p-Chloromercuribenzoate at 50 muM, which gave almost complete inhibition of the NADH-cytochrome c reductase fraction of the maize nitrate reductase, had no marked effect on the action of the inactivating enzyme. A reversible inactivation of the maize nitrate reductase has been shown to occur during incubation with NAD(P)H. In contrast to the action of the inactivating enzyme, it is the FADH2-nitrate reductase alone which is inactivated. No inactivation of the Neurospora nitrate reductase was produced by NAD(P)H alone and also in the presence of FAD. The lack of effect of the inactivating enzyme and NAD(P)H on the FADH2-nitrate reductase of Neurospora suggests some differences in its structure or conformation from that of the maize enzyme. A low level of cyanide (0.4 mu M) markedly enhanced the action of NAD(P)H on the maize enzyme; Cyanide at a higher level (6 mu M) did give inactivation of the Neurospora nitrate reductase in the presence of NADPH and FAD. The maize nitrate reductase, when partially inactivated by NADH and cyanide, was not altered as a substrate for the inactivating enzyme. The maize root inactivating enzyme was also shown to inactivate the nitrate reductase (NADH) in the pea leaf. It had no effect on the nitrate reductase from either Pseudomonas denitrificans or Nitrobacter agilis.


Evidence for phosphorylation of pancreatic islet pyruvate kinase.
Rabbit pancreatic islet cytosol catalyzes the calcium-activated phosphorylation by [gamma 32P]ATP of a protein with a molecular weight of 57,000 that is precipitated with antipyruvate kinase antibodies. We were unable to demonstrate that phosphorylation in the presence of calcium or cAMP had any immediate effect on rat pancreatic islet pyruvate kinase activity. This finding is consistent with our inability to confirm the finding of others that pancreatic islets contain phosphoenolpyruvate carboxykinase activity (Diabetes, 34:246, 1985). Since the carboxykinase catalyzes phosphoenolpyruvate formation and pyruvate kinase catalyzes essentially the opposite reaction, if the carboxykinase were present in the beta cell, pyruvate kinase would need to be inhibited to prevent recycling of phosphoenolpyruvate.


Complex benign horizontal canal positional vertigo: new perceptual management.
Horizontal semicircular canal site pathology of benign paroxysmal positional vertigo demonstrating three types of nystagmi on positional test were studied. We have attempted to design a protocol for its diagnosis and treatment. 320 patients of HSC-BPPV were subjected to two types of positional tests. Of these, patients with bilateral steady apogeotropic nysatgmus were treated with VAV modification of Semont's maneuver. Patients with unsteady or changing apo/geotropic signs were converted into steady geotropic ones by repetitive positional tests; followed by barbecue maneuver with forced prolong positioning. Overall 88% of patients had a total recovery. 92% of patients with geotropic nystagmus showed no symptoms after second maneuveral sitting. 85% of patients with apogeotropic nystagmus recovered fully after third maneuveral sitting. Correct identification of subtypes of HSC-BPPV is based on provoked nystagmus by positional tests. After locating the site and side on the basis of nystagmic pattern, physician can apply the appropriate PRM. II a.


Healthy Eating, Aerobic and Resistance Training in Youth (HEARTY): study rationale, design and methods.
The objective of the Healthy Eating Aerobic and Resistance Training in Youth (HEARTY) trial (ClinicalTrials.Gov # NCT00195858) was to examine the effects of resistance training, with and without aerobic training, on percent body fat in sedentary, post-pubertal overweight or obese adolescents aged 14-18 years. This paper describes the HEARTY study rationale, design and methods. After a 4-week supervised low-intensity exercise run-in period, 304 overweight or obese adolescents with a body mass index≥85th percentile for age and sex were randomized to 4 groups for 22 weeks (5 months): diet+aerobic exercise, diet+resistance exercise, diet+combined aerobic and resistance exercise, or a diet only waiting-list control. All participants received dietary counseling designed to promote healthy eating with a maximum daily energy deficit of -250 kcal. The primary outcome is percent body fat measured by Magnetic Resonance Imaging. Secondary outcomes include changes in anthropometry, regional body composition, resting energy expenditure, cardiorespiratory fitness, musculoskeletal fitness, cardiometabolic risk markers, and psychological health. To our knowledge, HEARTY is the largest clinical trial examining effects of aerobic training, resistance training, and combined aerobic and resistance training on changes in adiposity and cardiometabolic risk markers in overweight and obese adolescents. The findings will have important clinical implications regarding the role that resistance training should play in the management of adolescent obesity and its co-morbidities.


The Association between Patient Activation and Outcomes among Severely Mentally Ill Patients.
Little is known about the association between patient activation, health, service utilization, and cost among mental health (MH) patients. Patients aged 18 to 64 with schizophrenia (Sz, n = 43), bipolar disorder (BD, n = 59), or major depressive disorder (MDD, n = 34) completed the Patient Activation Measure for Mental Health (PAM-MH), the Colorado Symptom Index, demographic, socioeconomic, treatment, and social support questionnaire items. Average PAM-MH score indicated BD patients the most activated (66.6 ± 17.5), Sz (57.4 ± 10.4) less activated, and MDD the least activated (55.4 ± 14.6). The MDD cohort had the highest ($27,616 ± 26,229) and the BD had the lowest total annual healthcare cost ($18,312 ± 25,091). PAM-MH score was inversely correlated with healthcare costs and regression analysis showed a PAM-MH score × gender interaction. The strongest negative relationship between PAM and cost was for males. These analyses support the inverse association between PAM-MH and healthcare service utilization and cost.