The immune-related fatty acids are responsive to CO2 driven seawater acidification in a crustacean brine shrimp Artemia sinica.
The gradual increase of CO2 concentration in the atmosphere, absorbed by the ocean surface water through air to sea equilibration termed ocean acidification (OA), leads to the decline of pH in seawater. It is not clear so far how the composition of fatty acids, particular the immune-related, in marine crustacean and the subsequent energy supply in marine ecosystem are affected by OA. The brine shrimp Artemia sinica is an open and common feed that provide essential fatty acids for mariculture. In this study, the fatty acids profiles of brine shrimp cultured under different lower pH levels of CO2 driven seawater were investigated. The results showed a significant reduction of the proportion of total saturated fatty acids under the pH7.6 within one week. Meanwhile, the percentage of total monounsaturated fatty acids was significantly decreased at day 14 under pH7.8, and this percentage gave a significant increase of proportion within one week under pH7.6. Furthermore, the relative content of total polyunsaturated fatty acids (PUFAs) was found to be clearly increased with exposure to different seawater acidification at day 1, suggesting that the brine shrimp immune response was likely to be affected by acidified seawater as the PUFAs have been well known to be involved in immunomodulatory effects through alterations on cell membrane fluidity/lipid mediators and gene expression of cell signaling pathways. Notably, eicosapentaenoic acid and docosahexaenoic acid, which have essential effect on various physiological processes such as inflammatory cytokines production and cell structural stability, were strongly increased under two lower pH treatments within one week and with the significant increase at day 1 under pH7.6. These data clearly supported the hypothesis that OA might affect fatty acids composition, likely also the innate immunity, in crustacean and the subsequent energy transfer by food-chain system in the marine ecosystem.


The shed ectodomain of Nr-CAM stimulates cell proliferation and motility, and confers cell transformation.
Nr-CAM, a cell-cell adhesion molecule of the immunoglobulin-like cell adhesion molecule family, known for its function in neuronal outgrowth and guidance, was recently identified as a target gene of beta-catenin signaling in human melanoma and colon carcinoma cells and tissue. Retrovirally mediated transduction of Nr-CAM into fibroblasts induces cell motility and tumorigenesis. We investigated the mechanisms by which Nr-CAM can confer properties related to tumor cell behavior and found that Nr-CAM expression in NIH3T3 cells protects cells from apoptosis in the absence of serum by constitutively activating the extracellular signal-regulated kinase and AKT signaling pathways. We detected a metalloprotease-mediated shedding of Nr-CAM into the culture medium of cells transfected with Nr-CAM, and of endogenous Nr-CAM in B16 melanoma cells. Conditioned medium and purified Nr-CAM-Fc fusion protein both enhanced cell motility, proliferation, and extracellular signal-regulated kinase and AKT activation. Moreover, Nr-CAM was found in complex with alpha4beta1 integrins in melanoma cells, indicating that it can mediate, in addition to homophilic cell-cell adhesion, heterophilic adhesion with extracellular matrix receptors. Suppression of Nr-CAM levels by small interfering RNA in B16 melanoma inhibited the adhesive and tumorigenic capacities of these cells. Stable expression of the Nr-CAM ectodomain in NIH3T3 cells conferred cell transformation and tumorigenesis in mice, suggesting that the metalloprotease-mediated shedding of Nr-CAM is a principal route for promoting oncogenesis by Nr-CAM.


Systems-level analysis of human aging genes shed new light on mechanisms of aging.
Although studies over the last decades have firmly connected a number of genes and molecular pathways to aging, the aging process as a whole still remains poorly understood. To gain novel insights into the mechanisms underlying aging, instead of considering aging genes individually, we studied their characteristics at the systems level in the context of biological networks. We calculated a comprehensive set of network characteristics for human aging-related genes from the GenAge database. By comparing them with other functional groups of genes, we identified a robust group of aging-specific network characteristics. To find the structural basis and the molecular mechanisms underlying this aging-related network specificity, we also analyzed protein domain interactions and gene expression patterns across different tissues. Our study revealed that aging genes not only tend to be network hubs, playing important roles in communication among different functional modules or pathways, but also are more likely to physically interact and be co-expressed with essential genes. The high expression of aging genes across a large number of tissue types also points to a high level of connectivity among aging genes. Unexpectedly, contrary to the depletion of interactions among hub genes in biological networks, we observed close interactions among aging hubs, which renders the aging subnetworks vulnerable to random attacks and thus may contribute to the aging process. Comparison across species reveals the evolution process of the aging subnetwork. As the organisms become more complex, the complexity of its aging mechanisms increases and their aging hub genes are more functionally connected.


Phylogenetic analysis of vertebrate CXC chemokines reveals novel lineage specific groups in teleost fish.
In this study, we have identified 421 molecules across the vertebrate spectrum and propose a unified nomenclature for CXC chemokines in fish, amphibians and reptiles based on phylogenetic analysis. Expanding on earlier studies in teleost fish, lineage specific CXC chemokines that have no apparent homologues in mammals were confirmed. Furthermore, in addition to the two subgroups of the CXCL8 homologues known in teleost fish, a third group was identified (termed CXCL8_L3), as was a further subgroup of the fish CXC genes related to CXCL11. Expression of the CXC chemokines found in rainbow trout, Oncorhynchus mykiss, was studied in response to stimulation with inflammatory and antiviral cytokines, and bacterial. Tissue distribution analysis revealed distinct expression profiles for these trout CXC chemokines. Lastly three of the trout chemokines, including two novel fish specific CXC chemokines containing three pairs of cysteines, were produced as recombinant proteins and their effect on trout leucocyte migration studied. These molecules increased the relative expression of CD4 and MCSFR in migrated cells in an in vitro chemotaxis assay.


Apamin-sensitive, non-nitric oxide (NO) endothelium-dependent relaxations to bradykinin in the bovine isolated coronary artery: no role for cytochrome P450 and K+.
Since cytochrome P(450)-derived metabolites of arachidonic acid and K(+) have been implicated in endothelium-derived hyperpolarizing factor (EDHF)-dependent responses, the aim of this study was to determine whether such factors contribute to non-nitric oxide (NO), endothelium-dependent relaxation to bradykinin (BK) in bovine isolated coronary artery. In rings of artery contracted with U46619 and treated with indomethacin (3 microM) and N(G)-nitro-L-arginine (L-NOARG; 100 microM), relaxation to BK (0.01 nM-0.3 microM) was blocked by approximately 60% after inhibition of K(+) channels with either high extracellular K(+) (high [K(+)](o); 15 - 67 mM) or apamin (0.3 microM). Ouabain (1 microM), an inhibitor of Na(+)/K(+)-ATPase, decreased the sensitivity to BK without affecting the maximum response. In L-NOARG-treated rings, ouabain had no further effect on the relaxation to BK. An inhibitor of inward-rectifying K(+) channels, Ba(2+) (30 microM), had no effect on relaxations to BK in the absence or presence of either L-NOARG or ouabain. KCl (2.5 - 10 mM) elicited small relaxations ( approximately 20%) that were abolished by nifedipine (0.3 microM) and ouabain. Both the high [K(+)](o)/apamin-sensitive relaxation to BK, and the relaxation to the K(ATP) channel-opener, levcromakalim (0.6 microM), were unaffected by the cytochrome P(450) inhibitor, 7-ethoxyresorufin (10 microM), or by co-treatment with a phospholipase A(2) inhibitor, arachidonyl trifluoromethyl ketone (AACOCF(3); 3 microM) and a diacylglycerol (DAG)-lipase inhibitor, 1, 6-bis-(cyclohexyloximinocarbonylamino)-hexane (RHC 80267; 30 microM). The non-NO/high [K(+)](o)-insensitive, approximately 40% relaxation to BK was, however, abolished by these treatments. Therefore, neither cytochrome P(450)-derived metabolites of arachidonic acid nor K(+) appear to mediate the EDHF-like relaxation to BK (i.e the non-NO, high [K(+)](o)/apamin-sensitive component) in bovine coronary arteries. Cytochrome P(450)-derived metabolites may be released at higher BK concentrations to act in parallel with NO and the high [K(+)](o)/apamin-sensitive mechanism.


Parapapillary Deep-Layer Microvasculature Dropout in Glaucoma: Topographic Association With Glaucomatous Damage.
The purpose of this article was to compare the frequencies of the parapapillary deep-layer microvasculature dropout (MvD) detected by optical coherence tomography angiography in eyes with primary open-angle glaucoma (POAG) and healthy eyes and to determine the topographic correlation between the MvD and the glaucomatous retinal nerve fiber layer (RNFL) defect in POAG eyes. Microvasculature in the peripapillary deep-layer was evaluated in 150 POAG patients and 45 healthy controls using swept-source optical coherence tomography angiography to identify an MvD. Frequencies of MvDs were compared between the POAG and control groups. In POAG eyes with MvD, topographic correlation was assessed between the MvD and the RNFL defect defined based on the optical coherence tomography circumpapillary RNFL thickness measurement. MvD was observed as a sectoral filling defect in the parapapillary deep-layer in 53.9% (n = 62) of the POAG eyes, whereas none of the control eyes exhibited an MvD. POAG eyes with MvD had a thinner global RNFL (P < 0.001) and worse visual field mean deviation (P = 0.042) and were more myopic (P = 0.029), with axial length being longer (P = 0.046) than those without MvD. There was a good agreement between the circumferential extent and location of MvD and those of RNFL defect (95% limits of agreement of the difference ranged from -23.4 to 21.9° and -16.2 to 17.0°, respectively). MvD was identified in the parapapillary deep layer exclusively in POAG eyes at the location of glaucomatous damage using optical coherence tomography angiography. The finding suggests that peripapillary deep-layer circulation is directly related to the glaucomatous optic neuropathy.


Continuous Glucose Monitoring in the Intensive Care Unit During the COVID-19 Pandemic.
Real-time continuous glucose monitoring (rtCGM) in critically ill hospitalized patients holds promise; however, real-world data are needed. We placed Dexcom G6 CGM on intensive care unit (ICU) patients at Montefiore Medical Center with confirmed coronavirus disease 2019 (COVID-19) infection and glycemic variability. We analyzed inpatient CGM accuracy using point-of-care (POC) glucose-CGM matched pairs and included patients for analysis regardless of clinical status. We included 11 patients with CGM: 8 on continuous insulin infusion (CII), 8 on vasopressors, 8 intubated, 4 on high-dose glucocorticoids, 6 on renal replacement therapy, and 2 with anasarca. Accuracy was 12.58% for mean and 6.3% for median absolute relative difference. CGM reduced POC testing by ∼60% for patients on CII. In this real-world preliminary analysis of rtCGM during critical illness, we demonstrate early feasibility, considerable accuracy, and meaningful reduction in the frequency of POC glucose testing.


Is self-guided internet-based cognitive behavioural therapy (iCBT) harmful? An individual participant data meta-analysis.
Little is known about potential harmful effects as a consequence of self-guided internet-based cognitive behaviour therapy (iCBT), such as symptom deterioration rates. Thus, safety concerns remain and hamper the implementation of self-guided iCBT into clinical practice. We aimed to conduct an individual participant data (IPD) meta-analysis to determine the prevalence of clinically significant deterioration (symptom worsening) in adults with depressive symptoms who received self-guided iCBT compared with control conditions. Several socio-demographic, clinical and study-level variables were tested as potential moderators of deterioration. Randomised controlled trials that reported results of self-guided iCBT compared with control conditions in adults with symptoms of depression were selected. Mixed effects models with participants nested within studies were used to examine possible clinically significant deterioration rates. Thirteen out of 16 eligible trials were included in the present IPD meta-analysis. Of the 3805 participants analysed, 7.2% showed clinically significant deterioration (5.8% and 9.1% of participants in the intervention and control groups, respectively). Participants in self-guided iCBT were less likely to deteriorate (OR 0.62, p < 0.001) compared with control conditions. None of the examined participant- and study-level moderators were significantly associated with deterioration rates. Self-guided iCBT has a lower rate of negative outcomes on symptoms than control conditions and could be a first step treatment approach for adult depression as well as an alternative to watchful waiting in general practice.


Weak avalanche discrimination for gated-mode single-photon avalanche photodiodes.
The after-pulsing effect is a common problem that needs to be overcome for high-speed single-photon detection based on gated-mode single-photon avalanche photodiodes (SPADs). This paper presents a simple and practical method for suppression of the after-pulsing probability using an auxiliary signal to discriminate quite weak avalanches. The detection efficiency and after-pulse probability of an InGaAs/InP SPAD are investigated with a 10 MHz gating for conventional and proposed methods, and a sharp decrease of after-pulse probability is demonstrated with the application of the proposed method. At a gating frequency of 100 MHz, a detection efficiency of 10.4% is achieved with an after-pulse probability of 5.6% without dead time.


Expressions of positive emotion in women's college yearbook pictures and their relationship to personality and life outcomes across adulthood.
To test hypotheses about positive emotion, the authors examined the relationship of positive emotional expression in women's college pictures to personality, observer ratings, and life outcomes. Consistent with the notion that positive emotions help build personal resources, positive emotional expression correlated with the self-reported personality traits of affiliation, competence, and low negative emotionality across adulthood and predicted changes in competence and negative emotionality. Observers rated women displaying more positive emotion more favorably on several personality dimensions and expected interactions with them to be more rewarding; thus, demonstrating the beneficial social consequences of positive emotions. Finally, positive emotional expression predicted favorable outcomes in marriage and personal well-being up to 30 years later. Controlling for physical attractiveness and social desirability had little impact on these findings.


Effects of D-glucose concentration, D-fructose, and inhibitors of enzymes of the pentose phosphate pathway on the development and sex ratio of bovine blastocysts.
D-glucose at 5.6 mM reduces the progression of in vitro-produced (IVP) bovine embryos from the morula to the blastocyst stage and skews sex ratio towards males. Possibly, the presence of two transcriptionally active X-chromosomes compromises female embryo development through imbalance in glucose metabolism. Here, we have determined the threshold of embryo sensitivity to glucose, whether substitution of D-fructose for glucose reduces the selective loss of female embryos, and whether inhibition of an X-linked gene product, glucose 6-phosphate dehydrogenase (G6PD), normalizes sex ratio among bovine blastocysts. IVP zygotes were cultured in glucose-free medium to 72 hr post-insemination, at which time 8-cell embryos were selected for treatment and cultured until 144 hr post-insemination. Addition of 4 mM glucose at the 8-cell stage reduced (P < 0.05) the number of blastocyst that formed, whereas 2.5 and 1 mM glucose had no effect. Sex ratio in the presence of 4 and 2.5 mM glucose differed significantly from 0.5, while 1 mM glucose had no effect. D-fructose (5.6 mM) did not compromise development to blastocyst and did not alter the sex ratio of blastocysts that formed. When G6PD inhibitors (dehydroepiandrosterone: DHEA and 6-aminonicotinamide: 6-AN), which effectively inhibit the reduction of the NADPH-sensitive dye, brilliant cresyl blue (BCB) in bovine morulae, were added to the culture medium containing 4 mM glucose, embryo development was moderately reduced, but sex ratio skewing was corrected (with 6-AN) or lowered (with DHEA). In conclusion, glucose above 2.5 mM impairs bovine embryo development and increases sex ratio, most likely as a result of increased pentose-phosphate (PP) pathway activity in female embryos.


Rod hue biases for foveal stimuli on CRT displays.
Signals from rod photoreceptors bias (shift) the hues determined by cone photoreceptors for extrafoveal mesopic stimuli, creating green, blue, and red rod hue biases at long, middle, and short wavelengths, respectively. The fovea contains far fewer rods and S cones but may not be immune to rod hue biases. Here, we determine the biases found for mesopic foveal stimuli presented on a CRT display. The rod green bias was observed at unique yellow for all but one observer with 2° tests and persisted for most observers with 0.5° tests. The rod red bias typically seen at unique blue in extrafoveal studies was not apparent for either size of foveal test stimulus, and it was sometimes replaced by a rod green bias. The rod blue bias typically seen at unique green and unique red in extrafoveal studies was weak on average and inconsistent for both sizes of foveal test stimuli. Thus, small mesopic foveal stimuli permit rod influence on M- and L-cone color pathways but disadvantage rod influence on S-cone pathways, perhaps because of the sparseness of foveal S-cones. However, some observers did show idiosyncratic foveal rod hue biases that do not follow the general trends.


Transfusion of CXCR4-primed endothelial progenitor cells reduces cerebral ischemic damage and promotes repair in db/db diabetic mice.
This study investigated the role of stromal cell-derived factor-1α (SDF-1α)/CXC chemokine receptor 4 (CXCR4) axis in brain and endothelial progenitor cells (EPCs), and explored the efficacy of CXCR4 primed EPCs in treating ischemic stroke in diabetes. The db/db diabetic and db/+ mice were used in this study. Levels of plasma SDF-1α and circulating CD34+CXCR4+ cells were measured. Brain SDF-1α and CXCR4 expression were quantified at basal and after middle cerebral artery occlusion (MCAO). In in vitro study, EPCs were transfected with adenovirus carrying null (Ad-null) or CXCR4 (Ad-CXCR4) followed with high glucose (HG) treatment for 4 days. For pathway block experiments, cells were pre-incubated with PI3K inhibitor or nitric oxide synthase (NOS) inhibitor for two hours. The CXCR4 expression, function and apoptosis of EPCs were determined. The p-Akt/Akt and p-eNOS/eNOS expression in EPCs were also measured. In in vivo study, EPCs transfected with Ad-null or Ad-CXCR4 were infused into mice via tail vein. On day 2 and 7, the cerebral blood flow, neurologic deficit score, infarct volume, cerebral microvascular density, angiogenesis and neurogenesis were determined. We found: 1) The levels of plasma SDF-1α and circulating CD34+CXCR4+ cells were decreased in db/db mice; 2) The basal level of SDF-1α and MCAO-induced up-regulation of SDF-1α/CXCR4 axis were reduced in the brain of db/db mice; 3) Ad-CXCR4 transfection increased CXCR4 expression in EPCs and enhanced EPC colonic forming capacity; 4) Ad-CXCR4 transfection prevented EPCs from HG-induced dysfunction (migration and tube formation) and apoptosis via activation of PI3K/Akt/eNOS signal pathway; 4) Ad-CXCR4 transfection enhanced the efficacy of EPC infusion in attenuating infarct volume and promoting angiogenesis and neurogenesis. Our data suggest that Ad-CXCR4 primed EPCs have better therapeutic effects for ischemia stroke in diabetes than unmodified EPCs do.


Sexual reproduction in Candida lipolytica.
Candida lipolytica is a rather common yeast isolated more frequently from substrates containing lipids or proteins, such as dairy products, than from substrates rich in sugars. This species assimilates hydrocarbons and is currently being studied for its potential to convert petroleum into yeast cells for use in feeds and foods. We have found C. lipolytica to exist in nature primarily in the heterothallic haploid state. When appropriate strains of opposite sex are mixed on a suitable sporulation medium, conjugation occurs followed by the production of ascospores. Since heterothallism permits laboratory control of hybridization, this characteristic of C. lipolytica enhances the possibility of im proving its strains for technological uses.


Design of biomaterials for intracellular delivery of carbon monoxide.
Carbon monoxide (CO) is recognized as one of the most important gas signaling molecules involved in governing various therapeutic responses. Intracellular generation of CO is spatiotemporally controlled by catalytic reactions of heme oxygenases (HOs). Thus, the ability to control intracellular CO delivery with modulation of the CO-release rate in specific amounts and locations is expected to improve our fundamental understanding of the functions of CO and the development of clinical applications. For this purpose, CO-releasing molecules (CORMs) have been developed and investigated in vitro and in vivo. Most CORMs are based on transition metal carbonyl complexes. Recently, various biomaterials consisting of metal carbonyls with biomacromolecular scaffolds have been reported to improve the properties of bare metal carbonyls. In this mini-review, current progress in CO delivery, recent strategies for the development of CORMs, and future directions in this field are discussed.


Personal exposure to airborne particles and metals: results from the Particle TEAM study in Riverside, California.
The PTEAM Study was the first large-scale probability-based study of personal exposure to particles. Sponsored by the U.S. Environmental Protection Agency (EPA) and the Air Resources Board of California, it was carried out by the Research Triangle Institute (RTI) and the Harvard University School of Public Health (HSPH). HSPH designed and constructed a 4-lpm, battery-operated personal monitor for inhalable particles (PM10) that could be worn comfortably for up to 14 hours by persons from 10 to 70 years old. The monitor was worn for two consecutive 12-hour periods (day and night) during the fall of 1990 by 178 participants representing 139,000 nonsmoking residents of Riverside, California. Nearly identical monitors were employed to collect concurrent indoor and outdoor samples. The monitors were equipped with a different sampling nozzle to collect fine particles (PM2.5). Population-weighted daytime personal PM10 exposures averaged 150 +/- 9 (SE) micrograms/m3, compared to concurrent indoor and outdoor concentrations of 95 +/- 6 micrograms/m3. This suggested the existence of excess mass near the person, a "personal cloud" that appeared related to personal activities. Fourteen of 15 prevalent elements also were evaluated in the personal samples. The two major indoor sources of indoor particles were smoking and cooking; even in these homes, however, more than half of the indoor particles came from outdoors, and a substantial portion of the indoor particles were of undetermined indoor origin. Outdoor concentrations near the homes were well correlated with outdoor concentrations at the central site, supporting the idea of using the central site as an indicator of of ambient concentrations over a wider area. Indoor concentrations were only weakly correlated with outdoor concentrations, however, and personal exposures were even more poorly correlated with outdoor concentrations. Elemental profiles were obtained for environmental tobacco smoke (ETS) (major contributions from potassium and chlorine) and cooking emissions (aluminum, iron, calcium, and chlorine). These profiles can be used in future source apportionment studies.


Feasibility and acceptability of a proposed infant feeding intervention trial for the prevention of type I diabetes.
To determine the feasibility of a randomized double-blind controlled trial of an infant formula without intact cow's-milk protein for preventing type I diabetes in high-risk children. We surveyed 83 people who either were parents of a child with type I diabetes or were pregnant women with type I diabetes in the ambulatory diabetes and obstetrics clinics in a university hospital. After a written and verbal description of the cow's milk-diabetes hypothesis, participants were asked to sign a sham consent form. A questionnaire designed to explore factors affecting their decision to either sign or not sign the consent form, as well as infant-feeding patterns, was subsequently administered. Overall, 69.9% (95% confidence interval, 60.0-79.8%) consented to participation in the proposed randomized trial. The decision to consent was not affected by the degree of belief in the cow's milk-diabetes hypothesis, the child's risk of diabetes, the respondent's demographic data, or infant feeding habits. A randomized feeding intervention study is an acceptable and feasible way to determine whether avoidance of cow's-milk protein during the first 6 months of life prevents type I diabetes in North American children.


The Werner syndrome protein is involved in RNA polymerase II transcription.
Werner syndrome (WS) is a human progeroid syndrome characterized by the early onset of a large number of clinical features associated with the normal aging process. The complex molecular and cellular phenotypes of WS involve characteristic features of genomic instability and accelerated replicative senescence. The gene involved (WRN) was recently cloned, and its gene product (WRNp) was biochemically characterized as a helicase. Helicases play important roles in a variety of DNA transactions, including DNA replication, transcription, repair, and recombination. We have assessed the role of the WRN gene in transcription by analyzing the efficiency of basal transcription in WS lymphoblastoid cell lines that carry homozygous WRN mutations. Transcription was measured in permeabilized cells by [3H]UTP incorporation and in vitro by using a plasmid template containing the RNA polymerase II (RNA pol II)-dependent adenovirus major late promoter. With both of these approaches, we find that the transcription efficiency in different WS cell lines is reduced to 40-60% of the transcription in cells from normal individuals. This defect can be complemented by the addition of normal cell extracts to the chromatin of WS cells. Addition of purified wild-type WRNp but not mutated WRNp to the in vitro transcription assay markedly stimulates RNA pol II-dependent transcription carried out by nuclear extracts. A nonhelicase domain (a direct repeat of 27 amino acids) also appears to have a role in transcription enhancement, as revealed by a yeast hybrid-protein reporter assay. This is further supported by the lack of stimulation of transcription when mutant WRNp lacking this domain was added to the in vitro assay. We have thus used several approaches to show a role for WRNp in RNA pol II transcription, possibly as a transcriptional activator. A deficit in either global or regional transcription in WS cells may be a primary molecular defect responsible for the WS clinical phenotype.


Growth Performance and Characterization of Meat Quality of Broiler Chickens Supplemented with Betaine and Antioxidants under Cyclic Heat Stress.
Heat stress (HS) causes oxidative stress, which compromises broiler performance and meat quality. The aim of this study was to determine whether dietary antioxidants could be used as an amelioration strategy. Seventy-two day-old-male Ross-308 chicks were exposed to either thermoneutral or cyclical heat stress conditions. Diets were either control commercial diet (CON), CON plus betaine (BET), or with a combination of betaine, selenized yeast, and vitamin E (BET + AOX). Heat stress increased the rectal temperature (p < 0.001), respiration rate (p < 0.001), decreased blood pCO2 (p = 0.002), and increased blood pH (p = 0.02), which indicated the HS broilers had respiratory alkalosis. Final body weight was decreased by HS (p < 0.001), whereas it was improved with BET (p = 0.05). Heat stress reduced cooking loss (p = 0.007) and no effect on drip loss, while BET decreased the drip loss (p = 0.01). Heat stress reduced the myofibril fragmentation index (p < 0.001) and increased thiobarbituric acid reactive substances (p < 0.001), while these were improved with the combination of BET + AOX (p = 0.003). In conclusion, BET overall improved growth rates and product quality in this small university study, whereas some additional benefits were provided by AOX on product quality in both TN and HS broilers.


Antisense targeting of CXXC finger protein 1 inhibits genomic cytosine methylation and primitive hematopoiesis in zebrafish.
CXXC finger protein 1 (CFP1) binds to unmethylated CpG dinucleotides and is a component of the Set1 histone methyltransferase complex. Mice lacking CFP1 suffer a peri-implantation lethal phenotype, and CFP1-deficient embryonic stem cells are viable but unable to differentiate and exhibit a 60-80% decrease in genomic cytosine methylation. A zebrafish homolog of CFP1 has been identified, is approximately 70% similar to murine CFP1, and is widely expressed during development. Zebrafish embryos treated with a zCFP1 antisense morpholino oligonucleotide had little or no circulating red blood cells and exhibited abnormal yolk sac morphology at 48 h post-fertilization. Many of the antisense-treated zebrafish also exhibited cardiac edema, and 14% were dead at 24 h post-fertilization. Morphant zebrafish also exhibited elevated levels of apoptosis, particularly in the intermediate cell mass, the site of primitive erythropoiesis, as well as aberrations in vascular development. Genomic DNA isolated from morphant embryos exhibited a 60% reduction of global genomic cytosine methylation. A similar phenotype was observed with an independent zCFP1 antisense morpholino oligonucleotide, but not following injection of an unrelated control oligonucleotide. The morphant phenotype was rescued when mRNA encoding murine CFP1 was co-injected with the antisense oligonucleotide. Genomic data base analysis reveals the presence of a second version of zebrafish CFP1 (zCFP1b). However, the morphant phenotype observed following specific depletion of zCFP1 indicates that these related genes have nonredundant functions controlling normal zebrafish hematopoiesis and epigenetic regulation. These findings establish the importance of CFP1 during postgastrulation development.


Monitoring the Path to the Elimination of Infectious Diseases.
During the endgame of elimination programs, parasite populations may exhibit dynamical phenomena not typical of endemic disease. Particularly, monitoring programs for tracking infection prevalence may be hampered by overall rarity, the sporadic and unpredictable timing and location of outbreaks, and under-reporting. A particularly important problem for monitoring is determining the distance that must be covered to achieve the elimination threshold at an effective reproduction number less than one. In this perspective, we suggest that this problem may be overcome by measuring critical slowing down. Critical slowing down is a phenomenon exhibited by nonlinear dynamical systems in the vicinity of a critical threshold. In infectious disease dynamics, critical slowing down is expressed as an increase in the coefficient of variation and other properties of the fluctuations in the number of cases. In simulations, we show the coefficient of variation to be insensitive to under-reporting error and therefore a robust measurement of the approach to elimination. Additionally, we show that there is an inevitable delay between the time at which the effective reproduction number is reduced to below one and complete elimination is achieved. We urge that monitoring programs include dynamical properties such as critical slowing down in their metrics for measuring achievement and avoid withdrawing control activities prematurely.


Patterns of interphalangeal hand joint involvement of osteoarthritis among men and women: a British cohort study.
To characterize the pattern of involvement of osteoarthritis (OA) of the hand among men and women of the same age. Structured hand examinations were performed on 1,467 men and 1,519 women who were age 53 years and born in England, Scotland, or Wales during the first week of March 1946 (identified through the United Kingdom National Survey of Health and Development). OA at each joint site was characterized using a previously validated examination schedule. The interrelationship of involvement of different hand joints was analyzed by logistic regression and cluster analyses. There was clear evidence of polyarticular involvement in the hand joints of both the men and the women. Among the women, 161 subjects had >/=4 joints involved, compared with only 41 subjects expected in this category (P < 0.001). Among the men, 87 subjects were observed to have >/=4 joints involved, in contrast with only 7 subjects expected (P < 0.001). The pattern of hand joint involvement (characterized by clustering primarily by row and symmetric joint involvement, rather than clustering by ray) was found to be almost identical between the men and the women. This study confirms the existence of a polyarticular subset of OA among men that has characteristics similar to those of the variant observed among women. The data suggest that the genetic or metabolic influences underlying this particular variant of OA acts similarly in both sexes.


Diabetic kidney disease: a report from an ADA Consensus Conference.
The incidence and prevalence of diabetes mellitus have grown significantly throughout the world, due primarily to the increase in type 2 diabetes. This overall increase in the number of people with diabetes has had a major impact on development of diabetic kidney disease (DKD), one of the most frequent complications of both types of diabetes. DKD is the leading cause of end-stage renal disease (ESRD), accounting for approximately 50% of cases in the developed world. Although incidence rates for ESRD attributable to DKD have recently stabilized, these rates continue to rise in high-risk groups such as middle-aged African Americans, Native Americans, and Hispanics. The costs of care for people with DKD are extraordinarily high. In the Medicare population alone, DKD-related expenditures among this mostly older group were nearly $25 billion in 2011. Due to the high human and societal costs, the Consensus Conference on Chronic Kidney Disease and Diabetes was convened by the American Diabetes Association in collaboration with the American Society of Nephrology and the National Kidney Foundation to appraise issues regarding patient management, highlighting current practices and new directions. Major topic areas in DKD included 1) identification and monitoring, 2) cardiovascular disease and management of dyslipidemia, 3) hypertension and use of renin-angiotensin-aldosterone system blockade and mineralocorticoid receptor blockade, 4) glycemia measurement, hypoglycemia, and drug therapies, 5) nutrition and general care in advanced-stage chronic kidney disease, 6) children and adolescents, and 7) multidisciplinary approaches and medical home models for health care delivery. This current state summary and research recommendations are designed to guide advances in care and the generation of new knowledge that will meaningfully improve life for people with DKD.


Muscular dystrophy--reason for optimism?
Characterization of the mechanisms underlying various types of muscular dystrophy has been an outstanding triumph of molecular biology. Increasing clarification of the aberrant cellular processes responsible for these conditions may ultimately permit the development of effective means for molecular intervention, allowing correction of the abnormal cellular physiology that results in the dystrophic phenotype.


Gremlin Induces Ocular Hypertension in Mice Through Smad3-Dependent Signaling.
Transforming growth factor-β2 induces extracellular matrix (ECM) remodeling, which likely contributes to the defective function of the trabecular meshwork (TM) leading to glaucomatous ocular hypertension. Bone morphogenetic proteins (BMPs) inhibit these profibrotic effects of TGFβ2. The BMP antagonist gremlin is elevated in glaucomatous TM cells and increases IOP in an ex vivo perfusion culture model. The purpose of this study was to determine whether gremlin regulates ECM proteins in the TM, signals through the Smad3-dependent pathway, and induces ocular hypertension in mice. Ad5.Gremlin or Ad5.TGFβ2 was injected intravitreally into one eye of each mouse. Intraocular pressure measurements were taken using a TonoLab tonometer. Gremlin, TGFβ2, fibronectin (FN), and collagen-1 (Col-1) expression in the TM was determined by immunofluorescence, Western immunoblot, and quantitative (q)PCR analyses. Ad5.Gremlin or Ad5.TGFβ2 each caused significant IOP elevation in mice. Immunofluorescence and Western blot analysis demonstrated that gremlin and TGFβ2 reciprocally increased the expression of each other, and both increased FN expression in the TM and surrounding tissues. Ad5.Gremlin elevated IOP and increased Fn and Col-1 gene expression in the TM of Smad3 wild-type (WT) mice, but had no effect in Smad3 HET or Smad3 KO mice. Our results demonstrate that intravitreal injections of either Ad5.Gremlin or Ad5.TGFβ2 elevate IOP and upregulate the ECM protein FN in the TM of mice. These data show that gremlin signals through the Smad3-dependent pathway in the TM to elevate IOP. We determined for the first time gremlin's role in inducing ocular hypertension in an in vivo model system.


Substance abuse treatment need among older adults in 2020: the impact of the aging baby-boom cohort.
There is concern that as the baby boom population ages in the US, there will be a substantial increase in the number of older adults needing treatment for substance abuse problems. To address this concern, projections of future treatment need for older adults (defined as age 50 and older) were made. Using data from the National Household Survey on Drug Abuse, regression models including predictors of treatment need in 2000 and 2001 were developed. Treatment need was defined as having a DSM-IV alcohol or illicit drug use disorder in the past year. Regression parameters from these models were applied to the projected 2020 population to obtain estimates of the number of older adults needing treatment in 2020. The number of older adults in need of substance abuse treatment is estimated to increase from 1.7 million in 2000 and 2001 to 4.4 million in 2020. This is due to a 50 percent increase in the number of older adults and a 70 percent increase in the rate of treatment need among older adults. The aging baby boom cohort will place increasing demands on the substance abuse treatment system in the next two decades, requiring a shift in focus to address the special needs of an older population of substance abusers. There is also a need to develop improved tools for measuring substance use and abuse among older adults.


Bacopa monnieri extract enhances learning-dependent hippocampal long-term synaptic potentiation.
Bacopa monnieri has been used in Ayurvedic medicine as a memory enhancer for a long time; however, its direct effect on synaptic plasticity has not been investigated. To the best of our knowledge, this study is the first to report the effect of B. monnieri on long-term synaptic potentiation in acute hippocampal slices. Adult male Wistar rats were orally administered either sterile water or the ethanolic extract of B. monnieri for 60 days. The extracellular recording was performed to measure the field excitatory postsynaptic potential in the acute hippocampal slices of these rats. Our results showed that B. monnieri extract significantly increased long-term potentiation magnitude compared with the control group, whereas there was no change in basal synaptic transmission. The data support the beneficial mnemonic effect of B. monnieri, and suggest that this effect might be because of the increase of learning-associated synaptic machinery, resulting in the long-term potentiation enhancement and strengthening of hippocampal synapses, which plays a critical role in learning and memory formation.


FxkR provides the missing link in the fixL-fixK signal transduction cascade in Rhizobium etli CFN42.
Transcriptional control of the fixK gene in Rhizobium etli and R. leguminosarum bv. viciae is governed by a two-component signal transduction system that diverts from the conventional FixL-FixJ cascade that occurs in model rhizobia. Although a fixL gene, encoding a hybrid histidine kinase (hFixL), is present in R. etli, no fixJ, the cognate response regulator, has been identified. In this work, we present evidence that the pRet42f-located open reading frame RHE_PF00530 (fxkR) encodes a novel response regulator indispensable for fixKf activation under microaerobic growth. Moreover, results from complementation assays demonstrate that the activation of fixKf expression requires the presence of both hFixL and FxkR, and that the fxkR ortholog from R. leguminosarum bv. viciae is able to substitute for FxkR transcriptional control in R. etli. In addition, in these two organisms, hFixL- and FxkR-related proteins were identified in other bacteria, located in close proximity to a fixK-related gene. Using reporter fusions, site-directed mutagenesis, and electrophoretic mobility shift assays, we identified the FxkR binding site upstream from the transcriptional start site of fixKf. Similar to our previous observations for fixL and fixKf mutants, a null mutation in fxkR does not affect the symbiotic effectiveness of the strain. Thus, our findings reveal that FxkR is the long-standing missing key regulator that allows the transduction of the microaerobic signal for the activation of the FixKf regulon.


The Impact of Biofilm Formation on the Persistence of Candidemia.
This study aimed to determine the predictors of persistent candidemia and examine the impact of biofilm formation by Candida isolates in adult patients with candidemia. Of the adult patients with candidemia in Kaohsiung Chang Gung Memorial Hospital between January 2007 and December 2012, 68 case patients with persistent candidemia (repeated candidemia after a 3-day systemic antifungal therapy) and 68 control patients with non-persistent candidemia (Candida clearance from the bloodstream after a 3-day systemic antifungal therapy) were included based on propensity score matching and matching for the Candida species isolated. Biofilm formation by the Candida species was assessed in vitro using standard biomass assays. Presence of central venous catheters (CVCs) at diagnosis (adjusted odd ratio [AOR], 3.77; 95% confidence interval [CI], 1.09-13.00, p = 0.04), infection with higher biofilm forming strains of Candida species (AOR, 8.03; 95% CI, 2.50-25.81; p < 0.01), and receipt of suboptimal fluconazole doses as initial therapy (AOR, 5.54; 95% CI, 1.53-20.10; p < 0.01) were independently associated with persistent candidemia. Biofilm formation by Candida albicans, C. tropicalis, and C. glabrata strains was significantly higher in the case patients than in the controls. There were no significant differences in the overall mortality and duration of hospitalization between the two groups. Our data suggest that, other than presence of retained CVCs and use of suboptimal doses of fluconazole, biofilm formation was highly associated with development of persistent candidemia.


α-Ketoglutarate regulates acid-base balance through an intrarenal paracrine mechanism.
Paracrine communication between different parts of the renal tubule is increasingly recognized as an important determinant of renal function. Previous studies have shown that changes in dietary acid-base load can reverse the direction of apical α-ketoglutarate (αKG) transport in the proximal tubule and Henle's loop from reabsorption (acid load) to secretion (base load). Here we show that the resulting changes in the luminal concentrations of αKG are sensed by the αKG receptor OXGR1 expressed in the type B and non-A-non-B intercalated cells of the connecting tubule (CNT) and the cortical collecting duct (CCD). The addition of 1 mM αKG to the tubular lumen strongly stimulated Cl(-)-dependent HCO(3)(-) secretion and electroneutral transepithelial NaCl reabsorption in microperfused CCDs of wild-type mice but not Oxgr1(-/-) mice. Analysis of alkali-loaded mice revealed a significantly reduced ability of Oxgr1(-/-) mice to maintain acid-base balance. Collectively, these results demonstrate that OXGR1 is involved in the adaptive regulation of HCO(3)(-) secretion and NaCl reabsorption in the CNT/CCD under acid-base stress and establish αKG as a paracrine mediator involved in the functional coordination of the proximal and the distal parts of the renal tubule.


Exercise loading history and femoral neck strength in a sideways fall: A three-dimensional finite element modeling study.
Over 90% of hip fractures are caused by falls. Due to a fall-induced impact on the greater trochanter, the posterior part of the thin superolateral cortex of the femoral neck is known to experience the highest stress, making it a fracture-prone region. Cortical geometry of the proximal femur, in turn, reflects a mechanically appropriate form with respect to habitual exercise loading. In this finite element (FE) modeling study, we investigated whether specific exercise loading history is associated with femoral neck structural strength and estimated fall-induced stresses along the femoral neck. One hundred and eleven three-dimensional (3D) proximal femur FE models for a sideways falling situation were constructed from magnetic resonance (MR) images of 91 female athletes (aged 24.7±6.1years, >8years competitive career) and 20 non-competitive habitually active women (aged 23.7±3.8years) that served as a control group. The athletes were divided into five distinct groups based on the typical loading pattern of their sports: high-impact (H-I: triple-jumpers and high-jumpers), odd-impact (O-I: soccer and squash players), high-magnitude (H-M: power-lifters), repetitive-impact (R-I: endurance runners), and repetitive non-impact (R-NI: swimmers). The von Mises stresses obtained from the FE models were used to estimate mean fall-induced stresses in eight anatomical octants of the cortical bone cross-sections at the proximal, middle, and distal sites along the femoral neck axis. Significantly (p<0.05) lower stresses compared to the control group were observed: the H-I group - in the superoposterior (10%) and posterior (19%) octants at the middle site, and in the superoposterior (13%) and posterior (22%) octants at the distal site; the O-I group - in the superior (16%), superoposterior (16%), and posterior (12%) octants at the middle site, and in the superoposterior (14%) octant at the distal site; the H-M group - in the superior (13%) and superoposterior (15%) octants at the middle site, and a trend (p=0.07, 9%) in the superoposterior octant at the distal site; the R-I group - in the superior (14%), superoposterior (23%) and posterior (22%) octants at the middle site, and in the superoposterior (19%) and posterior (20%) octants at the distal site. The R-NI group did not differ significantly from the control group. These results suggest that exercise loading history comprising various impacts in particular is associated with a stronger femoral neck in a falling situation and may have potential to reduce hip fragility.


The fine structure of striated muscle; a comparison of insect flight muscle with vertebrate and invertebrate skeletal muscle.
The available evidence from phase contrast, polarization optical, and electron microscopic studies on vertebrate skeletal muscle, insect skeletal muscle, and dipteran flight muscle is interpreted as favoring the following general structure of striated muscle. A continuous array of filaments (actin) runs through all bands of the sarcomere. These are linked by an axially periodic system of transverse filamentous bridges. Myosin (and probably other substances) are localized in the A bands. The system of transverse bridges compensates the birefringence of actin and is thus responsible for the isotropy of the I band. Myosin is responsible for the birefringence of the A bands. On strong contraction, A band material migrates to the Z bands to form contraction bands. It is not yet certain whether this migration involves myosin or another A band component.


A freeze-dried fibrin disc as a biodegradable drug release matrix.
A fibrin clot loaded with soluble tetracycline (TET) was prepared and lyophilized to make discs of a size and shape to use as a drug delivery matrix. On subcutaneous implantation of these discs in mice, they were found to have degraded in 15 days as evidenced by gross and histological examination. The in vitro discharge kinetics of tetracycline from the disc into phosphate buffered saline (PBS) and human serum were compared. It was observed that the release rate of tetracycline from the matrix into serum remained steady from day 1 to day 12, maintaining sufficient concentration that may be required to control microbial growth in the medium. Two different concentrations of fibrinogen were used to fabricate discs denoted as FG200 and FG100, and in both cases the retention rate was comparable when the study medium was serum. In contrast, when suspended in PBS instead of serum, the delivery of the drug into the medium was found to be high for up to the 3rd day when a sharp decline in discharge was observed. The fibrinogen used is a factor that determines not only the longevity of discharge but also fibrinolysis. The degradation of the disc in vitro was visible when the discs were suspended in the buffer, and correspondingly fibrin degradation product (FDP) measured in the medium using an antibody-based assay system was high. Fibrin disc is haemostatic and biodegradable in vivo, and in vitro release of a small molecule at a controlled rate is demonstrated here. Hence, it may be a suitable candidate as a drug delivery implant for short-term use.


Effort-reward imbalance in the school setting: associations with somatic pain and self-rated health.
According to the workplace theory of effort-reward imbalance (ERI), individuals who perceive a lack of reciprocity between their effort spent at work and the rewards received in turn are at an increased risk of stress-related ill-health. It is also assumed that being overcommitted to work is linked to an increased risk of stress-related ill-health. This study applies the effort-reward imbalance model to the school setting. It aims to analyse the associations that effort-reward imbalance and overcommitment share with somatic pain and self-rated health among adolescents. Data are from the School Stress and Support Study (TriSSS), involving students in grades 8 and 9 (ages 14-16 years) in two schools in Stockholm, Sweden, during 2010 (n=403). Information on effort-reward imbalance and health outcomes was gathered from self-report questionnaires. An adjusted short version of ERI was used. Factor analysis showed that extrinsic effort, reward and overcommitment constitute three distinct dimensions. The designed measures demonstrated sound psychometric properties both for the full sample and for subgroups. Ordered logistic regressions were conducted. The analyses showed that low reward and higher overcommitment were associated with greater somatic pain and poorer self-rated health. Furthermore, effort-reward imbalance was linked with an elevated risk of somatic pain and poorer self-rated health. Students are more likely to experience stress-related ill-health when they perceive an imbalance between their effort and rewards. In addition, high overcommitment is associated with an increased risk of ill-health among students.


A Gold(III) Pincer Ligand Scaffold for the Synthesis of Binuclear and Bioconjugated Complexes: Synthesis and Anticancer Potential.
Cyclometalated (C^N^C)AuIII complexes bearing functionalized N-heterocyclic carbene (NHC) ligands provide a high-yielding, modular route to bioconjugated and binuclear complexes. This methodology has been applied to the synthesis of bioconjugated complexes presenting biotin and 17α-ethynylestradiol vectors, as well as to the synthesis of bimetallic AuIII /AuI complexes. The in vitro antiproliferative activities of these compounds against various cancer cells lines depend on the linker length, with the longer linker being the most potent. The estradiol conjugate AuC6 Estra proved to be more toxic against the estrogen receptor positive (ER+) cancer cells than against the ER- cancer cells and non-cancer cells. The bimetallic complex AuC6 Au was more selective for breast cancer cells with respect to a healthy cell standard than the monometallic complex AuNHC. The metal uptake study on cells expressing or not biotin and estrogen receptors revealed an improved and targeted delivery of gold for both the bioconjugated complexes AuC6 Biot and AuC6 Estra compared to the non-vectorised analogue AuNHC. The investigations of the interaction of the bioconjugates and bimetallic complexes with human telomeric G-quadruplex DNA using FRET-melting techniques revealed a reduced ability to stabilize this DNA structure with respect to the non-vectorised analogue AuNHC.


The MetaCyc database of metabolic pathways and enzymes and the BioCyc collection of pathway/genome databases.
The MetaCyc database (MetaCyc.org) is a comprehensive and freely accessible resource for metabolic pathways and enzymes from all domains of life. The pathways in MetaCyc are experimentally determined, small-molecule metabolic pathways and are curated from the primary scientific literature. With more than 1400 pathways, MetaCyc is the largest collection of metabolic pathways currently available. Pathways reactions are linked to one or more well-characterized enzymes, and both pathways and enzymes are annotated with reviews, evidence codes, and literature citations. BioCyc (BioCyc.org) is a collection of more than 500 organism-specific Pathway/Genome Databases (PGDBs). Each BioCyc PGDB contains the full genome and predicted metabolic network of one organism. The network, which is predicted by the Pathway Tools software using MetaCyc as a reference, consists of metabolites, enzymes, reactions and metabolic pathways. BioCyc PGDBs also contain additional features, such as predicted operons, transport systems, and pathway hole-fillers. The BioCyc Web site offers several tools for the analysis of the PGDBs, including Omics Viewers that enable visualization of omics datasets on two different genome-scale diagrams and tools for comparative analysis. The BioCyc PGDBs generated by SRI are offered for adoption by any party interested in curation of metabolic, regulatory, and genome-related information about an organism.


Reverse current pulse method to restore uniform concentration profiles in ion-selective membranes. 1. Galvanostatic pulse methods with decreased cycle time.
The applications of ion-selective electrodes (ISEs) have been broadened through the introduction of galvanostatic current pulse methods in potentiometric analysis. An important requirement in these applications is the restoration of the uniform equilibrium concentration profiles in the ISE membrane between each measurement. The simplest restoration method is zero-current relaxation, in which the membrane relaxes under open-circuit conditions in a diffusion-controlled process. This paper presents a novel restoration method using a reverse current pulse. An analytic model for this restoration method is derived to predict the concentration profiles inside ISE membranes following galvanostatic current pulses. This model allows the calculation of the voltage transients as the membrane voltage relaxes back toward its zero-current equilibrium value. The predicted concentration profiles and voltage transients are confirmed using spectroelectrochemical microscopy (SpECM). The reverse current restoration method described in this paper reduces the voltage drift and voltage error by 10-100 times compared to the zero-current restoration method. Therefore, this new method provides faster and more reproducible voltage measurements in most chronopotentiometric ISE applications, such as improving the detection limit and determining concentrations and diffusion coefficients of membrane species. One limitation of the reverse current restoration method is that it cannot be used in a few applications that require background electrolyte loaded membranes without excess of lipophilic cation exchanger.


More complex, less complicated? Explicit solvation of hydroxyl groups for the analysis of VCD spectra.
Solute-solvent interactions and in particular hydrogen bonding can significantly influence the appearance of vibrational spectra due to band shifts, intensity changes and band broadening. In VCD spectroscopy, solvation may also lead to sign changes and thus an overall drastic change in the spectral pattern. As the VCD spectral analysis relies heavily on the comparison with computed spectra, such solvent effects have to be accounted for in the calculations. For simple model systems with one stereocenter, we have previously shown for carboxylic acids and hydroxyl groups that considering solvation explicitly improves the match substantially. In the present study we evaluate if explicit solvation is always necessary and if larger, more complex molecules featuring several stereocenters show the same susceptibility to H-bonding induced spectral changes as the previously investigated model systems. We analyse the spectra of the diastereomeric pairs menthol/neomenthol and borneol/isoborneol and study both experimentally and computationally the influence of hydrogen bonding to dimethylsulfoxide-d6 (DMSO-d6) and acetonitrile-d3 (ACN-d3) on their VCD spectral signatures. Further chiral alcohols with tertiary hydroxyl group (terpinen-4-ol and cedrol) and more complex structures with multiple stereocenters (cholesterol) are investigated to show that solvent effects on the spectra become less pronounced. We related this to the increasing number of vibrational bands that are insensitive to solvation and thus overlap with actually affected modes. As a consequence, the analysis of the spectra does not require consideration of explicit solvation and in this respect becomes less complicated.


Willingness to pay as a measure of health benefits.
In this paper, we discuss the use of cost-benefit analysis (CBA) for evaluating new healthcare interventions, present the theoretical basis for the use of willingness to pay as a method for valuing benefits in a CBA and describe how to obtain willingness-to-pay (WTP) measures of health benefits and how to use these values in a CBA. We review selected economic studies on consumer demand and consumer surplus and studies presenting WTP estimates for healthcare interventions. The theoretical foundations of willingness to pay as a measure of commodity value are rooted in consumer demand theory. The area under the fixed income consumer demand curve represents the consumer's maximum willingness to pay for the commodity. We identify 3 types of potential benefits from a new healthcare intervention, namely patient benefits, option value and altruistic value, and suggest WTP questions for valuing different combinations of these benefits. We demonstrate how responses to these questions can be adjusted for income effects and incorporated into economic evaluations. We suggest that the lack of popularity of CBAs in the health area is related to the perceived difficulty in valuing health benefits as well as concern over how CBA incorporates the distribution of income. We show that health benefits can be valued using simple survey techniques and that these values can be adjusted to any desired income distribution.


Graduate student's guide to necessary skills for nonacademic conservation careers.
Graduate education programs in conservation science generally focus on disciplinary training and discipline-specific research skills. However, nonacademic conservation professionals often require an additional suite of skills. This discrepancy between academic training and professional needs can make it difficult for graduate students to identify the skills and experiences that will best prepare them for the conservation job market. We analyzed job advertisements for conservation-science positions and interviewed conservation professionals with experience hiring early-career conservation scientists to determine what skills employers of conservation professionals seek; whether the relative importance of skills varies by job sector (government, nonprofit, and private); and how graduate students interested in careers in conservation science might signal competency in key skills to potential employers. In job advertisements, disciplinary, interpersonal, and project-management skills were in the top 5 skills mentioned across all job sectors. Employers' needs for additional skills, like program leadership, conflict resolution and negotiation, and technical and information technology skills, varied across sectors. Our interview results demonstrated that some skills are best signaled to employers via experiences obtained outside thesis or dissertation work. Our findings suggest that graduate students who wish to be competitive in the conservation job market can benefit by gaining skills identified as important to the job sector in which they hope to work and should not necessarily expect to be competent in these skills simply by completing their chosen degree path.


[Forensic assessments from the Netherlands Institute of Forensic Psychiatry and Psychology in retrospect; applications of genetics and neuroscience, in 2000 and 2009].
Developments in neurosciences and genetics are relevant for forensic psychiatry. To find out whether and how genetic and neuroscientific applications are being used in forensic psychiatric assessments, and, if they are, to estimate to what extent new applications will fit in with these uses. We analysed 60 forensic psychiatric assessments from the Netherlands Institute of Forensic Psychiatry and Psychology, Pieter Baan Center, and 30 non-clinical assessments from 2000 and 2009. We found that (behavioral) genetic, neurological and neuropsychological applications played only a modest role in forensic psychiatric assessment and they represent different phases of the implementation process. Neuropsychological assessment already occupied a position of some importance, but needed to be better integrated. Applications from neurology were still being developed. Clinical genetic assessment was being used occasionally in order to diagnose a genetic syndrome with behavioral consequences. If further validated information becomes available in the future, it should be possible to integrate new research methods more fully into current clinical practice.


Dynamics and Diversity of Microbial Contamination in Poultry Bedding Materials Containing Parts of Medicinal Plants.
Microorganisms thriving in poultry bedding materials during their exploitation are involved in the development of several diseases and disfunctions of animals. They can also contaminate food products and pose risks to the environment and human health. This study provides an analysis of dynamics and diversity in microbiological contamination observed during the exploitation of poultry bedding materials containing parts of medicinal plants: Satureja hortensis, Origanum vulgare, Melissa officinalis, Salvia officinalis, and Thymus vulgaris, compared with standard types of beddings: straw chaff and straw pellets. The research was carried out in two 42-day experimental cycles involving in total 2400 broiler chickens. Each week, the total count of mesophilic bacteria, fungi and yeasts, the presumptive presence and count of Staphylococcus sp., Escherichia sp., Listeria sp., Salmonella sp., and Candida sp. were determined by culturing on selective media, along with pH and moisture measurements. After 35 days of the experiment, a reduction of the total count of mesophilic bacteria above 1 log compared to the control (11.86 vs. 13.02 log CFU/g) was observed. As the count of yeasts decreased after 21 days, an increase in the total count of bacteria was reported, which indicates a strong competition between microorganisms. The results improve our understanding of the temporal effects of using materials containing parts of medicinal plants on the microbial contamination in poultry litter.


Effects of endothelin-1 on endothelial cells in the porcine coronary artery.
The effects of endothelin-1 (ET-1) on endothelial cells from the porcine coronary artery were investigated using cascade bioassay and radioimmunoassay methods. For comparison, endothelial cells from the rabbit abdominal aorta were also used. Freshly isolated endothelial cells were perfused with Krebs' solution. Liberation of vasodilatory substances was detected under bioassay conditions by measuring the relaxing activity of the perfusate on endothelium-denuded strips of the porcine coronary artery contracted with ET-1. The perfusate in the absence of ET-1 slightly inhibited the contraction, suggesting that endothelial cells spontaneously release vasorelaxing substances. After application of ET-1, the perfusate-induced relaxation drastically increased. Oxyhemoglobin completely abolished the relaxation induced by perfusate from endothelial cells of the rabbit abdominal aorta but only marginally attenuated the relaxation induced by perfusate from endothelial cells of the porcine coronary artery. In contrast, indomethacin significantly attenuated the relaxation induced by perfusate from the endothelial cells of the porcine coronary artery. In endothelium-intact strips of the porcine coronary artery, ET-1 significantly increased the concentration of 6-ketoprostaglandin F1 alpha but did not modify the cellular concentration of either cAMP or cGMP. It is concluded that ET-1 augments the release of vasorelaxing factors from endothelial cells both in the porcine coronary artery and in the rabbit abdominal aorta and that the major vasorelaxing substance derived from endothelial cells may be different in these two blood vessels. Prostaglandin I2 in the former and nitric oxide in the latter are suggested as possible candidates. The vasorelaxation induced by prostaglandin I2 may not be mediated by cAMP-dependent mechanisms in the porcine coronary artery.


Nutritional requirements and growth characteristics of a biosurfactant-producingRhodococcus bacterium.
The nutritional requirements and growth characteristics of a biosurfactant-producingRhodococcus bacterium isolated from Kuwaiti soil were determined. Maximum cell yields (6.6 g/l) and biosurfactant production were achieved with a medium containing 2% (v/v)n-paraffin as a carbon and energy source, 0.2% lactose broth, optimal concentrations of nitrogen (nitrate), phosphorus, iron, magnesium and sodium sources, and minimal concentrations of potassium and trace element sources. The optimal pH was 6.8 for surfactant production and optimal temperature was 37°C. The biosurfactant produced after 16 to 33 h growth in a 7 I fermenter decreased both surface tension and interfacial tension of culture broth to below 27 and 1.8 mN/m, respectively, and was effective at critical micelle dilutions of 10(-3). Data on biosurfactant biosynthesis suggest that the product is produced as a primary metabolite and, therefore, could be produced effectively under continuous fermentation conditions.


Selective downregulation of the high molecular weight form of adiponectin in hyperinsulinemia and in type 2 diabetes: differential regulation from nondiabetic subjects.
Adiponectin is an adipocyte-specific secretory protein found in circulation in several different forms and is present at significantly lower levels in the plasma of diabetic patients compared with that of insulin-sensitive individuals. We wanted to test whether insulin per se is a contributing factor toward lower plasma adiponectin concentrations and, if so, whether the splanchnic bed contributes to this phenomenon. We sampled femoral artery and hepatic venous samples and measured the high-molecular weight (HMW) and low-molecular weight (LMW) fractions of adiponectin in 11 type 2 diabetic and 7 nondiabetic subjects matched for age, sex, and BMI during basal conditions and during a hyperglycemic (approximately 9.5 mmol/l) hyperinsulinemic (approximately 700 pmol/l) clamp. Under these conditions, total arterial adiponectin, HMW, and the ratio of HMW to total adiponectin all were lower (P < 0.01) in the diabetic versus nondiabetic subjects, whereas the LMW form did not significantly differ. Under hyperinsulinemic conditions, total adiponectin levels dropped, primarily due to a reduction of the HMW form, whereas LMW forms were not significantly affected. HMW adiponectin and the ratio of HMW to total adiponectin are lower in individuals with diabetes than in nondiabetic subjects. We conclude that HMW adiponectin is downregulated in hyperinsulinemia and type 2 diabetes.


Transcriptional Changes in Dorsal Spinal Cord Persist after Surgical Incision Despite Preemptive Analgesia with Peripheral Resiniferatoxin.
Peripheral nociceptors expressing the ion channel transient receptor potential cation channel, subfamily V, member 1, play an important role in mediating postoperative pain. Signaling from these nociceptors in the peri- and postoperative period can lead to plastic changes in the spinal cord and, when controlled, can yield analgesia. The transcriptomic changes in the dorsal spinal cord after surgery, and potential coupling to transient receptor potential cation channel, subfamily V, member 1-positive nociceptor signaling, remain poorly studied. Resiniferatoxin was injected subcutaneously into rat hind paw several minutes before surgical incision to inactivate transient receptor potential cation channel, subfamily V, member 1-positive nerve terminals. The effects of resiniferatoxin on postincisional measures of pain were assessed through postoperative day 10 (n = 51). Transcriptomic changes in the dorsal spinal cord, with and without peripheral transient receptor potential cation channel, subfamily V, member 1-positive nerve terminal inactivation, were assessed by RNA sequencing (n = 22). Peripherally administered resiniferatoxin increased thermal withdrawal latency by at least twofold through postoperative day 4, increased mechanical withdrawal threshold by at least sevenfold through postoperative day 2, and decreased guarding score by 90% relative to vehicle control (P < 0.05). Surgical incision induced 70 genes in the dorsal horn, and these changes were specific to the ipsilateral dorsal horn. Gene induction with surgical incision persisted despite robust analgesia from resiniferatoxin pretreatment. Many of the genes induced were related to microglial activation, such as Cd11b and Iba1. A single subcutaneous injection of resiniferatoxin before incision attenuated both evoked and nonevoked measures of postoperative pain. Surgical incision induced transcriptomic changes in the dorsal horn that persisted despite analgesia with resiniferatoxin, suggesting that postsurgical pain signals can be blocked without preventing transcription changes in the dorsal horn.


The effect of early-stage public health policies in the transmission of COVID-19 for South American countries.
The analysis of transmission dynamics is crucial to determine whether mitigation or suppression measures reduce the spread of coronavirus disease 2019 (COVID-19). This study sought to estimate the basic (R0 ) and time-varying (Rt ) reproduction number of COVID-19 and contrast the public health measures for ten South American countries. Data was obtained from the European Centre for Disease Prevention and Control. Country-specific R0 values during the first two weeks of the outbreak and Rt values after 90 days were estimated. Countries used a combination of isolation, physical distancing, quarantine, and community-wide containment measures to staunch the spread of COVID-19 at different points in time. R0 ranged from 1.52 (95% confidence interval: 1.13-1.99) in Venezuela to 3.83 (3.04-4.75) in Chile, whereas Rt after 90 days ranged from 0.71 (95% credible interval: 0.39-1.05) in Uruguay to 1.20 (1.19-1.20) in Brazil. Different R0 and Rt values may be related to the testing capacity of each country. R0 in the early phase of the outbreak varied across the South American countries. The public health measures adopted in the initial period of the pandemic appear to have reduced Rt over time in each country, albeit to different levels.


Genome-Scale Protein Microarray Comparison of Human Antibody Responses in Plasmodium vivax Relapse and Reinfection.
Large scale antibody responses in Plasmodium vivax malaria remains unexplored in the endemic setting. Protein microarray analysis of asexual-stage P. vivax was used to identify antigens recognized in sera from residents of hypoendemic Peruvian Amazon. Over 24 months, of 106 participants, 91 had two symptomatic P. vivax malaria episodes, 11 had three episodes, 3 had four episodes, and 1 had five episodes. Plasmodium vivax relapse was distinguished from reinfection by a merozoite surface protein-3α restriction fragment length polymorphism polymerase chain reaction (MSP3α PCR-RFLP) assay. Notably, P. vivax reinfection subjects did not have higher reactivity to the entire set of recognized P. vivax blood-stage antigens than relapse subjects, regardless of the number of malaria episodes. The most highly recognized P. vivax proteins were MSP 4, 7, 8, and 10 (PVX_003775, PVX_082650, PVX_097625, and PVX_114145); sexual-stage antigen s16 (PVX_000930); early transcribed membrane protein (PVX_090230); tryptophan-rich antigen (Pv-fam-a) (PVX_092995); apical merozoite antigen 1 (PVX_092275); and proteins of unknown function (PVX_081830, PVX_117680, PVX_118705, PVX_121935, PVX_097730, PVX_110935, PVX_115450, and PVX_082475). Genes encoding reactive proteins exhibited a significant enrichment of non-synonymous nucleotide variation, an observation suggesting immune selection. These data identify candidates for seroepidemiological tools to support malaria elimination efforts in P. vivax-endemic regions.


Calcium, dairy products, and risk of prostate cancer in a prospective cohort of United States men.
Intake of calcium and/or dairy products has been associated with increased risk of prostate cancer in some epidemiological studies. One potential biological mechanism is that high calcium intake down-regulates 1,25 dihydroxy vitamin D(3), which may increase cell proliferation in the prostate. We examined the association between calcium, dairy intake, and prostate cancer incidence in the Cancer Prevention Study II Nutrition Cohort, a prospective cohort of elderly United States adults. Participants in the study completed a detailed questionnaire on diet, medical history, and lifestyle at enrollment in 1992-1993. After excluding men with a history of cancer or incomplete dietary information, 65,321 men remained for analysis. During follow-up through August 31, 1999, we documented 3811 cases of incident prostate cancer. Multivariate-adjusted rate ratios (RRs) were calculated using Cox proportional hazards models. Total calcium intake (from diet and supplements) was associated with modestly increased risk of prostate cancer [RR = 1.2, 95% confidence interval (CI) = 1.0-1.6 for >or=2000 versus <700 mg/day, P trend = 0.02). High dietary calcium intake (>or=2000 versus <700 mg/day) was also associated with increased risk of prostate cancer (RR = 1.6, 95% CI = 1.1-2.3, P trend = 0.10), although moderate levels of dietary calcium were not associated with increased risk. Dairy intake was not associated with prostate cancer risk. The association between prostate cancer and total calcium intake was strongest for men who reported not having prostate-specific antigen testing before 1992 (RR = 1.5, 95% CI = 1.1-2.0, P trend < 0.01 for >or= 2000 mg/day of total calcium; RR = 2.1, 95% CI = 1.3-3.4 >or=2000 mg/day of dietary calcium, P trend = 0.04). Our results support the hypothesis that very high calcium intake, above the recommended intake for men, may modestly increase risk of prostate cancer.


Endoanal ultrasound compared to anorectal manometry for the evaluation of fecal incontinence: a study of the effect these tests have on clinical outcome.
Tests for evaluating incontinence include endoanal ultrasound (EUS) and anorectal manometry. We hypothesized that EUS would be superior to anorectal manometry in identifying the subset of patients with surgically correctable sphincter defects leading to an improvement in clinical outcome in these patients. The purpose of this study was to compare these 2 techniques to determine which is more predictive of outcome for fecal incontinence. Thirty-five unselected patients with fecal incontinence were prospectively studied with EUS and anorectal manometry to evaluate the internal anal sphincter (IAS) and external anal sphincter (EAS). EUS was performed with Olympus GFUM20 echoendoscope and a hypoechoic defect in the EAS or IAS was considered a positive test. Anorectal manometry was performed with a standard water-perfused catheter system. A peak voluntary squeeze pressure of < 60 mm Hg in women and 120 mm Hg in men was considered a positive test. All patients were administered the Cleveland Clinic Continence Grading Scale at baseline and at follow-up. Improvement in fecal control was defined as a 25% or greater decrease in continence score. EUS versus manometry were compared with subsequent surgical treatment and outcome. P-values were calculated using Fisher's exact test. Patients (n = 32; 31 females) were followed for a mean 25 months (range 13-46). Sixteen patients had improved symptoms (50%). There was no correlation between EUS or anorectal manometry sphincter findings and outcome. Seven of 14 (50%) patients who subsequently underwent surgery versus 9 of 18 (50%) without surgery improved (P = .578). In long-term follow-up, approximately half of patients improve regardless of the results of EUS or anorectal manometry, or whether surgery is performed.


Blockade of LAG3 enhances responses of tumor-infiltrating T cells in mismatch repair-proficient liver metastases of colorectal cancer.
Purpose: Liver metastasis develops in >50% of patients with colorectal cancer (CRC), and is a leading cause of CRC-related mortality. We aimed to identify which inhibitory immune checkpoint pathways can be targeted to enhance functionality of intra-tumoral T-cells in mismatch repair-proficient liver metastases of colorectal cancer (LM-CRC). Methodology: Intra-tumoral expression of multiple inhibitory molecules was compared among mismatch repair-proficient LM-CRC, peritoneal metastases of colorectal cancer (PM-CRC) and primary CRC. Expression of inhibitory molecules was also analyzed on leukocytes isolated from paired resected metastatic liver tumors, tumor-free liver tissues, and blood of patients with mismatch repair-proficient LM-CRC. The effects of blocking inhibitory pathways on tumor-infiltrating T-cell responses were studied in ex vivo functional assays. Results: Mismatch repair-proficient LM-CRC showed higher expression of inhibitory receptors on intra-tumoral T-cells and contained higher proportions of CD8+ T-cells, dendritic cells and monocytes than mismatch repair-proficient primary CRC and/or PM-CRC. Inhibitory receptors LAG3, PD-1, TIM3 and CTLA4 were higher expressed on CD8+ T-cells, CD4+ T-helper and/or regulatory T-cells in LM-CRC tumors compared with tumor-free liver and blood. Antibody blockade of LAG3 or PD-L1 increased proliferation and effector cytokine production of intra-tumoral T-cells isolated from LM-CRC in response to both polyclonal and autologous tumor-specific stimulations. Higher LAG3 expression on intra-tumoral CD8+ T-cells associated with longer progression-free survival of LM-CRC patients. Conclusion: Mismatch repair-proficient LM-CRC may be more sensitive to immune checkpoint inhibitors than mismatch repair-proficient primary CRC. Blocking LAG3 enhances tumor-infiltrating T-cell responses of mismatch repair-proficient LM-CRC, and therefore may be a new promising immunotherapeutic target for LM-CRC.


Bone mineral loss in young women with amenorrhoea.
To examine the impact of amenorrhoea on bone mineral density in women of reproductive age. Cross sectional study of 200 amenorrhoeic women compared with normally menstruating controls. Teaching hospital outpatient clinic specialising in reproductive medicine. 200 Women aged 16-40 with a past or current history of amenorrhoea from various causes and of a median duration of three years, and a control group of 57 age matched normal volunteers with no history of menstrual disorder. Bone mineral density in the lumbar spine (L1-L4) as measured by dual energy x ray absorptiometry. The amenorrhoeic group showed a mean reduction in bone mineral density of 15% (95% confidence interval 12% to 18%) as compared with controls (mean bone mineral density 0.89 (SD 0.12) g/cm2 v 1.05 (0.09) g/cm2 in controls). Bone loss was related to the duration of amenorrhoea and the severity of oestrogen deficiency rather than to the underlying diagnosis. Patients with a history of fracture had significantly lower bone density than those without a history of fracture. Ten patients had suffered an apparently atraumatic fracture. Amenorrhoea in young women should be investigated and treated to prevent bone mineral loss. Menopausal women with a past history of amenorrhoea should be considered to be at high risk of osteoporosis.


Beating the weekend trend: increased mortality in older adult traumatic brain injury (TBI) patients admitted on weekends.
Weekend admission is associated with mortality in cardiovascular emergencies and stroke but the effect of weekend admission for trauma is not well defined. We sought to determine whether differences in mortality outcomes existed for older adults with substantial head trauma admitted on a weekday versus over the weekend. Data from the 2006, 2007, and 2008 Nationwide Inpatient Sample were combined and head trauma admissions were isolated. Abbreviated injury scale (AIS) scores were calculated using ICDMAP-90 Software. Individuals aged 65 to 89 y with head AIS equal to 3 or 4 and no other region score <3 were included. Individual Charlson comorbidity scores were calculated and individuals with missing mortality, sex, or insurance data were excluded. Wilcoxon rank sum and Student t-tests compared demographics, length of stay, and total charges for weekday versus weekend admissions. The χ2 tests compared sex and head injury severity. Logistic regression modeled mortality adjusting for age, sex, injury severity, comorbidity, and insurance status. Of the 38,675 patients meeting criteria, 9937 (25.6%) were admitted on weekends. Mean age was similar (78.4 versus 78.4, P = 0.796) but more weekend admissions were female (51.6% versus 50.2%, P = 0.022). Weekend patients demonstrated slightly lower comorbidity (mean Charlson = 1.07 versus 1.14, P < 0.001) and head injury severity (58.3% versus 60.8% AIS = 4, P < 0.001). Median weekend length of stay was shorter (4 versus 5 d, P < 0.001). Weekend and weekday median total charges did not differ ($27,128 versus $27,703, respectively, P = 0.667). Proportional mortality was higher among weekend patients (9.3% versus 8.4%, P = 0.008). After adjustment, weekend patients demonstrated 14% increased odds of mortality (OR 1.14, 95% CI 1.05-1.23). Older adults with substantial head trauma admitted on weekends are less severely injured, carry less comorbidity, and generate similar total charges compared with those admitted on weekdays. However, after accounting for known risk confounders, weekend patients demonstrated 14% greater odds of mortality. Mechanisms behind this disparity must be determined and eliminated.


Registry and health insurance claims data in vascular research and quality improvement.
The expansion of procedures in multidisciplinary vascular medicine has sparked a controversy regarding measures of quality improvement. In addition to primary registries, the use of health insurance claims data is becoming of increasing importance. However, due to the fact that health insurance claims data are not collected for scientific evaluation but rather for reimbursement purposes, meticulous validation is necessary before and during usage in research and quality improvement matters. This review highlights the advantages and disadvantages of such data sources. A recent comprehensive expert opinion panel examined the use of health insurance claims data and other administrative data sources in medicine. Results from several studies concerning the validity of administrative data varied significantly. Validity of these data sources depends on the clinical relevance of the diagnoses considered. The rate of implausible information was 0.04 %, while the validity of the considered diagnoses varied between 80 and 97 % across multiple validation studies. A matching study between health insurance claims data of the third-largest German health insurance provider, DAK-Gesundheit, and a prospective primary registry of the German Society for Vascular Surgery demonstrated a good level of validity regarding the mortality of endovascular and open surgical treatment of abdominal aortic aneurysm in German hospitals. In addition, a large-scale international comparison of administrative data for the same disorder presented important results in treatment reality, which differed from those from earlier randomized controlled trials. The importance of administrative data for research and quality improvement will continue to increase in the future. When discussing the internal and external validity of this data source, one has to distinguish not only between its intended usage (research vs. quality improvement), but also between the included diseases and/or treatment procedures. Linkage between primary registry data and administrative data could be a reasonable solution to some current major issues of validity. .


Aluminum content of human semen: implications for semen quality.
A deterioration of human semen quality has been observed over recent decades. A possible explanation could be an increased exposure to environmental pollutants, including aluminum. Our aim was to measure the aluminum concentration in the semen of 62 patients and to carry out a preliminary evaluation on its impact on specific semen parameters. For each patient, semen analyses were performed according to WHO guidelines. A graphite furnace atomic absorption spectrometry method was used to determine semen aluminum concentration. A cytological analysis using an aluminum-specific fluor, lumogallion, was also performed. The mean aluminum concentration in human semen was 339 μg/L. Patients with oligozoospermia had a statistically higher aluminum concentration than others. No significant difference was observed for other semen parameters. Cytological analysis showed the presence of aluminum in spermatozoa. This study provided unequivocal evidence of high concentrations of aluminum in human semen and suggested possible implications for spermatogenesis and sperm count.


Water-borne fluoride and cortical bone mass: a comparison of two communities.
This study investigated the relationship between cortical bone mass in an older female population and their ingestion of fluoride from community water supplies. The study was conducted among lifelong female residents in Lordsburg (3.5 ppm fluoride) and Deming (0.7 ppm fluoride), NM. A total of 151 postmenopausal women ranging in age from 39 to 87 years took part; 69 were residents of the optimal-fluoride community, while the remaining 82 were residents of the high-fluoride community. Although bivariate analyses showed no difference in cortical bone mass between women in the two communities, with multiple regression analyses, significant predictors of bone mass (p less than 0.05) were weight, years since menopause, current estrogen supplementation, diabetes, and fluoride exposure status. Based on a model containing all of these variables, women living in the high-fluoride community had a bone mass ranging from 0.004 to 0.039 g/cm2 less than that of similar women living in the optimum-fluoride community. These results suggest that lifelong ingestion of water containing 3.5 ppm fluoride, compared with water containing 0.7 ppm fluoride, does not increase cortical bone mass in women of similar age, weight, and menopausal status. Under the conditions of this study, cortical bone mass might be reduced in a high-fluoride area.


External beam radiotherapy: a treatment option for massive haemoptysis caused by mycetoma.
Five patients with life threatening haemoptysis secondary to a mycetoma were treated with external beam radiotherapy (EBRT). External beam radiotherapy of 3.5 Gy was given once a week continuing for one fraction after the haemoptysis stopped. Three patients required 7 Gy, one required 10.5 Gy and the fifth patient required 14 Gy before the haemoptysis had completely stopped. Irradiation was successful in achieving haemostasis with no side effects being observed after treatment in all five patients. Radiation therapy is an effective modality of treatment for life-threatening haemoptysis due to a mycetoma with no significant acute or late side effects.


Endocytosis, signaling, and beyond.
The endocytic network comprises a vast and intricate system of membrane-delimited cell entry and cargo sorting routes running between biochemically and functionally distinct intracellular compartments. The endocytic network caters to the organization and redistribution of diverse subcellular components, and mediates appropriate shuttling and processing of materials acquired from neighboring cells or the extracellular milieu. Such trafficking logistics, despite their importance, represent only one facet of endocytic function. The endocytic network also plays a key role in organizing, mediating, and regulating cellular signal transduction events. Conversely, cellular signaling processes tightly control the endocytic pathway at different steps. The present article provides a perspective on the intimate relationships that exist between particular endocytic and cellular signaling processes in mammalian cells, within the context of understanding the impact of this nexus on integrated physiology.


Augmented vagal heart rate modulation in active hypoestrogenic pre-menopausal women with functional hypothalamic amenorrhoea.
Compared with eumenorrhoeic women, exercise-trained women with functional hypothalamic amenorrhoea (ExFHA) exhibit low heart rates (HRs) and absent reflex renin-angiotensin-system activation and augmentation of their muscle sympathetic nerve response to orthostatic stress. To test the hypothesis that their autonomic HR modulation is altered concurrently, three age-matched (pooled mean, 24 ± 1 years; mean ± S.E.M.) groups of women were studied: active with either FHA (ExFHA; n=11) or eumenorrhoeic cycles (ExOv; n=17) and sedentary with eumenorrhoeic cycles (SedOv; n=17). Blood pressure (BP), HR and HR variability (HRV) in the frequency domain were determined during both supine rest and graded lower body negative pressure (LBNP; -10, -20 and -40 mmHg). Very low (VLF), low (LF) and high (HF) frequency power spectra (ms(2)) were determined and, owing to skewness, log10-transformed. LF/HF ratio and total power (VLF + LF + HF) were calculated. At baseline, HR and systolic BP (SBP) were lower (P<0.05) and HF and total power were higher (P<0.05) in ExFHA than in eumenorrhoeic women. In all groups, LBNP decreased (P<0.05) SBP, HF and total power and increased (P<0.05) HR and LF/HF ratio. However, HF and total power remained higher (P<0.05) and HR, SBP and LF/HF ratio remained lower (P<0.05) in ExFHA than in eumenorrhoeic women, in whom measures did not differ (P>0.05). At each stage, HR correlated inversely (P<0.05) with HF. In conclusion, ExFHA women demonstrate augmented vagal yet unchanged sympathetic HR modulation, both at rest and during orthostatic stress. Although the role of oestrogen deficiency is unclear, these findings are in contrast with studies reporting decreased HRV in hypoestrogenic post-menopausal women.


Dietary intake and rural-urban migration in India: a cross-sectional study.
Migration from rural areas of India contributes to urbanisation and lifestyle change, and dietary changes may increase the risk of obesity and chronic diseases. We tested the hypothesis that rural-to-urban migrants have different macronutrient and food group intake to rural non-migrants, and that migrants have a diet more similar to urban non-migrants. The diets of migrants of rural origin, their rural dwelling sibs, and those of urban origin together with their urban dwelling sibs were assessed by an interviewer-administered semi-quantitative food frequency questionnaire. A total of 6,509 participants were included. Median energy intake in the rural, migrant and urban groups was 2731, 3078, and 3224 kcal respectively for men, and 2153, 2504, and 2644 kcal for women (p<0.001). A similar trend was seen for overall intake of fat, protein and carbohydrates (p<0.001), though differences in the proportion of energy from these nutrients were <2%. Migrant and urban participants reported up to 80% higher fruit and vegetable intake than rural participants (p<0.001), and up to 35% higher sugar intake (p<0.001). Meat and dairy intake were higher in migrant and urban participants than rural participants (p<0.001), but varied by region. Sibling-pair analyses confirmed these results. There was no evidence of associations with time in urban area. Rural to urban migration appears to be associated with both positive (higher fruit and vegetables intake) and negative (higher energy and fat intake) dietary changes. These changes may be of relevance to cardiovascular health and warrant public health interventions.


The ultrastructure of mouse lung; general architecture of capillary and alveolar walls.
The general architecture of capillary and alveolar walls of the mouse lung was studied by means of the electron microscope. In order to minimize tissue damage and to improve the cutting properties of embeddings, several modifications in the tissue processing methods were adopted. These modifications were: fixation by infusion, a prolonged time of dehydration, of impregnation, and of polymerization, the use of acetone for dehydration, ammonium sulfide treatment of the fixed and washed tissue, and an elevated (80 degrees C.) polymerization temperature combined with the use of prepolymerized methacrylate. The generally favorable effects of these modified methods upon preservation and cutting properties of embedded tissue are discussed. Both capillary endothelium and alveolar epithelium were found continuous and without pores. The endothelium was seen to be thinnest in those portions that were adjacent to alveolar air spaces. Two morphological "types" of alveolar epithelial cells were found. One protruded into the alveolar lumen with its thick portion containing the nucleus. The other was often located in a niche of the alveolar wall, and contained peculiar dark inclusions amidst numerous mitochondria. Both were attenuated at their periphery to form the thin epithelial layer. The layer between endothelium and epithelium was designated as basement membrane. It was seen to be generally thin and structureless, but was found thickened in some areas where it also contained collagen fibrils.


Repetitive DNA methylome analysis by small-scale and single-cell shotgun bisulfite sequencing.
Whole-genome shotgun bisulfite sequencing (WG-SBS) is currently the most powerful tool available for understanding genomewide cytosine methylation with single-base resolution; however, the high sequencing cost limits its widespread application, particularly for mammalian genomes. We mapped high- to low-coverage SBS short reads of mouse and human female developing germ cells to consensus sequences of repetitive elements that were multiplied in the respective host genome. This mapping strategy effectively identified active and evolutionarily young retrotransposon subfamilies and centromeric satellite repeats that were resistant to DNA demethylation during the investigated progressive stages of germ cell development. Notably, quantities of only tens of thousands of uniquely mapped reads provided sufficient sensitivity to allow for methylation analyses of multiple retrotransposons and satellite repeats in mice. Furthermore, we produced SBS results from single female murine germ cells by an improved multiplexing and amplification-free SBS method (scPBAT). The scPBAT results quantitatively provided ≥5× sequencing coverage for at least 30 repeats, and the individual methylation patterns detected were similar to the bulk cell-based results. Our single-cell methylome sequencing technique will allow researchers to investigate intergenic methylation characteristics from limited amounts of mammalian cells as well as cells from other organisms with genomic annotations.