470 million years of conservation of microsatellite loci among fish species.
Primers for 18 microsatellite loci originally isolated from whiting (Merlangius merlangus, n = 6), stickleback (Gasterosteus aculeatus, n = 5) and cod (Gadus morhua, n = 7) were tested across a panel of diverse fish species, representing the three principal superclasses and most principal superorders of fish, to examine conservation of microsatellite regions across distantly related taxa. Three methods were used. First, amplified fragments were analysed by Southern blotting using the relevant microsatellite motif probes. A total of 17 of the tested primer pairs gave a product in the expected size range in at least four of 11 tested species. Second, for two study loci the amplified polymerase chain reaction products were cloned and sequenced in five fish species to reveal a high level of conservation of the flanking and microsatellite sequences. Finally, the 17 loci successfully amplified in non-source species were tested for polymorphism in groups of unrelated individuals from nine species, in several cases revealing extensive polymorphism. Levels of polymorphism were generally high in species from which the loci were derived or among closely related species. The conservation of flanking sequences for particular microsatellite motifs over the span of fish evolution represented in the test species (470 million years) far exceeds that hitherto reported and lends support to the suggestion (derived from studies of whales and marine turtles) that the rate of base substitution in nuclear and mitochondrial sequences is lower in aquatic than terrestrial organisms. A further explanation could be that these sequences, although generally considered neutral, may play an important role in eukaryotic genomes, and may be under strong selective constraints. The study suggests that heterologous primers will be a ready source of polymorphic markers among fish species, but also indicates that caution should be used in cross-species comparisons of variability.


Oligodendrocytes express gap junction proteins connexin32 and connexin45.
Oligodendrocytes, the myelin-forming glia of brain, are connected by gap junctions in situ and in culture. Cultured oligodendrocytes from adult bovine and porcine brains were studied using immunocytochemical, molecular, and electrophysiological techniques in order to characterize the gap junction types. The expression of connexin32 was substantiated by the detection of low, but significant, signals using connexin-specific probes in Northern and Western blot analyses. Connexin43, which comprises gap junctions in astrocytes, was not detectable in pure oligodendrocytic cultures; mRNAs of connexin40 and connexin37 and connexin26 were also not detected. By means of two specific antibodies directed to the recently cloned connexin45 and by RT-PCR we were able to identify this connexin as a second oligodendrocytic gap junction protein. Whole cell voltage clamp recording provided evidence for electrical coupling between pairs of cultured oligodendrocytes (mean junctional conductance 3.9 nS, n = 38 pairs) and intracellular Lucifer Yellow injection indicated that oligodendrocytes were usually only weakly dye coupled, with spread generally being restricted to nearest neighbors. Unitary conductances ranged from > 20 to < 150 pS with modes of distribution at about 100 to 120pS and 40 to 20 pS, respectively. These unitary conductances are consistent with the channel events expected for connexin32 and connexin45. The low degree of functional coupling between oligodendrocytes in vitro corresponds with the low levels of connexin32 and connexin45 messenger RNAs and protein expression.


Positive T wave overshoot as a sign of ventricular enlargement.
A consecutive series of 86 patients with an inverted T wave showing terminal positivity (overshoot) of a specific pattern in the resting electrocardiogram were studied. Patients with bundle branch block or electrocardiographic evidence of acute infarction and those taking digoxin or a similar drug were excluded. In 67 patients the heart was examined by echocardiography and in a further two by direct inspection. Sixty six of the 69 patients had an abnormal thickness of the left (or right) ventricle or a calculated left ventricular mass greater than 200 g. Seven of the patients examined by echocardiography had clinically pure ischaemic heart disease; all showed evidence of left ventricular enlargement. In only 39 of the 63 patients with anatomical evidence of left ventricular hypertrophy or dilatation did the electrocardiogram satisfy the standard voltage criterion of left ventricular hypertrophy. In the absence of acute infarction, bundle branch block, or digitalisation positive T wave overshoot of the pattern described is a sign of increased ventricular mass.


Accurate and inexpensive prediction of the color optical properties of anthocyanins in solution.
The simulation of the color optical properties of molecular dyes in liquid solution requires the calculation of time evolution of the solute absorption spectra fluctuating in the solvent at finite temperature. Time-averaged spectra can be directly evaluated by combining ab initio Car-Parrinello molecular dynamics and time-dependent density functional theory calculations. The inclusion of hybrid exchange-correlation functionals, necessary for the prediction of the correct transition frequencies, prevents one from using these techniques for the simulation of the optical properties of large realistic systems. Here we present an alternative approach for the prediction of the color of natural dyes in solution with a low computational cost. We applied this approach to representative anthocyanin dyes: the excellent agreement between the simulated and the experimental colors makes this method a straightforward and inexpensive tool for the high-throughput prediction of colors of molecules in liquid solvents.


Intensely pleasurable responses to music correlate with activity in brain regions implicated in reward and emotion.
We used positron emission tomography to study neural mechanisms underlying intensely pleasant emotional responses to music. Cerebral blood flow changes were measured in response to subject-selected music that elicited the highly pleasurable experience of "shivers-down-the-spine" or "chills." Subjective reports of chills were accompanied by changes in heart rate, electromyogram, and respiration. As intensity of these chills increased, cerebral blood flow increases and decreases were observed in brain regions thought to be involved in reward/motivation, emotion, and arousal, including ventral striatum, midbrain, amygdala, orbitofrontal cortex, and ventral medial prefrontal cortex. These brain structures are known to be active in response to other euphoria-inducing stimuli, such as food, sex, and drugs of abuse. This finding links music with biologically relevant, survival-related stimuli via their common recruitment of brain circuitry involved in pleasure and reward.


Purification of IgG and albumin from human plasma by aqueous two phase system fractionation.
The current shortages in human plasma products at global levels justify the development of new, cost effective plasma fractionation methods. We have developed a fractionation process to obtain immunoglobulin G (IgG) and albumin-enriched fractions based on polymer-salt aqueous two phase system (ATPS). A small-scale (0.02 L) ATPS composed of polyethyleneglycol 3350 (PEG), potassium phosphate and sodium chloride, at pH 6.1, was evaluated and subjected to 50-fold scale-up (1 L). Further purification of the fractions was performed using caprylic acid precipitation and ion exchange chromatography. Similar yield and purity were obtained at both small and large scales. IgG precipitated in the PEG rich upper phase at 83% recovery and 2.75-fold purification factor. An 81% pure albumin fraction was obtained in the salt rich bottom phase with a 91% yield. After polishing, IgG was obtained at a recovery of 70% and a purity of 92%. Corresponding values for albumin were 91% and 90%. This IgG and albumin fractionation technology deserves further evaluation as it may represent a potential alternative to conventional plasma fractionation methods.


Introduction of the AmpliChip CYP450 Test to a South African cohort: a platform comparative prospective cohort study.
Adverse drug reactions and lack of therapeutic efficacy associated with currently prescribed pharmacotherapeutics may be attributed, in part, to inter-individual variability in drug metabolism. Studies on the pharmacogenetics of Cytochrome P450 (CYP) enzymes offer insight into this variability. The objective of this study was to compare the AmpliChip CYP450 Test® (AmpliChip) to alternative genotyping platforms for phenotype prediction of CYP2C19 and CYP2D6 in a representative cohort of the South African population. AmpliChip was used to screen for thirty-three CYP2D6 and three CYP2C19 alleles in two different cohorts. As a comparison cohort 2 was then genotyped using a CYP2D6 specific long range PCR with sequencing (CYP2D6 XL-PCR + Sequencing) platform and a PCR-RFLP platform for seven CYP2C19 alleles. Even though there was a low success rate for the AmpliChip, allele frequencies for both CYP2D6 and CYP2C19 were very similar between the two different cohorts. The CYP2D6 XL-PCR + Sequencing platform detected CYP2D6*5 more reliably and could correctly distinguish between CYP2D6*2 and *41 in the Black African individuals. Alleles not covered by the AmpliChip were identified and four novel CYP2D6 alleles were also detected. CYP2C19 PCR-RFLP identified CYP2C19*9,*15, *17 and *27 in the Black African individuals, with *2, *17 and *27 being relatively frequent in the cohort. Eliminating mismatches and identifying additional alleles will contribute to improving phenotype prediction for both enzymes. Phenotype prediction differed between platforms for both genes. Comprehensive genotyping of CYP2D6 and CYP2C19 with the platforms used in this study, would be more appropriate than AmpliChip for phenotypic prediction in the South African population. Pharmacogenetically important novel alleles may remain undiscovered when using assays that are designed according to Caucasian specific variation, unless alternate strategies are utilised.


Injectable graphene oxide/hydrogel-based angiogenic gene delivery system for vasculogenesis and cardiac repair.
The objective of this study was to develop an injectable and biocompatible hydrogel which can efficiently deliver a nanocomplex of graphene oxide (GO) and vascular endothelial growth factor-165 (VEGF) pro-angiogenic gene for myocardial therapy. For the study, an efficient nonviral gene delivery system using polyethylenimine (PEI) functionalized GO nanosheets (fGO) complexed with DNAVEGF was formulated and incorporated in the low-modulus methacrylated gelatin (GelMA) hydrogel to promote controlled and localized gene therapy. It was hypothesized that the fGOVEGF/GelMA nanocomposite hydrogels can efficiently transfect myocardial tissues and induce favorable therapeutic effects without invoking cytotoxic effects. To evaluate this hypothesis, a rat model with acute myocardial infarction was used, and the therapeutic hydrogels were injected intramyocardially in the peri-infarct regions. The secreted VEGF from in vitro transfected cardiomyocytes demonstrated profound mitotic activities on endothelial cells. A significant increase in myocardial capillary density at the injected peri-infarct region and reduction in scar area were noted in the infarcted hearts with fGOVEGF/GelMA treatment compared to infarcted hearts treated with untreated sham, GelMA and DNAVEGF/GelMA groups. Furthermore, the fGOVEGF/GelMA group showed significantly higher (p < 0.05, n = 7) cardiac performance in echocardiography compared to other groups, 14 days postinjection. In addition, no significant differences were noticed between GO/GelMA and non-GO groups in the serum cytokine levels and quantitative PCR based inflammatory microRNA (miRNA) marker expressions at the injected sites. Collectively, the current findings suggest the feasibility of a combined hydrogel-based gene therapy system for ischemic heart diseases using nonviral hybrid complex of fGO and DNA.


Physical Activity and Outcomes in Patients with Stage III Colon Cancer: A Correlative Analysis of Phase III Trial NCCTG N0147 (Alliance).
Background: Prior studies have supported an inverse association between physical activity and colon cancer risk and suggest that higher physical activity may also improve cancer survival. Among participants in a phase III adjuvant trial for stage III colon cancer, we assessed the association of physical activity around the time of cancer diagnosis with subsequent outcomes.Methods: Before treatment arm randomization (FOLFOX or FOLFOX + cetuximab), study participants completed a questionnaire including items regarding usual daily activity level and frequency of participation in recreational physical activity (N = 1,992). Using multivariable Cox models, we calculated HRs for associations of aspects of physical activity with disease-free (DFS) and overall survival (OS).Results: Over follow-up, 505 participants died and 541 experienced a recurrence. Overall, 75% of participants reported recreational physical activity at least several times a month; for participants who reported physical activity at least that often (vs. once a month or less), the HRs for DFS and OS were 0.82 [95% confidence interval (CI), 0.69-0.99] and 0.76 (95% CI, 0.63-0.93), respectively. There was no evidence of material effect modification in these associations by patient or tumor attributes, except that physical activity was more strongly inversely associated with OS in patients with stage T3 versus T4 tumors (Pinteraction = 0.03).Conclusions: These findings suggest that higher physical activity around the time of colon cancer diagnosis may be associated with more favorable colon cancer outcomes.Impact: Our findings support further research on whether colon cancer survival may be enhanced by physical activity. Cancer Epidemiol Biomarkers Prev; 27(6); 696-703. ©2018 AACR.


Open-Source High-Throughput Phenomics of Bacterial Promoter-Reporter Strains.
Open-source electronics are becoming more prevalent in biological sciences, enabling novel and unique means of data acquisition. Here, we present 3D-printed, open-source tools to acquire fluorescence phenotypes with high temporal resolution. Printed fluorescence imaging boxes (PFIboxes) cost approximately 200 US dollars to assemble, can be placed in incubators or hypoxic chambers, and accurately read high-density colony arrays of microorganisms. We demonstrate the utility of PFIboxes using a time course gene expression approach, examining global Escherichia coli promoter activity using a fluorescent reporter library across a diverse panel of 15 antibiotics, each at several concentrations. Many secondary and indirect effects were observed when E. coli was challenged with various drugs, including increased gene expression in carbon metabolism processes. Further, kinetic data acquisition enabled non-destructive time course gene expression, clustering of which revealed patterns of co-expression. In all, PFIboxes provide an open solution to gene expression, for about 2 US dollars per treatment condition, including technical replicates.


Precision, repeatability, and validation of the localization of cranial landmarks using computed tomography scans.
Computed tomography (CT) has brought to the craniofacial surgeon a three-dimensional representation of internal structures. CT scans provide visualization of anatomy for preoperative planning and postoperative evaluation. Beyond visualization, however, a CT scan enables assessment of measurements useful to clinicians and basic scientists. All measurement systems used with CT require the ability to accurately locate regions of interest on the image (i.e., areas, volumes, outlines, curves, surfaces, points). This study evaluates the precision and repeatability of locating anatomic landmarks in three dimensions on CT slice images, and validates these locations using an established measurement system. The average error of landmark position is always less than 0.5 mm and for some landmarks error is negligible. Repeatability studies show that less than 2% of the total variance in our data is due to measurement inaccuracy. Although data collected from CT scans are internally consistent, validation results caution the use of CT data in combination with data collected using calipers or other direct means of measurement.


Eimeria tenella: 14-3-3 protein interacts with telomerase.
Telomerase, consisting of telomerase RNA and telomerase reverse transcriptase (TERT), is responsible for the maintenance of the end of linear chromosomes. TERT, as the catalytic subunit of telomerase, plays a critical role in telomerase activity. Researches indicate TERT-associated proteins participate in the regulation of telomerase assembly, posttranslational modification, localization, and enzymatic function. Here, the telomerase RNA-binding domain of Eimeria tenella TERT (EtTRBD) was cloned into pGBKT7 and performed as the bait. α-Galactosidase assay showed that the bait plasmid did not activate Gal4 reporter gene. Further, we isolated an EtTRBD-associated protein, 14-3-3, by yeast two-hybrid screening using the constructed bait plasmid. To confirm the interaction, EtTRBD and 14-3-3 were expressed by prokaryotic and eukaryotic expression systems. Pull-down assays by purified proteins demonstrated a direct bind between EtTRBD and 14-3-3. Co-immunoprecipitation techniques successfully validated that 14-3-3 interacted with EtTRBD in 293T cells. The protein-protein interaction provides a starting point for more in-depth studies on telomerase and telomere regulation in E. tenella.


Interaction effects between total energy and macronutrient intakes and angiotensin-converting enzyme 1 ( ACE) I/D polymorphism on adiposity-related phenotypes in toddlers and preschoolers: the Growth, Exercise and Nutrition Epidemiological Study in preSchoolers: the GENESIS study. [corrected].
The aim of the present study was to investigate the interaction between the angiotensin-converting enzyme 1 (ACE) I/D polymorphism and energy and macronutrient intakes on adiposity-related phenotypes among toddlers and preschoolers. A representative sample of 2374 Greek children aged 1 to 5 years old was examined (Growth, Exercise and Nutrition Epidemiological Study in preSchoolers (GENESIS)). Dietary and anthropometric (i.e. BMI, waist circumference (WC)) assessments were carried out using standard procedures. DNA samples were obtained from 2102 children and were genotyped for the ACE I/D polymorphism. Among the entire population, 17 % were 'at risk of overweight' and a similar percentage were 'overweight'. The frequencies of the II, ID and DD genotypes were 16, 46 and 38 %, respectively. Significant interactions were found between the ACE I/D polymorphism and total energy intake on WC (P = 0.004 for interaction) and the ACE I/D polymorphism and protein intake on BMI and being overweight (P < 0.05 for interaction). Furthermore, it was found that the ACE I/D polymorphism may modify the effect of fat intake on WC and BMI, but this interaction disappeared after adjustment for additional potential confounders. Stratified analyses revealed that total energy is correlated with WC and protein intake is associated with BMI and being overweight only among carriers of the D-allele (i.e. DD or ID genotypes). These results suggest that the ACE I/D polymorphism may act as a modifying factor in the response of adiposity-related phenotypes to diet.


Comparative susceptibility of Deschutes River, Oregon, Tubifex tubifex populations to Myxobolus cerebralis.
Dams along the Deschutes River (DR) in central Oregon have blocked fish migration for over 40 years. Reestablishment of anadromous fish runs above the dams as part of a fish passage plan may introduce fish pathogens, such as Myxobolus cerebralis, the myxozoan parasite that causes salmonid whirling disease. This parasite is carried by adult salmon that stray into the DR system during their return to enzootic areas of the upper Columbia River basin, and it is now known to be established in at least one lower DR tributary. The life cycle of M. cerebralis involves two obligate hosts: a salmonid and the oligochaete worm Tubifex tubifex. To determine the likelihood of parasite establishment above the DR dams, we conducted benthic sediment surveys between 1999 and 2007 and found that T. tubifex had a patchy distribution and low relative abundance. Mitochondrial 16S ribosomal DNA gene analysis indicated that two lineages of T. tubifex (III and VI) were present both above and below the dams. Laboratory susceptibility studies to characterize differences in infection prevalence and parasite production between nine T. tubifex populations revealed that production varied considerably among exposed groups and was proportional to the number of lineage III worms present. Our results suggest that M. cerebralis could become established above the dams if infected fish are allowed passage into the upper DR system, but not all areas of the DR basin can be classified as having the same likelihood for parasite establishment, and the potential impact will be location dependent.


The effect of disinfectant agents on dimensional stability and surface roughness of a tissue conditioner material.
The aim of this study was to determine the effect of disinfectant procedures on the dimensional stability and surface quality of a tissue conditioner used as a functional impression material. A tissue conditioner (Visco-gel) used as a functional impression material was disinfected by immersion in 5.25% sodium hypochlorite or 2% glutaraldehyde solutions for 10 or 20 minutes, respectively. The control group consisted of specimens (n=10) that were not treated by disinfectants. For testing the dimensional stability, impressions of an aluminum edentulous arch with three reference points were made. After 24 hours of storage in distilled water, the specimens were treated by one of the disinfection procedures. Impressions were poured and, subsequently, 50 stone casts were measured with a Nikon profile projector. For surface roughness evaluation, disks that contained Visco-gel were pressed against glass slides. The disks were then stored in distilled water for 24 hours. After disinfection, 50 disk-shaped dental stone casts were prepared. Surface roughness values were determined using a profilometer. For each test, data were subjected to two-way analysis of variance (ANOVA) followed by Tukey's test (α = 0.05). The results of the dimensional stability test showed that immersion of Visco-gel impressions in 5.25% sodium hypochlorite solution or 2% glutaraldehyde solution did not significantly change the anteroposterior (AP) and cross-arch (CA) distances of the resultant stone casts when compared to the control group (P>0.05). The surface quality of dental stone casts formed by Visco-gel did not change significantly when impressions were immersed in the disinfectant solutions for either 10 or 20 minutes, or when stored in distilled water (P>0.05). The results showed that the disinfectant solutions used in this study did not have a significant effect on the surface quality and dimensional accuracy of Visco-gel as a functional impression material.


A temperature model for laser lithotripsy.
To derive and validate a mathematical model to predict laser-induced temperature changes in a kidney during kidney stone treatment. A simplified mathematical model to predict temperature change in the kidney for any given renal volume, irrigation flow rate, irrigation fluid temperature, and laser power was derived. We validated our model with matched in vitro experiments. Excellent agreement between the mathematical model predictions and laboratory data was obtained. The model obviates the need for repeated experimental validation. The model predicts scenarios where risk of renal tissue damage is high. With real-time knowledge of flow rate, irrigating fluid temperature and laser usage, safety warning levels could be predicted. Meanwhile, clinicians should be aware of the potential risk from thermal injury and take measures to reduce the risk, such as using room temperature irrigation fluid and judicious laser use.


Enzymatic labelling of snake venom phospholipase A2 toxins.
Almost all animal venoms contain secretory phospholipases A2 (PLA2s), 14 kDa disulfide-rich enzymes that hydrolyze membrane phospholipids at the sn-2 position, releasing lysophospholipids and fatty acids. These proteins, depending on their sequence, show a wide variety of biochemical, toxic and pharmacological effects and deserve to be studied for their numerous possible applications, and to improve antivenom drugs. The cellular localization and activity of a protein can be studied by conjugating it with a tag. In this work, we applied an enzymatic labelling method, using Streptomyces mobaraense transglutaminase, on three snake venom PLA2s: a recombinant neuro- and myotoxic group I PLA2 from Notechis scutatus scutatus, and two myotoxic group II PLA2s from Bothrops asper - one of them a natural catalytically inactive variant. We demonstrate that TGase can be used to produce active mono- or bi-derivatives of these three PLA2s modified at specific Lys residues, and that all three of these proteins, conjugated with fluorescent peptides, are internalized in primary myotubes.


Work-life imbalance and mental health among male and female employees in Switzerland.
To investigate the prevalence and mental health effects of an unequal work-life balance (WLB) including potential gender differences. A cross-sectional study based on a representative sample of the Swiss employed population aged 20 to 64 (women: n = 1661; men: n = 1591). Based on a single-item measure, more than every seventh employee in Switzerland indicated major difficulties combining work and private life. In certain socio-demographic categories, up to 30% showed such work-life conflict (WLC). For both genders, work-life imbalance turned out to be a risk factor affecting mental health. Employees with self-reported WLC presented a significantly higher relative risk for poor self-rated health (women: aOR = 2.6/men: aOR = 2.0), negative emotions and depression (aOR = 3.0/3.1), low energy and optimism (aOR = 2.1/1.6), fatigue (aOR = 2.4/2.6), and sleep disorders (aOR = 1.8/1.5) compared to employees with no WLC. Internationally, few data on the prevalence of WLC exist. In Switzerland, work-life imbalance is not a marginal phenomenon among the workforce and needs to be addressed as a notable public and mental health issue.


Societal Sentience: Constructions of the Public in Animal Research Policy and Practice.
The use of nonhuman animals as models in research and drug testing is a key route through which contemporary scientific knowledge is certified. Given ethical concerns, regulation of animal research promotes the use of less "sentient" animals. This paper draws on a documentary analysis of legal documents and qualitative interviews with Named Veterinary Surgeons and others at a commercial laboratory in the UK. Its key claim is that the concept of animal sentience is entangled with a particular imaginary of how the general public or wider society views animals. We call this imaginary societal sentience. Against a backdrop of increasing ethnographic work on care encounters in the laboratory, this concept helps to stress the wider context within which such encounters take place. We conclude that societal sentience has potential purchase beyond the animal research field, in helping to highlight the affective dimension of public imaginaries and their ethical consequences. Researching and critiquing societal sentience, we argue, may ultimately have more impact on the fate of humans and nonhumans in the laboratory than focusing wholly on ethics as situated practice.


Potential role for interleukin-28B genotype in treatment decision-making in recent hepatitis C virus infection.
Polymorphisms in the IL28B (interleukin-28B) gene region are important in predicting outcome following therapy for chronic hepatitis C virus (HCV) infection. We evaluated the role of IL28B in spontaneous and treatment-induced clearance following recent HCV infection. The Australian Trial in Acute Hepatitis C (ATAHC) was a study of the natural history and treatment of recent HCV, as defined by positive anti-HCV antibody, preceded by either acute clinical HCV infection within the prior 12 months or seroconversion within the prior 24 months. Factors associated with spontaneous and treatment-induced HCV clearance, including variations in IL28B, were assessed. Among 163 participants, 132 were untreated (n = 52) or had persistent infection (infection duration ≥26 weeks) at treatment initiation (n = 80). Spontaneous clearance was observed in 23% (30 of 132 participants). In Cox proportional hazards analysis (without IL28B), HCV seroconversion illness with jaundice was the only factor predicting spontaneous clearance (adjusted hazards ratio = 2.86; 95% confidence interval = 1.24, 6.59; P = 0.014). Among participants with IL28B genotyping (n = 102 of 163 overall and 79 of 132 for the spontaneous clearance population), rs8099917 TT homozygosity (versus GT/GG) was the only factor independently predicting time to spontaneous clearance (adjusted hazard ratio = 3.78; 95% confidence interval = 1.04, 13.76; P = 0.044). Participants with seroconversion illness with jaundice were more frequently rs8099917 TT homozygotes than other (GG/GT) genotypes (32% versus 5%, P = 0.047). Among participants adherent to treatment and who had IL28B genotyping (n = 54), sustained virologic response was similar among TT homozygotes (18 of 29 participants, 62%) and those with GG/GT genotype (16 of 25, 64%, P = 0.884). During recent HCV infection, genetic variations in IL28B region were associated with spontaneous but not treatment-induced clearance. Early therapeutic intervention could be recommended for individuals with unfavorable IL28B genotypes.


CD137 is expressed in follicular dendritic cell tumors and in classical Hodgkin and T-cell lymphomas: diagnostic and therapeutic implications.
CD137 (also known as 4-1BB and TNFRSF9) is a member of the tumor necrosis factor receptor superfamily. Originally identified as a costimulatory molecule expressed by activated T cells and NK cells, CD137 is also expressed by follicular dendritic cells, monocytes, mast cells, granulocytes, and endothelial cells. Anti-CD137 immunotherapy has recently shown promise as a treatment for solid tumors and lymphoid malignancies in preclinical models. We defined the expression of CD137 protein in both normal and neoplastic hematolymphoid tissue. CD137 protein is expressed by follicular dendritic cells in the germinal center and scattered paracortical T cells, but not by normal germinal-center B cells, bone marrow progenitor cells, or maturing thymocytes. CD137 protein is expressed by a select group of hematolymphoid tumors, including classical Hodgkin lymphoma, T-cell and NK/T-cell lymphomas, and follicular dendritic cells neoplasms. CD137 is a novel diagnostic marker of these tumors and suggests a possible target for tumor-directed antibody therapy.


Zebrafish neuromuscular junction: The power of N.
In the early 1950s, Katz and his colleagues capitalized on the newly developed intracellular microelectrode recording technique to investigate synaptic transmission. For study they chose frog neuromuscular junction (NMJ), which was ideally suited due to the accessibility and large size of the muscle cells. Paradoxically, the large size precluded the use of next generation patch clamp technology. Consequently, electrophysiological study of synaptic function shifted to small central synapses made amenable by patch clamp. Recently, however, the unique features offered by zebrafish have rekindled interest in the NMJ as a model for electrophysiological study of synaptic transmission. The small muscle size and synaptic simplicity provide the singular opportunity to perform in vivo spinal motoneuron-target muscle patch clamp recordings. Additional incentive is provided by zebrafish lines harboring mutations in key synaptic proteins, many of which are embryonic lethal in mammals, but all of which are able to survive well past synapse maturation in zebrafish. This mini-review will highlight features that set zebrafish NMJs apart from traditional NMJs. We also draw into focus findings that offer the promise of identifying features that define release sites, which serve to set the upper limit of transmitter release. Since its conception several candidates representing release sites have been proposed, most of which are based on distinctions among vesicle pools in their state of readiness for release. However, models based on distinctions among vesicles have become enormously complicated and none adequately account for setting an upper limit for exocytosis in response to an action potential (AP). Specifically, findings from zebrafish NMJ point to an alternative model, positing that elements other than vesicles per se set the upper limits of release.


A multiplex loop-mediated isothermal amplification assay for rapid detection of Bacillus cereus and Staphylococcus aureus.
Bacillus cereus (B. cereus) and Staphylococcus aureus (S. aureus) are major human food-borne pathogens that may produce a variety of toxins and cause diarrhea, food poisoning, and even death. In order to monitor and prevent the spread of these pathogens, a multiplex loop-mediated isothermal amplification (multi-LAMP) assay was developed to simultaneously and rapidly detect B. cereus and S. aureus. The sensitivity and specificity of the loop-mediated isothermal amplification (LAMP) reactions were determined via electrophoresis. The multi-LAMP showed 100% inclusivity and exclusivity, the sensitivity was 10 fg/μL and was 10 times more sensitive than that of polymerase chain reaction (PCR), the results were consistent with those of conventional PCR assay, and the entire assay should be finished within 40 min. This multi-LAMP assay was confirmed as a rapid and reliable diagnostic technique upon application for clinical samples and food samples. To our knowledge, this is the first study to report the application of multi-LAMP to detect B. cereus and S. aureus.


The use of cognitive testing to develop and evaluate CAHPS 1.0 core survey items. Consumer Assessment of Health Plans Study.
The main goal of the Consumer Assessments of Health Plans Study (CAHPS) is to develop an integrated set of tested, standardized surveys to obtain meaningful information from health plan enrollees about their experiences. The CAHPS project benefits from the complementary strengths of psychometric and cognitive testing. The CAHPS team conducted 150 cognitive interviews across three organizations in different geographic locations using multiple interview methods with different consumer populations. This article explains how cognitive testing was used in the CAHPS survey development process and shares the main findings from the cognitive interviews. A modified report format is more appropriate when asking about specific aspects of plan enrollees' experiences, whereas a rating format is useful for asking about overall assessments. Specifying a longer reference period is preferable to asking about the most recent visit when capturing experiences with care, because some respondents get frustrated when they cannot include experiences other than the most recent visit. Explicit screeners and tailored inapplicable response categories are beneficial in mail questionnaires, so people know that they should not answer questions about which they have no relevant experience. Cognitive testing was integral in the development and refinement of the CAHPS instrument. The cognitive testing findings contributed to an improved instrument that should capture consumers' health care and plan experiences with less response error than one not subjected to such testing. The cognitive testing process and findings can be useful to other researchers with similar survey development goals.


Clinical features of spinal and bulbar muscular atrophy.
Spinal and bulbar muscular atrophy is an X-linked motor neuron disease caused by a CAG repeat expansion in the androgen receptor gene. To characterize the natural history and define outcome measures for clinical trials, we assessed the clinical history, laboratory findings and muscle strength and function in 57 patients with genetically confirmed disease. We also administered self-assessment questionnaires for activities of daily living, quality of life and erectile function. We found an average delay of over 5 years from onset of weakness to diagnosis. Muscle strength and function correlated directly with serum testosterone levels and inversely with CAG repeat length, age and duration of weakness. Motor unit number estimation was decreased by about half compared to healthy controls. Sensory nerve action potentials were reduced in nearly all subjects. Quantitative muscle assessment and timed 2 min walk may be useful as meaningful indicators of disease status. The direct correlation of testosterone levels with muscle strength indicates that androgens may have a positive effect on muscle function in spinal and bulbar muscular atrophy patients, in addition to the toxic effects described in animal models.


Dysmyelination of auditory afferent axons increases the jitter of action potential timing during high-frequency firing.
Auditory neuropathies are linked to loss of temporal acuity of sound-evoked signals, which may be related to myelin loss. However, it is not known how myelin loss affects the waveform and temporal precision of action potentials (APs) in auditory CNS nerve terminals. Here we investigated the excitability of the calyx of Held nerve terminal in dysmyelinated auditory brainstems using the Long-Evans Shaker (LES) rat, a spontaneous mutant where compact myelin wrapping does not occur due to a genetic deletion of myelin basic protein. We found at relatively mature postnatal ages (15-17 d after birth) LES rat calyces showed prolonged spike latencies, indicative of a threefold reduction in the AP propagation velocity. Furthermore, LES rat afferent fiber-evoked APs showed a pronounced loss of temporal precision, even at low stimulation frequencies (10 Hz). While normal calyces were able to fire APs without failures at impressive rates of up to 1 kHz, LES calyces were unable to do so. Direct recordings of the presynaptic calyx terminal AP waveform revealed that myelin loss does not affect the AP spike upstroke and downstroke kinetics, but dysmyelination reduces the after-depolarization and enhances the fast after-hyperpolarization peak following the AP spike in the LES rat. Together these findings show that proper myelination is essential not only for fast AP propagation, but also for precise presynaptic AP firing that minimizes both spike jitter and failures, two characteristics critically important for the accurate processing of sound signals in the auditory brainstem.


Using a reporting system to protect the human rights of people living with HIV and key populations: a conceptual framework.
Human rights of people living with HIV (PLHIV) and key populations most affected by HIV are often violated, with negative implications for health outcomes. To facilitate access to justice in Ghana, a consortium of partners developed a web-based discrimination reporting system. The reporting system links the Commission on Human Rights and Administrative Justice (CHRAJ) to civil society organizations through case reporting, follow-up, and aggregated reported data use. A reporting system can form a critical part of a country's human rights protection network. In Ghana, the combination of a supportive legal framework, an institutional home, and engaged stakeholders have created a favorable environment for developing such a system. Other institutions may learn from Ghana's experience by adapting these lessons.


Analysis of power output time series in response to supramaximal exercise: an approach through dynamic factor analysis.
The responses to supramaximal exercise testing have been traditionally analyzed by means of standard parametric and nonparametric statistics. Unfortunately, these statistical approaches do not allow insight into the pattern of variation of a given parameter over time. The purpose of this study was to determine if the application of dynamic factor analysis (DFA) allowed discriminating different patterns of power output (PO), during supramaximal exercise, in two groups of children engaged in competitive sports: swimmers and soccer players. Data derived from Wingate testing were used in this study. Analyses were performed on epochs (30 s) of upper and lower body PO obtained from twenty two healthy boys (11 swimmers and 11 soccer players) age 11-12 years old. DFA revealed two distinct patterns of PO during Wingate. Swimmers tended to attain their peak PO (upper and lower body) earlier than soccer players. As importantly, DFA showed that children with a given pattern of upper body PO tend to perform similarly during lower body exercise.


Understanding the cryptic nature of Lassa fever in West Africa.
Lassa fever (LF) is increasingly recognized by global health institutions as an important rodent-borne disease with severe impacts on some of West Africa's poorest communities. However, our knowledge of LF ecology, epidemiology and distribution is limited, which presents barriers to both short-term disease forecasting and prediction of long-term impacts of environmental change on Lassa virus (LASV) zoonotic transmission dynamics. Here, we synthesize current knowledge to show that extrapolations from past research have produced an incomplete picture of the incidence and distribution of LF, with negative consequences for policy planning, medical treatment and management interventions. Although the recent increase in LF case reports is likely due to improved surveillance, recent studies suggest that future socio-ecological changes in West Africa may drive increases in LF burden. Future research should focus on the geographical distribution and disease burden of LF, in order to improve its integration into public policy and disease control strategies.


Integrating influenza antigenic dynamics with molecular evolution.
Influenza viruses undergo continual antigenic evolution allowing mutant viruses to evade host immunity acquired to previous virus strains. Antigenic phenotype is often assessed through pairwise measurement of cross-reactivity between influenza strains using the hemagglutination inhibition (HI) assay. Here, we extend previous approaches to antigenic cartography, and simultaneously characterize antigenic and genetic evolution by modeling the diffusion of antigenic phenotype over a shared virus phylogeny. Using HI data from influenza lineages A/H3N2, A/H1N1, B/Victoria and B/Yamagata, we determine patterns of antigenic drift across viral lineages, showing that A/H3N2 evolves faster and in a more punctuated fashion than other influenza lineages. We also show that year-to-year antigenic drift appears to drive incidence patterns within each influenza lineage. This work makes possible substantial future advances in investigating the dynamics of influenza and other antigenically-variable pathogens by providing a model that intimately combines molecular and antigenic evolution. DOI: http://dx.doi.org/10.7554/eLife.01914.001.


An Evaluation of Lysyl Oxidase-Derived Cross-Linking in Keratoconus by Liquid Chromatography/Mass Spectrometry.
Current literature contains scant information regarding the extent of enzymatic collagen cross-linking in the keratoconus (KC) cornea. The aim of the present study was to examine levels of enzymatic lysyl oxidase-derived cross-links in stromal collagen in KC tissue, and to correlate the cross-link levels with collagen fibril stability as determined by thermal denaturation temperature (Tm). Surgical KC samples (n = 17) and Eye-Bank control (n = 11) corneas of age 18 to 68 years were analyzed. The samples were defatted, reduced (NaBH4), hydrolyzed (6N HCl at 110°C for 18 hours), and cellulose enriched before analysis by C8 high-performance liquid chromatography equipped with parallel fluorescent and mass detectors in selective ion monitoring mode (20 mM heptafluorobutyric acid/methanol 70:30 isocratic at 1 mL/min). Nine different cross-links were measured, and the cross-link density was determined relative to collagen content (determined colorimetrically). The Tm was determined by differential scanning calorimetry. Cross-links detected were dihydroxylysinonorleucine (DHLNL), hydroxylysinonorleucine, lysinonorleucine (LNL), and histidinohydroxylysinonorleucine in both control and KC samples. Higher DHLNL levels were detected in KC, whereas the dominant cross-link, LNL, was decreased in KC samples. Decreased LNL levels were observed among KC ≤ 40 corneas. There was no difference in total cross-link density between KC samples and the controls. Pyridinolines, desmosines, and pentosidine were not detected. There was no notable correlation between cross-link levels with fibril instability as determined by Tm. Lower levels of LNL in the KC cornea suggest that there might be a cross-linking defect either in fibrillar collagen or the microfibrillar elastic network composed of fibrillin.


Daily bursts of biogenic cyanogen bromide (BrCN) control biofilm formation around a marine benthic diatom.
The spatial organization of biofilms is strongly regulated by chemical cues released by settling organisms. However, the exact nature of these interactions and the repertoire of chemical cues and signals that micro-organisms produce and exude in response to the presence of competitors remain largely unexplored. Biofilms dominated by microalgae often show remarkable, yet unexplained fine-scale patchy variation in species composition. Because this occurs even in absence of abiotic heterogeneity, antagonistic interactions might play a key role. Here we show that a marine benthic diatom produces chemical cues that cause chloroplast bleaching, a reduced photosynthetic efficiency, growth inhibition and massive cell death in naturally co-occurring competing microalgae. Using headspace solid phase microextraction (HS-SPME)-GC-MS, we demonstrate that this diatom exudes a diverse mixture of volatile iodinated and brominated metabolites including the natural product cyanogen bromide (BrCN), which exhibits pronounced allelopathic activity. Toxin production is light-dependent with a short BrCN burst after sunrise. BrCN acts as a short-term signal, leading to daily "cleaning" events around the algae. We show that allelopathic effects are H(2)O(2) dependent and link BrCN production to haloperoxidase activity. This strategy is a highly effective means of biofilm control and may provide an explanation for the poorly understood role of volatile halocarbons from marine algae, which contribute significantly to the atmospheric halocarbon budget.


Autologous stem cell transplantation in multiple myeloma: Post-transplant outcomes of Taiwan Blood and Marrow Transplantation Registry.
Multiple myeloma (MM) is a monoclonal plasma cell malignancy. The primary choice of treatment for MM is induction therapy followed by autologous stem cell transplantation (ASCT). This study aimed to analyze the treatment efficacy of ASCT in a Taiwanese cohort and evaluate possible prognostic factors. From the database of the Taiwan Blood and Marrow Transplantation registry, data on 396 patients with MM who underwent ASCT were reviewed. The average age of participants was 54.8 years, and there were more men than women (57.6% vs. 42.4%). Most patients were diagnosed with IgG-type myeloma (52.4%), followed by IgA-type (23.2%) and light-chain type (21.4%). Patients with Durie Salmon Staging System (DSS) III disease accounted for 61.9% of the study cohort, while 23.7% had stage II and 14.4% had stage I disease. The median progression-free survival (PFS) and overall survival (OS) after ASCT were 46.5 months and 70.4 months, respectively. DSS III was a poor prognostic factor affecting both PFS and OS with a duration of 35.9 months and 69.0 months, respectively, compared with the other two stages (p = 0.006 and p = 0.03, respectively). In addition, patients with better treatment response before ASCT had better PFS and OS compared with those who did not show a response (both p < 0.0001). The overall incidence of organ toxicities associated with transplantation was low. In conclusion, our cohort showed that myeloma patients with early DSS and better treatment response before ASCT had better long-term survival outcomes.


Interferon alpha2 recombinant and epidermal growth factor modulate proliferation and hypusine synthesis in human epidermoid cancer KB cells.
We previously found that interferon alpha2 recombinant (IFNalpha) increases the expression of epidermal growth factor receptor (EGF-R) in the human epidermoid cancer KB cell line. Here we report the effects of IFNalpha and epidermal growth factor (EGF) on KB cell cycle kinetics. IFNalpha (1000 i.u./ml) for 48 h decreased the S-phase fraction and diminished the expression of Ki67 and proliferating cell nuclear antigen on KB cells. Incubation of IFNalpha-treated KB cells with 10 nM EGF for 12 h reversed these effects. We then studied several biochemical markers of cell proliferation. Ornithine decarboxylase activity was decreased to about one-tenth by IFNalpha and partly restored by EGF. Hypusine is contained only in eukaryotic initiation factor 5A and its levels are correlated with cell proliferation. IFNalpha decreased hypusine synthesis by 75%; exposure of cells to EGF for 12 h restored hypusine synthesis almost completely. We also studied the effects of IFNalpha on the cytotoxicity of the recombinant toxin TP40, which inhibits elongation factor 2 through EGF-R binding and internalization. IFNalpha greatly enhanced the TP40-induced inhibition of protein synthesis in KB cells. In conclusion, IFNalpha, which affects protein synthesis machinery and increases EGF-R expression, enhances the tumoricidal activity of TP40 and hence could be useful in the setting of anti-cancer therapy.


Endless frequency shifting of optical frequency comb lines.
The functional principle of a novel technique for frequency shifting lines of an optical frequency comb is demonstrated. The underlying principle is to shift the carrier frequency by changing the carrier phase within the time span between subsequent pulses of a mode-locked laser used as comb generator. This universal frequency shifter does not require intrusion into the comb generator and provides high agility for arbitrary temporal frequency evolutions.


Effects of prenatal androgens on rhesus monkeys: a model system to explore the organizational hypothesis in primates.
After proposing the organizational hypothesis from research in prenatally androgenized guinea pigs (Phoenix, C.H., Goy, R.W., Gerall, A.A., Young, W.C., 1959. Organizational action of prenatally administered testosterone propionate on the tissues mediating mating behavior in the female guinea pig. Endocrinology 65, 369-382.), the same authors almost immediately extended the hypothesis to a nonhuman primate model, the rhesus monkey. Studies over the last 50 years have verified that prenatal androgens have permanent effects in rhesus monkeys on the neural circuits that underlie sexually dimorphic behaviors. These behaviors include both sexual and social behaviors, all of which are also influenced by social experience. Many juvenile behaviors such as play, mounting, and vocal behaviors are masculinized and/or defeminized, and aspects of adult sexual behavior are both masculinized (e.g. approaches, sex contacts, and mounts) and defeminized (e.g. sexual solicits). Different behavioral endpoints have different periods of maximal susceptibility to the organizing actions of prenatal androgens. Aromatization is not important, as both testosterone and dihydrotestosterone are equally effective in rhesus monkeys. Although the full story of the effects of prenatal androgens on sexual and social behaviors in the rhesus monkey has not yet completely unfolded, much progress has been made. Amazingly, a large number of the inferences drawn from the original 1959 study have proved applicable to this nonhuman primate model.


A conditional entropy minimization criterion for dimensionality reduction and multiple kernel learning.
Reducing the dimensionality of high-dimensional data without losing its essential information is an important task in information processing. When class labels of training data are available, Fisher discriminant analysis (FDA) has been widely used. However, the optimality of FDA is guaranteed only in a very restricted ideal circumstance, and it is often observed that FDA does not provide a good classification surface for many real problems. This letter treats the problem of supervised dimensionality reduction from the viewpoint of information theory and proposes a framework of dimensionality reduction based on class-conditional entropy minimization. The proposed linear dimensionality-reduction technique is validated both theoretically and experimentally. Then, through kernel Fisher discriminant analysis (KFDA), the multiple kernel learning problem is treated in the proposed framework, and a novel algorithm, which iteratively optimizes the parameters of the classification function and kernel combination coefficients, is proposed. The algorithm is experimentally shown to be comparable to or outperforms KFDA for large-scale benchmark data sets, and comparable to other multiple kernel learning techniques on the yeast protein function annotation task.


Acid-responsive H2-releasing Fe nanoparticles for safe and effective cancer therapy.
Hydrogen therapy is an emerging and promising strategy for treatment of inflammation-related diseases owing to the excellent bio-safety of hydrogen molecules (H2), but is facing a challenge that the H2 concentration at the local disease site is hardly accumulated because of its high diffusibility and low solubility, limiting the efficacy of hydrogen therapy. Herein, we propose a nanomedicine strategy of imaging-guided tumour-targeted delivery and tumour microenvironment-triggered release of H2 to address this issue, and develop a kind of biocompatible carboxymethyl cellulose (CMC)-coated/stabilized Fe (Fe@CMC) nanoparticle with photoacoustic imaging (PAI), tumour targeting and acid responsive hydrogen release properties for cancer therapy. The Fe@CMC nanoparticles have demonstrated high intratumoural accumulation capability, high acid responsiveness, excellent PAI performance, selective cancer-killing effect and high bio-safety in vitro and in vivo. Effective inhibition of tumour growth is achieved by intravenous injection of the Fe@CMC nanoparticles, and the selective anti-cancer mechanism of Fe@CMC is discovered to be originated from the energy metabolism homeostasis regulatory function of the released H2. The proposed nanomedicine-mediated hydrogen therapy strategy will open a new window for precise, high-efficacy and safe cancer treatment.


Gender differences in joint biomechanics during walking: normative study in young adults.
The effect of gender on specific joint biomechanics during gait has been largely unexplored. Given the perceived, subjective, and temporal differences in walking between genders, we hypothesized that quantitative analysis would reveal specific gender differences in joint biomechanics as well. Sagittal kinematic (joint motion) and kinetic (joint torque and power) data from the lower limbs during walking were collected and analyzed in 99 young adult subjects (49 females), aged 20 to 40 years, using an optoelectronic motion analysis and force platform system. Kinetic data were normalized for both height and weight. Female and male data were compared graphically and statistically to assess differences in all major peak joint kinematic and kinetic values. Females had significantly greater hip flexion and less knee extension before initial contact, greater knee flexion moment in pre-swing, and greater peak mechanical joint power absorption at the knee in pre-swing (P < 0.0019 for each parameter). Other differences were noted (P < 0.05) that were not statistically significant when accounting for multiple comparisons. These gender differences may provide new insights into walking dynamics and may be important for both clinical and research studies in motivating the development of separate biomechanical reference databases for males and females.


The RNAi Inheritance Machinery of Caenorhabditis elegans.
Gene silencing mediated by dsRNA (RNAi) can persist for multiple generations in Caenorhabditis elegans (termed RNAi inheritance). Here we describe the results of a forward genetic screen in C. elegans that has identified six factors required for RNAi inheritance: GLH-1/VASA, PUP-1/CDE-1, MORC-1, SET-32, and two novel nematode-specific factors that we term here (heritable RNAi defective) HRDE-2 and HRDE-4 The new RNAi inheritance factors exhibit mortal germline (Mrt) phenotypes, which we show is likely caused by epigenetic deregulation in germ cells. We also show that HRDE-2 contributes to RNAi inheritance by facilitating the binding of small RNAs to the inheritance Argonaute (Ago) HRDE-1 Together, our results identify additional components of the RNAi inheritance machinery whose conservation provides insights into the molecular mechanism of RNAi inheritance, further our understanding of how the RNAi inheritance machinery promotes germline immortality, and show that HRDE-2 couples the inheritance Ago HRDE-1 with the small RNAs it needs to direct RNAi inheritance and germline immortality.


Obesity and symptoms and quality of life indicators of individuals with disabilities.
Health risks of obesity are well known, but effects of obesity on health-related quality of life (HRQOL) have not been well-studied in people with physical disabilities. We examined the association between obesity and HRQOL in people with disabilities relative to the general US population. We hypothesized (a) overall, individuals with disabilities will report worse HRQOL than the general US population and (b) obese individuals with disabilities will report worse HRQOL than non-obese individuals. Individuals with muscular dystrophy, multiple sclerosis, post-polio syndrome, and spinal cord injury (N = 1849) completed Patient Reported Outcomes Measurement Information System (PROMIS) measures of fatigue, pain interference, physical and social function, depression, sleep disturbance, and sleep-related impairment. Participants were classified as obese or non-obese based on self-reported weight and height (BMI) and/or waist circumference (WC). PROMIS T-scores were compared to norms and between obesity groups. Mean BMI was 26.4 kg/m(2) with 23.4% classified as obese. Mean WC was 37.5 inches (males) and 34.0 inches (females); 26.4% reported abdominal obesity. Based on BMI and/or WC, 33.3% (n = 616) were classified obese. Compared to PROMIS norms, obese individuals reported worse HRQOL on all domains (p < 0.0001). Compared to non-obese individuals, obese individuals reported worse functioning on all domains except depression (p < 0.01). Obese individuals with MS and MD reported worse outcomes than non-obese counterparts. Obesity in people with physical disabilities is associated with poorer HRQOL. More research is needed to inform clinicians in identifying obese patients and helping them achieve healthy weight, reduce symptom burden, and improve QOL.


Daily fluctuation in negative affect for family caregivers of individuals with dementia.
The study examined associations of intrinsic fluctuation in daily negative affect (i.e., depression and anger) with adult day service (ADS) use, daily experiences, and other caregiving characteristics. This was an 8-day diary of 173 family caregivers of individuals with dementia. Multilevel models with common within-person variance were fit first to show average associations between daily stressors and mean level of daily affect. Then multilevel models with heterogeneous within-person variance were fit to test the hypotheses on associations between ADS use, daily experiences, and intrinsic fluctuation in daily affect. The study showed that, when the sum of ADS days was greater than average, there was a stabilizing effect of ADS use on caregivers' within-person fluctuation in negative affect. Moreover, fewer daily stressors and greater-than-average daily care-related stressors, more positive events, not being a spouse, greater-than-average duration of caregiving, and less-than-average dependency of individuals with dementia on activities of daily living were associated with less fluctuation. Better sleep quality was associated with less intrinsic fluctuation in anger; and younger age and more years of education were associated with less intrinsic fluctuation in daily depression. Because emotional stability has been argued as an aspect of emotional well-being in the general populations, intrinsic fluctuation of emotional experience was suggested as an outcome of evidence-based interventions for family caregivers.


How Much Does Education Improve Intelligence? A Meta-Analysis.
Intelligence test scores and educational duration are positively correlated. This correlation could be interpreted in two ways: Students with greater propensity for intelligence go on to complete more education, or a longer education increases intelligence. We meta-analyzed three categories of quasiexperimental studies of educational effects on intelligence: those estimating education-intelligence associations after controlling for earlier intelligence, those using compulsory schooling policy changes as instrumental variables, and those using regression-discontinuity designs on school-entry age cutoffs. Across 142 effect sizes from 42 data sets involving over 600,000 participants, we found consistent evidence for beneficial effects of education on cognitive abilities of approximately 1 to 5 IQ points for an additional year of education. Moderator analyses indicated that the effects persisted across the life span and were present on all broad categories of cognitive ability studied. Education appears to be the most consistent, robust, and durable method yet to be identified for raising intelligence.


The patchwork mouse phenotype: implication for melanocyte replacement in the hair follicle.
Mice homozygous for the recessive patchwork (pwk) mutation are characterized by a variegated pigment pattern with a mixture of unpigmented and normally pigmented hairs. The pigmented hair bulbs contain functional melanocytes. By contrast, the unpigmented hair bulbs contain no melanocytes. This lack results from the death of melanoblasts in the hair follicle at the end of embryogenesis. Here, we report that melanoblasts and melanocytes are found in the epidermis of pwk/pwk mice. Furthermore, these epidermal pigment cells are able to colonize new hair follicles after skin wounding. Despite the presence of epidermal pigment cells with a colonization potential, a follicle that had produced an unpigmented hair produces a new unpigmented hair during the successive hair growth cycles. This hair color continuity is also true for the pigmented hair follicles. Thus, in normal conditions, the hair acts as an independent functional unit as regards its pigment cells population.


Sleep Duration, Sleep Quality, and Markers of Subclinical Arterial Disease in Healthy Men and Women.
Short and long sleep duration are associated with increased risk of clinical cardiovascular events, but the association between sleep duration and subclinical cardiovascular disease is not well established. We examined the association between sleep duration and sleep quality with coronary artery calcification (CAC) and with brachial-ankle pulse wave velocity (PWV) in a large sample of young and middle-aged asymptomatic adults. We conducted a cross-sectional study of adult men and women who underwent a health checkup examination, including assessment of sleep duration and quality and coupled with either CAC (n=29 203) or brachial-ankle PWV (n=18 106). The multivariate-adjusted CAC score ratios (95% confidence interval) comparing sleep durations of ≤5, 6, 8, and ≥9 hours with 7 hours of sleep were 1.50 (1.17-1.93), 1.34 (1.10-1.63), 1.37 (0.99-1.89), and 1.72 (0.90-3.28), respectively (P for quadratic trend=0.002). The corresponding average differences in brachial-ankle PWV were 6.7 (0.75-12.6), 2.9 (-1.7 to 7.4), 10.5 (4.5-16.5), and 9.6 (-0.7 to 19.8) cm/s, respectively (P for quadratic trend=0.019). Poor subjective sleep quality was associated with CAC in women but not in men, whereas the association between poor subjective sleep quality and brachial-ankle PWV was stronger in men than in women. In this large study of apparently healthy men and women, extreme sleep duration and poor subjective sleep quality were associated with increased prevalence of CAC and higher PWV. Our results underscore the importance of an adequate quantity and quality of sleep to maintain cardiovascular health.


Energy cost of common activities in children and adolescents.
The Compendium of Energy Expenditures for Youth assigns MET values to a wide range of activities. However, only 35% of activity MET values were derived from energy cost data measured in youth; the remaining activities were estimated from adult values. To determine the energy cost of common activities performed by children and adolescents and compare these data to similar activities reported in the compendium. Thirty-two children (8-11 years old) and 28 adolescents (12-16 years) completed 4 locomotion activities on a treadmill (TRD) and 5 age-specific activities of daily living (ADL). Oxygen consumption was measured using a portable metabolic analyzer. In children, measured METs were significantly lower than compendium METs for 3 activities [basketball, bike riding, and Wii tennis (1.1-3.5 METs lower)]. In adolescents, measured METs were significantly lower than compendium METs for 4 ADLs [basketball, bike riding, board games, and Wii tennis (0.3-2.5 METs lower)] and 3 TRDs [2.24 m · s(-1), 1.56 m · s(-1), and 1.34 m · s(-1) (0.4-0.8 METs lower)]. The Compendium of Energy Expenditures for Youth is an invaluable resource to applied researchers. Inclusion of empirically derived data would improve the validity of the Compendium of Energy Expenditures for Youth.


HTA of medical devices: Challenges and ideas for the future from a European perspective.
Health Technology Assessment (HTA) of Medical devices (MDs) and MD-based procedures can be challenging due to the unique features and particularities of this group of technologies, such as device-operator interaction. The aim of this study was to (1) clarify, and supplement earlier findings on European HTA institutions' structural, procedural and methodological characteristics with regard to the assessment of MDs and to (2) capture the institutions' perceptions regarding challenges and future trends. Semi-structured telephone interviews with 16 representatives from leading European HTA institutions were performed between April and July 2015. Summative and directed content analysis was used for the analysis, which is reported according to the COREQ checklist. Findings from the analysis of the interviews were manifold and partially confirmed what has been noted in the literature (e.g. scarce evidence; identifying relevant studies challenging due to more incremental innovations). Additional themes emerged that can be important for future considerations by HTA institutions and policy-makers alike (e.g. areas for future research; need for specific tools). The collective opinion of 16 European HTA institutions on MD evaluation could provide ideas to ameliorate the current regulatory situation beyond the modified EU regulation and start broader, more in-depth methodological discussions around the issue. Interviewed experts seem to agree that new approaches such as coverage with evidence development as some countries already introduced could help to overcome the problem with scarce evidence.


Toxicological response of Sprague Dawley rats from inhalation exposure to perfluorooctane sulfonyl fluoride (POSF).
Perfluorooctane sulfonyl fluoride (POSF) was a volatile starting material in the production of perfluorooctane sulfonate (PFOS), a stable surfactant that has been extensively studied due to its ubiquitous environmental distribution and slow clearance in humans. Because the inhalation toxicity of POSF on repeated exposure has not been previously reported, the current study evaluated the inhalation toxicity of POSF at 30, 100, and 300ppm (v/v) in rats for up to 13 weeks with a four-week recovery period. The extent of PFOS formation was also measured because POSF hydrolyzed to form PFOS. In addition, detailed urinalysis and examination of the urinary bladder were included to determine if factors associated with the development of bladder cancer were present. Exposure to POSF at 300ppm was associated with reduction in body weight-gain, necrosis of laryngeal cartilage, increased lung and bronchi weight with septal thickening, and changes in alveolar macrophages. The microscopic observations in larynx and lung are consistent with likely hydrolysis of POSF to form hydrogen fluoride (HF). Exposure to POSF at 100 and 300ppm was associated with increased relative liver weight, hepatocellular hypertrophy (except for females exposed to 100ppm POSF), and lowering of serum cholesterol (male only). After 13 weeks of exposure to 30, 100, or 300ppm POSF, serum PFOS concentration approximated 7, 35, or 100μg/mL, respectively. Approximately 0.1% of inhaled POSF was converted to PFOS. No changes indicative of bladder effects were observed in these rats exposed to POSF at any dose.


De novo mutations in epileptic encephalopathies.
Epileptic encephalopathies are a devastating group of severe childhood epilepsy disorders for which the cause is often unknown. Here we report a screen for de novo mutations in patients with two classical epileptic encephalopathies: infantile spasms (n = 149) and Lennox-Gastaut syndrome (n = 115). We sequenced the exomes of 264 probands, and their parents, and confirmed 329 de novo mutations. A likelihood analysis showed a significant excess of de novo mutations in the ∼4,000 genes that are the most intolerant to functional genetic variation in the human population (P = 2.9 × 10(-3)). Among these are GABRB3, with de novo mutations in four patients, and ALG13, with the same de novo mutation in two patients; both genes show clear statistical evidence of association with epileptic encephalopathy. Given the relevant site-specific mutation rates, the probabilities of these outcomes occurring by chance are P = 4.1 × 10(-10) and P = 7.8 × 10(-12), respectively. Other genes with de novo mutations in this cohort include CACNA1A, CHD2, FLNA, GABRA1, GRIN1, GRIN2B, HNRNPU, IQSEC2, MTOR and NEDD4L. Finally, we show that the de novo mutations observed are enriched in specific gene sets including genes regulated by the fragile X protein (P < 10(-8)), as has been reported previously for autism spectrum disorders.


PSYCHOMETRIC PROPERTIES OF THE ALABAMA PARENTING QUESTIONNAIRE ADAPTED TO FAMILIES OF CHILEAN PRESCHOOLERS.
The Alabama Parenting Questionnaire (APQ) is a well-known tool to assess empirically identified aspects of positive and negative parenting practices. This study evaluates the psychometric properties of an adapted version of the APQ for its use with parents of children between 2 and 6 years of age in Chile. The participants were 557 parents of children aged 2 to 6 years. A confirmatory factor analysis showed that the best fit was obtained by a four-factor model (positive reinforcement, parental involvement, inconsistency of disciplinary practices, and punitive practices). The invariance analysis for this model by sex and social composition was positive. Disciplinary inconsistency and punitive practices were correlated with externalized and internalized behaviors in children. Results suggest that this adaptation of the APQ may result in a useful tool for clinical and research purposes in this age group.


Depot-specific differences in adipose tissue gene expression in lean and obese subjects.
Intra-abdominal and subcutaneous adipose tissue display important metabolic differences that underlie the association of visceral, but not subcutaneous, fat with obesity-related cardiovascular and metabolic problems. Because the molecular mechanisms contributing to these differences are not yet defined, we compared by reverse transcription-polymerase chain reaction the expression of 15 mRNAs that encode proteins of known importance in adipocyte function in paired omental and subcutaneous abdominal biopsies. No difference in mRNA expression between omental and subcutaneous adipose tissue was observed for hormone sensitive lipase, lipoprotein lipase, 6-phosphofructo-1-kinase, insulin receptor substrate 1, p85alpha regulatory subunit of phosphatidylinositol-3-kinase, and Rad. Total amount of insulin receptor expression was significantly higher in omental adipose tissue. Most of this increase was accounted for by expression of the differentially spliced insulin receptor lacking exon 11, which is considered to transmit the insulin signal less efficiently than the insulin receptor with exon 11. Perhaps consistent with a less efficient insulin signaling, a twofold reduction in GLUT4, glycogen synthase, and leptin mRNA expression was observed in omental adipose tissue. Finally peroxisome proliferator activated receptor-gamma (PPAR-gamma) mRNA levels were significantly lower in visceral adipose tissue in subjects with a BMI <30 kg/m2, but not in obese subjects, indicating that relative PPAR-gamma expression is increased in omental fat in obesity. This suggests that altered expression of PPAR-gamma might play a role in adipose tissue distribution and expansion.


In vivo antitumoral activity of stem pineapple (Ananas comosus) bromelain.
Stem bromelain (EC 188.8.131.52) is a major cysteine proteinase, isolated from pineapple ( Ananas comosus) stem. Its main medicinal use is recognized as digestive, in vaccine formulation, antitumoral and skin debrider for the treatment of burns. To verify the identity of the principle in stem fractions responsible for the antitumoral effect, we isolated bromelain to probe its pharmacological effects. The isolated bromelain was obtained from stems of adult pineapple plants by buffered aqueous extraction and cationic chromatography. The homogeneity of bromelain was confirmed by reverse phase HPLC, SDS-PAGE and N-terminal sequencing. The in vivo antitumoral/antileukemic activity was evaluated using the following panel of tumor lines: P-388 leukemia, sarcoma (S-37), Ehrlich ascitic tumor (EAT), Lewis lung carcinoma (LLC), MB-F10 melanoma and ADC-755 mammary adenocarcinoma. Intraperitoneal administration of bromelain (1, 12.5, 25 mg/kg), began 24 h after tumor cell inoculation in experiments in which 5-fluorouracil (5-FU, 20 mg/kg) was used as positive control. The antitumoral activity was assessed by the survival increase (% survival index) following various treatments. With the exception of MB-F10 melanoma, all other tumor-bearing animals had a significantly increased survival index after bromelain treatment. The largest increase ( approximately 318 %) was attained in mice bearing EAT ascites and receiving 12.5 mg/kg of bromelain. This antitumoral effect was superior to that of 5-FU, whose survival index was approximately 263 %, relative to the untreated control. Bromelain significantly reduced the number of lung metastasis induced by LLC transplantation, as observed with 5-FU. The antitumoral activity of bromelain against S-37 and EAT, which are tumor models sensitive to immune system mediators, and the unchanged tumor progression in the metastatic model suggests that the antimetastatic action results from a mechanism independent of the primary antitumoral effect.


Transcutaneous Auricular Vagus Nerve Stimulation.
Invasive vagus nerve stimulation (VNS) is an approved treatment for drug-resistant epilepsy. Besides recognized clinical efficacy in about 60% of patients, there are major drawbacks such as invasiveness and common side effects including hoarseness, sore throat, shortness of breath, and coughing. Invasive VNS applies electrical stimulation to the left cervical branch of the vagus nerve and excites thick-myelinated afferent nerve fibers. Peripheral vagus nerve afferent volley initiates brainstem activity in the nucleus of the solitary tract and provokes typical brainstem and cerebral activation patterns that mediate the anticonvulsive mode of action. Whereas invasive VNS is an established neuromodulatory treatment in drug-resistant epilepsy, transcutaneous VNS (tVNS) of the auricular branch of the vagus nerve is suggested to be an alternative access path to the same neuronal network without invasiveness. Preclinical and clinical studies indicate that especially the cymba conchae of the auricle is selectively supplied by the auricular branch of the vagus nerve. Recent anatomical data demonstrate existence and quantity of thick-myelinated afferent nerve fibers of the left auricular branch of the vagus nerve that carries 21% of thick-myelinated afferent nerve fibers counted in the left thoracic vagus nerve in humans. Projection of auricular branch of the vagus nerve afferents from the auricle to the nucleus of the solitary tract is known from histochemical and electrophysiological experiments in rodents and confirmed in humans by functional imaging. Cerebral activation patterns triggered by invasive and tVNS resemble each other in appearance. Clinical trials in patients address safety and performance of tVNS and provide evidence for application in drug-resistant epilepsy.


A Recurrent Probabilistic Neural Network with Dimensionality Reduction Based on Time-series Discriminant Component Analysis.
This paper proposes a probabilistic neural network (NN) developed on the basis of time-series discriminant component analysis (TSDCA) that can be used to classify high-dimensional time-series patterns. TSDCA involves the compression of high-dimensional time series into a lower dimensional space using a set of orthogonal transformations and the calculation of posterior probabilities based on a continuous-density hidden Markov model with a Gaussian mixture model expressed in the reduced-dimensional space. The analysis can be incorporated into an NN, which is named a time-series discriminant component network (TSDCN), so that parameters of dimensionality reduction and classification can be obtained simultaneously as network coefficients according to a backpropagation through time-based learning algorithm with the Lagrange multiplier method. The TSDCN is considered to enable high-accuracy classification of high-dimensional time-series patterns and to reduce the computation time taken for network training. The validity of the TSDCN is demonstrated for high-dimensional artificial data and electroencephalogram signals in the experiments conducted during the study.


Preliminary efficacy and feasibility of embedding high intensity interval training into the school day: A pilot randomized controlled trial.
Current physical activity and fitness levels among adolescents are low, increasing the risk of chronic disease. Although the efficacy of high intensity interval training (HIIT) for improving metabolic health is now well established, it is not known if this type of activity can be effective to improve adolescent health. The primary aim of this study is to assess the effectiveness and feasibility of embedding HIIT into the school day. A 3-arm pilot randomized controlled trial was conducted in one secondary school in Newcastle, Australia. Participants (n = 65; mean age = 15.8(0.6) years) were randomized into one of three conditions: aerobic exercise program (AEP) (n = 21), resistance and aerobic exercise program (RAP) (n = 22) and control (n = 22). The 8-week intervention consisted of three HIIT sessions per week (8-10 min/session), delivered during physical education (PE) lessons or at lunchtime. Assessments were conducted at baseline and post-intervention to detect changes in cardiorespiratory fitness (multi-stage shuttle-run), muscular fitness (push-up, standing long jump tests), body composition (Body Mass Index (BMI), BMI-z scores, waist circumference) and physical activity motivation (questionnaire), by researchers blinded to treatment allocation. Intervention effects for outcomes were examined using linear mixed models, and Cohen's d effect sizes were reported. Participants in the AEP and RAP groups had moderate intervention effects for waist circumference (p = 0.024), BMI-z (p = 0.037) and BMI (not significant) in comparison to the control group. A small intervention effect was also evident for cardiorespiratory fitness in the RAP group.


Deficiency of type I interferon contributes to Sle2-associated component lupus phenotypes.
Studies in mice and humans have implicated type I interferon (IFN-I) in the pathogenesis of lupus. Given that the locus for IFN-I is positioned within the Sle2 murine lupus susceptibility interval on chromosome 4, we undertook this study to investigate whether differences in IFN-I levels might potentially contribute to the phenotypes ascribed to this locus. IFN-I, anti-IFN-I, isotype control antibody, or phosphate buffered saline was administered to C57BL/6 and B6.Sle2 mice, and the serologic and cellular phenotypes were studied. In addition, B6.Sle2 mice were examined for structural and expression polymorphisms in the IFN-I gene. In both B6.Sle2 congenic mice and C57BL/6 control mice, antibody-mediated blockade of IFN-I augmented serum autoantibody levels and boosted B1a cell numbers. Administration of IFN-I ameliorated these 2 features previously attributed to this disease locus. Importantly, compared with B6 controls, B6.Sle2 mice had reduced levels of IFN-I in their sera and cell culture supernatants, following stimulation. Although several sequence polymorphisms were noted in the Sle2 alleles of various IFN-I genes, it was not established whether any of the noted sequence variations were causally related to the observed phenotypes. Unexpectedly, reduction of IFN-I levels reproduced the serologic and cellular phenotypes previously associated with the Sle2 lupus susceptibility interval. Placing these findings in the context of other studies, the effect of IFN-I on systemic autoimmunity appears to be far more complex than originally perceived.


Inhibition of striatal dopamine release by CB1 receptor activation requires nonsynaptic communication involving GABA, H2O2, and KATP channels.
The main psychoactive component of marijuana, Delta9-tetrahydrocannabinol (THC), acts in the CNS via type 1 cannabinoid receptors (CB1Rs). The behavioral consequences of THC or synthetic CB1R agonists include suppression of motor activity. One explanation for movement suppression might be inhibition of striatal dopamine (DA) release by CB1Rs, which are densely localized in motor striatum; however, data from previous studies are inconclusive. Here we examined the effect of CB1R activation on locally evoked DA release monitored with carbon-fiber microelectrodes and fast-scan cyclic voltammetry in striatal slices. Consistent with previous reports, DA release evoked by a single stimulus pulse was unaffected by WIN55,212-2, a cannabinoid receptor agonist. However, when DA release was evoked by a train of stimuli, WIN55,212-2 caused a significant decrease in evoked extracellular DA concentration ([DA]o), implicating the involvement of local striatal circuitry, with similar suppression seen in guinea pig, rat, and mouse striatum. Pulse-train evoked [DA]o was not altered by either AM251, an inverse CB1R agonist, or VCHSR1, a neutral antagonist, indicating the absence of DA release regulation by endogenous cannabinoids with the stimulation protocol used. However, both CB1R antagonists prevented and reversed suppression of evoked [DA]o by WIN55,212-2. The effect of WIN55,212-2 was also prevented by picrotoxin, a GABAA receptor antagonist, and by catalase, a metabolizing enzyme for hydrogen peroxide (H2O2). Furthermore, blockade of ATP-sensitive K+ (KATP) channels by tolbutamide or glybenclamide prevented the effect of WIN55,212-2 on DA release. Together, these data indicate that suppression of DA release by CB1R activation within striatum occurs via a novel nonsynaptic mechanism that involves GABA release inhibition, increased generation of the diffusible messenger H2O2, and activation of KATP channels to inhibit DA release. In addition, the findings suggest a possible physiological substrate for the motor effects of cannabinoid agonist administration.


Communication about sexual issues: mothers, fathers, and friends.
To identify the content, characteristics, and comfort level of discussions about sexuality held between mothers and their early adolescent children and to determine the extent to which the conversations predicted sexual values and initiation of sexual intercourse of the adolescent. This was part of a larger study to evaluate the impact of personal family characteristics on human immunodeficiency virus risk-reduction behavior among low-income predominately African-American adolescents, ages 13-15 years. Adolescents attending a metropolitan community-based afterschool program and their mothers were invited to participate in a 1-hour interview. Mothers and adolescents were interviewed separately. The interview included questions about the type of information related to sexuality that adolescents discussed with mothers, fathers, and friends; in addition, mothers were asked what topics they discussed with their adolescents. Four hundred five adolescents and 382 mothers participated. Some mothers had more than one adolescent in the study. The results showed that both male and female adolescents were more likely to discuss sexual topics with their mothers than their fathers. Male adolescents were more likely than female adolescents to discuss sex-based topics with their fathers. Both male and female adolescents were less likely to discuss sex-based topics with their friends than with their mothers, but more likely to discuss these topics with their friends than their fathers. Content of conversations of male adolescents was fairly consistent among mothers, fathers, and friends, and sexually transmitted disease/acquired immune deficiency syndrome and condom use were popular topics of discussion. Female adolescents tended to talk about the menstrual cycle with their mothers, sexual abstinence with their fathers, and sexual intercourse with their friends. Adolescents who reported a greater number of topics discussed with their mothers were more likely not to have initiated sexual intercourse and to have conservative values, whereas adolescents who reported a greater number of topics discussed with their friends were more likely to report the initiation of intercourse and more "liberal" sexual values. Both male and female adolescents were most comfortable discussing sexual issues with their friends. Male adolescents were less comfortable talking to mothers, but more comfortable talking to their fathers than were females. Mothers were likely to report feeling very comfortable talking about almost all discussion areas. Fathers' comfort level was not measured, as they were not directly questioned. Early adolescence (13-15 years old) is characterized by more sex-based discussions with mothers than friends or fathers. Daughters and sons discuss different topics with their fathers, although discussion by both genders with fathers is limited. If an adolescent talks more with the mother about sexual issues than with friends, he/she is less likely to initiate sexual intercourse and more likely to have conservative values. This points to the importance of fostering good communication and comfort between parents and adolescents about sexual issues.


Evening primrose oil and celecoxib inhibited pathological angiogenesis, inflammation, and oxidative stress in adjuvant-induced arthritis: novel role of angiopoietin-1.
Rheumatoid arthritis is a chronic inflammatory disease characterized by overproduction of inflammatory mediators along with undermined oxidative defensive mechanisms. Pathological angiogenesis was found to play a critical role in the progression of this disease. The current study was carried out to evaluate the anti-angiogenic, anti-inflammatory, and anti-oxidant effects of evening primrose oil (EPO), rich in gamma linolenic acid (GLA), either alone or in combination with aspirin or celecoxib, on adjuvant-induced arthritis. Arthritis was induced by subcutaneous injection of complete Freund's adjuvant (CFA) in the right hind paw of male albino rats. All treatments were administered orally from day 0 (EPO, 5 g/kg b.w.) or day 4 (celecoxib, 5 mg/kg; aspirin, 150 mg/kg) till day 27 after CFA injection. In the arthritic group, the results revealed significant decrease in the body weight and increase in ankle circumference, plasma angiopoietin-1 (ANG-1) and tumor necrosis factor-alpha (TNF-α) levels. Anti-oxidant status was suppressed as manifested by significant decline in reduced glutathione content along with decreased enzymatic activity of superoxide dismutase and increased lipid peroxidation. Oral administration of EPO exerted normalization of body weight, ANG-1, and TNF-α levels with restoration of activity as shown by reduced malondialdehyde levels. Moreover, histopathological examination demonstrated that EPO significantly reduced the synovial hyperplasia and inflammatory cells invasion in joint tissues, an effect that was enhanced by combination with aspirin or celecoxib. The joint use of GLA-rich natural oils, which possess anti-angiogenic, anti-inflammatory, and anti-oxidant activities, with traditional analgesics represents a promising strategy to restrain the progression of rheumatoid arthritis.


Oxidative stress and mitochondrial dysfunction in fibromyalgia.
Fibromyalgia (FM) is a chronic pain syndrome with unknown etiology and pathophysiology. Recent studies have shown some evidence demonstrating that oxidative stress may have a role in the pathophysiology of FM. Furthermore, it is controversial the role of mitochondria in the oxidant imbalance documented in FM. Signs and symptoms associated with muscular alteration and mitochondrial dysfunction, including oxidative stress, have been observed in patients with FM. To this respect, Coenzyme Q10 (CoQ10) deficiency, an essential electron carrier in the mitochondrial respiratory chain and a strong antioxidant, alters mitochondria function and mitochondrial respiratory complexes organization and leading to increased ROS generation. Recently have been showed CoQ10 deficiency in blood mononuclear cells in FM patients, so if the hypothesis that mitochondrial dysfunction is the origin of oxidative stress in FM patients is demonstrated, could help to understand the complex pathophysiology of this disorder and may lead to development of new therapeutic strategies for prevention and treatment of this disease.


Inhibition of endoglin-GIPC interaction inhibits pancreatic cancer cell growth.
Endoglin, a 180-kDa disulfide-linked homodimeric transmembrane receptor protein mostly expressed in tumor-associated endothelial cells, is an endogenous binding partner of GAIP-interacting protein, C terminus (GIPC). Endoglin functions as a coreceptor of TβRII that binds TGFβ and is important for vascular development, and consequently has become a compelling target for antiangiogenic therapies. A few recent studies in gastrointestinal stromal tumor (GIST), breast cancer, and ovarian cancer, however, suggest that endoglin is upregulated in tumor cells and is associated with poor prognosis. These findings indicate a broader role of endoglin in tumor biology, beyond angiogenic effects. The goal of our current study is to evaluate the effects of targeting endoglin in pancreatic cancer both in vitro and in vivo. We analyzed the antiproliferative effect of both RNAi-based and peptide ligand-based inhibition of endoglin in pancreatic cancer cell lines, the latter yielding a GIPC PDZ domain-targeting lipopeptide with notable antiproliferative activity. We further demonstrated that endoglin inhibition induced a differentiation phenotype in the pancreatic cancer cells and sensitized them against conventional chemotherapeutic drug gemcitabine. Most importantly, we have demonstrated the antitumor effect of both RNAi-based and competitive inhibitor-based blocking of endoglin in pancreatic cancer xenograft models in vivo. To our knowledge, this is the first report exploring the effect of targeting endoglin in pancreatic cancer cells.


Study protocol: a phase III randomised, double-blind, parallel arm, stratified, block randomised, placebo-controlled trial investigating the clinical effect and cost-effectiveness of sertraline for the palliative relief of breathlessness in people with chronic breathlessness.
Breathlessness remains a highly prevalent and distressing symptom for many patients with progressive life-limiting illnesses. Evidence-based interventions for chronic breathlessness are limited, and there is an ongoing need for high-quality research into developing management strategies for optimal palliation of this complex symptom. Previous studies have suggested that selective serotonin reuptake inhibitors such as sertraline may have a role in reducing breathlessness. This paper presents the protocol for a large, adequately powered randomised study evaluating the use of sertraline for chronic breathlessness in people with progressive life-limiting illnesses. A total of 240 participants with modified Medical Research Council Dyspnoea Scale breathlessness of level 2 or higher will be randomised to receive either sertraline or placebo for 28 days in this multisite, double-blind study. The dose will be titrated up every 3 days to a maximum of 100 mg daily. The primary outcome will be to compare the efficacy of sertraline with placebo in relieving the intensity of worst breathlessness as assessed by a 0-100 mm Visual Analogue Scale. A number of other outcome measures and descriptors of breathlessness as well as caregiver assessments will also be recorded to ensure adequate analysis of participant breathlessness and to allow an economic analysis to be performed. Participants will also be given the option of continuing blinded treatment until either study data collection is complete or net benefit ceases. Appropriate statistical analysis of primary and secondary outcomes will be used to describe the wealth of data obtained. Ethics approval was obtained at all participating sites. Results of the study will be submitted for publication in peer-reviewed journals and the key findings presented at national and international conferences. ACTRN12610000464066.


A Quantitative Analysis of Undisclosed Conflicts of Interest in Pharmacology Textbooks.
Disclosure of potential conflicts of interest (CoI) is a standard practice for many biomedical journals but not for educational materials. The goal of this investigation was to determine whether the authors of pharmacology textbooks have undisclosed financial CoIs and to identify author characteristics associated with CoIs. The presence of potential CoIs was evaluated by submitting author names (N = 403; 36.3% female) to a patent database (Google Scholar) as well as a database that reports on the compensation ($USD) received from 15 pharmaceutical companies (ProPublica's Dollars for Docs). All publications (N = 410) of the ten highest compensated authors from 2009 to 2013 and indexed in Pubmed were also examined for disclosure of additional companies that the authors received research support, consulted, or served on speaker's bureaus. A total of 134 patents had been awarded (Maximum = 18/author) to textbook authors. Relative to DiPiro's Pharmacotherapy: A Pathophysiologic Approach, contributors to Goodman and Gilman's Pharmacological Basis of Therapeutics and Katzung's Basic and Clinical Pharmacology were more frequently patent holders (OR = 6.45, P < .0005). Female authors were less likely than males to have > 1 patent (OR = 0.15, P < .0005). A total of $2,411,080 USD (28.3% for speaking, 27.0% for consulting, and 23.9% for research), was received by 53 authors (Range = $299 to $310,000/author). Highly compensated authors were from multiple fields including oncology, psychiatry, neurology, and urology. The maximum number of additional companies, not currently indexed in the Dollars for Docs database, for which an author had potential CoIs was 73. Financial CoIs are common among the authors of pharmacology and pharmacotherapy textbooks. Full transparency of potential CoIs, particularly patents, should become standard procedure for future editions of educational materials in pharmacology.