Influence of gender, obesity, and muscle lipase activity on intramyocellular lipids in sedentary individuals.
Obesity and type 2 diabetes are associated with elevated intramyocellular lipids (IMCLs) and insulin resistance. We tested the hypothesis that skeletal muscle lipases activity could influence IMCL content (including diacylglycerol and ceramides). The present study included 48 subjects with a wide range of age (19-68 yr) and body mass index (20-45 kg/m(2)) who underwent skeletal muscle biopsy, dual-energy x-ray absorptiometry and a hyperinsulinemic euglycemic clamp. Insulin sensitivity by hyperinsulinemic clamp, and intramyocellular triacylglycerol (IMTG), diacylglycerol (DAG), and ceramides content, and triacylglycerol and diacylglycerol hydrolase activities were measured in biopsies of vastus lateralis. IMCL was measured by (1)H-magnetic resonance spectroscopy in a subgroup of 25 subjects. Multivariate regression analyses were performed to identify the main predictors of IMCL. Body fat was the main predictor of IMTG independently of the method and the type of muscle; IMTG concentration was higher in females vs. males and obese vs. nonobese subjects. Muscle DAG and ceramides concentrations were elevated in obese and type 2 diabetic subjects and were not related to body fat and fasting free fatty acids, whereas a direct association with the ratio of diacylglycerol hydrolase to triacylglycerol hydrolase activity (an index of incomplete triacylglycerol hydrolysis) was observed, which explained 54 and 38% of the variance in DAG and ceramides (P < 0.001), respectively. DAG content was the main determinant of insulin resistance. These data suggest that intramyocellular DAG is an independent predictor of insulin resistance in humans and that its levels correlate with lipolytic enzymes activity in skeletal muscle but not with markers of adiposity.


Elective high frequency oscillatory ventilation versus conventional ventilation for acute pulmonary dysfunction in preterm infants.
Respiratory failure due to lung immaturity is a major cause of mortality in preterm infants. Although the use of intermittent positive pressure ventilation (IPPV) in neonates with respiratory failure saves lives, its use is associated with lung injury and chronic lung disease (CLD). Conventional IPPV is provided at 30-80 breaths per minute, while a newer form of ventilation called high frequency oscillatory ventilation (HFOV) provides 'breaths' at 10 - 15 cycles per second. This has been shown to result in less lung injury in experimental studies. The objective of this review is to determine the effect of the elective use of high frequency oscillatory ventilation (HFOV) as compared to conventional ventilation (CV) in preterm infants who are mechanically ventilated for respiratory distress syndrome (RDS), on the incidence of chronic lung disease, mortality and other complications associated with prematurity and assisted ventilation. Searches were made of the Oxford Database of Perinatal Trials, MEDLINE, EMBASE, previous reviews including cross references, abstracts, conferences and symposia proceedings, expert informants, journal hand searching by the Cochrane Collaboration, mainly in the English language. The search was updated in April 2007. Randomised controlled trials comparing HFOV and CV in preterm or low birth weight infants with pulmonary dysfunction, mainly due to RDS, who were given IPPV. Randomisation and commencement of treatment needed to be as soon as possible after the start of IPPV and usually in the first 12 hours of life. The methodological quality of each trial was independently reviewed by the various authors. The standard effect measures are relative risk (RR) and risk difference (RD). From 1/RD the number needed to treat (NNT) to produce one outcome were calculated. For all measures of effect, 95% confidence intervals were used. In subgroup analyses the 99% CIs are also given for summary RRs in the text. Meta-analysis was performed using a fixed effects model. Where heterogeneity was over 50%, the random effects RR is also given. Fifteen eligible studies of 3,585 infants were included. Meta-analysis comparing HFOV with CV revealed no evidence of effect on mortality at 28 - 30 days of age or at approximately term equivalent age. These results were consistent across studies and in subgroup analyses. The effect of HFOV on CLD in survivors at term equivalent gestational age was inconsistent across studies and the reduction was of borderline significance overall. Subgroups of trials showed a significant reduction in CLD with HFOV when high volume strategy for HFOV was used, when piston oscillators were used for HFOV, when lung protective strategies for CV were not used, when randomisation occurred at two to six hours of age, and when inspiratory:expiratory ratio of 1:2 was used for HFOV. In the meta-analysis of all trials, pulmonary air leaks occurred more frequently in the HFOV group. In some studies, short-term neurological morbidity with HFOV was found, but this effect was not statistically significant overall. The subgroup of two trials not using a high volume strategy with HFOV found increased rates of Grade 3 or 4 intraventricular haemorrhage and of periventricular leukomalacia. An adverse effect of HFOV on long-term neurodevelopment was found in one large trial but not in the five other trials that reported this outcome. The rate of retinopathy of prematurity is reduced overall in the HFOV group. There is no clear evidence that elective HFOV offers important advantages over CV when used as the initial ventilation strategy to treat preterm infants with acute pulmonary dysfunction. There may be a small reduction in the rate of CLD with HFOV use, but the evidence is weakened by the inconsistency of this effect across trials and the overall borderline significance. Future trials on elective HFOV should target those infants who are at most risk of CLD (extremely preterm infants), compare different strategies for generating HFOV and CV, and report important long-term neurodevelopmental outcomes.


[Laser microdissection for biology and medicine].
For routine extraction of DNA, RNA, proteins and metabolites, small tissue pieces are placed into lysing solution. These tissue pieces in general contain different cell types. For this reason, lysate contains components of different cell types, which complicates the interpretation of molecular analysis results. The laser microdissection allows overcoming this trouble. The laser microdissection is a method to procure tissue samples contained defined cell subpopulations, individual cells and even subsellular components under direct microscopic visualization. Collected samples can be undergone to different downstream molecular assays: DNA analysis, RNA transcript profiling, cDNA library generation and gene expression analysis, proteomic analysis and metabolite profiling. The laser microdissection has wide applications in oncology (research and routine), cellular and molecular biology, biochemistry and forensics. This paper reviews the principles of different laser microdissection instruments, examples of laser microdissection application and problems of sample preparation for laser microdissection.


[C7GC4]4 association into supra molecular i-motif structures.
The self-associative properties of cytidine-rich oligonucleotides into symmetrical i-motif tetramers give to these oligonucleotides the capacity of forming supramolecular structures (sms) that have potential applications in the nanotechnology domain. In order to facilitate sms formation, oligonucleotides containing two cytidine stretches of unequal length (C(n)XC(m)) separated by a non-cytidine spacer were synthesized. They were designed to associate into a tetramer including an i-motif core built by intercalation of the C.C(+) pairs of the longer C stretch with the two dangling non-intercalated strands of the shorter C stretch at each end. Gel filtration chromatography shows that the non-intercalated C-rich ends give to this structure the capacity of forming extremely stable sms. Using C(7)GC(4) as a model, we find that the sms formation rate varies as the oligonucleotide concentration and increases at high temperature. Competitively with the tetramer involved in sms elongation, C(n)XC(m) oligonucleotides form i-motif dimers that compete with sms elongation. The dimer stability is strongly reduced when the pH is moved away from the cytidine pK. This results in an equilibrium shift towards the tetramer and in the acceleration of the sms formation rate. The chromatograms of the sms formed by C(7)GC(4) indicate a broad distribution. In a 1.5 mM solution incubated at 37 degrees C, the equilibrium distribution is centered on a molecular weight corresponding to the assembly of nine tetramers and the upper limit corresponds to 80 tetramers. The lifetime of this structure is about 4 days at 40 degrees C, pH 4.6.


Multiplexing schemes for generic SNP genotyping assays.
Association studies in populations relate genomic variation among individuals with medical condition. Key to these studies is the development of efficient and affordable genotyping techniques. Generic genotyping assays are independent of the target SNPs and offer great flexibility in the genotyping process. Efficient use of such assays calls for identifying sets of SNPs that can be interrogated in parallel under constraints imposed by the genotyping technology. In this paper, we study problems arising in the design of genotyping experiments using generic assays. Our problem formulation deals with two main factors that affect the genotyping cost: the number of assays used and the number of PCR reactions required for sample preparation. We prove that the resulting computational problems are hard, but provide approximate and heuristic solutions to these problems. Our algorithmic approach is based on recasting the multiplexing problems as partitioning and packing problems on a bipartite graph. We tested our algorithmic approaches on an extensive collection of synthetic data and on data that was simulated using real SNP sequences. Our results show that the algorithms achieve near-optimal designs in many cases and demonstrate the applicability of generic assays to SNP genotyping.


Development of a microfluidics biosensor for agarose-bead immobilized Escherichia coli bioreporter cells for arsenite detection in aqueous samples.
Contamination with arsenic is a recurring problem in both industrialized and developing countries. Drinking water supplies for large populations can have concentrations much higher than the permissible levels (for most European countries and the United States, 10 μg As per L; elsewhere, 50 μg As per L). Arsenic analysis requires high-end instruments, which are largely unavailable in developing countries. Bioassays based on genetically engineered bacteria have been proposed as suitable alternatives but such tests would profit from better standardization and direct incorporation into sensing devices. The goal of this work was to develop and test microfluidic devices in which bacterial bioreporters could be embedded, exposed and reporter signals detected, as a further step towards a complete miniaturized bacterial biosensor. The signal element in the biosensor is a nonpathogenic laboratory strain of Escherichia coli, which produces a variant of the green fluorescent protein after contact to arsenite and arsenate. E. coli bioreporter cells were encapsulated in agarose beads and incorporated into a microfluidic device where they were captured in 500 × 500 μm(2) cages and exposed to aqueous samples containing arsenic. Cell-beads frozen at -20 °C in the microfluidic chip retained inducibility for up to a month and arsenic samples with 10 or 50 μg L(-1) could be reproducibly discriminated from the blank. In the 0-50 μg L(-1) range and with an exposure time of 200 minutes, the rate of signal increase was linearly proportional to the arsenic concentration. The time needed to reliably and reproducibly detect a concentration of 50 μg L(-1) was 75-120 minutes, and 120-180 minutes for a concentration of 10 μg L(-1).


Alpha-Glucosidase Inhibitors Alter Gut Microbiota and Ameliorate Collagen-Induced Arthritis.
Acarose is an anti-diabetic drug and exhibits anti-arthritic effects. We hypothesized that acarbose influences the gut microbiota to affect the course of arthritis and tested this hypothesis in a collagen-induced arthritis (CIA) murine model. Acarbose in drinking water was administered via gastric gavage started prior to or at the time of CIA induction. Gut microbiota were evaluated with 16S rRNA gene sequencing from fecal pellets collected prior to arthritis induction, during onset of arthritis, and after treatment. Immune response was evaluated by measuring changes in T helper-17 (Th17) and T regulatory (Treg) cells in the spleen and intestine, as well as serum cytokine levels. Before induction of CIA, acarbose significantly reduced the incidence of arthritis and attenuated clinical severity of arthritis. The frequency of Th17 cells was significantly decreased in the intestinal lamina propria in acarbose treated mice. Mice that were treated with acarbose showed significantly increased CD4+CD25+Foxp3+ Treg cells with elevation of Helios and CCR6. A remarkable alteration in microbial community was observed in acarbose treated mice. Bacterial diversity and richness in mice with arthritis were significantly lower than those in acarbose treated groups. The frequency of Firmicutes was significantly reduced after arthritis onset but was restored after treatment with acarbose. The frequency of Lactobacillus, Anaeroplasma, Adlercreutzia, RF39 and Corynebacterium was significantly higher in control groups than in acarbose treated, while Oscillospira, Desulfovibrio and Ruminococcus enriched in acarbose treated group. Miglitol, another α-glucosidase inhibitor showed a similar but less potent anti-arthritic effect to that of acarbose. These data demonstrate that acarbose alleviated CIA through regulation of Th17/Treg cells in the intestinal mucosal immunity, which may have resulted from the impact of acarbose on gut microbial community. Inexpensive antidiabetic drugs with an excellent safety profile are potentially useful for managing rheumatoid arthritis.


Acute kidney injury during extracorporeal life support in cardiogenic shock: Does flow matter?
This study examines the role of extracorporeal life support flow in the development of acute kidney injury in cardiogenic shock. We performed a retrospective analysis of 465 patients placed on extracorporeal life support at our institution between January 2015 and December 2020 for cardiogenic shock. Flow index was calculated by dividing mean flow by body surface. Stages of acute kidney injury were determined according to Kidney Disease: Improving Global Outcomes (KDIGO) organization guidelines. There were 179 (38.5%) patients who developed acute kidney injury, 63.1% of which were classified as Stage 3--the only subgroup associated with 1-year mortality (hazard ratio = 2.03, p < .001). Risk of kidney injury increased up to a flow index of 1.6 L/min/m2, and kidney injury was more common among patients with flow index greater than 1.6 L/min/m2 (p = .034). Those with kidney injury had higher baseline lactate levels (4.4 vs 3.1, p = .04), and Stage 3 was associated wit higher baseline creatinine (p < .001). In our cohort, kidney injury was common and Stage 3 kidney injury was associated with worse outcomes compared to other stages. Low flow was not associated with increased risk of kidney injury. Elevated baseline lactate and creatinine among patients with acute kidney injury suggest underlying illness severity, rather than flow, may influence kidney injury risk.


Metastatic Melanoma: Treatment and Survival in the US after the Introduction of Ipilimumab and Vemurafenib.
The 5-year survival of metastatic melanoma is < 18%. Historically, treatment options were limited. In 2011, 2 new agents were approved. We re-abstracted the medical records of a random sample (n = 520) of metastatic melanoma patients who had been diagnosed in 2011 and reported to population-based registries in the U.S. We also queried their treating physicians. Factors associated with treatment and survival were assessed using logistic and Cox proportional hazards regressions, respectively. 21.4% of patients received no treatment, 20.8% received ipilimumab and 57.5% of patients with BRAF-positive tumors received vemurafenib/dabrafenib. Receipt of ipilimumab was less likely among patients of 75 years or older (vs. < 55 years: odds ratio (OR) 0.32; 95% confidence interval (CI) 0.15-0.66) and patients without private/military insurance. 46.8% of patients received BRAF testing. Receipt of BRAF testing was less likely among patients of 65 years or more and uninsured patients (OR 0.22; 95% CI 0.07-0.65). Receipt of ipilimumab was associated with better survival during the first 18 months after diagnosis (hazard ratio (HR) 0.66; 95% CI 0.51-0.84) and vemurafenib/dabrafenib with better survival during the first 10 months after diagnosis (HR 0.51; 95% CI 0.36-0.73). The initial dissemination of ipilimumab and vemurafenib/dabrafenib was limited. Additional research is needed to investigate the apparent lack of long-term survival benefit from these agents.


Validation of Whole-Slide Imaging for Histolopathogical Diagnosis: Current State.
Rapid advances in informatics and technological improvements have led to the development of high-throughput whole-slide imaging (WSI) scanners able to produce high-quality digital images, which allow achieving a correct diagnosis of the biopsies using virtual viewers. This technology is currently prepared to be introduced in the departments of pathology for routine diagnosis. The aim of this review is to analyze the current evidence regarding the use of WSI in primary or routine diagnosis in the different subspecialties of pathology. An increasing number of studies have shown almost perfect inter- and intraobserver agreement between the diagnoses obtained with WSI and the classical diagnoses based on conventional light microscopy. The only exception seems to be cytology, which still requires some technological development. Although validation studies are needed in some areas of pathology, growing evidence indicates that WSI is a reliable tool for routine diagnosis. Pathologists have a positive perception of the ergonomics of the workstations, the low magnification of WSI and the possibility of making annotations and measurements. WSI can be used from any device and anywhere, thereby providing great opportunities for teleconsultation. New technologies such as the recognition of histopathology patterns using image analysis may facilitate diagnosis and improve the reproducibility among pathologists in the future.


Ischemic Stroke: Treatments to Improve Neuronal Functional Recovery in vitro.
The understanding of the neurophysiological processes that occur in the areas that surround the core of a brain infarct is crucial for the creation of new therapies and treatments to improve neuronal recovery. The present study aims to demonstrate that both rodent and human neuronal networks lose their activity under low oxygen conditions and that electrical stimulation can increase the probability of recovery. Hypoxia was induced in rodent and human neurons and the effects of electrical stimulation were assessed in the rat cultures. The results obtained show that neuronal activation, in the form of electrical stimulation, has the potential to maintain the networks at higher levels of activity and, therefore, to improve cell survival. This study will open the way for new treatment strategies based on brain-stimulation to enhance neuronal recovery and will be of large relevance for patients, families, and society.


Flap complications and thrombophilia: an evidence-based model and cost analysis for preoperative screening.
Preoperative screening for thrombophilias in free flap candidates may be cost-effective. We developed a model for thrombogenic flap complications using reported thrombophilia prevalences and thromboembolic risk ratios, as well as free flap complication rates from our institution. We performed a break-even and incremental cost-effective ratio analysis for several screening and intervention scenarios. Our thrombotic free flap complication rate is 4.9%. A full thrombophilia screen breaks even when the cost of complication exceeds $57 000 per patient; a limited screen breaks even at $39 000, and a scenario in which all patients undergo chemoprophylaxis breaks even at $49 000. Incremental cost-effective ratio analyses estimate a cost per avoided flap complication of $33 638 for a full panel scenario, $15 617 for a limited panel scenario and $25 455 for an all therapy scenario. Our analyses show that preoperative thrombophilia screening may be a cost-effective measure for the prevention of free flap thrombotic complications.


From the Past to the Future: Considering the Value and Limits of Evolutionary Prediction.
The complex interplay of the multiple genetic processes of evolution and the ecological contexts in which they proceed frustrates detailed identification of many of the states of populations, both past and future, that may be of interest. Prediction of rates of adaptation, in the sense of change in mean fitness, into the future would, however, valuably inform expectations for persistence of populations, especially in our era of rapid environmental change. Heavy investment in genomics and other molecular tools has fueled belief that those approaches can effectively predict adaptation into the future. I contest this view. Genome scans display the genomic footprints of the effects of natural selection and the other evolutionary processes over past generations, but it remains problematic to predict future change in mean fitness via genomic approaches. Here, I advocate for a direct approach to prediction of rates of ongoing adaptation. Following an overview of relevant quantitative genetic approaches, I outline the promise of the fundamental theorem of natural selection for the study of the adaptive process. Empirical implementation of this concept can productively guide efforts both to deepen scientific insight into the process of adaptation and to inform measures for conserving the biota in the face of rapid environmental change.


Neuronal influence on hormone release from anglerfish islet cells.
Pancreatic islets in anglerfish (AF) are macroscopic collections of nearly pure endocrine cells that are densely innervated. Immunohistochemical staining for neurotransmitter biosynthetic enzymes revealed noradrenergic and cholinergic innervation of AF islets. An in vitro preparation of perifused dispersed AF islet cells was developed to study nutrient and neural control of islet hormone secretion. Glucose stimulated insulin and somatostatin-14 (SS-14) secretion in a dose-dependent manner, and 16.7 mM glucose inhibited glucagon secretion. In 2 mM glucose, norepinephrine and isoproterenol stimulated glucagon and SS-14 release. Isoproterenol stimulated insulin secretion, and norepinephrine stimulated or inhibited insulin release, depending on the concentration. Clonidine potently inhibited glucose-stimulated insulin secretion but stimulated glucagon release. Methacholine, a muscarinic cholinergic agonist, stimulated insulin, glucagon, and SS-14 release. The control of AF hormone release by neurotransmitter agonists in vitro was similar to that in higher vertebrate species. Therefore we used this tissue preparation to study postsynaptic interactions between glucose and neurotransmitters in islets.


Oral healthcare delivery in institutionalised older people: A health-economic evaluation.
This health-economic evaluation aimed to assess the cost-effectiveness of a number of alternatives for preventive and curative oral health care in institutionalised older people in Flanders. A six-state Markov model was used to compare expected costs and healthy oral years (HOYs) of four alternatives: (1) usual care; (2) on-site preventive care; (3) on-site preventive care + curative care in the community; and (4) on-site preventive care + on-site curative care. A healthcare payer perspective was adopted, and the time horizon was 10 years. Deterministic and probabilistic sensitivity analyses were performed. Incremental cost-effectiveness ratios (ICERs) of alternatives 2, 3 and 4 (all compared to alternative 1) were as follows: (2) 7944 €/HOY gained; (3) 1576 €/HOY gained; and (4) 1132 €/HOY gained. Hence, alternatives 2 and 3 were not cost-effective compared to alternative 4. The probability that oral care interventions are more effective and cost-saving than usual care was <3% for all three interventions. For institutionalised older people, on-site solutions for preventive and curative oral health care might be the most cost-effective alternative. It should be kept in mind that on-site solutions require large initial investment and that the advanced age of the population and the high costs of oral health care make it unlikely that these interventions would become cost-saving, even in the long term.


Tld1 is a regulator of triglyceride lipolysis that demarcates a lipid droplet subpopulation.
Cells store lipids in the form of triglyceride (TG) and sterol ester (SE) in lipid droplets (LDs). Distinct pools of LDs exist, but a pervasive question is how proteins localize to and convey functions to LD subsets. Here, we show that the yeast protein YDR275W/Tld1 (for TG-associated LD protein 1) localizes to a subset of TG-containing LDs and reveal it negatively regulates lipolysis. Mechanistically, Tld1 LD targeting requires TG, and it is mediated by two distinct hydrophobic regions (HRs). Molecular dynamics simulations reveal that Tld1's HRs interact with TG on LDs and adopt specific conformations on TG-rich LDs versus SE-rich LDs in yeast and human cells. Tld1-deficient yeast display no defect in LD biogenesis but exhibit elevated TG lipolysis dependent on lipase Tgl3. Remarkably, overexpression of Tld1, but not LD protein Pln1/Pet10, promotes TG accumulation without altering SE pools. Finally, we find that Tld1-deficient cells display altered LD mobilization during extended yeast starvation. We propose that Tld1 senses TG-rich LDs and regulates lipolysis on LD subpopulations.


Role of interleukin polymorphisms in gastric cancer: "Pros and cons".
Helicobacter pylori (H. pylori) infection is the leading cause of gastric cancer worldwide. Infection with this bacterium causes a chronic active immune response that persists for the life of the host. The combination of bacterial factors, environmental insults, and the host immune response drives the initiation and progression of mucosal atrophy, metaplasia, and dysplasia toward GC. Among the host factors, IL-1 gene cluster polymorphisms (IL-1B encoding IL-1β and IL-1RN encoding IL-1ra, its naturally occurring receptor antagonist) play a decisive role in modulating the risk of developing hypochlorhydria, gastric atrophy and GC in the presence of H. pylori infection. In particular, one single nucleotide polymorphism in the IL-1B promoter (IL-1B-511C⁄T), and the short allele of a 86-bp variable number of tandem repeats polymorphism in the IL-1RN second intron (IL-1RN*2) are associated with an increased risk for GC. However this hypothesis is still to be fully confirmed. This review focuses on the divergent results obtained by several epidemiological and functional in vitro and in vivo studies and show that IL-1 genotyping has still no role in the clinical management of patients with H. pylori infection.


Anaerobic culture in podiatric practice.
Although few podiatric infections are anaerobic, it is important to isolate, identify, and test all of the various types of bacteria for sensitivity. Dr. Lantor did not write this synopsis to analyze the procedures mentioned, but to outline some of the signs and symptoms associated with anaerobic growths and to make foot surgeons aware of the need for anaerobic culture techniques.


A human mannose-binding protein is an acute-phase reactant that shares sequence homology with other vertebrate lectins.
Mannose-binding proteins have been isolated from the liver of rats and humans and subsequently been found in the serum of rats, rabbits, and humans. We report the isolation of cDNA clones isolated from a human liver cDNA library that encodes a human mannose-binding protein. The primary structure has three domains: (a) an NH2-terminal cysteine-rich segment of 19 amino acids which appears to be involved in the formation of interchain disulfide bonds that would stabilize multimeric forms of the protein; (b) a collagen-like region consisting of 19 repeats of the sequence Gly-x-y; and (c) a COOH-terminal putative carbohydrate-binding domain consisting of 148 residues. This human mannose-binding protein bears 51% overall homology (allowing three gaps) with a rat mannose-binding protein C and 48% homology (allowing seven gaps) with a rat mannose-binding protein A. Like these homologous rat proteins, the human mannose-binding protein COOH-terminal sequences are homologous to the carbohydrate recognition portion of several other lectin-like proteins including mammalian hepatic receptors, an insect-soluble hemolymph, and a sea urchin lectin found in coelomic fluid. The apoproteins of dog and human surfactant and the human lymphocyte IgE Ec receptor have not been shown to have lectin-like properties, yet by homology are members of this family of lectin-like proteins. The human mannose-binding protein is preceded by a typical hydrophobic signal sequence and its hepatic secretion is induced as part of the acute-phase response consistent with its probable role in host defense.


Evidence for a Ca2+ pool associated with secretory granules in rat submandibular acinar cells.
Intracellular Ca2+ stores in rat submandibular acinar cells were characterized using the Ca(2+)-sensitive fluorescent indicator fura 2 and the radiotracer 45Ca2+. Acetylcholine induced a rapid Ca2+ release from a store sensitive to inositol 1,4,5-trisphosphate (IP3) and to thapsigargin (TG). After this store was presumably depleted, ionomycin caused a further increase in cytosolic free Ca2+ concentration ([Ca2+]i), suggesting the presence of an IP3-insensitive Ca2+ release from a store that is more extensive and heterogeneous than the IP3-sensitive one and includes a small mitochondrial component. After both of these stores had been discharged, exposure to monensin caused an additional release of Ca2+ from a third store. This store appears to be associated with secretory granules, since Ca2+ release was significantly reduced when degranulation was induced by isoprenaline. This third store appears to be insensitive to IP3, discharges Ca2+ when the pH gradient across the limiting membrane is collapsed with monensin and only in the presence of both ionomycin and monensin. Ca2+ release from this store is not by Na+/Ca2+ exchange, since simply altering [Na+]i did not cause significant Ca2+ release. In permeabilized cells, IP3 and TG released approx. 35% of 45Ca2+, and ionomycin released an additional 57%, whereas monensin only caused a small additional release, suggesting that only IP3- and ionomycin-sensitive stores are loaded with 45Ca2+ under these conditions. The absence of significant isotope uptake into the ionomycin+monensin-sensitive store may result from a low rate of tracer accumulation or from the lack of Ca2+ pumps in the store. The pattern of response was similar in the presence and absence of mitochondrial inhibitors, indicating that the store is not located in mitochondria. In summary, these results suggest that a substantial IP3-insensitive Ca2+ store is present in secretory granules in rat submandibular acinar cells.


Diagnostic utility of fluid biomarkers in multiple system atrophy: a systematic review and meta-analysis.
Multiple system atrophy (MSA) is an adult onset, fatal neurodegenerative disease. However, no reliable biomarker is currently available to guide clinical diagnosis and help to determine the prognosis. Thus, a comprehensive meta-analysis is warranted to determine effective biomarkers for MSA and provide useful guidance for clinical diagnosis. A comprehensive literature search was made of the PubMed, Embase, Cochrane and Web of Science databases for relevant clinical trial articles for 1984-2019. Two review authors examined the full-text records, respectively, and determined which studies met the inclusion criteria. We estimated the mean difference, standard deviation and 95% confidence intervals. A total of 28 studies and 11 biomarkers were included in our analysis. Several biomarkers were found to be useful to distinguish MSA patients from healthy controls, including the reduction of phosphorylated tau, α-synuclein (α-syn), 42-amino-acid form of Aβ and total tau (t-tau), the elevation of neurofilament light-chain protein (NFL) in cerebrospinal fluid, the elevation of uric acid and reduction of homocysteine and coenzyme Q10 in plasma. Importantly, α-syn, NFL and t-tau could be used to distinguish MSA from Parkinson's disease (PD), indicating that these three biomarkers could be useful biomarkers in MSA diagnosis. The findings of our meta-analysis demonstrated diagnostic biomarkers for MSA. Moreover, three biomarkers could be used in differential diagnosis of MSA and PD. The results could be helpful for the early diagnosis of MSA and the accuracy of MSA diagnosis.


Supporting patients with long-term conditions in the community: Evaluation of the Greater Manchester Community Pharmacy Care Plan Service.
The Greater Manchester Community Pharmacy Care Plan (GMCPCP) service provided tailored care plans to help adults with one or more qualifying long-term condition (hypertension, asthma, diabetes and COPD) to achieve health goals and better self-management of their long-term conditions. The service ran between February and December 2017. The aim of this study was to investigate the impact of the service on patient activation, as measured by the PAM measure (primary outcome). Secondary outcomes included quality of life (EQ-5D-5L, EQ-VAS), medication adherence (MARS-5), NHS resource use and costs, systolic and diastolic blood pressure, HDL cholesterol ratio levels and body mass index (BMI). A before and after design was used, with follow-up at 6-months. A questionnaire was distributed at follow-up and telephone interviews with willing participants were used to investigate patient satisfaction with the service. The study was approved by the University of Manchester Research Ethics Committee. Quantitative data were analysed in SPSS v22 (IBM). A total of 382 patients were recruited to the service; 280 (73%) remained at follow-up. Ten patients were interviewed and 43 completed the questionnaire. A total of 613 goals were set; mean of 1.7 goals per patient. Fifty percent of goals were met at follow-up. There were significant improvements in PAM, EQ-5D-5L and EQ-VAS scores and significant reductions in systolic blood pressure, BMI and HDL cholesterol ratio at follow-up. Mean NHS service use costs were significantly lower at follow-up; with a mean decrease per patient of £236.43 (±SD £968.47). The mean cost per patient for providing the service was £203.10, resulting in potential cost-savings of £33.33 per patient (SD ± 874.65). Questionnaire respondents reported high levels of satisfaction with the service. This study suggests that the service is acceptable to patients and may lead to improvements in health outcomes and allows for modest cost savings. Limitations of the study included the low response rate to the patient questionnaire.


Effect of oral misoprostol, alone or in combination with aglepristone, on mid-term pregnancy termination in cats.
This study aimed to determine the efficacy and safety of oral misoprostol (MIS) administration in the induction of mid-term pregnancy termination in cats. Twenty-eight cats that were pregnant for 30-40 days were allocated to four groups. The aglepristone (AGL) group (n = 7) received 10 mg/kg SC aglepristone q24h for two consecutive days. In the AGL+MIS group (n = 7), AGL (as administered in the AGL group) and MIS (200 µg/cat PO q12h until the start of abortion) were administered. The MIS200 (n = 7) and MIS400 groups (n = 7) received MIS (200 or 400 µg/cat misoprostol, respectively) alone PO q12h until the start of abortion. Blood samples were collected at the start of treatment (d0), 4 days after the start of treatment (d4) and on the day of complete abortion/end of administration (dA/d7). The efficacy of the treatment was 71.4% in the AGL group, 100% in the AGL+MIS group, 0% in MIS200 group and 57.4% in MIS400 group (P = 0.004). No significance was found in relation to the interval from treatment to the start/end of abortion and the duration of abortion in all groups. The most observed side effect was vomiting in both groups administered MIS, particularly in the MIS400 group (56.7%). Progesterone (P4) concentrations were reduced during the abortion, but not to basal levels, in all groups. P4 concentrations were significantly lower at dA/d7 in the MIS400 group compared with the AGL and AGL+MIS groups (P = 0.002). The results obtained from this study showed that low doses of MIS do not induce abortions in cats but increase the effect of AGL. Although higher doses could terminate pregnancies, this also causes intense unwanted side effects. Therefore, the use of MIS alone as an abortifacient in cats is not recommended. For mid-term pregnancy termination in cats, the combination of misoprostol and aglepristone provides a more effective abortifacient than using either of them alone.


Crystallization within Intermediate Amorphous Phases Determines the Polycrystallinity of Nanoparticles from Coprecipitation.
Intense research on nanocrystals synthesized in solution is motivated by their original physical properties, which are determined by their sizes and shapes on various scales. However, morphology control on the nanoscale is limited by our understanding of crystallization, which is challenged by the now well-established prevalence of noncrystalline intermediates. In particular, the impact of such intermediates on the final sizes and crystal quality remains unclear because the characterization of their evolution on the nanometer and millisecond scales with nonperturbative analyses has remained a challenge. Here we use in situ X-ray scattering to show that the nucleation and growth of YVO4:Eu nanocrystals is spatially restrained within amorphous, nanometer-scaled intermediates. The reactivity and size of these amorphous intermediates determine (i) the mono versus polycrystalline character of final crystals and (ii) the size of final crystals. This implies that designing amorphous intermediates themselves that form in <6 ms is one of the keys to controlled bottom-up syntheses of optimized nanoparticles.


THE ASSOCIATION OF EPIRETINAL MEMBRANE WITH MACULAR HOLE FORMATION AFTER RHEGMATOGENOUS RETINAL DETACHMENT REPAIR.
To describe the clinical and optical coherence tomography findings associated with the development of full-thickness macular holes after rhegmatogenous retinal detachment (RRD) repair. Retrospective, interventional case series. All patients who developed full-thickness macular holes after successful RRD repair from 3 clinical practices were reviewed. All cases of combined/simultaneous full-thickness macular hole and RRD were excluded. The main outcome measure was the presence of an epiretinal membrane at time of diagnosis of macular hole. Twenty-five full-thickness macular holes were diagnosed after successful retinal detachment repair. Surgical approach to RRD repair included pneumatic retinopexy (6, 24%), scleral buckle alone (5, 20%), pars plana vitrectomy only (8, 32%), and combined scleral buckle and pars plana vitrectomy (6, 24%). The preceding RRD involved the macula in 19 patients (76%) before the formation of the macular hole. The median time to full-thickness macular hole diagnosis after RRD repair was 63 days (range, 4-4,080 days). An epiretinal membrane was present in all 25 (100%) macular holes. Two macular holes (8%) spontaneously closed, whereas the other 23 (92%) were successfully closed with a single surgical procedure. Mean visual acuity improved by approximately 5 lines to 20/72 (range, 20/20 to counting fingers at 1 foot) from 20/240 (range, 20/30 to hand motions) after macular hole repair (P < 0.0001). Full-thickness macular hole formation can occur after all types of RRD repair and is associated with an epiretinal membrane. The epiretinal membrane may play a role in the pathogenesis of secondary macular hole formation after RRD repair.


Impact of coronary computed tomographic angiography results on patient and physician behavior in a low-risk population.
The impact of screening coronary computed tomographic angiography (CCTA) on physician and patient behavior is unclear. We studied asymptomatic patients from a health-screening program. Our study population comprised 1000 patients who underwent CCTA as part of a prior study and a matched control group of 1000 patients who did not. We assessed medication use, secondary test referrals, revascularizations, and cardiovascular events at 90 days and 18 months. A total of 215 patients in the CCTA group had coronary atherosclerosis (CCTA positive). Medication use was increased in the CCTA-positive group compared with both the CCTA-negative (no atherosclerosis) and control groups at 90 days (statin use, 34% vs 5% vs 8%, respectively; aspirin use, 40% vs 5% vs 8%, respectively), and 18 months (statin use, 20% vs 3% vs 6%, respectively; aspirin use, 26% vs 3% vs 6%, respectively). After multivariable risk adjustment, the odds ratios for statin and aspirin use in the CCTA-positive group at 18 months were 3.3 (95% confidence interval [CI], 1.3-8.3) and 4.2 (95% CI, 1.8-9.6), respectively. At 90 days, in the total CCTA group vs controls, there were more secondary tests (55 [5%] vs 22 [2%]; P < .001) and revascularizations (13 [1%] vs 1 [0.1%]; P < .001). One cardiovascular event occurred in each group over 18 months. An abnormal screening CCTA result was predictive of increased aspirin and statin use at 90 days and 18 months, although medication use lessened over time. Screening CCTA was associated with increased invasive testing, without any difference in events at 18 months. Screening CCTA should not be considered a justifiable test at this time.


Molecular cloning and characterization of buffalo alpha(s1)-casein gene.
Buffaloes in Indian subcontinent play an important role as the producer of milk and milk products. The alpha(s1)-casein constitutes 38% of the total milk proteins. The present study was carried out to characterize the gene in Murrah breed of Riverine buffalo. Buffalo alpha(s1)-casein cDNA was synthesized by RT-PCR, then cloned using pDRIVE-cloning vector and sequenced. The sequencing revealed that the size of alpha(s1)-casein cDNA was of 645 bp with GC content of 45.58%. The alpha(s1)-casein gene coded 214 amino acids precursor with a signal peptide of 15 amino acid residues. The similarity of buffalo alpha(s1)-casein mRNA sequence with that of cattle, goat, sheep, pig, camel, equine and human were estimated as 97.2, 93, 92.3, 57.2, 59.5, 55.9 and 46.6%, respectively. A similar trend was observed when compared amino acid sequences of these species. In the phylogenetic trees, constructed from the data of the alpha(s1)-casein mRNA as well as protein sequences, it has been observed that buffalo, cattle, goat and sheep formed a cluster with a closer relationship between cattle and buffalo followed by goat and sheep.


Pseudosphingobacterium domesticum gen. nov., sp. nov., isolated from home-made compost.
A bacterial strain, DC-186(T), isolated from home-made compost, was characterized for its phenotypic and phylogenetic properties. The isolate was a Gram-negative rod that was able to grow at 15-36 degrees C and pH 5.5-8.0. Strain DC-186(T) was positive in tests for catalase, oxidase and beta-galactosidase activities and aesculin hydrolysis. The predominant fatty acids were the summed feature C(16 : 1)/iso-C(15 : 0) 2-OH (42 %) and iso-C(15 : 0) (26 %), the major respiratory quinone was menaquinone-7 and the genomic DNA G+C content was 42 mol%. 16S rRNA gene sequence analysis and phenetic characterization indicated that this organism belongs to the phylum Bacteroidetes and revealed its affiliation to the family Sphingobacteriaceae. Of recognized taxa, strain DC-186(T) was most closely related to Sphingobacterium daejeonense (90 % sequence similarity) based on 16S rRNA gene sequence analysis. The low 16S rRNA gene sequence similarity with other recognized taxa and the identification of distinctive phenetic features for this isolate support the definition of a new genus within the family Sphingobacteriaceae. The name Pseudosphingobacterium domesticum gen. nov., sp. nov. is proposed, with strain DC-186(T) (=CCUG 54353(T)=LMG 23837(T)) as the type strain.


[Neuroticism in the elderly. The utility of the shortened DPQ-scales].
This article reports on the relation between aging and personal adjustment. Current personality scales are not developed for older persons. Scales contain items which are not valid for an aging population and contain too many items for administration in older populations. As part of the Longitudinal Aging Study Amsterdam (LASA) Neuroticism in older persons was measured with a shortened version of the Inadequacy (IN) and Social Inadequacy (SI) scales of the Dutch Personality Questionnaire (DPQ). The utility of these shortened scales was assessed based on internal consistency, inter-item correlations, test-retest reliability and factor analysis. The consistency of the personality dimension Neuroticism was assessed based on cohort-differences and a 6-year longitudinal comparison. The research-population contained 2118 respondents at baseline, aged between 55 and 85 years, 49% were male and they were not living in an institution. The shortened scales appeared to be reliable and valid instruments to measure Neuroticism in the elderly. The gaining of time due to the administration of the shortened scales enlarges the feasibility of the scales for measuring Neuroticism in older persons. Results showed no significant age-difference on the IN-scale, but revealed a significant difference on the SI-scale (p < .01). The 65+ elderly (65-74 and 75-85) have higher scores on Social Inadequacy than the youngest elderly (55-64). Longitudinal analyses showed an interaction between age at baseline and the stability and change of the level of Neuroticism. On both scales the youngest age-group showed a significant decline in mean level of Neuroticism (p < .01). The mean level of Social Inadequacy in the oldest age-group showed an increase during the 6-year follow-up period (p < .05). However, the differences were very small. Future research is needed to assess the effect of related variables on Neuroticism in older persons.


Enhancing individual placement and support (IPS) - Supported employment: A Type 1 hybrid design randomized controlled trial to evaluate virtual reality job interview training among adults with severe mental illness.
Individual Placement and Support (IPS) is the evidence-based model of supported employment that increases employment rates in adults with severe mental illness (SMI). Although IPS is largely successful, over 80% of adults with SMI remain unemployed. An enhancement to high fidelity IPS could be an evidence-based job interview training component. To meet this training need, our group recently completed a series of randomized controlled efficacy trials funded by the National Institute of Mental Health to develop and test virtual reality job interview training (VR-JIT) in a lab setting. The results demonstrated that the intervention was efficacious at helping trainees improve their job interview skills and receive job offers within six months of completing VR-JIT compared to non-trainees. The overarching goal of this study is to evaluate the effectiveness of VR-JIT as an enhancement to IPS when delivered in a large community-based mental health service provider via a randomized controlled trial and initial process evaluation. Our aims are to: evaluate whether IPS services-as-usual in combination with VR-JIT, compared to IPS services-as-usual alone, enhances IPS outcomes for adults with SMI; evaluate mechanisms of employment outcomes and psychological distress; and conduct a multilevel, multidisciplinary, and mixed-method process evaluation of VR-JIT adoption and implementation to assess the acceptability, scalability, generalizability, and affordability of VR-JIT.


Characterization of virologic failure after an initially successful 48-week course of antiretroviral therapy in HIV/AIDS outpatients treated in Santos, Brazil.
We characterized the virologic failure after an initially successful 48-week course of antiretroviral therapy among HIV/AIDS patients in a retrospective cohort study involving patients from Santos, Brazil. Patients with plasma HIV RNA below 500 copies/mL for 48 weeks were included. Variables analyzed included gender, age, level of education, marital status, mode of HIV acquisition, viral load, and CD4 cell count upon admission. There were 4,909 patients registered with the clinic, of which 669 patients met all the inclusion criteria (41.6% female and 58.4% male). Only 27.5% of the patients maintained undetectable viral loads during up to one year of follow-up. After 48 weeks, virologic failure occurred earlier in females and in patients first treated with an antiretroviral regimen other than highly active antiretroviral therapy. Patients who were married or had a steady partner experienced virologic failure later than did those who were separated or widowed. The percentage of public health clinic patients who maintain undetectable viral loads for a period of over a year is much lower than that observed among patients enrolled in clinical trials. Females, individuals in unstable relationships, single individuals and widowed individuals should be given special attention in order to improve durability of viral suppression.


Effect of hamycin on plasma cholesterol of rat.
Hamycin, a polyene antibiotic, when administered orally with food or by a cannula or administered intraperitoneally produced a dose-dependent hypocholesterolemic effect on normal albino rats. A 50% decrease occurred in serum cholesterol level at 25-50 mg/kg oral dose of hamycin. No further decrease occurred in serum cholesterol level with increase in the oral dosage. Decrease in the serum cholesterol occurring with 0.5-1.0 mg/kg, ip dose of the antibiotic was similar to that observed with oral hamycin and suggest that the effect was possibly mediated through mechanisms other than those involving prevention of absorption of cholesterol through the gastrointestinal tract.


Impact of the gyral geometry on the electric field induced by transcranial magnetic stimulation.
The spatial extent of the effects of transcranial magnetic stimulation (TMS) on neural tissue is only coarsely understood. One key problem is the realistic calculation of the electric field induced in the brain, which proves difficult due to the complex gyral folding pattern that results in an inhomogeneous conductivity distribution within the skull. We used the finite element method (FEM) together with a high-resolution volume mesh of the human head to better characterize the field induced in cortical gray matter (GM). The volume mesh was constructed from T1-weighted structural magnetic resonance images to allow for an anatomically accurate modeling of the gyrification pattern. Five tissue types were taken into account, corresponding to skin, skull, cerebrospinal fluid (CSF) including the ventricles as well as cortical gray and white matter. We characterized the effect of the current direction on the electric field distribution in GM. Importantly, the field strength in GM was increased by up to 51% when the induced currents were perpendicular to the local gyrus orientation. This effect was mainly restricted to the gyral crowns and lips, but did not extend into the sulcal walls. As a result, the focality of the fields induced in GM was increased. This enhancement effect might in part underlie the dependency of stimulation thresholds on coil orientation, as commonly observed in TMS motor cortex studies. In contrast to the clear-cut effects of the gyrification pattern on the induced field strength, current directions were predominantly influenced by the CSF-skull boundary.


The Potential Role of Awe for Depression: Reassembling the Puzzle.
Recently, interest in the unique pathways linking discrete positive emotions to specific health outcomes has gained increasing attention, but the role of awe is yet to be elucidated. Awe is a complex and transformative emotion that can restructure individuals' mental frames so deeply that it could be considered a therapeutic asset for major mental health major issues, including depression. Despite sparse evidence showing a potential connection between depression and awe, this link has not been combined into a proposal resulting in specific intervention guidelines. The aim of this perspective was three-fold: (i) to provide a new unifying model of awe's functioning-the Matryoshka model; (ii) to show systematic and explicit connections between this emotion and depression; and (iii) to suggest specific guidelines of intervention utilizing the potential therapeutic role of awe for mental health, specifically for depression. This theoretical endeavor in its entirety has been framed within the health domain.


Answering head and neck cancer questions: An assessment of ChatGPT responses.
To examine and compare ChatGPT versus Google websites in answering common head and neck cancer questions. Commonly asked questions about head and neck cancer were obtained and inputted into both ChatGPT-4 and Google search engine. For each question, the ChatGPT response and first website search result were compiled and examined. Content quality was assessed by independent reviewers using standardized grading criteria and the modified Ensuring Quality Information for Patients (EQIP) tool. Readability was determined using the Flesch reading ease scale. In total, 49 questions related to head and neck cancer were included. Google sources were on average significantly higher quality than ChatGPT responses (4.2 vs 3.6, p = 0.005). According to the EQIP tool, Google and ChatGPT had on average similar response rates per criterion (24.4 vs 20.5, p = 0.09) while Google had a significantly higher average score per question than ChatGPT (13.8 vs 11.7, p < 0.001) According to the Flesch reading ease scale, ChatGPT and Google sources were both considered similarly difficult to read (33.1 vs 37.0, p = 0.180) and at a college level (14.3 vs 14.2, p = 0.820.) CONCLUSION: ChatGPT responses were as challenging to read as Google sources, but poorer quality due to decreased reliability and accuracy in answering questions. Though promising, ChatGPT in its current form should not be considered dependable. Google sources are a preferred resource for patient educational materials.


Spreading of methylation along DNA.
Mouse DNA methyltransferase is able to catalyse the transfer of a methyl group to certain CG-containing single-stranded oligonucleotides. The presence of a methylcytosine is required for efficient transfer. This methylcytosine may or may not be on the same oligonucleotide as that containing the accepting CG dinucleotide. When the accepting CG dinucleotide forms part of an unmethylated CG dinucleotide pair, its accepting activity is dramatically reduced. This provides the potential for methylation to spread along the DNA when it is rendered single-stranded at replication. It could also help to maintain fully methylated CG islands and asymmetrically methylated sites.


Elinogrel: pharmacological principles, preclinical and early phase clinical testing.
Antiplatelet drug therapy represents the cornerstone of treatment for cardiovascular atherothrombotic disease processes. Dual antiplatelet therapy with aspirin and oral ADP-receptor antagonists such as clopidogrel are currently the standard care for prevention of ischemic events in patients with acute coronary syndrome and who are undergoing percutaneous coronary intervention. However, despite the clinical benefit associated with clopidogrel therapy, this drug has several limitations, including a broad interindividual response variability, drug-drug interactions, slow onset of action and irreversible platelet inhibition, emphasizing the need for novel P2Y(12)-receptor antagonists. Elinogrel (PRT060128) is a reversible, potent and competitive inhibitor of the P2Y(12) receptor with a fast onset and offset of action that can be administered by both oral and intravenous routes and rapidly achieves near complete platelet inhibition. Preclinical and early-phase clinical testing have shown promising results with this novel compound, which awaits further testing in outcome-driven clinical trials. This article provides an overview of the current level of knowledge regarding elinogrel, focusing on its pharmacologic properties and preclinical and early-phase clinical development.


A plant glutamate decarboxylase containing a calmodulin binding domain. Cloning, sequence, and functional analysis.
Molecular procedures have been applied to isolate plant calmodulin-binding proteins. A petunia cDNA expression library was screened with 35S-labeled recombinant calmodulin as a probe, and a cDNA coding for a Ca(2+)-dependent calmodulin-binding protein was isolated. The deduced amino acid sequence of the petunia protein (500 amino acid residues, 58 kDa) has 67% overall amino acid sequence similarity to glutamate decarboxylase (GAD) from Escherichia coli (466 amino acid residues, 53 kDa). The recombinant protein expressed in E. coli cells displays GAD activity, i.e. catalyzes the conversion of glutamic acid to gamma-aminobutyric acid and binds calmodulin, whereas E. coli GAD does not bind calmodulin. The calmodulin binding domain in the petunia GAD was mapped by binding truncated forms of GAD immobilized on nitrocellulose membranes to recombinant petunia 35S-calmodulin as well as to biotinylated bovine calmodulin and by binding truncated forms of GAD to calmodulin-Sepharose columns. The calmodulin binding domain in petunia GAD is part of a carboxyl end extension that is not present in E. coli GAD. Polyclonal antibodies raised against the recombinant petunia GAD detect a single protein band from plant extracts of gel mobility identical to that of the recombinant GAD. Moreover, the plant protein binds calmodulin in vitro. This is the first report of the isolation of a GAD gene from plants and of a calmodulin-binding GAD from any organism. Our results raise the possibility that intracellular Ca2+ signals via calmodulin are involved in the regulation of gamma-aminobutyric acid synthesis in plants.


Adverse health effects related to tobacco smoke exposure in a cohort of three-year olds.
To analyse the importance of mothers' smoking during pregnancy and/or environmental tobacco smoke (ETS) exposure in early childhood for children's health and well-being at the age of 3 years. Four groups from a population based cohort (n=8850) were compared: children with nonsmoking mother during pregnancy and nonsmoking parents at the age of 3 years (n=7091); children with only foetal exposure (n=149); children exposed only postnatally (n=895) and children exposed both pre- and postnatally (n=595). Odds ratios and 95% confidence intervals were calculated. Children exposed both pre- and postnatally had more wheezing (1.14; 1.07-1.21) and rhinitis (1.16; 1.06-1.26), used more cough-mixture (1.07; 1.01-1.14) and broncodilatating drugs (1.08; 1.02-1.15) and suffered more from excessive crying (1.31; 1.13-1.51) and irritability (1.27; 1.09-1.48) compared to children with nonsmoking parents. Children exposed only postnatally had more rhinitis (1.24; 1.12-1.37), used more cough-mixture (1.14; 1.05-1.29) and suffered more from poor sleep (1.26; 1.07-1.47) than children of nonsmoking parents. Children with prenatal exposure only used more broncodilatating drugs (1.45; 1.03-2.04) and suffered more from poor sleep (2.06; 1.09-3.87). Health differences, small but significant, indicate that prenatal and/or postnatal ETS exposure alone, or in combination, seems to interfere with child health, supporting the importance of zero tolerance. However, as most smoking parents in Sweden try to protect their children from ETS exposure, the results also might indicate that protective measures are worthwhile.


Development of a kinematic 3D carpal model to analyze in vivo soft-tissue interaction across multiple static postures.
We developed a subject-specific kinematic model to analyze in vivo soft-tissue interaction in the carpus in static, unloaded postures. The bone geometry was extracted from a reference computed tomography volume image. The soft-tissue geometry, including cartilage and ligament tissues, was computationally modeled based on kinematic constraints; the constraints were extracted from multiple computed tomography scans corresponding to different carpal postures. The data collected in vivo was next coupled with numerical simulation in order to analyze the role of soft-tissues in different postures. The resulting model extends the state of biomechanical modeling by incorporating soft-tissue constraints across the carpus range of motion, while successfully using only physiological constraints. The model results suggest that soft-tissue wrapping constraints have substantial impact on carpus stability.


Failure to replicate an excess of the long dopamine D4 exon III repeat polymorphism in ADHD in a family-based study.
The DRD4 exon III repeat polymorphism has been associated in adults with Novelty Seeking personality traits and in children with attention deficit hyperactivity disorder (ADHD) in some but not all studies. In a previous report we failed to observe preferential transmission of the long DRD4 repeat in ADHD compared to the haplotype relative risk (HRR) derived control group in a group of 49 triads (both parents and ADHD child) recruited in the Jerusalem area. In the current study we independently recruited an additional group of 49 triads from a different geographical location (Petak Tikvah) in Israel but having a similar ethnic background. In contrast to previous findings from a number of groups, in the current study an excess of the long DRD4 alleles was observed in the HRR control group compared to the ADHD subjects (Likelihood ratio = 5.50, P = 0. 02). In the expanded Israeli group of 98 triads so-far examined for the DRD4 repeat polymorphism there is an excess of the long alleles in the HRR control group (Likelihood ratio = 3.81, P = 0.05). These results attest to the complexity of ADHD inheritance and the likelihood that genetic heterogeneity characterizes this disorder especially across ethnic and cultural boundaries.


Bio-Bentall and hybrid arch frozen elephant trunk repair for acute type A aortic dissection with malperfusion.
The extent of repair in patients with acute type A aortic dissection is often determined by factors such as entry tear location, aortic anatomy, malperfusion and team expertise. The hybrid arch frozen elephant trunk, which has become an established technique to extend the distal acute type A aortic dissection repair, is particularly useful in malperfusion; however, it remains technically challenging and is associated with increased duration of circulatory arrest and risks of spinal cord ischaemia. Proximal dissection flap extension often determines repairability versus replacement of the aortic root. We present a case report highlighting the proximal and distal extent of repair in a patient with a known ascending aortic aneurysm presenting with an acute type A aortic dissection, with malperfusion, undergoing a successful bio-Bentall procedure and hybrid arch frozen elephant trunk repair.


GAMUT: GPU accelerated microRNA analysis to uncover target genes through CUDA-miRanda.
Non-coding sequences such as microRNAs have important roles in disease processes. Computational microRNA target identification (CMTI) is becoming increasingly important since traditional experimental methods for target identification pose many difficulties. These methods are time-consuming, costly, and often need guidance from computational methods to narrow down candidate genes anyway. However, most CMTI methods are computationally demanding, since they need to handle not only several million query microRNA and reference RNA pairs, but also several million nucleotide comparisons within each given pair. Thus, the need to perform microRNA identification at such large scale has increased the demand for parallel computing. Although most CMTI programs (e.g., the miRanda algorithm) are based on a modified Smith-Waterman (SW) algorithm, the existing parallel SW implementations (e.g., CUDASW++ 2.0/3.0, SWIPE) are unable to meet this demand in CMTI tasks. We present CUDA-miRanda, a fast microRNA target identification algorithm that takes advantage of massively parallel computing on Graphics Processing Units (GPU) using NVIDIA's Compute Unified Device Architecture (CUDA). CUDA-miRanda specifically focuses on the local alignment of short (i.e., ≤ 32 nucleotides) sequences against longer reference sequences (e.g., 20K nucleotides). Moreover, the proposed algorithm is able to report multiple alignments (up to 191 top scores) and the corresponding traceback sequences for any given (query sequence, reference sequence) pair. Speeds over 5.36 Giga Cell Updates Per Second (GCUPs) are achieved on a server with 4 NVIDIA Tesla M2090 GPUs. Compared to the original miRanda algorithm, which is evaluated on an Intel Xeon firstname.lastname@example.com GHz CPU, the experimental results show up to 166 times performance gains in terms of execution time. In addition, we have verified that the exact same targets were predicted in both CUDA-miRanda and the original miRanda implementations through multiple test datasets. We offer a GPU-based alternative to high performance compute (HPC) that can be developed locally at a relatively small cost. The community of GPU developers in the biomedical research community, particularly for genome analysis, is still growing. With increasing shared resources, this community will be able to advance CMTI in a very significant manner. Our source code is available at https://sourceforge.net/projects/cudamiranda/.


Discrete photonics resonator in coupled waveguide arrays.
We demonstrate both theoretically and experimentally that discrete diffraction resonance can be designed, fabricated, and successfully probed in functionalized - guidonic - coupled waveguide arrays. We evidence that double-barrier patterning of the coupling creates wavelength-independent angular tunnel resonance in the transmitted and the reflected intensity of light beams freely propagating in the plane of the array. Transmission peaks obtained are associated with resonant excitation of the engineered array bound supermodes of the functionalized array, in agreement with accurate and practical numerical modeling based on extended coupled-mode theory. The linear operation of the guidonic resonant tunneling double barrier makes up an original resonator for discrete photonics, suitable for all-optical control of light.


Influence of light and spawn quantity on the growth of Nigerian mushroom Pleurotus tuber-regium.
The effects of light and quantity of spawn on the sporophore and sclerotial yields of Pleurotus tuber-regium, cultivated on cotton wastes, rice straw, cocoyam peel, comcob, groundnut shell and sawdusts of Mansonia altissima, Khaya ivorensis and Boscia angustifolia were observed. The organism had sporophore and sclerotial yield values of 36.8 and 27.6 g kg(-1) waste, respectively, in cotton waste, at light quantum of 695 lux. There was a highly significant increase in yield of sclerotia (188.0 g kg(-1) waste), in total darkness, while malformed fruitbodies (sporophores) were produced in all the substrates under the same condition. Increasing the quantity of spawn from 5 to 30% reduced the period of spawn run from 13 to 6, 15 to 8 and 24 to 17 days, respectively in P. tuber-regium fruitbodies grown in cotton waste, rice straw and sawdust of B. angustifolia, with yield values of 38.0 and 20.0 g kg(-1) waste in cotton waste and rice straw. The optimal spawn levels forsclerotia formation in the two wastes were 10 and 5%, respectively. The mushroom did not produce sclerotia in corn cob and groundnut shell when exposed to light. However, maximal yield values of 286.8 and 288.4 g kg(-1) waste were obtained in both substrates in total darkness.


Optical coherence refractometry.
We introduce a novel approach to refractometry using a low coherence interferometer at multiple angles of incidence. We show that for plane parallel samples it is possible to measure their phase refractive index rather than the group index that is usually measured by interferometric methods. This is a significant development because it enables bulk refractive index measurement of scattering and soft samples, not relying on surface measurements that can be prone to error. Our technique is also noncontact and compatible with in situ refractive index measurements. Here, we demonstrate this new technique on a pure silica test piece and a highly scattering resin slab, comparing the results with standard critical angle refractometry.


Lentinan protects cardiomyocytes against hypoxia-induced injury by regulation of microRNA-22/Sirt1.
Myocardial ischemia is a serious disease which threatens human's life. Lentinan (LEN) possesses multiple biological properties: anticancer, antibacterial, antiviral and antioxidant effects. Our study investigated the effects of LEN on hypoxia-stimulated cardiomyocytes and the underlying mechanism. Primary neonatal rat ventricular cardiomyocytes (PNCM) were isolated from neonate rat pups. PNCM and H9c2 cells were stimulated by hypoxia and treated by LEN. Cell viability and apoptosis were detected by cell counting kit-8 and flow cytometry, respectively. Moreover, apoptotic factors were examined by western blot. Phosphatidylinositol 3'-kinase (PI3K)/protein kinase B (AKT) and β-catenin pathways related proteins were analyzed by western blot. Furthermore, the expression of microRNA-22 (miR-22) was detected by qRT-PCR. Altered expression of miR-22 and silenced information regulator 1 (Sirt1) was achieved by transfection. The relationship between miR-22 and Sirt1 was verified by luciferase assay. We found that LEN promoted cell viability and decreased apoptosis which led to the contrary results with what hypoxia induced. Moreover, LEN decreased the ratio of Bax to Bcl-2 and the level of cleaved caspase-3, as well as activated PI3K/AKT and β-catenin. LEN decreased the expression of miR-22 which was upregulated by hypoxia. miR-22 overexpression broken the promoting effects led by LEN. Moreover, Sirt1 was verified to be a target of miR-22. Silence of Sirt1 led to the opposite results with LEN. In conclusion, LEN relieved hypoxia-induced cellular injuries evidenced by increasing viability and decreasing apoptosis via down-regulation of miR-22, which was accompanied by activation of PI3K/AKT and β-catenin pathways. Highlights Lentinan alleviates hypoxia-induced injuries of PNCM and H9c2 cells; microRNA-22 expression is decreased by lentinan; Lentinan reduces hypoxia-induced injury by microRNA-22 downregulation; Lentinan regulates PI3K/AKT and Wnt/β-catenin by regulation of microRNA-22/Sirt1.


Prevention and treatment of diabetic retinopathy: evidence from large, randomized trials. The emerging role of fenofibrate.
Diabetic retinopathy (DR) remains a leading cause of preventable vision loss, despite advances in diabetes care. The burden of DR is likely to increase as the evolving pandemic of type 2 diabetes progresses. Tight control of blood glucose levels and blood pressure are essential for preventing or arresting the development of diabetic retinopathy, but are often difficult to achieve, and DR thus develops in a high proportion of patients. Current treatments for DR such as laser photocoagulation, intravitreous injections of corticosteroids or anti-vascular endothelial growth factor (VEGF) agents are indicated for advanced DR and have significant adverse effects. Therefore, new pharmacological treatments for the early stages of DR are needed. The Action to Control Cardiovascular Risk in Diabetes (ACCORD) trial included a lipid management arm, in which patients satisfying additional inclusion criteria for the atherogenic dyslipidemia phenotype were randomly assigned to fenofibrate or placebo, each with a statin. In the ACCORD-EYE substudy, randomization to fenofibrate was associated with a significant reduction in the risk of progression of DR. These data confirm and extend the results of the Fenofibrate Intervention and Event Lowering in Diabetes (FIELD) study, in which type 2 diabetes patients randomized to fenofibrate benefitted from a significantly lower incidence of laser treatment for retinopathy, progression of retinopathy or a composite measure of retinopathy outcomes. The results of ACCORD-EYE, together with those of FIELD, identify a place for fenofibrate for the prevention of retinopathy alongside intensive management of traditional risk factors, such as hyperglycemia and high blood pressure.


Hydatidosis: experience with hepatic and pulmonary hydatid disease.
From 1989-1992, 35 cases comprising of 20 hepatic and 15 pulmonary hydatid cysts were seen. Four patients with pulmonary hydatid disease had previous or concomitant hepatic hydatidosis compared to no concomitant pulmonary hydatid disease in the hepatic group. Hepatic hydatid disease had a greater tendency to be right sides, infected and calcified. Anaphylactic reactions occurred in 3 of 20 patients with hepatic hydatid disease against none in 15 patients with pulmonary disease. Management consisted of evacuation, drainage and obliteration of the residual cavity by capittonage in the lung and omentoplasty in the liver. Two patients, one hepatic and one with pulmonary hydatid disease developed infection of the residual cavity. Bronchocystic fistula occurred in 4 pulmonary and cystobiliary fistula in 2 hepatic hydatid disease patients. One patient with severe pleuropulmonary hydatidosis required a thoracoplasty. No recurrences have been noted in either group over a mean follow-up of 11 months (range 3 weeks- 3 years).


Development and validation of a next-generation sequencing assay with open-access analysis software for detecting resistance-associated mutations in CMV.
Cytomegalovirus (CMV) resistance testing by targeted next-generation sequencing (NGS) allows for the simultaneous analysis of multiple genes. We developed and validated an amplicon-based Ion Torrent NGS assay to detect CMV resistance mutations in UL27, UL54, UL56, and UL97 and compared the results to standard Sanger sequencing. NGS primers were designed to generate 83 overlapping amplicons of four CMV genes (~10 kb encompassing 138 mutation sites). An open-access software plugin was developed to perform read alignment, call variants, and interpret drug resistance. Plasmids were tested to determine NGS error rate and minor variant limit of detection. NGS limit of detection was determined using the CMV WHO International Standard and quantified clinical specimens. Reproducibility was also assessed. After establishing quality control metrics, 185 patient specimens previously tested using Sanger were reanalyzed by NGS. The NGS assay had a low error rate (<0.05%) and high accuracy (95%) for detecting CMV-associated resistance mutations present at ≥5% in contrived mixed populations. Mutation sites were reproducibly sequenced with 40× coverage when plasma viral loads were ≥2.6 log IU/mL. NGS detected the same resistance-associated mutations identified by Sanger in 68/69 (98.6%) specimens. In 16 specimens, NGS detected 18 resistance mutations that Sanger failed to detect; 14 were low-frequency variants (<20%), and six would have changed the drug resistance interpretation. The NGS assay showed excellent agreement with Sanger and generated high-quality sequence from low viral load specimens. Additionally, the higher resolution and analytic sensitivity of NGS potentially enables earlier detection of antiviral resistance.


Diastolic Electrocardiographic Parameters Predict Implantable Device Detected Asymptomatic Atrial Fibrillation.
Atrial fibrillation is the most common clinically significant arrhythmia. It is now established that atrial high-rate episodes are highly correlated with atrial fibrillation. To investigate the relation between diastolic electrocardiographic parameters and subclinical atrial fibrillation detected by cardiac implantable electronic devices. Ccross-sectional study. A total of 203 patients who had a dual-chamber, rate-modulated pacing pacemaker implanted due to sinus node dysfunction were prospectively enrolled in this study. Atrial high-rate episodes were defined as any lasting more than 5 min with an atrial rate of ≥220 beats per minute during the previous year. Patient groups were categorized on the basis of pacemaker interrogation as the absence of atrial high-rate episodes [atrial high-rate episodes (-)] and the presence of atrial high-rate episodes [atrial high-rate episodes (+)]. Episodes related to atrial over sensing were excluded. Twelve-lead surface electrocardiography was independently analyzed by two experienced readers for the measurement of diastolic electrocardiography parameters. Among 203 patients (mean age: 67.5±9.1, 60.1% male), 51 (25.1%) with atrial high-rate episodes were defined as group 1 and 152 (74.9%) without atrial high-rate episodes were defined as group 2. Both groups were similar in terms of demographic characteristics and cardiovascular risk factors. Tend-Q and Tend-P were significantly longer in group 2. PQ interval was statistically longer in group 1. Corrected QT interval was significantly longer in group 1. Diastolic electrocardiography index, heart rate and PQ and QT intervals were the only independent predictors of atrial high-rate episodes in patients with dual pacemakers in multivariate analysis. Abnormal diastolic electrocardiography parameters are powerful predisposing factors for the initiation of incident atrial fibrillation. Diastolic electrocardiography parameters and a novel diastolic index predict atrial high-rate episodes. Evaluating these parameters enables clinicians to identify patients who are at high risk and who may benefit from prophylactic treatment.


Acute identification of cranial burst fracture: comparison between CT and MR imaging findings.
Scalp swelling associated with cranial burst fracture, a widely diastatic skull fracture of infants associated with dural laceration and acute cerebral extrusion, may be confused with that of a simple subgaleal hematoma. Both conditions can also be associated with hemorrhagic shock. We sought to improve the early evaluation of infants believed to have sustained cranial burst fracture by including MR imaging, since this study clearly delineates the dural-cortical interface, the site of injury. Seven infants aged 1 through 11 months who sustained cranial burst fractures, all initially imaged with skull radiography and CT, were studied or treated from 1992 through 1996. MR imaging was obtained after resuscitation and stabilization. Surgery or autopsy confirmed MR findings (dural laceration and extracalvarial cerebral tissue) in all seven infants. MR imaging allows early diagnosis of skull fracture associated with acute cerebral extrusion.


The histone deacetylase inhibitor MS-275 interacts synergistically with fludarabine to induce apoptosis in human leukemia cells.
Interactions between the novel benzamide histone deacetylase (HDAC) inhibitor MS-275 and fludarabine were examined in lymphoid and myeloid human leukemia cells in relation to mitochondrial injury, signal transduction events, and apoptosis. Prior exposure of Jurkat lymphoblastic leukemia cells to a marginally toxic concentration of MS-275 (e.g., 500 nM) for 24 h sharply increased mitochondrial injury, caspase activation, and apoptosis in response to a minimally toxic concentration of fludarabine (500 nM), resulting in highly synergistic antileukemic interactions and loss of clonogenic survival. Simultaneous exposure to MS-275 and fludarabine also led to synergistic effects, but these were not as pronounced as observed with sequential treatment. Similar interactions were noted in the case of (a) other human leukemia cell lines (e.g., U937, CCRF-CEM); (b) other HDAC inhibitors (e.g., sodium butyrate); and (c) other nucleoside analogues (e.g., 1-beta-D-arabinofuranosylcytosine, gemcitabine). Potentiation of fludarabine lethality by MS-275 was associated with acetylation of histones H3 and H4, down-regulation of the antiapoptotic proteins XIAP and Mcl-1, enhanced cytosolic release of proapoptotic mitochondrial proteins (e.g., cytochrome c, Smac/DIABLO, and apoptosis-inducing factor), and caspase activation. It was also accompanied by the caspase-dependent down-regulation of p27(KIP1), cyclins A, E, and D(1), and cleavage and diminished phosphorylation of retinoblastoma protein. However, increased lethality of the combination was not associated with enhanced fludarabine triphosphate formation or DNA incorporation and occurred despite a slight reduction in the S-phase fraction. Prior exposure to MS-275 attenuated fludarabine-mediated activation of MEK1/2, extracellular signal-regulated kinase, and Akt, and enhanced c-Jun NH(2)-terminal kinase phosphorylation; furthermore, inducible expression of constitutively active MEK1/2 or Akt significantly diminished MS-275/fludarabine-induced lethality. Combined exposure of cells to MS-275 and fludarabine was associated with a significant increase in generation of reactive oxygen species; moreover, both the increase in reactive oxygen species and apoptosis were largely attenuated by coadministration of the free radical scavenger L-N-acetylcysteine. Finally, prior administration of MS-275 markedly potentiated fludarabine-mediated generation of the proapoptotic lipid second messenger ceramide. Taken together, these findings indicate that the HDAC inhibitor MS-275 induces multiple perturbations in signal transduction, survival, and cell cycle regulatory pathways that lower the threshold for fludarabine-mediated mitochondrial injury and apoptosis in human leukemia cells. They also provide insights into possible mechanisms by which novel, clinically relevant HDAC inhibitors might be used to enhance the antileukemic activity of established nucleoside analogues such as fludarabine.


Protein profiling of mouse livers with peroxisome proliferator-activated receptor alpha activation.
Peroxisome proliferator-activated receptor alpha (PPARalpha) is important in the induction of cell-specific pleiotropic responses, including the development of liver tumors, when it is chronically activated by structurally diverse synthetic ligands such as Wy-14,643 or by unmetabolized endogenous ligands resulting from the disruption of the gene encoding acyl coenzyme A (CoA) oxidase (AOX). Alterations in gene expression patterns in livers with PPARalpha activation were delineated by using a proteomic approach to analyze liver proteins of Wy-14,643-treated and AOX(-/-) mice. We identified 46 differentially expressed proteins in mouse livers with PPARalpha activation. Up-regulated proteins, including acetyl-CoA acetyltransferase, farnesyl pyrophosphate synthase, and carnitine O-octanoyltransferase, are involved in fatty acid metabolism, whereas down-regulated proteins, including ketohexokinase, formiminotransferase-cyclodeaminase, fructose-bisphosphatase aldolase B, sarcosine dehydrogenase, and cysteine sulfinic acid decarboxylase, are involved in carbohydrate and amino acid metabolism. Among stress response and xenobiotic metabolism proteins, selenium-binding protein 2 and catalase showed a dramatic approximately 18-fold decrease in expression and a modest approximately 6-fold increase in expression, respectively. In addition, glycine N-methyltransferase, pyrophosphate phosphohydrolase, and protein phosphatase 1D were down-regulated with PPARalpha activation. These observations establish proteomic profiles reflecting a common and predictable pattern of differential protein expression in livers with PPARalpha activation. We conclude that livers with PPARalpha activation are transcriptionally geared towards fatty acid combustion.


Head positioning for high +Gz loads: an analysis of the techniques used by F/A-18 pilots.
A study was undertaken among Royal Australian Air Force fighter pilots to determine whether, as an occupational group, they had developed any particular protective head positioning strategies in order to minimize their risk of sustaining a +Gz-induced neck injury during air combat maneuvering. There were 42 F/A-18 Hornet pilots who responded to an anonymous questionnaire which asked about head positioning strategies. Of these, 29 pilots reported using a protective strategy. Several pilots reported using several different techniques. The most popular strategy reported was positioning the head prior to the application of +Gz (13 respondents). Eleven pilots reported using aircraft structures to wedge or brace their heads against. The results of this study show that fighter pilots who are regularly exposed to high +Gz forces develop an individualized approach to protecting their cervical spines from +Gz-induced injury.


Lysolecithin as feed additive enhances collagen expression and villus length in the jejunum of broiler chickens.
Adding lysolecithin to feed has reportedly improved the performance of broiler chickens. Lysolecithin is generated by phospholipase catalyzed hydrolysis of lecithin. The enzymatic reaction converts various phospholipids into the corresponding lysophospholipids, with lysophosphatidylcholine (LPC) one of the primary products. Here we compared supplementation with a commercial lysolecithin (Lysoforte®) with comparable levels of highly purified LPC for effects on broilers. Despite no differences in weight gain during the starter period, we discovered a significant increase in average villus length with lysolecithin and an increase in villus width with purified LPC. High-throughput gene expression microarray analyses revealed many more genes were regulated in the epithelium of the jejunum by lysolecithin compared to purified LPC. The most up-regulated genes and pathways were for collagen, extracellular matrix, and integrins. Staining sections of the jejunum with Picrosirius Red confirmed the increased deposition of collagen fibrils in the villi of broilers fed lysolecithin, but not purified LPC. Thus, lysolecithin elicits gene expression in the intestinal epithelium, leading to enhanced collagen deposition and villus length. Purified LPC alone as a supplement does not mimic these responses. Feed supplementation with lysolecithin triggers changes in the intestinal epithelium with the potential to improve overall gut health and performance.


Adiponectin and all-cause mortality in elderly people with type 2 diabetes.
To assess the association between serum adiponectin level and all-cause mortality in people with type 2 diabetes. Because of the insulin-sensitizing, anti-inflammatory, and antiatherogenic effects of adiponectin, we hypothesized that higher adiponectin level would be associated with lower all-cause mortality. A total of 609 men and women aged 72 ± 6.3 years with type 2 diabetes and information on total and high molecular weight adiponectin were followed for a median of 5 years. The longitudinal association between adiponectin and all-cause mortality was analyzed with Cox proportional hazards models with time from adiponectin measurement to death as the time-to-event variable. Analyses were adjusted for demographic variables and significant diabetes parameters, significant cardiovascular parameters, and significant diabetes medications. Total and high molecular weight adiponectin were highly correlated. The highest adiponectin quartile was strongly associated with higher all-cause mortality compared with the lowest quartile (hazard ratio = 4.0 [95% CI: 1.7-9.2]) in the fully adjusted model. These results did not change in analyses stratified by sex and thiazolidinedione use, after exclusion of people who died within one year of adiponectin measurement, or when change in weight before adiponectin measurement was considered. Contrary to our hypothesis, higher adiponectin level was related to higher all-cause mortality. This association was not explained by confounding by other characteristics, including medications or preceding weight loss.


Identifying migrations in marine fishes through stable-isotope analysis.
The isotopic composition of many elements varies across both land and ocean surfaces in a predictable fashion. These stable-isotope ratios are transferred into animal tissues, potentially providing a powerful natural geospatial tag. To date, most studies using stable isotopes as geolocators in marine settings have focussed on mammals and seabirds conducting large ocean-basin scale migrations. An increasing understanding of isotopic variation in the marine environment, and improved sampling and analytical techniques, however, means that stable isotopes now hold genuine promise as a natural geolocation tag in marine fishes. Here, the theoretical background underpinning the use of stable isotopes of C, N and O in otolith, scale and muscle tissues as geolocation tools in the marine environment is reviewed, and examples of their applications are provided.


Reliability of Cyclotorsion measurements using Scanning Laser Ophthalmoscopy imaging in healthy subjects: the CySLO study.
To evaluate inter-rater and intra-rater reliability of objective cyclotorsion measurements obtained in healthy subjects using the Heidelberg Spectralis spectral domain-optical coherence tomography (SD-OCT) device. In this prospective methodological study, 32 healthy subjects (14 men, 18 women; aged 21-64 years) were enrolled, 31 right eyes were examined and their foveo-papillary angles (FPAs) were measured. The retinal nerve fibre layer (RNFL) programme by Heidelberg Spectralis (Heidelberg Engineering, Heidelberg, Germany) was used to measure cyclotorsion based on the in-built algorithm for the measurement of the FPA on the scanning laser ophthalmoscopy (SLO) image. Repeated scans of the fixating and non-fixating right eye without and with eye tracker each were obtained by three different examiners. The mean FPA measured overall by all three examiners was 6.6°±2.8°. The inter-rater reliability of the measured FPAs using the linear mixed effects model is estimated as rhointer=0.8803. The intra-rater reliability is estimated as rhointra=0.9589. Repeatability of objective cyclotorsion measurements within and between observers using the Heidelberg Spectralis OCT is very high. SD-OCT/SLO imaging is a reliable imaging technique to measure and study cyclotorsion. NCT02631369, Results.


Biosensor Förster resonance energy transfer detection by the phasor approach to fluorescence lifetime imaging microscopy.
We present here the phasor approach to biosensor Förster resonance energy transfer (FRET) detection by fluorescence lifetime imaging microscopy (FLIM) and show that this method of data representation is robust towards biosensor design as well as the fluorescence artifacts inherent to the cellular environment. We demonstrate this property on a series of dual and single chain biosensors, which report the localization of Rac1 and RhoA activity, whilst performing concomitant ratiometric FRET analysis on the acquired FLIM data by the generalized polarization (GP) approach. We then evaluate and compare the ability of these two methods to quantitatively image biosensor FRET signal as a function of time and space. We find that with lifetime analysis in the phasor plot each molecular species is transformed into a two-dimensional coordinate system where independent mixtures of fluorophores can be distinguished from changes in lifetime due to FRET. This enables the fractional contribution of the free and bound state of a dual chain biosensor or the low and high FRET species of a single chain biosensor to be quantified in each pixel of an image. The physical properties intrinsic to each biosensor design are also accurately characterized by the phasor analysis; thus, this method could be used to inform biosensor optimization at the developmental stage. We believe that as biosensors become more sophisticated and are multiplexed with other fluorescent molecular tools, biosensor FRET detection by the phasor approach to FLIM will not only become imperative to their use but also their advancement.


Non-insulin pharmacological therapy in pregnancy.
During pregnancy, when glycaemic levels remain uncontrolled, despite lifestyle modification, pharmacotherapy is advised, usually insulin which has been the gold standard for treatment. Recent studies however suggest that certain oral anti diabetic agents (OADs) may be safe and acceptable alternatives. There may be potential advantages for the use of metformin over insulin in GDM with respect to maternal weight gain and neonatal outcomes. However, as metformin crosses the placenta, its use during pregnancy raises concerns regarding potential adverse effects on the mother and foetus. Glibenclamide, a second generation sulfonylurea, more effective in glycaemic control in women with GDM has a lower treatment failure rate than metformin but there is lack of long term follow up data. Even though generally well tolerated, some studies report higher rates of preeclampsia, macrosomia, neonatal jaundice, neonatal hypoglycaemia and longer stay in neonatal care unit.


Prediction of B-cell epitopes using evolutionary information and propensity scales.
Development of computational tools that can accurately predict presence and location of B-cell epitopes on pathogenic proteins has a valuable application to the field of vaccinology. Because of the highly variable yet enigmatic nature of B-cell epitopes, their prediction presents a great challenge to computational immunologists. We propose a method, BEEPro (B-cell epitope prediction by evolutionary information and propensity scales), which adapts a linear averaging scheme on 16 properties using a support vector machine model to predict both linear and conformational B-cell epitopes. These 16 properties include position specific scoring matrix (PSSM), an amino acid ratio scale, and a set of 14 physicochemical scales obtained via a feature selection process. Finally, a three-way data split procedure is used during the validation process to prevent over-estimation of prediction performance and avoid bias in our experiment results. In our experiment, first we use a non-redundant linear B-cell epitope dataset curated by Sollner et al. for feature selection and parameter optimization. Evaluated by a three-way data split procedure, BEEPro achieves significant improvement with the area under the receiver operating curve (AUC) = 0.9987, accuracy = 99.29%, mathew's correlation coefficient (MCC) = 0.9281, sensitivity = 0.9604, specificity = 0.9946, positive predictive value (PPV) = 0.9042 for the Sollner dataset. In addition, the same parameters are used to evaluate performance on other independent linear B-cell epitope test datasets, BEEPro attains an AUC which ranges from 0.9874 to 0.9950 and an accuracy which ranges from 93.73% to 97.31%. Moreover, five-fold cross-validation on one benchmark conformational B-cell epitope dataset yields an accuracy of 92.14% and AUC of 0.9066. Compared with other current models, our method achieves a significant improvement with respect to AUC, accuracy, MCC, sensitivity, specificity, and PPV. Thus, we have shown that an appropriate combination of evolutionary information and propensity scales with a support vector machine model can significantly enhance the prediction performance of both linear and conformational B-cell epitopes.


Effects of superovulated heifer diet type and quantity on relative mRNA abundances and pyruvate metabolism in recovered embryos.
This study investigated the effects of quantity and type of diet fed to superovulated donor heifers on molecular and metabolic indices of embryonic development. These effects included the relative abundances of mRNAs for the alpha 1 subunit of Na/K-ATPase and the antioxidant enzyme Cu/Zn-SOD, as well as pyruvate utilization in bovine morulae and blastocysts developed in vivo. Heifers were fed a daily ration of either grass silage and a citrus-beet pulp-based concentrate or grass silage and a barley-based concentrate for 116 days, both at 3 kg per day or ad libitum. In embryos derived from heifers fed the pulp-based diets, the relative abundances of the transcripts were not affected by either day of collection or quantity of diet. In embryos derived from heifers fed the barley-based diets, the relative abundances of the Na/K-ATPase transcripts were also not changed by these main effects, while the relative abundances of the Cu/Zn-SOD transcripts were affected by day of collection and by the quantity of diet. Pyruvate metabolism was affected by day of collection, and was significantly increased in day 8 embryos compared with day 7 and day 6 embryos. Diet quantity did not affect pyruvate utilization, whereas diet type did increase pyruvate metabolism in the barley group when compared with the pulp group. The results of this study show for the first time that molecular and metabolic variations may exist in embryos derived in vivo and developed in donor heifers on nutritional regimens differing in type and quantity. Differences in embryos collected on different developmental days may be attributed to varying cell numbers. Alterations in the relative abundances of the Cu/Zn-SOD transcripts and pyruvate metabolism caused by the quantity of diet fed to the donor animal were likely to have been due to alterations in metabolic end products that accumulate in reproductive tract fluids, whereas differences in embryonic metabolism caused by type of diet are related to the composition of the diet. These findings characterize embryos produced in vivo at the molecular level, indicating that the molecular markers used in the present study can differentiate between populations of embryos produced under different nutritional regimens and determine conditions conductive to the production of good quality embryos.