A Frameshift in CSF2RB Predominant Among Ashkenazi Jews Increases Risk for Crohn's Disease and Reduces Monocyte Signaling via GM-CSF.
Crohn's disease (CD) has the highest prevalence in Ashkenazi Jewish populations. We sought to identify rare, CD-associated frameshift variants of high functional and statistical effects. We performed exome sequencing and array-based genotype analyses of 1477 Ashkenazi Jewish individuals with CD and 2614 Ashkenazi Jewish individuals without CD (controls). To validate our findings, we performed genotype analyses of an additional 1515 CD cases and 7052 controls for frameshift mutations in the colony-stimulating factor 2-receptor β common subunit gene (CSF2RB). Intestinal tissues and blood samples were collected from patients with CD; lamina propria leukocytes were isolated and expression of CSF2RB and granulocyte-macrophage colony-stimulating factor-responsive cells were defined by adenomatous polyposis coli (APC) time-of-flight mass cytometry (CyTOF analysis). Variants of CSF2RB were transfected into HEK293 cells and the expression and functions of gene products were compared. In the discovery cohort, we associated CD with a frameshift mutation in CSF2RB (P = 8.52 × 10(-4)); the finding was validated in the replication cohort (combined P = 3.42 × 10(-6)). Incubation of intestinal lamina propria leukocytes with granulocyte-macrophage colony-stimulating factor resulted in high levels of phosphorylation of signal transducer and activator of transcription (STAT5) and lesser increases in phosphorylation of extracellular signal-regulated kinase and AK straining transforming (AKT). Cells co-transfected with full-length and mutant forms of CSF2RB had reduced pSTAT5 after stimulation with granulocyte-macrophage colony-stimulating factor, compared with cells transfected with control CSF2RB, indicating a dominant-negative effect of the mutant gene. Monocytes from patients with CD who were heterozygous for the frameshift mutation (6% of CD cases analyzed) had reduced responses to granulocyte-macrophage colony-stimulating factor and markedly decreased activity of aldehyde dehydrogenase; activity of this enzyme has been associated with immune tolerance. In a genetic analysis of Ashkenazi Jewish individuals, we associated CD with a frameshift mutation in CSF2RB. Intestinal monocytes from carriers of this mutation had reduced responses to granulocyte-macrophage colony-stimulating factor, providing an additional mechanism for alterations to the innate immune response in individuals with CD.


Policy implications of complementary and alternative medicine use in Australia: data from the National Health Survey.
The objective of this study was to investigate the drivers of complementary and alternative medicine (CAM) use in the general population in Australia and to identify key policy implications. The National Health Survey 2007/2008, a representative survey of the Australian population, provides information on CAM use (practitioners and products) in the last 12 months. All adult respondents (N=15,779) aged 18 years or older are included in this study. Logistic regression is employed to determine the effect of socio-economic, condition-specific, health behavior variables, and private health insurance status on CAM use. In addition to socio-economic variables known to affect CAM use, individuals who have a chronic condition, particularly a mental health condition, are more likely to use CAM. There does not appear to be a correlation between CAM use and more frequent General Practitioner use; however, ancillary private health insurance is correlated with a greater likelihood of CAM use, as expected. The Australian government does not currently intervene in the CAM market in a systematic way. CAM is clearly considered to be a legitimate and important component of health care for many Australians, despite the limited availability of clinical evidence for its efficacy and safety. Policy interventions may include the regulation of CAM products, practitioners, and information as well as providing subsidies for cost-effective modalities.


Comparison of corneal endothelial cell measurements by two non-contact specular microscopes.
Measurement of corneal endothelial cell density is important both for clinical diagnosis as well as clinical studies. Since endothelial cell loss is considered irreversible in humans, even small changes in endothelial cell density are relevant. Therefore it is important to know whether different instruments for endothelial cell density measurements give the same results and can thus be used interchangeably. In this study we compare corneal endothelial cell density and morphometry measurements from two widely used non-contact specular microscopes, the Topcon SP3000P and Konan Noncon Robo SP8000. Endothelial cell measurements were performed with both the Topcon SP3000P and Konan Noncon Robo SP8000 on 34 eyes of 18 consecutive patients of our cornea clinics with poor image quality being the only exclusion criterion. Images were obtained using the auto-focussing method and manual cell selection. Endothelial cell density (ECD), hexagonal cell ratio (HEX) and coefficient of value (CV) of the endothelial cell layer were calculated by the instruments' built-in software. ECD values calculated by the Konan were systematically higher than Topcon values: in 94 % of eyes Konan gave a higher value than Topcon, leading to a mean difference in ECD between the instruments of 187 cells/mm(2) (P < 0.001 in paired Wilcoxon test). HEX showed a broad range of values and differed greatly with only weak correlation between the two instruments. CV values for Konan mostly exceeded Topcon values, and only showed a weak correlation between the two instruments as well. Values for ECD between the Konan and the Topcon do correlate well, but the ECDs calculated by the Konan are systematically higher than Topcon values. Both HEX and CV vary greatly and do not correlate sufficiently. Thus we recommend not to use the Konan and the Topcon instrument interchangeably.


Evidence-based management of acute myocardial infarction in the elderly--current perspectives.
Cardiovascular disease accounts for significant morbidity and mortality in the elderly. Despite several, large cardiovascular clinical trials, data to guide therapy in this growing population subset are relatively limited. This review focuses on treatment approaches and recommendations for the management of elderly patients with acute myocardial infarction (MI) obtained from subgroup analyses from major clinical trials.Treatment options for acute MI in the elderly have changed dramatically since the 1990s. Reperfusion therapy by primary percutaneous coronary intervention has superseded the use of thrombolytic therapy for the treatment of acute ST-elevation myocardial infarction (STEMI). Clinical trial data have demonstrated that even transferring patients to facilities that have primary angioplasty capabilities is better than thrombolytic therapy, if the anticipated transfer time is of acceptable duration. Additionally, adjunctive use of the intravenous glycoprotein (GP) receptor antagonist, abciximab, during primary angioplasty affords a reduction in the composite primary end point of death, reinfarction, and target vessel revascularization, with much of the benefit derived from the latter. Thrombolytic therapy, barring any contraindication, must be used when mechanical revascularization is not available; however, the risk for complications in the elderly is higher, especially for those 75 years and older. Studies investigating the use of thrombolytics plus GP receptor antagonists with and without percutaneous coronary intervention show questionable benefit in the elderly.


Learning to design synergetic computers with an extended symmetric diffusion network.
This article proposes an extended symmetric diffusion network that is applied to the design of synergetic computers. The state of a synergetic computer is translated to that of order parameters whose dynamics is described by a stochastic differential equation. The order parameter converges to the Boltzmann distribution, under some condition on the drift term, derived by the Fokker-Planck equation. The network can learn the dynamics of the order parameters from a nonlinear potential. This property is necessary to design the coefficient values of the synergetic computer. We propose a searching function for the image processing executed by the synergetic computer. It is shown that the image processing with the searching function is superior to the usual image-associative function of synergetic computation. The proposed network can be related, as a special case, to the discrete-state Boltzmann machine by some transformation. Finally, the extended symmetric diffusion network is applied to the estimation problem of an entire density function, as well as the proposed searching function for the image processing.


Fabrication and assessment of an electrospun polymeric microfiber-based platform under bulk flow conditions with rapid and efficient antigen capture.
This study investigated the fabrication and proof of concept design demonstrating rapid and highly sensitive antigen capture utilizing electrospun polystyrene (PS) microfiber mat substrates paired with vacuum pump pressurization to induce bulk flow. In comparison with conventional flat PS surfaces used for immunoassay purposes, this system optimizes the increased surface area of the electrospun polystyrene (ESPS) fiber mat substrates and the accelerated propagation of the antigen through the detection platform by using a vacuum pump to enable efficient and rapid antigen capture. The novelty of this work was demonstrated through a parametric study detailing how a fiber substrate can capture antigen sensitively and at high speeds. In terms of sensitivity, the current system is comparable to the conventionally used flat PS substrates. Additionally, the amount of antigen captured on a flat PS substrate in 60 minutes was surpassed in under 5 seconds when utilizing the ESPS-vacuum system. Three-dimensional ESPS fiber mats were then noted as a comparison between Damkohler numbers and between flat PS and ESPS-vacuum systems. The bulk flow of the ESPS-vacuum system allows for a Damkohler number of 0.37 indicating a balance between the flow rate and the reaction rate as opposed to a PS flat platform of 5.80 × 104 which illustrates a diffusion rate limited system. Finally, the overall ESPS-vacuum system was tested for its immunoassay capability. A sandwich fluorescence-based immunoassay was performed on both PS flat-diffusion and ESPS-vacuum systems. The ESPS-vacuum system indicated a wider detection range capability from 5 to 1000 ng mL-1 in comparison with the PS flat-diffusion system at 5 to 100 ng mL-1.


Rapid constriction of the selectivity filter underlies C-type inactivation in the KcsA potassium channel.
C-type inactivation is a time-dependent process observed in many K+ channels whereby prolonged activation by an external stimulus leads to a reduction in ionic conduction. While C-type inactivation is thought to be a result of a constriction of the selectivity filter, the local dynamics of the process remain elusive. Here, we use molecular dynamics (MD) simulations of the KcsA channel to elucidate the nature of kinetically delayed activation/inactivation gating coupling. Microsecond-scale MD simulations based on the truncated form of the KcsA channel (C-terminal domain deleted) provide a first glimpse of the onset of C-type inactivation. We observe over multiple trajectories that the selectivity filter consistently undergoes a spontaneous and rapid (within 1-2 µs) transition to a constricted conformation when the intracellular activation gate is fully open, but remains in the conductive conformation when the activation gate is closed or partially open. Multidimensional umbrella sampling potential of mean force calculations and nonequilibrium voltage-driven simulations further confirm these observations. Electrophysiological measurements show that the truncated form of the KcsA channel inactivates faster and greater than full-length KcsA, which is consistent with truncated KcsA opening to a greater degree because of the absence of the C-terminal domain restraint. Together, these results imply that the observed kinetics underlying activation/inactivation gating reflect a rapid conductive-to-constricted transition of the selectivity filter that is allosterically controlled by the slow opening of the intracellular gate.


Interventions to improve adherence to antiretroviral therapy: a systematic review and network meta-analysis.
High adherence to antiretroviral therapy is crucial to the success of HIV treatment. We evaluated comparative effectiveness of adherence interventions with the aim of informing the WHO's global guidance on interventions to increase adherence. For this systematic review and network meta-analysis, we searched for randomised controlled trials of interventions that aimed to improve adherence to antiretroviral therapy regimens in populations with HIV. We searched Cochrane Central Register of Controlled Trials, Embase, and MEDLINE for reports published up to July 16, 2015, and searched major conference abstracts from Jan 1, 2013, to July 16, 2015. We extracted data from eligible studies for study characteristics, interventions, patients' characteristics at baseline, and outcomes for the study populations of interest. We used network meta-analyses to compare adherence and viral suppression for all study settings (global network) and for studies in low-income and middle-income countries only (LMIC network). We obtained data from 85 trials with 16 271 participants. Short message service (SMS; text message) interventions were superior to standard of care in improving adherence in both the global network (odds ratio [OR] 1·48, 95% credible interval [CrI] 1·00-2·16) and in the LMIC network (1·49, 1·04-2·09). Multiple interventions showed generally superior adherence to single interventions, indicating additive effects. For viral suppression, only cognitive behavioural therapy (1·46, 1·05-2·12) and supporter interventions (1·28, 1·01-1·71) were superior to standard of care in the global network; none of the interventions improved viral response in the LMIC network. For the global network, the time discrepancy (whether the study outcome was measured during or after intervention was withdrawn) was an effect modifier for both adherence to antiretroviral therapy (coefficient estimate -0·43, 95% CrI -0·75 to -0·11) and viral suppression (-0·48; -0·84 to -0·12), suggesting that the effects of interventions wane over time. Several interventions can improve adherence and viral suppression; generally, their estimated effects were modest and waned over time. WHO.


Selection of organisms for the co-evolution-based study of protein interactions.
The prediction and study of protein interactions and functional relationships based on similarity of phylogenetic trees, exemplified by the mirrortree and related methodologies, is being widely used. Although dependence between the performance of these methods and the set of organisms used to build the trees was suspected, so far nobody assessed it in an exhaustive way, and, in general, previous works used as many organisms as possible. In this work we asses the effect of using different sets of organism (chosen according with various phylogenetic criteria) on the performance of this methodology in detecting protein interactions of different nature. We show that the performance of three mirrortree-related methodologies depends on the set of organisms used for building the trees, and it is not always directly related to the number of organisms in a simple way. Certain subsets of organisms seem to be more suitable for the predictions of certain types of interactions. This relationship between type of interaction and optimal set of organism for detecting them makes sense in the light of the phylogenetic distribution of the organisms and the nature of the interactions. In order to obtain an optimal performance when predicting protein interactions, it is recommended to use different sets of organisms depending on the available computational resources and data, as well as the type of interactions of interest.


Reconfiguration of yeast 40S ribosomal subunit domains by the translation initiation multifactor complex.
In the process of protein synthesis, the small (40S) subunit of the eukaryotic ribosome is recruited to the capped 5' end of the mRNA, from which point it scans along the 5' untranslated region in search of a start codon. However, the 40S subunit alone is not capable of functional association with cellular mRNA species; it has to be prepared for the recruitment and scanning steps by interactions with a group of eukaryotic initiation factors (eIFs). In budding yeast, an important subset of these factors (1, 2, 3, and 5) can form a multifactor complex (MFC). Here, we describe cryo-EM reconstructions of the 40S subunit, of the MFC, and of 40S complexes with MFC factors plus eIF1A. These studies reveal the positioning of the core MFC on the 40S subunit, and show how eIF-binding induces mobility in the head and platform and reconfigures the head-platform-body relationship. This is expected to increase the accessibility of the mRNA channel, thus enabling the 40S subunit to convert to a recruitment-competent state.


The Experiences of Young Liver Patients Transferring From Children's to Adult Services and Their Support Needs for a Successful Transition.
The period of transition from pediatric to adult services represents a time when young people need support, information, and appropriate care in order to successfully move. It is a period that is associated with nonadherence and disengagement with care. To explore the experiences of young liver transplant recipients transitioning to adult services and determine what they require in order to achieve a successful move. The research also explored the possibility of using a mobile phone application (app) as a tool to support transition. Qualitative approach using novel arts-based focus groups and one-to-one interviews. Twenty-one young people aged 16 to 25 years, 16 health-care professionals involved in their care, and 7 young people as follow-up. Participants used services provided by the 3 liver centers in England (Leeds, Birmingham, and London). Data highlighted the variability of transition pathways in England for young people moving from child to adult health services. The results showed that they required clear information regarding transition processes including specific medical information and that there was a shortfall in such information. Support was required in the form of a designated transition coordinator or similar specialist who could act as a point of reference and guidance throughout the process. Transitions needed to be individualized and based upon transition readiness rather than age, although the research showed that age cut-offs were still used. Young people welcomed apps to provide information, reminders, contacts, and connections. Future research should explore the efficacy of such apps.


Overt attention in contextual cuing of visual search is driven by the attentional set, but not by the predictiveness of distractors.
Two experiments examined biases in selective attention during contextual cuing of visual search. When participants were instructed to search for a target of a particular color, overt attention (as measured by the location of fixations) was biased strongly toward distractors presented in that same color. However, when participants searched for targets that could be presented in 1 of 2 possible colors, overt attention was not biased between the different distractors, regardless of whether these distractors predicted the location of the target (repeating) or did not (randomly arranged). These data suggest that selective attention in visual search is guided only by the demands of the target detection task (the attentional set) and not by the predictive validity of the distractor elements. (PsycINFO Database Record


Systems biology in inflammatory bowel diseases: on the way to precision medicine.
Inflammatory bowel diseases (IBD) are chronic and recurrent inflammatory disorders of the gastrointestinal tract. The elucidation of their etiopathology requires complex and multiple approaches. Systems biology has come to fulfill this need in approaching the pathogenetic mechanisms of IBD and its etiopathology, in a comprehensive way, by combining data from different scientific sources. In combination with bioinformatics and network medicine, it uses principles from computer science, mathematics, physics, chemistry, biology, medicine and computational tools to achieve its purposes. Systems biology utilizes scientific sources that provide data from omics studies (e.g., genomics, transcriptomics, etc.) and clinical observations, whose combined analysis leads to network formation and ultimately to a more integrative image of disease etiopathogenesis. In this review, we analyze the current literature on the methods and the tools utilized by systems biology in order to cover an innovative and exciting field: IBD-omics.


Resolving the variable genome and epigenome in human disease.
The individual human genome and epigenome are being defined at unprecedented resolution by current advances in sequencing technologies with important implications for human disease. This review uses examples relevant to clinical practice to illustrate the functional consequences of genetic and epigenetic variation. The insights gained from genome-wide association studies are described together with current efforts to understand the role of rare variants in common disease, set in the context of recent successes in Mendelian traits through the application of whole exome sequencing. The application of functional genomics to interrogate the genome and epigenome, build up an integrated picture of the regulatory genomic landscape and inform disease association studies is discussed, together with the role of expression quantitative trait mapping and analysis of allele-specific gene expression.


Ba2+ ions evoke two kinetically distinct patterns of exocytosis in chromaffin cells, but not in neurohypophysial nerve terminals.
The coupling between divalent cations and exocytosis of large dense-cored vesicles (LDCV) was studied with capacitance-detection techniques in nerve terminals of the rat neurohypophysis (NHP) and bovine chromaffin cells. Ba2+ substitution for Ca2+ produced kinetically distinct responses in the two preparations. In NHP terminals, Ba2+ ions behave as weak substitutes for Ca2+. Exocytotic events occur principally during depolarizing pulses, i.e., events are "stimulus-coupled" to Ba2+ entry through voltage-gated Ca2+ channels. Stimulus-coupled exocytosis apparently requires elevated submembrane cation concentrations that dissipate rapidly on hyperpolarization-induced Ca(2+)-channel closure. Intracellular dialysis of NHP terminals with Ba2+ does not evoke exocytosis, nor does it interfere with depolarization-evoked Ca2+ influx and exocytosis. In chromaffin cells, Ba2+ ions evoke a small quantity of stimulus-coupled secretion, but the dominant response is an additional pronounced poststimulus capacitance increase that outlasts channel closures by 20-50 sec. "Stimulus-decoupled" exocytosis is slow (approximately 25-40 fF/sec) compared with Ca(2+)-evoked stimulus-coupled exocytosis (approximately 1000 fF/sec). Decoupled secretion is not attributable to Ba2+ displacement of intracellular Ca2+ ions, because it is insensitive to 10 mM EGTA or thapsigargin. Slow exocytosis is initiated by inclusion of Ba2+ ions in the recording pipette and continues steadily for 5-12 min, producing a total increase of several thousand fF, which ultimately doubles or triples the original cell-surface area. We propose that two pathways of regulated exocytosis with distinct kinetics and divalent cation sensitivity exist in chromaffin cells. Only a single kinetic pattern is detected in NHP terminals, suggesting that mechanisms for secretion are not universally distributed in excitable cells.


Distribution of neurotensin/neuromedin N mRNA in rat forebrain: unexpected abundance in hippocampus and subiculum.
We have used in situ hybridization to determine the regional distribution of mRNA encoding the neurotensin/neuromedin N (NT/N) precursor in the forebrain of the adult male rat. Cells containing NT/N mRNA are widely distributed in the forebrain. These areas include the septum, bed nucleus of the stria terminalis, preoptic area, hypothalamus, amygdala, accumbens nucleus, caudate-putamen, and piriform and retrosplenial cortex. In general, the regional distribution of NT/N mRNA corresponds to the previously determined distribution of neurotensin-immunoreactive cell bodies; however, several notable exceptions were observed. The most striking difference occurs specifically in the CA1 region of the hippocampus, where intense labeling is associated with the pyramidal cell layer despite the reported absence of neurotensin-immunoreactive cells in this region. Analysis of microdissected tissue by S1 nuclease protection assay confirmed the abundance of authentic NT/N mRNA in CA1. A second major discrepancy between NT/N mRNA abundance and neurotensin-immunoreactivity occurs in the intensely labeled subiculum, a region that contains only scattered neurotensin-immunoreactive cells in the adult. These results suggest that, in specific regions of the forebrain, NT/N precursor is processed to yield products other than neurotensin. In addition, these results provide an anatomical basis for studying the physiological regulation of NT/N mRNA levels in the forebrain.


A Replication Study for the Association of rs11190870 With Curve Severity in Adolescent Idiopathic Scoliosis in Japanese.
Case-only study. The aim of this study was to confirm the association of rs11190870 with adolescent idiopathic scoliosis (AIS) severity in Japanese patients with AIS. Although the association of rs11190870 with AIS susceptibility is replicated in multiple ethnics, the association of rs11190870 with curve severity is controversial. Since the previous studies are of small, we performed a replication study using far larger number of patients than previous studies. A total of 1860 Japanese patients with AIS who had reached skeletal maturity or undergone surgical fusion were included in the study. We evaluated the association between rs11190870 and AIS progression for the entire group, and then for patients grouped according to a severe curve (a Cobb angle of ≥40°) or mild curve (a Cobb angle <30°). Because braces could affect the results of the present study, patients in the mild-curve group were divided according to whether or not they had worn a brace. We then evaluated associations between rs11190870 genotype and curve severity in these groups. The mean Cobb angles were 54.8° ± 12.1° in the severe-curve group and 24.4° ± 4.0° in the mild-curve group. The difference in rs11190870 risk-allele frequency between the severe- and mild-curve groups was evaluated. No significant differences were observed. We then examined the association of rs11190870 risk-allele frequency between patients in the mild- and severe-curve groups using the χ test for three models, and found a marginal association between rs11190870 and curve severity in the dominant model (P = 0.035, odds ratio = 1.51). We found no association between rs11190870 and curve severity using the criteria of previous study. However, we found a marginal association between rs11190870 and curve severity. Large-scale replication studies that consider skeletal maturity and brace history, including replication studies in other ethnic groups, would be helpful for clarifying the association. 4.


Multidrug-resistant and extended-spectrum beta-lactamase-producing uropathogens in children in Bhaktapur, Nepal.
The emergence of multidrug-resistant (MDR) and extended-spectrum beta-lactamase (ESBL)-producing uropathogens has complicated the treatment of urinary tract infections (UTI). Paediatric UTI is a common illness, which if not treated properly, may lead to acute and long-term complications, such as renal abscess, septicaemia, and renal scarring. This study aimed to determine the prevalence of MDR and ESBL-producing uropathogens among children. During the study period (April 2017-April 2018), midstream urine samples were collected following aseptic procedures from children < 16 years in Siddhi Memorial Hospital. Standard culture and biochemical tests were performed to identify uropathogens and antimicrobial susceptibility test was done by modified Kirby-Bauer disc diffusion method following Clinical and Laboratory Standard Institute (CLSI) guidelines. ESBL-producing uropathogens were screened by ceftazidime (30 μg) and cefotaxime (30 μg) discs, and confirmed by the combination disc tests: ceftazidime + clavulanic acid (30/10 μg) or cefotaxime + clavulanic acid (30/10 μg) as recommended by CLSI. We processed 5545 non-repeated urine samples from the children with symptoms of UTI. A significant growth of uropathogens was observed in 203 samples (3.7%). The median age of the children was 24 months (interquartile range (IQR), 12-53 months). Escherichia coli (n = 158, 77.8%) and Klebsiella pneumoniae (n = 30, 14.8%) were common among the uropathogens. Among them, 80.3% were resistant to amoxycillin and 51.2% were resistant to cotrimoxazole. Most of them were susceptible to amikacin, nitrofurantoin, and ofloxacin. MDR was detected in 34.5% (n = 70/203) and ESBL producers in 24.6% (n = 50/203) of them. The proportion of MDR isolates was higher in children < 5 years (n = 59/153, 38.6%) than children ≥ 5 years (n = 11/50, 22%) (P = 0.03). Nitrofurantoin, ofloxacin, and amikacin can be used for the empirical treatment for UTI in children in Bhaktapur, Nepal. MDR and ESBL-producing uropathogens are prevalent; this warrants a continuous surveillance of antimicrobial resistance.


Successful awake proning is associated with improved clinical outcomes in patients with COVID-19: single-centre high-dependency unit experience.
The SARS-CoV-2 can lead to severe illness with COVID-19. Outcomes of patients requiring mechanical ventilation are poor. Awake proning in COVID-19 improves oxygenation, but on data clinical outcomes is limited. This single-centre retrospective study aimed to assess whether successful awake proning of patients with COVID-19, requiring respiratory support (continuous positive airways pressure (CPAP) or high-flow nasal oxygen (HFNO)) on a respiratory high-dependency unit (HDU), is associated with improved outcomes. HDU care included awake proning by respiratory physiotherapists. Of 565 patients admitted with COVID-19, 71 (12.6%) were managed on the respiratory HDU, with 48 of these (67.6%) requiring respiratory support. Patients managed with CPAP alone 22/48 (45.8%) were significantly less likely to die than patients who required transfer onto HFNO 26/48 (54.2%): CPAP mortality 36.4%; HFNO mortality 69.2%, (p=0.023); however, multivariate analysis demonstrated that increasing age and the inability to awake prone were the only independent predictors of COVID-19 mortality. The mortality of patients with COVID-19 requiring respiratory support is considerable. Data from our cohort managed on HDU show that CPAP and awake proning are possible in a selected population of COVID-19, and may be useful. Further prospective studies are required.


Identification of a muscle-specific enhancer within the 5'-flanking region of the human myoglobin gene.
A 2-kilobase fragment from the 5'-flanking region of the human myoglobin gene extending from -2038 to +7 relative to the cap site regulates expression of a heterologous reporter gene in a cell-specific and developmentally regulated manner. Functional analyses of 5' and internal deletions indicate that sequences located between -261 and -205 are essential for muscle-specific expression in cooperation with the myoglobin core promoter. A 167-base pair fragment containing these sequences (-371 to -205) enhances expression from myoglobin core promoter elements in a manner that is independent of its orientation and position relative to the cap site. When linked to the herpes simplex virus thymidine kinase promoter, this 167-base pair fragment of the myoglobin gene also enhances expression in differentiated myotubes but not in undifferentiated myoblasts or fibroblasts. Nucleotide sequences within the region that is essential for enhancing activity (-261 to -205) are conserved in other mammalian myoglobin genes (seal, mouse) and resemble sequences within control regions of other genes that are expressed selectively in striated myocytes. These data indicate that the 5'-flanking region of the human myoglobin gene contains an enhancer-like element that is important for transcriptional activation during myocyte differentiation.


Synergy testing of FDA-approved drugs identifies potent drug combinations against Trypanosoma cruzi.
An estimated 8 million persons, mainly in Latin America, are infected with Trypanosoma cruzi, the etiologic agent of Chagas disease. Existing antiparasitic drugs for Chagas disease have significant toxicities and suboptimal effectiveness, hence new therapeutic strategies need to be devised to address this neglected tropical disease. Due to the high research and development costs of bringing new chemical entities to the clinic, we and others have investigated the strategy of repurposing existing drugs for Chagas disease. Screens of FDA-approved drugs (described in this paper) have revealed a variety of chemical classes that have growth inhibitory activity against mammalian stage Trypanosoma cruzi parasites. Aside from azole antifungal drugs that have low or sub-nanomolar activity, most of the active compounds revealed in these screens have effective concentrations causing 50% inhibition (EC50's) in the low micromolar or high nanomolar range. For example, we have identified an antihistamine (clemastine, EC50 of 0.4 µM), a selective serotonin reuptake inhibitor (fluoxetine, EC50 of 4.4 µM), and an antifolate drug (pyrimethamine, EC50 of 3.8 µM) and others. When tested alone in the murine model of Trypanosoma cruzi infection, most compounds had insufficient efficacy to lower parasitemia thus we investigated using combinations of compounds for additive or synergistic activity. Twenty-four active compounds were screened in vitro in all possible combinations. Follow up isobologram studies showed at least 8 drug pairs to have synergistic activity on T. cruzi growth. The combination of the calcium channel blocker, amlodipine, plus the antifungal drug, posaconazole, was found to be more effective at lowering parasitemia in mice than either drug alone, as was the combination of clemastine and posaconazole. Using combinations of FDA-approved drugs is a promising strategy for developing new treatments for Chagas disease.


TrawlerWeb: an online de novo motif discovery tool for next-generation sequencing datasets.
A strong focus of the post-genomic era is mining of the non-coding regulatory genome in order to unravel the function of regulatory elements that coordinate gene expression (Nat 489:57-74, 2012; Nat 507:462-70, 2014; Nat 507:455-61, 2014; Nat 518:317-30, 2015). Whole-genome approaches based on next-generation sequencing (NGS) have provided insight into the genomic location of regulatory elements throughout different cell types, organs and organisms. These technologies are now widespread and commonly used in laboratories from various fields of research. This highlights the need for fast and user-friendly software tools dedicated to extracting cis-regulatory information contained in these regulatory regions; for instance transcription factor binding site (TFBS) composition. Ideally, such tools should not require prior programming knowledge to ensure they are accessible for all users. We present TrawlerWeb, a web-based version of the Trawler_standalone tool (Nat Methods 4:563-5, 2007; Nat Protoc 5:323-34, 2010), to allow for the identification of enriched motifs in DNA sequences obtained from next-generation sequencing experiments in order to predict their TFBS composition. TrawlerWeb is designed for online queries with standard options common to web-based motif discovery tools. In addition, TrawlerWeb provides three unique new features: 1) TrawlerWeb allows the input of BED files directly generated from NGS experiments, 2) it automatically generates an input-matched biologically relevant background, and 3) it displays resulting conservation scores for each instance of the motif found in the input sequences, which assists the researcher in prioritising the motifs to validate experimentally. Finally, to date, this web-based version of Trawler_standalone remains the fastest online de novo motif discovery tool compared to other popular web-based software, while generating predictions with high accuracy. TrawlerWeb provides users with a fast, simple and easy-to-use web interface for de novo motif discovery. This will assist in rapidly analysing NGS datasets that are now being routinely generated. TrawlerWeb is freely available and accessible at: http://trawler.erc.monash.edu.au .


Dissociable changes in spike and wave discharges following exposure to injected cannabinoids and smoked cannabis in Genetic Absence Epilepsy Rats from Strasbourg.
There is significant interest in the use of cannabinoids for the treatment of many epilepsies including absence epilepsy (AE). Genetic Absence Epilepsy Rats from Strasbourg (GAERS) model many aspects of AE including the presence of spike-and-wave discharges (SWDs) on electroencephalogram (EEG) and behavioral comorbidities, such as elevated anxiety. However, the effects of cannabis plant-based phytocannabinoids have not been tested in GAERS. Therefore, we investigated how SWDs in GAERS are altered by the two most common phytocannabinoids, Δ9 -tetrahydrocannabinol (THC) and cannabidiol (CBD), and exposure to smoke from two different chemovars of cannabis. Animals were implanted with bipolar electrodes in the somatosensory cortex and EEGs were recorded for 2 hr. Injected THC (1-10 mg/kg, i.p.) dose-dependently increased SWDs to over 200% of baseline. In contrast, CBD (30-100 mg/kg, i.p.) produced a ~50% reduction in SWDs. Exposure to smoke from a commercially available chemovar of high-THC cannabis (Mohawk, Aphria Inc.) increased SWDs whereas a low-THC/high-CBD chemovar of cannabis (Treasure Island, Aphria Inc.) did not significantly affect SWDs in GAERS. Pre-treatment with a CB1R antagonist (SR141716A) did not prevent the high-THC cannabis smoke from increasing SWDs, suggesting that the THC-mediated increase may not be CB1R-dependent. Plasma concentrations of THC and CBD were similar to previously reported values following injection and smoke exposure. Compared to injected CBD, it appears Treasure Island did not increase plasma levels sufficiently to observe an anti-epileptic effect. Together these experiments provide initial evidence that acute phytocannabinoid administration exerts the biphasic modulation of SWDs and may differentially impact patients with AE.


Participation in social activities among adolescents with an autism spectrum disorder.
Little is known about patterns of participation in social activities among adolescents with an autism spectrum disorder (ASD). The objectives were to report nationally representative (U.S.) estimates of participation in social activities among adolescents with an ASD, to compare these estimates to other groups of adolescents with disabilities, and examine correlates of limited social participation. We analyzed data from wave 1 of the National Longitudinal Transition Study 2, a large cohort study of adolescents enrolled in special education. Three comparison groups included adolescents with learning disabilities, mental retardation, and speech/language impairments. Adolescents with an ASD were significantly more likely never to see friends out of school (43.3%), never to get called by friends (54.4%), and never to be invited to social activities (50.4%) when compared with adolescents from all the other groups. Correlates of limited social participation included low family income and having impairments in conversational ability, social communication, and functional cognitive skills. Compared with prior research, our study significantly expands inquiry in this area by broadening the range of social participation indicators examined, increasing the external validity of findings, focusing on the under-studied developmental stage of adolescence, and taking an ecological approach that included many potential correlates of social participation. There were notable differences in social participation by income, a dimension of social context seldom examined in research on ASDs.


Cost effectiveness of liraglutide in type II diabetes: a systematic review.
As novel treatments for type II diabetes enter the market, there is a need to assess their long-term clinical and economic outcomes against currently available treatment alternatives. Objective compilation and evaluation of current pharmacoeconomic evidence can assist payers and decision makers in determining the appropriate place in therapy of a new medication. Our objective was to review the existing pharmacoeconomic literature evaluating the cost effectiveness and overall costs of treatment associated with liraglutide in type II diabetes. Medical literature indexed in MEDLINE, EMBASE, PsycINFO, CINAHL, and EconLit through 1 June 2014 was searched. Full-text, English-language cost-effectiveness, cost-utility, and other cost analyses in type II diabetes that compared liraglutide to one or more anti-diabetic agents were included. Initial screening was based on relevance of titles and abstracts followed by examination of the study methods of each remaining manuscript. Studies conducting original pharmacoeconomic analyses were chosen for inclusion. Articles meeting the inclusion criteria were retrieved, and information on the study design and results was abstracted. Abstracted data elements were chosen and assessed based on the authors' experience as well as criteria set forth by the International Society for Pharmacoeconomics and Outcomes Research (ISPOR) Health Economic Evaluation Publication Guidelines Task Force. Additionally, reported incremental cost-effectiveness ratios (ICERs) and selected sensitivity analysis results were converted to $US, year 2012 values, in order to facilitate comparison across studies. A total of six cost studies and seven cost-utility studies were identified for inclusion. Across cost studies, liraglutide treatment resulted in costs ranging from a loss of $US2,730 (liraglutide 1.8 mg vs. sitagliptin; pharmacy costs only) over a 1-year time horizon to a savings of $US9,367 (liraglutide 1.8 mg vs. glimepiride; diabetes-related complication costs only) over a 30-year time horizon. Cost-utility analysis results reported base-case ICERs ranging from $US15,774 (vs. glimepiride) to $US40,128 (vs. rosiglitazone) per quality-adjusted life-year (QALY) ($US, year 2012) for liraglutide 1.2 mg and $US8,497 (vs. exenatide) to $US66,031 (vs. rosiglitazone)/QALY ($US, year 2012) for liraglutide 1.8 mg. Estimates were most sensitive to variations in time horizon and cardiovascular complication rates. Based on frequently cited, country-specific cost-utility thresholds, liraglutide was determined to have a probability of being cost effective of between 58 % (liraglutide 1.8 mg vs. sitagliptin) and 93 % (liraglutide 1.2 mg vs. glimepiride). Weaknesses of included studies related primarily to study model inputs that assumed long-term morbidity and mortality benefits in favor of liraglutide based on improvements in clinical biomarkers observed in short-term clinical trials. The exclusion of drug acquisition costs in two identified cost studies as well as the assumed lifetime duration of treatment with liraglutide in several cost-utility studies were also identified as weaknesses. The authors' review was limited by the possibility of incomplete literature retrieval, unintended omission of relevant data elements, and comparison of costs and ICERs generated from healthcare systems from differing countries. The current literature presents liraglutide as a cost-effective adjunct treatment for type II diabetes that may also be associated with a reduction in diabetes-related complication costs; however, ICER values are largely dependent on assumptions regarding the benefits of long-term liraglutide treatment and the time horizon of the analysis. Real-world use may make liraglutide unattractive from a payer and policy-maker perspective.


Retrofitting SBR systems to nutrient removal in sensitive tourist areas.
Retrofitting of existing SBR systems for nutrient removal is evaluated and defined for small communities in sensitive coastal areas, with seasonal fluctuations in wastewater quantity and quality. The proposed approach is developed by means of basic process stoichiometry and verified using ASM2d. The efficiency of retrofitting is found to rely on the delicate balance between the overall sludge age, the initial settled sludge volume in the reactor, and the ratio of the initial volume to the feed volume in each cycle, a parameter corresponding to the recycle ratio in continuous systems.


Studies of the life-history and energetics of marine and brackish-water nematodes : I. Demography of Monhystera disjuncta at different temperature and feeding conditions.
Aspects of the demography of Monhystera disjuncta were investigated at different temperatures (in agnotobiotic cultures) and in different feeding conditions (monoxenic cultures with different bacterial strains, and different densities in the feeding suspension with one strain). Embryonic development time, minimum generation time, egg deposition rate and adult longevity depend on temperature, quality and quantity of food offered. Body mass at maturity is an allometric function of food density. It is shown that a previously inferred selectivity in food uptake is an artifact of culture conditions. pH buffering and addition of sterols permit culture of the species on a wide variety of bacterial strains. M. disjuncta is less well adapted to take advantage of high food density than are nematodes from polysaprobic environments. The animals channel surplus energy intake into a larger body mass, without being able to increase their rate of population growth accordingly.


Profiling of the cell surface proteome.
The in depth-mining of the proteome necessitates the comprehensive analysis of proteins in individual subcellular compartments to uncover interesting patterns of protein expression that include assessment of protein location, trafficking and of post-translational modifications that are location specific. One of the compartments of substantial interest from a diagnostic and therapeutic point of view is the plasma membrane which contains intrinsic membrane proteins and other proteins expressed on the cell surface. Technologies are currently available for the comprehensive profiling of the cell surface proteome that rely on protein tagging of intact cells. Studies are emerging that point to unexpected patterns of expression of specific proteins on the cell surface, with a common occurrence of proteins previously considered to occur predominantly in other compartments, notably the endoplasmic reticulum. The profiling of the cell surface and plasma membrane proteomes will likely provide novel insights and uncover disease related alterations.


Behavioral studies with anxiolytic drugs. IV. Serotonergic involvement in the effects of buspirone on punished behavior of pigeons.
Interactions of the nonbenzodiazepine anxiolytic, buspirone, with serotonin (5-HT) were studied using behavioral and neurochemical procedures. Punished responding was studied in pigeons as this behavior is a generally acknowledged preclinical predictor of anxiolytic activity and because buspirone increases punished responding of pigeons with greater potency and efficacy than in other species. Keypeck responses were maintained under either fixed-interval or fixed-ratio schedules of food presentation; every 30th response produced a brief electric shock and suppressed responding (punishment). Buspirone (0.1-5.6 mg/kg i.m.) produced dose-related increases in punished responding which reached a maximum at 1 mg/kg. A serotonin agonist, MK-212 (0.01 mg/kg), antagonized whereas the 5-HT antagonist, cyproheptadine (0.01 mg/kg), potentiated the effects of buspirone without having behavioral effects of their own. The characteristics of [3H]-5-HT binding in pigeon brain membranes were similar to results reported in mammalian brain. Neither buspirone, MJ-13805 (gepirone, a related analog), nor MJ-13653 (a buspirone metabolite), significantly affected [3H]-5-HT binding and none of the compounds appreciably inhibited uptake of [3H]-5-HT into pigeon cerebral synaptosomes. Hill coefficients significantly less than unity for all drugs except 5-HT suggested multiple serotonergic binding sites for buspirone and analogs. Buspirone and MJ-13805 (1 nM) inhibited [3H]ketanserin binding (a measure of 5-HT2 binding sites) in pigeon cerebrum with Ki values above 10(-6) M. The number of [3H]ketanserin binding sites was estimated to be 109 fmol/mg of protein in pigeon cerebrum compared to 400 fmol/mg of protein in rat cerebrum.(ABSTRACT TRUNCATED AT 250 WORDS)


Molecular hydrogen inhalation attenuates postoperative cognitive impairment in rats.
Postoperative cognitive decline is a major clinical problem with high morbidity and mortality after surgery. Many studies have found that molecular hydrogen (H2) has significant neuroprotection against acute and chronic neurological injury by regulating inflammation and apoptosis. In this study, we hypothesized that H2 treatment could ameliorate the development of cognitive impairment following surgery. Adult male rats were subjected to stabilized tibial fracture operation under anesthesia. Two percent of H2 was inhaled for 3 h beginning at 1 h after surgery. Separate cohorts of rats were tested for cognitive function with fear conditioning and the Y-maze test, or euthanized to assess blood-brain barrier integrity, and systemic and hippocampal proinflammatory cytokine and caspase-3 activity. Surgery-challenged animals showed significant cognitive impairment evidenced by a decreased percentage of freezing time and an increased number of learning trials on days 1, 3, and 7 after operation, which were significantly improved by H2 treatment. Furthermore, H2 treatment significantly ameliorated the increase in serum and hippocampal proinflammatory cytokines tumor necrosis factor-α, interleukin-1β, interleukin-6, and high-mobility group protein 1 in surgery-challenged animals. Moreover, H2 treatment markedly improved blood-brain barrier integrity and reduced caspase-3 activity in the hippocampus of surgery-challenged animals. These findings suggest that H2 treatment could significantly mitigate surgery-induced cognitive impairment by regulating inflammation and apoptosis.


The association of high-density lipoprotein cholesterol with cancer incidence in type II diabetes: a case of reverse causality?
Low high-density lipoprotein cholesterol (HDL-C) and type II diabetes are associated with an increased risk for cancer. Patients with type II diabetes typically have low HDL-C; however, the association between HDL-C and cancer has not been examined in this population. A total of 11,140 patients with type II diabetes were followed for a median of 5 years. Cox proportional hazard models were used to assess the association between baseline HDL-C and risk of cancer incidence and cancer death, with adjustments made for potential confounders. To explore the possibility of reverse causation, analyses were repeated for the cancers occurring in the first and second halves of follow-up. Six hundred and ninety-nine patients developed cancer, with 48% occurring within the first half of follow-up. For every 0.4 mmol/L lower baseline HDL-C, there was a 16% higher risk of cancer [HR 1.16; 95% confidence interval (CI), 1.06-1.28; P = 0.0008] and cancer death (HR 1.16; 95% CI, 1.01-1.32; P = 0.03). After adjustment for confounding, the higher risk remained significant for cancer (adjusted HR 1.10; 95% CI, 1.00-1.22; P = 0.05) but not for cancer death (adjusted HR 1.08; 95% CI, 0.93-1.25; P = 0.31). The association was driven by cancers occurring within the first half of follow-up (adjusted HR 1.22; 95% CI, 1.05-1.41; P = 0.008) as no significant association was found between HDL-C and cancer in the second half of follow-up. Low HDL-C is associated with cancer risk in patients with type II diabetes. However, this association may be explained by confounding and reverse causation. HDL-C is not a risk factor for cancer in type II diabetes.


A scoping review of undergraduate ambulatory care education.
Since a disproportionate amount of medical education still occurs in hospitals, there are concerns that medical school graduates are not fully prepared to deliver efficient and effective care in ambulatory settings to increasingly complex patients. To understand the current extent of scholarship in this area. A scoping review was conducted by searching electronic databases and grey literature sources for articles published between 2001 and 2011 that identified key challenges and models of practice for undergraduate teaching of ambulatory care. Relevant articles were charted and assigned key descriptors, which were mapped onto Canadian recommendations for the future of undergraduate medical education. Most of the relevant articles originated in the United States, Australia, or the United Kingdom. Recommendations related to faculty development, learning contexts and addressing community needs had numerous areas of scholarly activity while scholarly activity was lacking for recommendations related to inter-professional practice, the use of technology, preventive medicine, and medical leadership. Systems should be established to support education and research collaboration between medical schools to develop best practices and build capacity for change. This method of scoping the field can be applied using best practices and recommendations in other countries.


Secreted protein acidic and rich in cysteine (SPARC) suppresses angiogenesis by down-regulating the expression of VEGF and MMP-7 in gastric cancer.
Secreted protein acidic and rich in cysteine (SPARC) is a glycoprotein that functions to inhibit angiogenesis, proliferation, and invasion in different types of cancer. The ability of SPARC to modulate neovascularisation is believed to be mediated in part by its ability to modulate the expression of vascular endothelial growth factor (VEGF) and matrix metalloproteinases (MMPs). In this study, we aimed to determine the effect of SPARC expression in gastric cancer cells on proliferation and angiogenesis in vitro and in vivo. We evaluated expression of SPARC in seven human gastric cancer cell lines. Then we established a stably transfected SPARC overexpressed cell line (BGC-SP) and a stably transfected SPARC knock-down cell line (HGC-sh). The effect of SPARC overexpression and SPARC silencing was studied by examining capillary formation of HUVECs in vitro and a dorsal skin-fold chamber model in vivo. Quantitative real-time PCR and western blotting were performed to detect if the expressions of VEGF and MMP-7 were modulated by SPARC expression. To further determine the effect of SPARC expression on angiogenesis in vivo, xenograft models were established and microvessel density (MVD) of different clones were detected by immunohistochemistry. Endogenous SPARC overexpression inhibited the expression of VEGF and MMP-7, as well as the angiogenesis induced by BGC-SP cells. Correspondingly, SPARC silencing increased the expression of VEGF and MMP-7, as well as the angiogenesis induced by HGC-sh cells. Elevated angiogenesis induced by SPARC silencing in HGC-sh cells was decreased when VEGF was neutralised by antibodies, and MMP-7 was knocked down in vitro. SPARC suppresses angiogenesis of gastric cancer by down-regulating the expression of VEGF and MMP-7.


Assessment of the consistency among three drug compendia in listing and ranking of drug-drug interactions.
Inconsistent information about drug-drug interactions can cause variations in prescribing, and possibly increase the incidence of morbidity and mortality. The aim of this study was to assess whether there is an inconsistency in drug-drug interaction listing and ranking in three authoritative, freely accessible online drug information sources: The British National Formulary; The Compendium about Drugs Licensed for Use in the United Kingdom (the Electronic Medicines Compendium) and the Compendium about Drugs Licensed for Use in the United States (the DailyMed). Information on drug-drug interactions for thirty drugs which have a high or medium potential for interactions have been selected for analysis. In total, 1971 drug-drug interactions were listed in all three drug information sources, of these 992 were ranked as the interactions with the potential of clinical significance. Comparative analysis identified that 63.98% of interactions were listed in only one drug information source, and 66.63% of interactions were ranked in only one drug information source. Only 15.12% listed and 11.19% ranked interactions were identified in all three information sources. Intraclass correlation coefficient indicated a weak correlation among the three drug information sources in listing (0.366), as well as in ranking drug interactions (0.467). This study showed inconsistency of information on drug-drug interaction for the selected drugs in three authoritative, freely accessible online drug information sources. The application of a uniform methodology in assessment of information, and then the presentation of information in a standardized format is required to prevent and adequately manage drug-drug interactions.


Hypertrichosis cubiti (hairy elbow syndrome): a clue to a malformation syndrome.
Hypertrichosis cubiti (HC) or hairy elbow syndrome (OMIM # 139600) consists of a localised form of long vellus hair on the extensor surfaces of the distal third of the upper arm and the proximal third of the forearm bilaterally, or occasionally on other parts of the body. In the 28 cases reported in the literature so far the elbow hair abnormality was either isolated or associated with short stature or other physical abnormalities. Most of these cases were sporadic, but autosomal dominant as well as autosomal recessive inheritance patterns have been postulated. We report on three unrelated girls (aged 7 to 11 years) of whom one presented with excess hair in the elbows alone and the other two had associated abnormalities including short stature, dysmorphic facial features and mental retardation. The literature on this subject has been reviewed and the authors focus on cases of HC with associated anomalies. A pathogenic explanation by somatic mosaicism is proposed.


Control of TRPC and store-operated channels by protein kinase C.
TRPC channels are widely expressed among cells and are believed to play important roles in receptor-mediated Ca2+ signalling. We determined that the function of TRPC channels is highly regulated by protein kinase C (PKC). Application of diacylglycerol (DAG) or elevated endogenous DAG resulting from either DAG-lipase or DAG-kinase inhibition, completely prevented TRPC5 or TRPC4 activation in both HEK293 cells and DT40 cells. This inhibitory action of DAG on TRPC5 and TRPC4 channels was clearly mediated by PKC, in distinction to the stimulatory action of DAG on TRPC3 which was PKC-independent. PKC activation totally blocked TRPC3 channel-activated in response to OAG, and was restored by PKC-blockade. PKC-inhibition resulted in decreased TRPC3 channel deactivation. Store-operated Ca2+ entry in response to PLC-coupled receptor activation but not store-depletion per se, was substantially reduced by OAG or DAG-lipase inhibition in a PKC-dependent manner. The results reveal that each TRPC subtype is strongly inhibited by DAG-induced PKC activation reflecting a likely universal feedback control on TRPCs. The profound yet distinct control by PKC and DAG on the activation of TRPC channel subtypes may be the basis of a spectrum of regulatory phenotypes of expressed TRPC channels.


Specific dose-dependent effects of ethane 1,2-dimethanesulfonate in rat and mouse Leydig cells and non-steroidogenic cells on programmed cell death.
The mechanism by which ethane 1,2-dimethanesulfonate (EDS) selectively kills Leydig cells is poorly understood. To characterize further the cell-specific actions of EDS, we studied biochemical and morphological changes during apoptosis in different Leydig cell and non-steroidogenic cell models. Rat testicular and H540 tumor Leydig cells were killed by 1-2 mM EDS, whereas 20 mM EDS were required for MA-10 cells. This higher concentration of EDS was also necessary for activation of apoptosis in non-steroidogenic Chinese hamster ovary cells, whereas COS-1 monkey kidney cells were resistant. These variable effects of EDS on apoptosis were independent of new protein synthesis and, interestingly, could be delayed by co-incubation with dibutyrl cyclic AMP. Along with cell death, we also observed chromosomal fragmentation and other hallmarks indicative of apoptosis as evidenced by DNA laddering and fluorescent microscopy. Time-lapse photography with a confocal microscope showed that the time of onset, duration and even the sequence of apoptotic events between individual H540 cells was heterogeneous. When the dose of EDS was gradually increased from 2 to 10 mM, the proportion of cells showing normal apoptotic features gradually decreased. Intriguingly, treatment with 10 mM EDS did not result in death for most cells and was marked by an absence of DNA laddering and ultrastructural features of apoptosis and necrosis. However, incubation with 20 mM EDS resulted in necrosis.These results demonstrated that the effects of EDS on cell survival are not specific to Leydig cells, that different cell types have different sensitivities to EDS and that stimulation of the cAMP pathway may mitigate EDS action. The data obtained with H540 cells further revealed that EDS can induce two types of programmed cell death.


Coffee Consumption and All-Cause, Cardiovascular, and Cancer Mortality in an Adult Mediterranean Population.
We assessed the association between usual coffee consumption and all-cause, cardiovascular (CV), and cancer mortality in an adult population in Spain, taking into account both the amount and type of coffee consumed. We used baseline data on coffee consumption and other personal variables, and the number of deaths during an 18-year follow-up period, for 1567 participants aged 20 years and older from the Valencia Nutrition Study in Spain. Total, caffeinated, and decaffeinated coffee consumption was assessed using a validated food frequency questionnaire. Cox regression models were used to estimate adjusted hazard ratios (HRs) and 95% confidence intervals (CIs). During the 18-year follow-up period, 317 died; 115 due to CV disease and 82 due to cancer. Compared with no-consumption, the consumption of ≤1 cup per day and >1 cup per day of coffee was associated with a lower risk of all-cause mortality, HR = 0.73 (95% CI: 0.56-0.97) and HR 0.56 (95% CI: 0.41-0.77), respectively. A lower cancer mortality was observed among drinkers of more than 1 cup per day compared with nondrinkers, HR 0.41 (95% CI 0.20-0.86). Regarding the type of coffee, only the overall consumption of caffeinated coffee was associated with lower all-cause mortality at 12 and 18 years of follow-up, HR = 0.66 (95% CI:0.46-0.94) and HR = 0.59 (95% CI: 0.44-0.79), respectively. In conclusion, this study suggests that the moderate consumption of coffee, particularly caffeinated coffee (range 1-6.5 cups per day), is associated with a lower all-cause and cancer mortality after a long follow-up period. No significant association was found between coffee consumption and CVD mortality.


[The pathway of vasopressin as a pharmacological target in nephrology: a narrative review].
ADH is a hormone secreted by neurohypophysis that plays different roles based on the target organ. At the renal level, this peptide is capable of causing electrolyte-free water absorption, thus playing a key role in the hydro-electrolytic balance. There are pathologies and disorders that jeopardize this balance and, in this field, ADH receptor inhibitors such as Vaptans could play a key role. By inhibiting the activation pathway of vasopressin, they are potentially useful in euvolemic and hypervolemic hypotonic hyponatremia. However, clinical trials in heart failure have not given favourable results on clinical outcomes. Even in SIADH, despite their wide use, there is no agreement by experts on their use. Since vaptans inhibit the cAMP pathway in tubular cells, their use has been proposed to inhibit cystogenesis. A clinical trial has shown favourable effects on ADPKD progression. Because vaptans have been shown to be effective in models of renal cysts disorders other than ADPKD, their use has been proposed in diseases such as nephronophthisis and recessive autosomal polycystic disease. Other possible uses of vaptans could be in kidney transplantation and cardiorenal syndrome. Due to the activity of ADH in coagulation and haemostasis, ADH's activation pathway by Desmopressin Acetate could be a useful strategy to reduce the risk of bleeding in biopsies in patients with haemorrhagic risk.


Autosomal Dominant Polycystic Kidney Disease: Clinical Assessment of Rapid Progression.
Autosomal dominant polycystic kidney disease (ADPKD) causes the development of renal cysts and leads to a decline in renal function. Limited guidance exists in clinical practice on the use of tolvaptan. A decision algorithm from the European Renal Association-European Dialysis and Transplant Association (ERA-EDTA) Working Groups of Inherited Kidney Disorders and European Renal Best Practice (WGIKD/ERBP) has been proposed to identify candidates for tolvaptan treatment; however, this algorithm has not been assessed in clinical practice. Eighteen-month cross-sectional, unicenter, observational study assessing 305 consecutive ADPKD patients. The ERA-EDTA WGIKD/ERBP algorithm with a stepwise approach was used to assess rapid progression (RP). Subsequently, expanded criteria based on the REPRISE trial were applied to evaluate the -impact of extended age (≤55 years) and estimated glomerular filtration rate (eGFR; ≥25 mL/min/1.73 m2). Historical eGFR decline, indicative of RP, was fulfilled in 26% of 73 patients who were candidates for RP assessment, mostly aged 31-55 years. Further tests including ultrasound and MRI measurements of kidney volume plus genetic testing enabled the evaluation of the remaining patients. Overall, 15.7% of patients met the criteria for rapid or likely RP using the algorithm, and the percentage increased to 27% when extending age and eGFR. The ERA-EDTA WGIKD/ERBP algorithm provides a valuable means of identifying in routine clinical practice patients who may be eligible for treatment with tolvaptan. The impact of a new threshold for age and eGFR may increase the percentage of patients to be treated.


Optimal peeling order for pedigrees with incomplete genotypic information.
The likelihood approach is common in linkage analysis of large extended pedigrees. Various peeling procedures, based on the conditional independence of separate parts of a pedigree, are typically used for likelihood calculations. A peeling order may significantly affect the complexity of such calculations, particularly for pedigrees with loops or when many pedigrees members have unknown genotypes. Several algorithms have been proposed to address this problem for pedigrees with loops. However, the problem has not been solved for pedigrees without loops until now. In this paper, we suggest a new graph theoretic algorithm for optimal selection of peeling order in zero-loop pedigrees with incomplete genotypic information. It is especially useful when multiple likelihood calculation is needed, for example, when genetic parameters are estimated or linkage with multiple marker loci is tested. The algorithm can be easily introduced into the existing software packages for linkage analysis based on the Elston-Stewart algorithm for likelihood calculation. The algorithm was implemented in a software package PedPeel, which is freely available at http://mga.bionet.nsc.ru/nlru/.


Removal of heavy metals from contaminated soil by electrodialytic remediation enhanced with organic acids.
The soil from an industrial area in Algeria was contaminated with Cr (8370 mg kg-1), Ni (1135 mg kg-1) and zinc (1200 mg kg-1). The electrodialytic remediation of this soil was studied using citric acid and EDTA as facilitating agents. 0.1 M citric acid or EDTA was added directly to the soil before it was introduced in an electrodialytic cell in an attempt to enhance the heavy metal solubility in the interstitial fluid. The more acidic pH in the soil when citric acid was used as the facilitating agent was not enough to mobilize and remove the metals from the soil. Only 7.2% of Ni and 6.7% of Zn were removed from the soil in the test with citric acid. The best results were found with EDTA, which was able to solubilize and complex Zn and Ni forming negatively charged complexes that were transported and accumulated in the anolyte. Complete removal was observed for Ni and Zn in the electrodialytic treatment with EDTA. Minor amounts of Cr were removed with both EDTA and citric acid.


Evolution of Darwin's finches and their beaks revealed by genome sequencing.
Darwin's finches, inhabiting the Galápagos archipelago and Cocos Island, constitute an iconic model for studies of speciation and adaptive evolution. Here we report the results of whole-genome re-sequencing of 120 individuals representing all of the Darwin's finch species and two close relatives. Phylogenetic analysis reveals important discrepancies with the phenotype-based taxonomy. We find extensive evidence for interspecific gene flow throughout the radiation. Hybridization has given rise to species of mixed ancestry. A 240 kilobase haplotype encompassing the ALX1 gene that encodes a transcription factor affecting craniofacial development is strongly associated with beak shape diversity across Darwin's finch species as well as within the medium ground finch (Geospiza fortis), a species that has undergone rapid evolution of beak shape in response to environmental changes. The ALX1 haplotype has contributed to diversification of beak shapes among the Darwin's finches and, thereby, to an expanded utilization of food resources.


Spatio-temporal reconstruction of dPET data using complex wavelet regularisation.
Traditionally, dynamic PET studies reconstruct temporally contiguous PET images using algorithms which ignore the inherent consistency between frames. We present a method which imposes a regularisation constraint based on wavelet denoising. This is achieved efficiently using the Dual Tree Complex Wavelet Transform (DT-CWT) of Kingsbury, which has many important advantages over the traditional discrete wavelet transform: shift invariance, implicit measure of local phase, and directional selectivity. In this paper, we apply the decomposition to the full spatio-temporal volume and use it for the reconstruction of dynamic (spatio-temporal) PET data. Instead of using traditional wavelet thresholding schemes we introduce a locally defined and empirically-determined Cross Scale regularisation technique. We show that wavelet based regularisation has the potential to produce superior reconstructions and examine the effect various levels of boundary enhancement have on the overall images. We demonstrate that wavelet-based spatio-temporally regularised reconstructions have superior performance over conventional Gaussian smoothing in simulated and clinical experiments. We find that our method outperforms conventional methods in terms of signal-to-noise ratio (SNR) and Mean Square Error (MSE), and removes the need to post-smooth the reconstruction.


Speciation in Western Scrub-Jays, Haldane's rule, and genetic clines in secondary contact.
Haldane's Rule, the tendency for the heterogametic sex to show reduced fertility in hybrid crosses, can obscure the signal of gene flow in mtDNA between species where females are heterogametic. Therefore, it is important when studying speciation and species limits in female-heterogametic species like birds to assess the signature of gene flow in the nuclear genome as well. We studied introgression of microsatellites and mtDNA across a secondary contact zone between coastal and interior lineages of Western Scrub-Jays (Aphelocoma californica) to test for a signature of Haldane's Rule: a narrower cline of introgression in mtDNA compared to nuclear markers. Our initial phylogeographic analysis revealed that there is only one major area of contact between coastal and interior lineages and identified five genetic clusters with strong spatial structuring: Pacific Slope, Interior US, Edwards Plateau (Texas), Northern Mexico, and Southern Mexico. Consistent with predictions from Haldane's Rule, mtDNA showed a narrower cline than nuclear markers across a transect through the hybrid zone. This result is not being driven by female-biased dispersal because neutral diffusion analysis, which included estimates of sex-specific dispersal rates, also showed less diffusion of mtDNA. Lineage-specific plumage traits were associated with nuclear genetic profiles for individuals in the hybrid zone, indicating that these differences are under genetic control. This study adds to a growing list of studies that support predictions of Haldane's Rule using cline analysis of multiple loci of differing inheritance modes, although alternate hypotheses like selection on different mtDNA types cannot be ruled out. That Haldane's Rule appears to be operating in this system suggests a measure of reproductive isolation between the Pacific Slope and interior lineages. Based on a variety of evidence from the phenotype, ecology, and genetics, we recommend elevating three lineages to species level: A. californica (Pacific Slope); A. woodhouseii (Interior US plus Edwards Plateau plus Northern Mexico); A. sumichrasti (Southern Mexico). The distinctive Edwards Plateau population in Texas, which was monophyletic in mtDNA except for one individual, should be studied in greater detail given habitat threat.


[Cost-effectiveness of addiction care].
A large number of interventions are available for the treatment of addiction. Professionals need to know about the effectiveness and cost-effectiveness of interventions so they can prioritise appropriate interventions for the treatment of addiction. To provide an overview of the scientific literature on the cost-effectiveness of addiction treatment for alcohol- and drug-abusers. We searched the databases Medline and Centre for Reviews and Dissemination. To be relevant for our study, articles had to focus on interventions in the health-care setting, have a Western context and have a health-related outcome measure such as quality adjusted life years (QALY). Twenty-nine studies met our inclusion criteria: 15 for alcohol and 14 for drugs. The studies on alcohol addiction related mainly to brief interventions. They proved to be cost-saving or had a favourable incremental cost-effectiveness ratio (ICER), remaining below the threshold of € 20,000 per QALY. The studies on drug addiction all involved pharmacotherapeutic interventions. In the case of 10 out of 14 interventions, the ICER was less than € 20,000 per QALY. Almost all of the interventions studied were cost-saving or cost-effective. Many studies consider only health-care costs. Additional research, for instance using a social cost-benefit analysis, could provide more details about the costs of addiction and about the impact that an intervention could have in these/the costs.


In vitro and in vivo models analyzing von Hippel-Lindau disease-specific mutations.
Mutations in the von Hippel-Lindau (VHL) tumor suppressor gene cause tissue-specific tumors, with a striking genotype-phenotype correlation. Loss of VHL expression predisposes to hemangioblastoma and clear cell renal cell carcinoma, whereas specific point mutations predispose to pheochromocytoma, polycythemia, or combinations of hemangioblastoma, renal cell carcinoma, and/or pheochromocytoma. The VHL protein (pVHL) has been implicated in many cellular activities including the hypoxia response, cell cycle arrest, apoptosis, and extracellular matrix remodeling. We have expressed missense pVHL mutations in Vhl(-/-) murine embryonic stem cells to test genotype-phenotype correlations in euploid cells. We first examined the ability of mutant pVHL to direct degradation of the hypoxia inducible factor (HIF) subunits HIF1alpha and HIF2alpha. All mutant pVHL proteins restored proper hypoxic regulation of HIF1alpha, although one VHL mutation (VHL(R167Q)) displayed impaired binding to Elongin C. This mutation also failed to restore HIF2alpha regulation. In separate assays, these embryonic stem cells were used to generate teratomas in immunocompromised mice, allowing independent assessment of the effects of specific VHL mutations on tumor growth. Surprisingly, teratomas expressing the VHL(Y112H) mutant protein displayed a growth disadvantage, despite restoring HIFalpha regulation. Finally, we observed increased microvessel density in teratomas derived from Vhl(-/-) as well as VHL(Y112H), VHL(R167Q), and VHL(R200W) embryonic stem cells. Together, these observations support the hypothesis that pVHL plays multiple roles in the cell, and that these activities can be separated via discrete VHL point mutations. The ability to dissect specific VHL functions with missense mutations in a euploid model offers a novel opportunity to elucidate the activities of VHL as a tumor suppressor.


A wearable freestanding electrochemical sensing system.
To render high-fidelity wearable biomarker data, understanding and engineering the information delivery pathway from epidermally retrieved biofluid to a readout unit are critical. By examining the biomarker information delivery pathway and recognizing near-zero strained regions within a microfluidic device, a strain-isolated pathway to preserve biomarker data fidelity is engineered. Accordingly, a generalizable and disposable freestanding electrochemical sensing system (FESS) is devised, which simultaneously facilitates sensing and out-of-plane signal interconnection with the aid of double-sided adhesion. The FESS serves as a foundation to realize a system-level design strategy, addressing the challenges of wearable biosensing, in the presence of motion, and integration with consumer electronics. To this end, a FESS-enabled smartwatch was developed, featuring sweat sampling, electrochemical sensing, and data display/transmission, all within a self-contained wearable platform. The FESS-enabled smartwatch was used to monitor the sweat metabolite profiles of individuals in sedentary and high-intensity exercise settings.


Reversal of learning deficits in a Tsc2+/- mouse model of tuberous sclerosis.
Tuberous sclerosis is a single-gene disorder caused by heterozygous mutations in the TSC1 (9q34) or TSC2 (16p13.3) gene and is frequently associated with mental retardation, autism and epilepsy. Even individuals with tuberous sclerosis and a normal intelligence quotient (approximately 50%) are commonly affected with specific neuropsychological problems, including long-term and working memory deficits. Here we report that mice with a heterozygous, inactivating mutation in the Tsc2 gene (Tsc2(+/-) mice) show deficits in learning and memory. Cognitive deficits in Tsc2(+/-) mice emerged in the absence of neuropathology and seizures, demonstrating that other disease mechanisms are involved. We show that hyperactive hippocampal mammalian target of rapamycin (mTOR) signaling led to abnormal long-term potentiation in the CA1 region of the hippocampus and consequently to deficits in hippocampal-dependent learning. These deficits included impairments in two spatial learning tasks and in contextual discrimination. Notably, we show that a brief treatment with the mTOR inhibitor rapamycin in adult mice rescues not only the synaptic plasticity, but also the behavioral deficits in this animal model of tuberous sclerosis. The results presented here reveal a biological basis for some of the cognitive deficits associated with tuberous sclerosis, and they show that treatment with mTOR antagonists ameliorates cognitive dysfunction in a mouse model of this disorder.


Social networks, social participation and self-perceived health among older people in transitional Kosovo.
A number of studies proved that social networks and social participation have beneficial health effects in western countries. However, the evidence from southeast European region is scant. We aimed to assess the extent of social networks and social participation and their relationship with self-perceived health status among older people in post-war Kosovo. A nationwide cross-sectional study was conducted in Kosovo in 2011 including a representative sample of 1890 individuals aged ≥65 years (949 men, mean age 73 ± 6 years; 941 women, mean age 74 ± 7 years; response rate: 83%). Social networks were assessed by means of number of friends and family members that participants had contacts with, whereas social participation by involvement in social groupings/organizations. Information on self-perceived health status and demographic and socioeconomic characteristics was also collected. Overall, 93% of study participants reported that they had at least weekly contacts with more than one family member, and 97% reported daily contacts with their respective friends. Conversely, only 14% of participants reported engagement with social groupings. Generally, individuals who had contacts with friends and/or engaged with social organizations reported a better health status. Our findings point to strong family ties in this patriarchal society. Conversely, levels of social participation were considerably lower in Kosovo compared with the western European countries. The low participation levels in social groupings and their putative deleterious health effects should raise the awareness of policymakers to improve the conditions and increase the degree of social participation among older people in transitional Kosovo.


Chromosomal translocation master genes, mouse models and experimental therapeutics.
Molecular biologists have elucidated general principles about chromosomal translocations by cloning oncogenes or fusion genes at chromosomal translocation junctions. These genes invariably encode intracellular proteins and in acute cancers, often involve transcription and developmental regulators, which are master regulators of cell fate (e.g. LMO2 which is involved in acute leukaemia). Chromosomal translocations are usually associated with specific cell types. The reason for this close association is under investigation using mouse models. We are trying to emulate the cell-specific consequences of chromosomal translocations in mice using homologous recombination in embryonic stem cells to generate de novo chromosomal translocations or to mimic the consequence of these translocations. In addition, chromosomal translocation genes and their products are important targets for therapy. We have designed new therapeutic strategies which include antigen-specific recruitment of endogenous cellular pathways to affect cellular viability and a novel structured form of antisense to ablate the function of fusion mRNAs. We will evaluate these procedures in the mouse models of chromosomal translocations and the long term aim is to perfect rapid procedures for characterizing patient-specific chromosomal translocations to tailor therapy to individual patients.


Enhanced material purity and resolution via synchronized laser assisted electron beam induced deposition of platinum.
We introduce a laser assisted electron beam induced deposition (LAEBID) process which is a nanoscale direct write synthesis method that integrates an electron beam induced deposition process with a synchronized pulsed laser step to induce thermal desorption of reaction by-products. Localized, spatially overlapping electron and photon pulses enable the thermal desorption of the reaction by-product while mitigating issues associated with bulk substrate heating, which can shorten the precursor residence time and distort pattern fidelity due to thermal drift. Current results demonstrate purification of platinum deposits (reduced carbon content by ~50%) with the addition of synchronized laser pulses as well as a significant reduction in deposit resistivity. Measured resistivities from platinum LAEBID structures (4 × 10(3)μΩ cm) are nearly 4 orders of magnitude lower than standard EBID platinum structures (2.2 × 10(7)μΩ cm) from the same precursor and are lower than the lowest reported EBID platinum resistivity with post-deposition annealing (1.4 × 10(4)μΩ cm). Finally the LAEBID process demonstrates improved deposit resolution by ~25% compared to EBID structures under the conditions investigated in this work.


Possible Benefit of Dietary Carnosine towards Depressive Disorders.
Many stress-related and depressive disorders have been shown to be associated with one or more of the following; shortened telomeres, raised cortisol levels and increased susceptibility to age-related dysfunction. It is suggested here that insufficient availability of the neurological peptide, carnosine, may provide a biochemical link between stress- and depression-associated phenomena: there is evidence that carnosine can enhance cortisol metabolism, suppress telomere shortening and exert anti-aging activity in model systems. Dietary supplementation with carnosine has been shown to suppress stress in animals, and improve behaviour, cognition and well-being in human subjects. It is therefore proposed that the therapeutic potential of carnosine dietary supplementation towards stress-related and depressive disorders should be examined.


Speed discrimination in the far monocular periphery: A relative advantage for interocular comparisons consistent with self-motion.
Some animals with lateral eyes (such as bees) control their navigation through the 3D world using velocity differences between the two eyes. Other animals with frontal eyes (such as primates, including humans) can perceive 3D motion based on the different velocities that a moving object projects upon the two retinae. Although one type of 3D motion perception involves a comparison between velocities from vastly different (monocular) portions of the visual field, and the other involves a comparison within overlapping (binocular) portions of the visual field, both compare velocities across the two eyes. Here we asked whether human interocular velocity comparisons, typically studied in the context of binocularly overlapping vision, operate in the far lateral (and hence, monocular) periphery and, if so, whether these comparisons were accordant with conventional interocular motion processing. We found that speed discrimination was indeed better between the two eyes' monocular visual fields, as compared to within a single eye's (monocular) visual field, but only when the velocities were consistent with commonly encountered motion. This intriguing finding suggests that mechanisms sensitive to relative motion information on opposite sides of an animal may have been retained, or at some point independently achieved, as the eyes became frontal in some animals.


Detection of 10 sweeteners in various foods by liquid chromatography/tandem mass spectrometry.
The analytical method for sweeteners in various food matrixes is very important for food quality control and regulation enforcement. A simple and rapid method for the simultaneous determination of 10 sweeteners [acesulfame potassium (ACS-K), aspartame (ASP), cyclamate (CYC), dulcin (DUL), glycyrrhizic acid (GA), neotame (NEO), neohesperidin dihydrochalcone (NHDC), saccharin (SAC), sucralose (SCL), and stevioside (STV)] in various foods by liquid chromatography/tandem mass chromatography (LC-MS/MS) was developed. The chromatographic separation was performed on a Phenomenex Luna Phenyl-Hexyl (5 μm, 4.6 mm × 150 mm) column with gradient elution of 10 mM ammonium acetate in water and 10 mM ammonium acetate in methanol. The recoveries of the 10 sweeteners were between 75% and 120%, and the coefficients of variation were less than 20%. The limits of quantification were 0.5 μg/kg for NHDC and SCL. For the other sweeteners, the limits of quantification were 0.1 μg/kg. Compared to the traditional high-performance liquid chromatography method, the LC-MS/MS method could provide better sensitivity, higher throughput, enhanced specificity, and more sweeteners analyzed in a single run. The samples included 27 beverages (16 alcoholic and 11 nonalcoholic beverages) and 15 pickled foods (1 pickled pepper, 3 candies, and 11 candied fruits). Two remanufactured wines were found to contain 7.2, 8.5 μg/g SAC and 126.5, 123 μg/g CYC, respectively. ACS-K, ASP, SCL, and NEO were detected in five beverages and drinks. The pickled peppers and candied fruits were found to contain SAC, GA, CYC, ASP, STV, NEO, and ACS-K. The wine with sweeteners detected was remanufactured wine, not naturally fermented wine. Therefore, the ingredient label for the sweeteners of remanufactured wine should be regulated by the proper authority for inspection of sweeteners.


Comparative study of chemical and biochemical properties of different melon cultivars: standard, hybrid, and grafted melons.
Chemical and biochemical properties of standard, hybrid, and grafted melons cultivated under the same agricultural conditions in adjacent fields in the Cumra region of Turkey were investigated and compared based on pH, Brix, antioxidant activity, total phenolics, ascorbic acid, individual phenolics, sugar, and organic acid values. Seventeen different phenolic constituents were quantified by reverse phase high-performance liquid chromatography (RP-HPLC). The highest phenolic acid variability and content were detected in the standard melon. Sugar and organic acid compositions of melon cultivars were tested by capillary electrophoresis, and significant differences in types and contents of individual sugars and organic acids were determined among the cultivars. Standard Cinikiz Cumra melons had the highest ascorbic acid, total phenolics, and total sugar contents. The fructose/glucose ratio increased three times in grafted melon as compared with standard melon. While sugar alcohol mannitol existed in the standard and hybrid cultivars, this constituent disappeared in the grafted types. Citric acid found in the standard cultivar was not detected in the hybrid and grafted types. Consequently, it was concluded that the nutritional value of melons changed by the application of hybridization, grafting, or standard (open pollinated) production methods. The standard melon was found to have the highest score in terms of taste, because of its highest sweetness and sourness. It was also found preferable because of its high antioxidant activity, total phenolic and ascorbic acid contents.


Heterogeneous vancomycin resistance in Staphylococcus aureus: a review of epidemiology, diagnosis, and clinical significance.
To provide an overview of heterogeneously glycopeptide intermediate Staphylococcus aureus, its epidemiology, methods of diagnosis, and clinical relevance. Literature was retrieved in December 2009 through Academic Search Premier, Alt-Health Watch, MEDLINE, OVID, and PubMed, using the search terms heterogeneous glycopeptide-intermediate Staphylococcus aureus, hGISA, hVISA, vancomycin-intermediate Staphylococcus aureus, GISA, and VISA (from 2003 to December 2009). Additional references were obtained through review of publication citations. All articles retrieved through the literature search and reference review were evaluated for inclusion in this review. Heterogeneously glycopeptide-intermediate S. aureus (hGISA) has been shown to significantly complicate treatment for patients with bacteremia and often escapes detection in clinical laboratories. Population analysis profile area under the curve ratio (PAP-AUC) is considered to be the gold standard of hGISA detection; however, it is labor-intensive, expensive, and not feasible in a clinical setting. The Etest macromethod is a reasonable alternative to PAP, yielding high sensitivity and specificity, and has potential for routine use in clinical laboratories. Due to these difficulties in detection, an exact prevalence is difficult to obtain though, based on available data, it appears that approximately 5-15% of isolates display this phenotype. hGISA is associated with prolonged bacteremia, high bacterial load infections, prolonged antibiotic therapy, prolonged hospitalization, treatment failure, and potential for increased mortality. Overall, hGISA is associated with a poor clinical course in patients with bacteremia. Information is still lacking regarding the actual prevalence of hGISA as well as the extent of its clinical impact. A uniform method of diagnosis must be established in order to better account for all cases of hGISA. More studies must be conducted to assess clinical outcomes of hGISA.


Pre-treatment magnetic resonance enterography findings predict the response to TNF-alpha inhibitors in Crohn's disease.
Identifying predictors of therapeutic response is the cornerstone of personalised medicine. To identify predictors of long-term healing of severe inflammatory lesions based on magnetic resonance enterography (MRE) findings in patients with Crohn's disease (CD) treated with tumour necrosis factor alpha (TNF-α) inhibitors. This prospective longitudinal single-centre study included patients with clinically active CD requiring treatment with TNF-α inhibitors with at least one intestinal segment with a severe inflammatory lesion detected by MRE (segmental MaRIA ≥11). MRE data were obtained at baseline, and at weeks 14 and 46. The primary endpoint was healing of severe inflammatory lesions (MaRIA <11) in each segment. The secondary endpoint was healing of all severe inflammatory lesions on a per-patient analysis. We included 58 patients with 86 intestinal segments with severe inflammatory lesions. At week 46, healing of severe lesions was found in 51/86 (59.3%) segments, and complete healing of inflammatory lesions in all segments was found in 28/58 (48.6%) patients. Multivariable analysis found baseline-negative predictors of long-term healing of severe inflammation were ileal (as opposed to colonic) location (OR 0.00, [0.00-0.56] P = 0.002) and presence of creeping fat on MRE (OR 0.00 [0.00-0.57]; P = 0.001). Persistence of segmental MaRIA score >10.6 at week 14 was a negative predictor of healing at week 46 (OR 0.3 [0.04--0.38]; P < 0.001). In patients with CD, the absence of creeping fat detected at baseline MRE and location of severe inflammatory lesions are clinically relevant predictors of long-term healing of severe inflammation under treatment with TNF-α inhibitors.


Clinical outcome of bi-unicompartmental knee arthroplasty for both medial and lateral femorotibial arthritis: a systematic review-is there proof of concept?
Unicompartmental knee arthroplasty (UKA) is a well-accepted treatment for isolated unicompartmental osteoarthritis (OA) of the knee. In previous literature, it has been suggested that bi-unicompartmental knee arthroplasty (bi-UKA) which uses two UKA implants in both the medial and lateral compartments of the same knee is a feasible and viable option for the treatment of knee OA. Given the advantages of UKA treatment, it is warranted to review the literature of bi-UKA and discuss the evidence in terms of implant selection, indications, surgical techniques, and outcomes, respectively. Following the PRISMA guidelines, PubMed, Medline, Embase, CINAHL, Web of Science, and Cochrane Library were searched for studies presenting outcome of bi-UKA. Studies were included if they reported clinical outcomes using two unicompartmental prostheses for both medial and lateral femorotibial arthritis. Studies with the addition of patellofemoral arthroplasty or concomitant soft-tissue reconstruction and those not published in English were excluded. In the early literature, the procedure of bi-UKA were performed for very severe OA and rheumatoid arthritis, but indications have evolved to reflect a more contemporary case-mix of knee OA patients. Both mobile and fixed bearing implants have been used, with the latter being the most frequent choice. A medial parapatellar approach for incision and arthrotomy has been the most frequently used technique. The present review found a promising clinical outcome of both simultaneous and staged bi-UKA although the number of long-term follow-up studies was limited. Both simultaneous and staged bi-UKA has demonstrated good functional outcomes. However, the volume and level of evidence in general is low for studies captured in this review, and the data on long-term outcomes remain limited. The present review indicates that bi-UKA is a feasible and viable surgical option for bicompartmental femorotibial OA in carefully selected patients.


Common oxytocin receptor gene variant interacts with rejection sensitivity to influence cortisol reactivity during negative evaluation.
The study tested the hypothesis that variation in the oxytocin receptor gene (OXTR rs53576) and self-report of rejection sensitivity are associated with adrenocortical reactivity to social stress. Participants (N=94; 36.17% male; Mage=20.18yrs; 62.8% Caucasian) completed a writing task on a salient problem in society, provided self-report via questionnaire of rejection sensitivity, and were then informed that a committee of peers would evaluate their written comments. Participants received either scripted praise or criticism as a way to induce social evaluative threat. Saliva was collected before and after the stress task and assayed for cortisol. Results revealed that cortisol levels increased in participants with moderate levels of social rejection sensitivity-inferred by rs53576 genotype and reported rejection sensitivity-while cortisol levels decreased in participants with high and low levels of social rejection sensitivity. Our findings suggest a curvilinear relationship between social rejection sensitivity and cortisol reactivity in the context of social rejection, warranting further consideration in future studies.