Broad-Spectrum Kinase Profiling in Live Cells with Lysine-Targeted Sulfonyl Fluoride Probes.
Protein kinases comprise a large family of structurally related enzymes. A major goal in kinase-inhibitor development is to selectively engage the desired kinase while avoiding myriad off-target kinases. However, quantifying inhibitor interactions with multiple endogenous kinases in live cells remains an unmet challenge. Here, we report the design of sulfonyl fluoride probes that covalently label a broad swath of the intracellular kinome with high efficiency. Protein crystallography and mass spectrometry confirmed a chemoselective reaction between the sulfonyl fluoride and a conserved lysine in the ATP binding site. Optimized probe 2 (XO44) covalently modified up to 133 endogenous kinases, efficiently competing with high intracellular concentrations of ATP. We employed probe 2 and label-free mass spectrometry to quantify intracellular kinase engagement by the approved drug, dasatinib. The data revealed saturable dasatinib binding to a small subset of kinase targets at clinically relevant concentrations, highlighting the utility of lysine-targeted sulfonyl fluoride probes in demanding chemoproteomic applications.


Imaging Correlates of Memory and Concussion History in Retired National Football League Athletes.
To our knowledge, this is the first study to show an association between concussion, cognition, and anatomical structural brain changes across the age spectrum in former National Football League athletes. To assess the relationship of hippocampal volume, memory performance, and the influence of concussion history in retired National Football League athletes with and without mild cognitive impairment (MCI). This retrospective cohort study assessed differences between groups, mean hippocampal volumes, and memory performance by computing age quintiles based on group-specific linear regression models corrected for multiple comparisons for both athletes and control participants. The study was conducted starting in November 2010 and is ongoing at a research center in the northern region of Texas. This current analysis was conducted from October 9, 2013, to August 21, 2014. Participants included 28 retired National Football League athletes, 8 of whom had MCI and a history of concussion, 21 cognitively healthy control participants, and 6 control participants with MCI without concussion. Hippocampal volume, age, California Verbal Learning Test scores, and the number of grade 3 (G3) concussions. In addition, the number of games played was examined as an objective variable pertaining to football history. The mean (SD) age was 58.1 (13) years for the 28 former athletes and 59.0 (12) years for the 27 control participants. Retired athletes with concussion history but without cognitive impairment had normal but significantly lower California Verbal Learning Test scores compared with control participants (mean [SD], 52.5 [8] vs 60.24 [7]; P = .002); those with a concussion history and MCI performed worse (mean [SD], 37 [8.62]) compared with both control participants (P < .001) and athletes without memory impairment (P < .001). Among the athletes, 17 had a G3 concussion and 11 did not. Older retired athletes with at least 1 G3 concussion had significantly smaller bilateral hippocampal volumes compared with control participants at the 40th age percentile (left, P = .04; right, P = .03), 60th percentile (left, P = .009; right, P = .01), and 80th percentile (left, P = .001; right, P = .002) and a smaller right hippocampal volume compared with athletes without a G3 concussion at the 40th percentile (P = .03), 60th percentile (P = .02), and 80th percentile (P = .02). Athletes with a history of G3 concussion were more likely to have MCI (7 of 7) compared with retired athletes without a history of G3 concussion (1 of 5) older than 63 years (P = .01). In addition, the left hippocampal volume in retired athletes with MCI and concussion was significantly smaller compared with control participants with MCI (P = .03). Prior concussion that results in loss of consciousness is a risk factor for increased hippocampal atrophy and the development of MCI. In individuals with MCI, hippocampal volume loss appears greater among those with a history of concussion.


A nonsecreted variant of interleukin-4 is associated with apoptosis: implication for the T helper-2 polarization in HIV infection.
We report the detection of an interleukin-4 (IL-4) variant whose expression is tightly associated with deprivation apoptosis. It is detected with the 8D4 anti-IL-4 monoclonal antibody (mAb) not only in T helper-2 (Th2) but also in Th1 clones, and primary T cells, and it is a nonsecreted molecule. It is not expressed during primary necrosis. Our data suggest that de novo IL-4 transcription of an alternative IL-4 mRNA (IL-4 delta(13)) is induced during deprivation apoptosis. In HIV-infected patients, increased expression of IL-4 in T cells is highly correlated to increased apoptosis, restricted to 8D4 reactivity (r(2) = 0.84 between % 8D4-8(+) and % 7- amino-actinomycin D-positive [7-AAD(+)] peripheral T cells, P <.0001), and associated with disease progression. The particular reactivity of apoptotic T cells to 8D4 mAb may explain some discordances among studies analyzing the Th1/Th2 balance in HIV infection and questions the function of this intracellular type 2 signal.


Optoelectronic data filter for selection and projection.
Database processing, like the majority of nonnumerical applications, exhibits a high degree of functional parallelism but does not require complex operations; therefore it is amenable to optical solutions. The architecture of an optoelectronic filter that is capable of performing selection and projection operations on a two-dimensional data array in a relational database environment is presented. The system receives input from a parallel optical memory, one page at a time, and performs logic operations by using optoelectronic smart pixels based on heterostructure phototransistors and vertical-cavity surfaceemitting lasers. Combinations of AND and XOR gates are used to realize row-lcolumn masking and comparisons of input data against user-supplied search arguments. The main goal of the filter is to reduce the effective data rate between the highly parallel optical storage and the low input data rate conventional electronic computer, thus efficiently interfacing currently available photonic and electronic technologies.


Calling Where It Counts: Subordinate Pied Babblers Target the Audience of Their Vocal Advertisements.
For territorial group-living species, opportunities to reproduce on the natal territory can be limited by a number of factors including the availability of resources within a territory, access to unrelated individuals, and monopolies on reproduction by dominant group members. Individuals looking to reproduce are therefore faced with the options of either waiting for a breeding opportunity to arise in the natal territory, or searching for reproductive opportunities in non-natal groups. In the cooperatively breeding Southern pied babbler, Turdoides bicolor, most individuals who achieve reproductive success do so through taking up dominant breeding positions within non-natal groups. For subordinate pied babblers therefore, searching for breeding opportunities in non-natal groups is of primary importance as this represents the major route to reproductive success. However, prospecting (where individuals leave the group to search for reproductive opportunities within other groups) is costly and individuals rapidly lose weight when not part of a group. Here we demonstrate that subordinate pied babblers adopt an alternative strategy for mate attraction by vocal advertisement from within their natal territories. We show that subordinates focus their calling efforts on the edges of their territory, and specifically near boundaries with neighbouring groups that have potential breeding partners (unrelated individuals of the opposite sex). In contrast to prospecting, calling individuals showed no body mass loss associated with this behaviour, suggesting that calling from within the group may provide a 'cheap' advertisement strategy. Additionally, we show that subordinates use information regarding the composition of neighbouring groups to target the greatest number of potential mating partners.


Vitamin D3 Prevents Calcium-Induced Progression of Early-Stage Prostate Tumors by Counteracting TRPC6 and Calcium Sensing Receptor Upregulation.
Active surveillance has emerged as an alternative to immediate treatment for men with low-risk prostate cancer. Accordingly, identification of environmental factors that facilitate progression to more aggressive stages is critical for disease prevention. Although calcium-enriched diets have been speculated to increase prostate cancer risk, their impact on early-stage tumors remains unexplored. In this study, we addressed this issue with a large interventional animal study. Mouse models of fully penetrant and slowly evolving prostate tumorigenesis showed that a high calcium diet dramatically accelerated the progression of prostate intraepithelial neoplasia, by promoting cell proliferation, micro-invasion, tissue inflammation, and expression of acknowledged prostate cancer markers. Strikingly, dietary vitamin D prevented these calcium-triggered tumorigenic effects. Expression profiling and in vitro mechanistic studies showed that stimulation of PC-3 cells with extracellular Ca2+ resulted in an increase in cell proliferation rate, store-operated calcium entry (SOCE) amplitude, cationic channel TRPC6, and calcium sensing receptor (CaSR) expression. Notably, administration of the active vitamin D metabolite calcitriol reversed all these effects. Silencing CaSR or TRPC6 expression in calcium-stimulated PC3 cells decreased cell proliferation and SOCE. Overall, our results demonstrate the protective effects of vitamin D supplementation in blocking the progression of early-stage prostate lesions induced by a calcium-rich diet. Cancer Res; 77(2); 355-65. ©2016 AACR.


Insanity findings and evaluation practices: A state-wide review of court-ordered reports.
Evaluations of legal sanity are some of the most complex and consequential mental health evaluations that forensic clinicians perform for the courts. Thus, there is strong reason to monitor the wide-scale process and conclusions of sanity evaluations. In this study, we review 1,111 court-ordered sanity evaluation reports submitted by 74 evaluators in Virginia from the first year after the state initiated an oversight system that allowed for such comprehensive review. Overall, the base rate of insanity findings was 16.9%, although base rates of insanity findings among individual evaluators varied from 0% to 50%. Similarly, most evaluators cited the cognitive (rather than volitional) criteria of the insanity defense as the basis for their insanity findings, although evaluators varied in their patterns of citing these underlying insanity criteria. Our review revealed other trends in practice, such as the rarity of psychological testing (2% of cases) and the frequency of conveying conclusions in "ultimate issue" format (76%). Overall, findings reveal that a majority of reports seem reasonably consistent with practice guidelines, but also reveal some idiosyncratic practices or patterns that suggest there is opportunity for improvement.


Mineral Formation in the Larval Zebrafish Tail Bone Occurs via an Acidic Disordered Calcium Phosphate Phase.
Both in vivo and ex vivo observations support the hypothesis that bone mineral formation proceeds via disordered precursor phases. The characteristics of the precursor phases are not well defined, but octacalcium phosphate-like, amorphous calcium phosphate-like, and HPO42--enriched phases were detected. Here we use in vivo Raman spectroscopy and high-resolution wide-angle X-ray diffraction (WAXD) to characterize and map at 2 μm resolution the mineral phases in the rapidly forming tail fin bones of living zebrafish larvae and zebrafish larvae immediately after sacrifice, respectively. Raman spectroscopy shows the presence of an acidic disordered calcium phosphate phase with additional characteristic features of HPO42- at the bone-cell interface. The complexity in the position and shape of the ν1 PO4 peak viewed by in vivo Raman spectroscopy emphasizes the heterogeneity of the mineral during bone formation. WAXD detects an additional isolated peak, appearing alone or together with the characteristic diffraction pattern of carbonated hydroxyapatite. This unidentified phase is located at the interface between the mature bone and the surrounding tissue, similar to the location at which the disordered phase was observed by Raman spectroscopy. The variable peak positions and profiles support the notion that this is an unstable disordered precursor phase, which conceivably crystallized during the X-ray diffraction measurement. Interestingly, this precursor phase is co-aligned with the c-axes of the mature bone crystals and thus is in intimate relation with the surrounding collagen matrix. We conclude that a major disordered precursor mineral phase containing HPO42- is part of the deposition pathway of the rapidly forming tail fin bones of the zebrafish.


Methods uncovering usability issues in medication-related alerting functions: results from a systematic review.
This paper aims at listing the methods used to evaluate the usability of medication-related alerting functions and at knowing what type of usability issues those methods allow to detect. A sub-analysis of data from this systematic review has been performed. Methods applied in the included papers were collected. Then, included papers were sorted in four types of evaluation: "expert evaluation", "user- testing/simulation", "on site observation" and "impact studies". The types of usability issues (usability flaws, usage problems and negative outcomes) uncovered by those evaluations were analyzed. Results show that a large set of methods are used. The largest proportion of papers uses "on site observation" evaluation. This is the only evaluation type for which every kind of usability flaws, usage problems and outcomes are detected. It is somehow surprising that, in a usability systematic review, most of the papers included use a method that is not often presented as a usability method. Results are discussed about the opportunity to provide usability information collected after the implementation of the technology during their design process, i.e. before their implementation.


Art activities and qigong exercise for the well-being of older adults in nursing homes in Indonesia: a randomized controlled trial.
Objective: This study aims to investigate the effect of an integrated intervention of art activities and Qigong exercise on the well-being of older adults in nursing homes in Indonesia.Method: We employed a randomized controlled trial with 4 specific groups, i.e. art, Qigong, integration of art and Qigong, and control group. A total of 267 participants aged 50 years or older were recruited from 9 nursing homes in Jakarta, Indonesia. The participants were randomly allocated to one of the four groups, attending two intervention sessions per week for eight weeks (16 sessions), lasting 90 minutes each. Measurements were administered at baseline (T0) and post-intervention (T1). The primary outcome was well-being (WHOQOL-Bref) and secondary outcomes were satisfaction with life (SWLS), depression (BDI-II), and health status (SF-36).Results: The art intervention had a significant positive effect on well-being, in particular in the domain of social relations. It also led to a decrease in depressive symptoms, as did the integration intervention. No significant effects were visible in the Qigong group nor in the integrated intervention compared to either art or Qigong alone.Conclusion: Interventions such as art programs and an integration of art and Qigong may give psychological benefits to older adults. Yet, results of the study need to be interpreted with caution and need to be replicated. A qualitative approach would be welcome to get an in-depth understanding of why art intervention is especially beneficial. (Trial registration: Clinicaltrials.gov NCT02957773, registered 28 September 2016).


Molecular mechanisms of ezetimibe-induced attenuation of postprandial hypertriglyceridemia.
Postprandial hypertriglyceridemia (PHTG) has been shown repeatedly to be associated with metabolic syndrome and atherosclerotic cardiovascular diseases. We have recently reported that ezetimibe inhibits PHTG in patients with type IIb hyperlipidemia. Ezetimibe was also reported to atten-uate PHTG in combination with low-dose statins in patients with obesity or metabolic syndrome. We reported CD36-deficient (CD36KO) mice as a new model for PHTG, in which the synthesis of chylomicron (CM) in the small intestines is enhanced. In the current study, we investigated the effect of ezetimibe on PHTG in this mouse model of metabolic syndrome. Wild-type (WT) mice fed a western diet, and CD36KO mice fed a normal chow diet, respectively, were treated for 3 weeks with and without ezetimibe, followed by an evaluation of triglyceride (TG) concentrations by enzymatic method and by high performance liquid chromatogra-phy (HPLC) as well as those of and apolipoprotein (Apo) B-48 in plasma and intestinal lymph after oral fat loading with olive oil. Intestinal mucosa was also harvested to evaluate the transcriptional regulation of the genes involved in the intestinal production of ApoB-containing lipoproteins. Ezetimibe dramatically reduced PHTG in both WT and CD36KO mice. HPLC analysis of plasma showed that the decrease in TG content in CM and CM remnants-sized particles contributed to this suppression, suggesting that CM production in the small intestines might be reduced after ezetimibe treatment. Intestinal lymph was collected after oral fat loading in ezetimibe-treated and non-treated mice. Both TG content and ApoB-48 mass were decreased in ezetimibe-treated mice. The quantitative RT-PCR of intestinal mucosa showed down-regulation of the mRNA expression of FATP4 and ApoB in both groups along with FABP2, DGAT1, DGAT2 and SCD1 in WT mice at postprandial state after ezetimibe treatment. Ezetimibe alone reduces PHTG by blocking both the absorption of cholesterol and the intracellular trafficking and metabolism of long-chain fatty acids in enterocytes, resulting in the reduction of the formation of ApoB-48 which is necessary for the ApoB48-containing lipoprotein production in the small intestines.


Radiological manifestations of cardiovocal syndrome.
Cardiovocal syndrome is a rare clinical diagnosis that signifies vocal cord palsy associated with various cardiovascular diseases. The radiological manifestations of six cases of clinically diagnosed cardiovocal syndrome are reviewed retrospectively. The common clinical presentation of these six cases was hoarseness that was due to left vocal cord palsy. By means of chest X-rays (CXR) and cardioangiography, four cases were respectively diagnosed to have atrial septal defect (n = 1), ventricular septal defect (n = 1) and mitral stenosis (n = 2), all of which were associated with a common radiological finding of dilatation of pulmonary trunk. The other two cases were separately diagnosed to have proximal descending aortic saccular aneurysm (n = 1) by means of CXR and angiography, and fusiform ascending aortic aneurysm (n = 1) by computed tomography and magnetic resonance image. All these radiological presentations of either aortic aneurysm or dilated pulmonary trunk were associated with encroachment of aorticopulmonary window for which the left recurrent laryngeal nerve was presumably compressed with the result of left vocal cord palsy. Hoarseness that is caused by these underlying cardiovascular diseases is correctable after treatment of the underlying diseases. Hence, clinicians and radiologists should pay attention to the radiological manifestations of patients whose chief complaints are hoarseness and cardiovocal syndrome.


Nurse-Guided Internet-Delivered Cognitive Behavioral Therapy for Insomnia in General Practice: Results from a Pragmatic Randomized Clinical Trial.
Guidelines recommend cognitive behavioral therapy for insomnia (CBT-I) as the first line of treatment for insomnia in general practice, but CBT-I is rarely available. Nurse-guided Internet-delivered CBT-I might be a solution to improve access to care. We aimed to determine the effectiveness of nurse-guided Internet-delivered CBT-I (I-CBT-I) on insomnia severity experienced by patients in general practice. Nurse-guided I-CBT-I ("i-Sleep") was compared to care-as-usual (and I-CBT-I after 6 months) in 15 participating general practices among 134 patients (≥18 years old) with clinical insomnia symptoms. Assessments took place at 8, 26 and 52 weeks. Primary outcome was self-reported insomnia severity (Insomnia Severity Index) at 8 weeks. Secondary outcomes were sleep diary indices, depression and anxiety symptoms (Hospital Anxiety and Depression Scale), fatigue, daytime consequences of insomnia, sleep medication and adverse events. Two thirds of the 69 intervention patients (n = 47; 68%) completed the whole intervention. At the posttest examination, there were large significant effects for insomnia severity (Cohen's d =1.66), several sleep diary variables (wake after sleep onset, number of awakenings, terminal wakefulness, sleep efficiency, sleep quality) and depression. At 26 weeks there were still significant effects on insomnia severity (d = 1.02) and on total sleep time and sleep efficiency. No significant effects were observed for anxiety, fatigue, daily functioning or sleep medication. No adverse events were reported. Nurse-guided I-CBT-I effectively reduces insomnia severity among general practice patients. I-CBT-I enables general practitioners to offer effective insomnia care in accordance with the clinical guidelines.


Nasal toxicity of manganese sulfate and manganese phosphate in young male rats following subchronic (13-week) inhalation exposure.
Growing evidence suggests that nasal deposition and transport along the olfactory nerve represents a route by which inhaled manganese and certain other metals are delivered to the rodent brain. The toxicological significance of olfactory transport of manganese remains poorly defined. In rats, repeated intranasal instillation of manganese chloride results in injury to the olfactory epithelium and neurotoxicity as evidenced by increased glial fibrillary acidic protein (GFAP) concentrations in olfactory bulb astrocytes. The purpose of the present study was to further characterize the nasal toxicity of manganese sulfate (MnSO(4)) and manganese phosphate (as hureaulite) in young adult male rats following subchronic (90-day) exposure to air, MnSO(4) (0.01, 0.1, and 0.5 mg Mn/m(3)), or hureaulite (0.1 mg Mn/m(3)). Nasal pathology, brain GFAP levels, and brain manganese concentrations were assessed immediately following the end of the 90-day exposure and 45 days thereafter. Elevated end-of-exposure olfactory bulb, striatum, and cerebellum manganese concentrations were observed following MnSO(4) exposure to > or = 0.01, > or = 0.1, and 0.5 mg Mn/m(3), respectively. Exposure to MnSO(4) or hureaulite did not affect olfactory bulb, cerebellar, or striatal GFAP concentrations. Exposure to MnSO(4) (0.5 mg Mn/m(3)) was also associated with reversible inflammation within the nasal respiratory epithelium, while the olfactory epithelium was unaffected by manganese inhalation. These results confirm that high-dose manganese inhalation can result in nasal toxicity (irritation) and increased delivery of manganese to the brain; however, we could not confirm that manganese inhalation would result in altered brain GFAP concentrations.


Multi-region hemispheric specialization differentiates human from nonhuman primate brain function.
The human behavioral repertoire greatly exceeds that of nonhuman primates. Anatomical specializations of the human brain include an enlarged neocortex and prefrontal cortex (Semendeferi et al. in Am J Phys Anthropol 114:224-241, 2001), but regional enlargements alone cannot account for these vast functional differences. Hemispheric specialization has long believed to be a major contributing factor to such distinctive human characteristics as motor dominance, attentional control and language. Yet structural cerebral asymmetries, documented in both humans and some nonhuman primate species, are relatively minor compared to behavioral lateralization. Identifying the mechanisms that underlie these functional differences remains a goal of considerable interest. Here, we investigate the intrinsic connectivity networks in four primate species (humans, chimpanzees, baboons, and capuchin monkeys) using resting-state fMRI to evaluate the intra- and inter- hemispheric coherences of spontaneous BOLD fluctuation. All three nonhuman primate species displayed lateralized functional networks that were strikingly similar to those observed in humans. However, only humans had multi-region lateralized networks, which provide fronto-parietal connectivity. Our results indicate that this pattern of within-hemisphere connectivity distinguishes humans from nonhuman primates.


Aspirin and Risk of Subarachnoid Hemorrhage: Systematic Review and Meta-Analysis.
Recent studies have suggested that the use of low-dose aspirin may reduce the risk of aneurysmal subarachnoid hemorrhage (aSAH). We aimed to evaluate any association between aspirin use and risk of aSAH based on the literature, and whether this is influenced by duration or frequency of aspirin use. A search of electronic databases was done from inception to September 2016. For each study, data on risk of aSAH in aspirin versus nonaspirin users were used to generate odds ratios and 95% confidence intervals, and combined using inverse variance-weighted averages of logarithmic odds ratios in a random-effects models. From 7 included studies, no significant difference was noted between aspirin use of any duration or frequency and nonaspirin users (odds ratio, 1.00; 95% confidence interval, 0.81-1.24; P=0.99). We found a significant association between short-term use of aspirin (<3 months) and the risk of aSAH (odds ratio, 1.61; 95% confidence interval, 1.20-2.18; P=0.002). No significant difference was found in terms of risk of aSAH for 3 to 12 months, 1 to 3 years, and >3 years of durations of use. No significant association was found between infrequent aspirin use (≤2× per week) or frequent use (≥3× per week) with risk of aSAH. Current evidence suggests that short-term (<3 months) use of aspirin is associated with increased risk of aSAH. Limitations include substantial heterogenity of the included studies. The role of long-term aspirin in reducing risk of aSAH remains unclear and ideally should be addressed by an appropriately designed randomized controlled trial.


Cutaneous burn increases apoptosis in the gut epithelium of mice.
Gut mucosal homeostasis depends on a balance between cell proliferation and cell death. After cutaneous burn injury, gut mucosal weight has been shown to decrease. This decrease in weight was paradoxically associated with an increase in gut proliferative factors. For mucosal weight to decrease in the presence of increased proliferation, there must be an even greater increase in cell death. We postulate that cutaneous burn injury causes an increase in gut epithelial cell death primarily by apoptosis. We produced a 30% full-thickness scald burn in the dorsum of anesthetized male C57BL6 mice and collected the proximal small bowel at 12, 24, 36, 48, and 60 hours after injury. Sham burned animals served as controls. Apoptosis and proliferation were measured by immunohistochemical assays (terminal deoxyuridine nick-end labeling for apoptosis and proliferative cell nuclear antigen assay for proliferation). Apoptosis was also measured by ELISA for cytoplasmic histone-associated DNA fragments. Mucosal height was determined on histologic sections. The two groups were compared at each time point using Wilcoxon two-sample test and t-test with Bonferroni's correction where appropriate. The percentage of apoptotic cells (number of cells stained by terminal deoxyuridine nick-end labeling per 100 villus cells) was significantly higher at 12, 24, and 48 hours after injury. This increase was corroborated by an increase in the ELISA at 12 hours. Proliferation as measured by immunostaining for proliferative cell nuclear antigen significantly increased at 12, 24, 48, and 60 hours. Mucosal height as a gross measure of mucosal atrophy was not different between the groups. We have shown an increase in apoptosis coupled with an increase in proliferation after a burn injury. These results imply an increase in cell turnover in the gut epithelial cells after a burn. Impaired bowel function has been demonstrated repeatedly after burn injury, and this increase in cell turnover may be related.


EAHP European Statements baseline survey 2015: methodology.
The European Statements of Hospital Pharmacy are a set of hospital pharmacy practice standards published by the European Association of Hospital Pharmacists (EAHP) for European health systems to ensure safe, effective and optimal use of medicines in collaboration with multidisciplinary teams. Keele University was commissioned to conduct a survey among European hospital pharmacists to establish a baseline to assess awareness of the Statements and to identify any barriers to their implementation. The survey was conducted from January 2015 to March 2015, spanning 16 languages and 34 countries. The survey consisted of three sections: ▸ Section A: general questions about the participant's hospital pharmacy, such as workforce skill-mix and number of beds served; ▸ Section B: questions about the current activity of pharmacists around each statement; ▸ Section C: questions about the hospital's readiness and ability to implement the statements. In section B, a value was allocated to each response to rate the degree to which they were able to comply with each statement (where 1=never able to comply, 5=always complied). In section C, they were asked to what degree they agreed with the question (1 for strongly disagree, 5 for strongly agree). A response of 1 or 2 was deemed to indicate some difficulty in complying with that statement-a 'negative response'. Where this was the case, the participant was given the option to provide a free text response explaining the reasons for this difficulty. The full results are discussed in a subsequent paper (see page 69). Overall, the response rate was 18%, but the variation in this was marked. 22 of the 34 countries had a response rate of over 30%. The methods used for this survey results informs the methodology and scope of future EAHP surveys Recommendations for future surveys are as follows: ▸ Keep the survey short and easy to complete (to within 20 min); ▸ Specifically enquire for each question if capacity and capability are the key barriers to implementation; ▸ Construct survey response options for each question to identify barriers other than capacity and capability; ▸ Identify the key drivers for change in countries where implementation has occurred or is occurring; ▸ Compile better intelligence about the number of surveys sent out in the first place (as those countries with a low response rate sent out lots of invites to participate which may be unrealistic); ▸ A named person (country coordinator) to send out invite survey link; ▸ Weekly reminders should be sent out by the named person (country coordinator).


Calciphylaxis: Approach to Diagnosis and Management.
Calciphylaxis is a rare disorder of poor prognosis that can lead to intense, painful lesions involving the skin and subcutaneous tissue. Although mostly described in dialysis patients, it can affect patients with normal kidney function. The diagnosis of calciphylaxis is complicated by the absence of a gold standard marker of disease such as a clear histopathological finding. Late diagnosis and advanced lesions can significantly shorten life expectancy. Calciphylaxis wounds can have a major influence on the quality of life of patients, usually due to the immense unbearable pain these patients suffer from. The management of calciphylaxis mainly comprises aggressive wound care and symptomatic management. Therapeutic options are few and far between and are limited to off-label uses. Recent understanding of the pathogenesis of lesions has enabled development of novel therapeutic options, some of which are being studied in clinical trials (sodium thiosulfate, vitamin K). Vascular calcification and thrombosis underlie development of these lesions and research has been aimed at studying drugs that counteract such processes. Future research is required to establish clear causal pathways and improve on the treatment options currently available to patients.


Methyl Donor Deficiency Blocks Colorectal Cancer Development by Affecting Key Metabolic Pathways.
Our understanding of the role of folate one-carbon metabolism in colon carcinogenesis remains incomplete. Previous studies indicate that a methyl donor-deficient (MDD) diet lacking folic acid, choline, methionine, and vitamin B12 is associated with long-lasting changes to the intestinal epithelium and sustained tumor protection in Apc-mutant mice. However, the metabolic pathways by which the MDD diet affects these changes are unknown. Colon samples harvested from ApcΔ14/+ mice fed the MDD diet for 18 weeks were profiled using a GC-MS and LC-MS/MS metabolomics platform. Random forest and pathway analyses were used to identify altered metabolic pathways, and associated gene expression changes were analyzed by RT-PCR. Approximately 100 metabolites affected by the MDD diet were identified. As expected, metabolites within the methionine cycle, including methionine (-2.9-fold, P < 0.001) and betaine (-3.3-fold, P < 0.001), were reduced. Elevated homocysteine (110-fold, P < 0.001) was associated with increased flux through the transsulfuration pathway. Unexpectedly, levels of deoxycholic acid (-4.5-fold, P < 0.05) and several other secondary bile acids were reduced. There were also unexpected reductions in the levels of carnitine (-2.0-fold, P < 0.01) and a panel of acylcarnitines involved in fatty acid β-oxidation. Finally, metabolites involved in redox balance, including ascorbate and hypotaurine, were found to be persistently elevated. These findings provide clues to the molecular changes underlying MDD-mediated tumor protection and identify regulatable metabolic pathways that may provide new targets for colon cancer prevention and treatment. IMPLICATIONS: Metabolomic profiling reveals molecular changes underlying MDD-induced tumor protection and may provide new targets for colorectal cancer prevention and treatment.


Cyp27c1 Red-Shifts the Spectral Sensitivity of Photoreceptors by Converting Vitamin A1 into A2.
Some vertebrate species have evolved means of extending their visual sensitivity beyond the range of human vision. One mechanism of enhancing sensitivity to long-wavelength light is to replace the 11-cis retinal chromophore in photopigments with 11-cis 3,4-didehydroretinal. Despite over a century of research on this topic, the enzymatic basis of this perceptual switch remains unknown. Here, we show that a cytochrome P450 family member, Cyp27c1, mediates this switch by converting vitamin A1 (the precursor of 11-cis retinal) into vitamin A2 (the precursor of 11-cis 3,4-didehydroretinal). Knockout of cyp27c1 in zebrafish abrogates production of vitamin A2, eliminating the animal's ability to red-shift its photoreceptor spectral sensitivity and reducing its ability to see and respond to near-infrared light. Thus, the expression of a single enzyme mediates dynamic spectral tuning of the entire visual system by controlling the balance of vitamin A1 and A2 in the eye.


Treating Medicaid patients with hepatitis C: clinical and economic impact.
To estimate change in chronic hepatitis C virus (HCV) disease and the economic burden associated with comprehensive treatment of the chronic HCV-infected Medicaid population. Decision-analytic Markov model. Treatment-naïve patients with genotype 1 chronic HCV were followed over a lifetime horizon from the third-party payer perspective. Patients entered the model insured under Medicaid and were treated under state-specific restrictions by Metavir fibrosis stage (base case) or all treated (all-patient strategy) with an approved all-oral regimen (ledipasvir/sofosbuvir [LDV/SOF] for 8 weeks or 12 weeks, depending on cirrhosis status, viral load, and state-specific LDV/SOF restrictions). Untreated patients were assumed to age into Medicare at 65 years, where they were treated with LDV/SOF without restriction by fibrotic stage. The sustained virologic response (SVR) rate of the current Medicaid LDV/SOF restriction strategy was 75.2% versus 95.9% if all LDV/SOF-eligible patients were treated under Medicaid. Treating all eligible Medicaid patients with LDV/SOF, regardless of fibrotic stage, was projected to result in 36,752 fewer cases of cirrhosis; 1739 fewer liver transplants; 8169 fewer cases of hepatocellular carcinoma; 16,173 fewer HCV-related deaths; 0.84 additional life-years per patient; and 1.03 additional quality-adjusted life-years per patient. Treating all Medicaid patients with chronic HCV using LDV/SOF resulted in a 39.4% ($3.8 billion) savings and decreased the proportion of total costs attributable to downstream costs of care to 18.3%. A "treat all" strategy in a Medicaid population resulted in superior SVRs, substantial reductions in downstream negative clinical outcomes, and considerable cost savings. Current restrictive state policies regarding HCV treatment in Medicaid populations must be reassessed in light of these data.


HIV testing preferences in Tanzania: a qualitative exploration of the importance of confidentiality, accessibility, and quality of service.
HIV counseling and testing (HCT), an effective preventive strategy and an entry point for care, remains under-utilized in Tanzania. Limited uptake of HCT, despite the widespread availability of varied testing options, suggests that existing options may not align well with population preferences for testing. Between October and December 2011, we conducted an exploratory study in the Kilimanjaro Region to develop a conceptual framework for understanding which characteristics of HIV testing are associated with preferences for testing. Forty individuals (55% women, 53% never having tested) participated in in-depth interviews and focus groups to identify factors that influence whether and where people test for HIV. A variety of discrete characteristics of testing venues, test providers, and testing procedures (e.g. distance to testing, counselor experience, type of HIV test, and availability of antiretroviral therapy) mapped conceptually to three domains: confidentiality of testing and test results, quality of HCT, and accessibility and availability of ancillary services. We noted heterogeneous preferences and demonstrate that while some test characteristics overlap and reinforce across multiple domains, others demand clients to make trade-offs between domains. Testing decisions appear to be influenced by an array of often inter-linked factors across multiple domains, including quality, confidentiality, and accessibility; perceptions of these factors varied greatly across participants and across available testing options. HCT interventions that jointly target barriers spanning the three domains have the potential to increase uptake of HIV testing and deserve further exploration.


Risk factors for encephalopathy and mortality during melarsoprol treatment of Trypanosoma brucei gambiense sleeping sickness.
This paper reviews the incidence of, and risk factors for, drug-induced encephalopathy and mortality (from all causes) during treatment with melarsoprol of 1083 patients with Trypanosoma brucei gambiense sleeping sickness in Nioki hospital (Zaire) between 1983 and 1990. Sixty-four patients (5.9%) developed encephalopathy and 62 (5.7%) died: 43 from reactive encephalopathy and 19 from other causes. Univariate and multivariate analyses showed that the administration of prednisolone reduced significantly the incidence of encephalopathy and mortality during treatment, especially in patients with trypanosomes observed in the cerebrospinal fluid (CSF) and/or with a CSF white blood cell (WBC) count of 100 or more per mm3. The risk of encephalopathy was associated more strongly with the CSF WBC count than with the presence of CSF trypanosomes. In the subgroup of patients with a CSF WBC count of 100 or more mm3, changing the melarsoprol regimen to 3 series of 3 injections instead of 3 series of 4 injections halved the mortality rate during treatment. Treatment of patients who do develop reactive encephalopathy with the heavy metal chelator dimercaprol, in addition to intravenous steroids and anticonvulsants, may be harmful. The data suggest that a further reduction of the total dose of melarsoprol may decrease toxicity without jeopardizing efficacy.


Identification of the Mamestra configurata (Lepidoptera: Noctuidae) peritrophic matrix proteins and enzymes involved in peritrophic matrix chitin metabolism.
The peritrophic matrix (PM) is essential for insect digestive system physiology as it protects the midgut epithelium from damage by food particles, pathogens, and toxins. The PM is also an attractive target for development of new pest control strategies due to its per os accessibility. To understand how the PM performs these functions, the molecular architecture of the PM was examined using genomic and proteomic approaches in Mamestra configurata (Lepidoptera: Noctuidae), a major pest of cruciferous oilseed crops in North America. Liquid chromatography-tandem mass spectrometry analyses of the PM identified 82 proteins classified as: (i) peritrophins, including a new class with a CBDIII domain; (ii) enzymes involved in chitin modification (chitin deacetylases), digestion (serine proteases, aminopeptidases, carboxypeptidases, lipases and α-amylase) or other reactions (β-1,3-glucanase, alkaline phosphatase, dsRNase, astacin, pantetheinase); (iii) a heterogenous group consisting of polycalin, REPATs, serpin, C-Type lectin and Lsti99/Lsti201 and 3 novel proteins without known orthologs. The genes encoding PM proteins were expressed predominantly in the midgut. cDNAs encoding chitin synthase-2 (McCHS-2), chitinase (McCHI), and β-N-acetylglucosaminidase (McNAG) enzymes, involved in PM chitin metabolism, were also identified. McCHS-2 expression was specific to the midgut indicating that it is responsible for chitin synthesis in the PM, the only chitinous material in the midgut. In contrast, the genes encoding the chitinolytic enzymes were expressed in multiple tissues. McCHS-2, McCHI, and McNAG were expressed in the midgut of feeding larvae, and NAG activity was present in the PM. This information was used to generate an updated model of the lepidopteran PM architecture.


Effects of a POEA surfactant system (Genamin T-200(®)) on two life stages of the Pacific oyster, Crassostrea gigas.
Surfactants used in herbicide formulations are generally considered inert with no toxic effects on animals. Polyethoxylated tallow amines (POEAs) are non-ionic surfactants used in many herbicide formulations to promote the penetration of the active matter into plant cuticles. The present study aimed to assess the toxicity of a POEA surfactant system, the Genamin T-200®, on two larval stages of the Pacific oyster, Crassostrea gigas. The embryotoxicity of Genamin T-200® was quantified after 36 hr of exposure, considering both arrested development and abnormalities in D-shaped larvae. The ability of pediveliger larvae to metamorphose was studied after 24 hr exposure to Genamin T-200®. According to the European toxicity classification, the present results suggest that Genamin T-200® could be considered very toxic to embryo larval development, with an EC₅₀ of 262 µg/l, and toxic to metamorphosis processes with an EC₅₀ of 3,027 µg/l. The high toxicity of glyphosate-based formulations compared to the active ingredient and its by-product appears to be due primarily to surfactants.


The protease inhibitor, N-acetyl-L-leucyl-L-leucyl-leucyl-L-norleucinal, decreases the pool of major histocompatibility complex class I-binding peptides and inhibits peptide trimming in the endoplasmic reticulum.
N-acetyl-L-leucyl-L-leucyl-L-norleucinal, (LLnL), which inhibits proteasomes in addition to other proteases, was found to prolong the association of major histocompatibility complex class I molecules with the transporters associated with antigen processing (TAP), and to slow their transport out of the endoplasmic reticulum (ER). LLnL induced a reversible accumulation of ubiquitinated proteins and changed the spectrum of peptides bound by class I molecules. These effects can probably be attributed to proteasome inhibition. Unexpectedly, in the TAP-deficient cell line .174, the rate of intracellular transport of human histocompatibility leukocyte antigen (HLA) A2 was also reduced by LLnL, and the generation of most HLA-A2-associated signal sequence peptides was inhibited. The inhibition of HLA-A2 transport in .174 cells was found to be less sensitive to LLnL than in wild-type cells, and a similar difference was found for a second protease inhibitor, benzyloxycarbonyl-L-leucyl-L-leucyl-L-phenylalanilal. These data suggest that under some conditions such inhibitors can block trimming of peptides by an ER peptidase in addition to inhibiting cytosolic peptide generation.


Perceptions of mental illness among Muslim general practitioners in South Africa.
Mental health literacy on the part of medical practitioners is an important component of mental healthcare. General practitioners (GPs) are typically the first doctors consulted by a person who is ill. Exploration of their perceptions regarding mental illness, aetiological issues and treatment is important. To investigate perceptions of mental illness in a sample of ten South African Muslim GPs (five male, five female) in the Lenasia area (Johannesburg, South Africa). Using a qualitative approach, semi-structured interviews were conducted with each GP. The questionnaire encompassed 37 questions relating to the context in which the GPs practised, perceptions of mental illness, understanding of religion and culture, and treatment of mental illness (including aspects of spiritual illness). Thematic content analysis was used to analyse the data. Six dominant themes were identified, namely GPs' understanding of mental illness and its causation; stigma, secrecy and somatisation; the beneficial effects of religion in mental illnesses; perceptions of spiritual illnesses; collaboration with traditional healers; and collaboration with psychiatrists and psychologists. Greater awareness regarding the stigmatisation of mental illness is needed. Furthermore, it is important that healthcare professionals have an understanding of religious and cultural taxonomies of illness as well as the use of traditional healing as a mode of treatment. Participants identified a need for increased collaboration between healthcare professionals, including traditional healers.


Molecular profiling of pancreatic adenocarcinoma and chronic pancreatitis identifies multiple genes differentially regulated in pancreatic cancer.
The molecular basis of pancreatic cancer is not understood. Previous attempts to determine the specific genes expressed in pancreatic cancer have been hampered by similarities between adenocarcinoma and chronic pancreatitis. In the current study, microarrays (Affymetrix) were used to profile gene expression in pancreatic adenocarcinoma (10), pancreatic cancer cell lines (7), chronic pancreatitis (5), and normal pancreas (5). Molecular profiling indicated a large number of genes differentially expressed between pancreatic cancer and normal pancreas but many fewer differences between pancreatic cancer and chronic pancreatitis, likely because of the shared stromal influences in the two diseases. To specifically identify genes expressed in neoplastic epithelium, we selected genes more highly expressed (>2-fold, p < 0.01) in adenocarcinoma compared with both normal pancreas and chronic pancreatitis and which were also highly expressed in pancreatic cancer cell lines. This strategy yielded 158 genes, of which 124 were not previously associated with pancreatic cancer. Quantitative-reverse transcription-PCR for two molecules, S100P and 14-3-3sigma, validated the microarray data. Support for the success of the neoplastic cell gene expression identification strategy was obtained by immunocytochemical localization of four representative genes, 14-3-3sigma, S100P, S100A6, and beta4 integrin, to neoplastic cells in pancreatic tumors. Thus, comparisons between pancreatic adenocarcinoma, pancreatic cancer cell lines, normal pancreas, and chronic pancreatitis have identified genes that are selectively expressed in the neoplastic epithelium of pancreatic adenocarcinoma. These data provide new insights into the molecular pathology of pancreatic cancer that may be useful for detection, diagnosis, and treatment.


Origin of repopulating cells after localized bone marrow depletion.
The restoration of marrow in a mechanically depleted segment of rabbit femur is locally determined and apparently initiated by cells normally resident in bone. This conclusion follows from results of two types of radiation experiments: local x-irradiation of the femur shaft immediately before depopulation and x-irradiation of the total body with exception of the femur diaphysis which was depopulated either just before or after irradiation. In contrast to the rapid initiation of marrow restoration in an unirradiated femur, there was little regeneration during the first 3 weeks in an irradiated femur. Recovery of the shielded depopulated femur in the rabbit that otherwise received total-body irradiation was similar to that in the unirradiated animal. Hence, it would appear that the seeding of circulating hemic stem cells is not essential to repopulation and that cells with hematopoietic potential are present in osseous tissue.


Contribution of choline-binding proteins to cell surface properties of Streptococcus pneumoniae.
Nonspecific interactions related to physicochemical properties of bacterial cell surfaces, such as hydrophobicity and electrostatic charge, are known to have important roles in bacterium-host cell encounters. Streptococcus pneumoniae (pneumococcus) expresses multiple, surface-exposed, choline-binding proteins (CBPs) which have been associated with adhesion and virulence. The purpose of this study was to determine the contribution of CBPs to the surface characteristics of pneumococci and, consequently, to learn how CBPs may affect nonspecific interactions with host cells. Pneumococcal strains lacking CBPs were derived by adapting bacteria to a defined medium that substituted ethanolamine for choline. Such strains do not anchor CBPs to their surface. Cell surface hydrophobicity was tested for the wild-type and adapted strains by using a biphasic hydrocarbon adherence assay, and electrostatic charge was determined by zeta potential measurement. Adherence of pneumococci to human-derived cells was assessed by fluorescence-activated cell sorter analysis. Strains lacking both capsule and CBPs were significantly more hydrophobic than nonencapsulated strains with a normal complement of CBPs. The effect of CBPs on hydrophobicity was attenuated in the presence of capsule. Removal of CBPs conferred a greater electronegative net surface charge than that which cells with CBPs possessed, regardless of the presence of capsule. Strains that lack CBPs were poorly adherent to human monocyte-like cells when compared with wild-type bacteria with a full complement of CBPs. These results suggest that CBPs contribute significantly to the hydrophobic and electrostatic surface characteristics of pneumococci and may facilitate adherence to host cells partially through nonspecific, physicochemical interactions.


Light trapping in thin-film solar cells with randomly rough and hybrid textures.
We study light-trapping in thin-film silicon solar cells with rough interfaces. We consider solar cells made of different materials (c-Si and μc-Si) to investigate the role of size and nature (direct/indirect) of the energy band gap in light trapping. By means of rigorous calculations we demonstrate that the Lambertian Limit of absorption can be obtained in a structure with an optimized rough interface. We gain insight into the light trapping mechanisms by analysing the optical properties of rough interfaces in terms of Angular Intensity Distribution (AID) and haze. Finally, we show the benefits of merging ordered and disordered photonic structures for light trapping by studying a hybrid interface, which is a combination of a rough interface and a diffraction grating. This approach gives a significant absorption enhancement for a roughness with a modest size of spatial features, assuring good electrical properties of the interface. All the structures presented in this work are compatible with present-day technologies, giving recent progress in fabrication of thin monocrystalline silicon films and nanoimprint lithography.


Binding Modes and Selectivity of Ruthenium Complexes to Human Telomeric DNA G-Quadruplexes.
Metal complexes constitute an important class of DNA binders. In particular, a few ruthenium polyazaaromatic complexes are attractive as "light switches" because of their strong luminescence enhancement upon DNA binding. In this paper, a comprehensive study on the binding modes of several mononuclear and binuclear ruthenium complexes to human telomeric sequences, made of repeats of the d(TTAGGG) fragment is reported. These DNA sequences form G-quadruplexes (G4s) at the ends of chromosomes and constitute a relevant biomolecular target in cancer research. By combining spectroscopy experiments and molecular modelling simulations, several key properties are deciphered: the binding modes, the stabilization of G4 upon binding, and the selectivity of these complexes towards G4 versus double-stranded DNA. These results are rationalized by assessing the possible deformation of G4 and the binding free energies of several binding modes via modelling approaches. Altogether, this comparative study provides fundamental insights into the molecular recognition properties and selectivity of Ru complexes towards this important class of DNA G4s.


A novel multiplex real-time PCR assay for the concurrent detection of hepatitis A, B and C viruses in patients with acute hepatitis.
A novel multiplex real-time PCR assay for concurrent detection of hepatitis viruses was evaluated for its clinical performance in screening patients with acute hepatitis. A total of 648 serum samples were collected from patients with acute symptoms of hepatitis. Concurrent detection of nucleic acids of HAV, HBV and HCV was performed using the Magicplex™ HepaTrio Real-time Detection test. Serum nucleic acid levels of HBV and HCV were also quantified by the Cobas® AmpliPrep/Cobas® TaqMan® (CAP/CTM) HBV and HCV tests. Patients' medical records were also reviewed. Concordance rates between the results from the HepaTrio and the CAP/CTM tests for the detection of HBV and HCV were 94.9% (k = 0.88) and 99.2% (k = 0.98), respectively. The cycle threshold values with the HepaTrio test were also correlated well with the levels of HBV DNA (r = -0.9230) and HCV RNA (r = -0.8458). The sensitivity and specificity of the HepaTrio test were 93.8% and 98.2%, respectively, for detecting HBV infection, and 99.1% and 100.0%, respectively, for HCV infection. For the HepaTrio test, 21 (3.2%) cases were positive for both HBV and HCV. Among the positive cases, 6 (0.9%) were true coinfections. This test also detected 18 (2.8%) HAV positives. The HepaTrio test demonstrated good clinical performance and produced results that agreed well with those of the CAP/CTM assays, especially for the detection of HCV. This assay was also able to detect HAV RNA from anti-HAV IgM-positive individuals. Therefore, this new multiplex PCR assay could be useful for the concurrent detection of the three hepatitis viruses.


The level of TSH appeared favourable in maintaining bone mineral density in postmenopausal women.
Since the positive role of thyrotropin (TSH) in bone remodeling has been recently emphasized, this cross-section study is aimed to evaluate the association of bone status with the level of TSH and free thyroxine (FT4) in the cohort of postmenopausal women after long-term treatment of thyroid disorders and age matched controls. Urinary calcium (dUCa) and serum level of TSH, FT4 and of bone turnover markers (BTMs) such as alkaline phosphatase (ALP), osteocalcin (OC), cross linked N-telopeptide of type 1 collagen (NTx) as well as lumbar spine L 1-4 (BMD-L) and femoral hip (BMD-F) mineral density were determined in 113 postmenopausal women consisting of 42 patients with Graves disease treated by carbimazole, 32 patients with thyroid cancer treated with L-thyroxine and 39 age matched women without any thyroid and osteological disorders. For statistical evaluation t-test, Pearson's correlation coefficient and linear multiple regression were used. To compare the association of TSH versus FT4 with BMD and BTMs the pooled cohort of all 113 women was divided in two groups in terms of TSH level: 1. 34 women with low TSH (>or=0.50 mU/l); 2. 79 women with normal TSH (0.51-4.3 mU/l). In spite of significantly higher FT4 level, the Group 2 with normal TSH level had significantly higher BMD-L and BMD-F (p<0.001) and, in contrast, significantly lower urinary dUCa, ALP, OC (all at p<0.001) and NTx (p<0.01) as compared to the Group 1 with low TSH level. Linear multiple regression showed highly significant influence of TSH on BMD-L and BMD-F0 (p<0.001) independent of age, FT4 and body mass index, while that of FT4 was not significant. The strength of linear interrelation between all variables used was finally tested by Pearson's correlation coefficient (Table 3) which was highly positive for TSH with BMD-F and BMD-L, but highly negative for TSH with serum NTx, OC, ALP) and urinary calcium (dUCa). In contrast, no significant correlation was found between the level of FT4 and BMD. Irrespectively of FT4 level, postmenopausal women with normal TSH level showed a favorable bone status as compared to these with low level of TSH which is consistent with the view that TSH itself possibly participates in playing a favorable role in influencing the bone mineral density in adult women.


Quantifying Microsatellite Mutation Rates from Intestinal Stem Cell Dynamics in Msh2-Deficient Murine Epithelium.
Microsatellite sequences have an enhanced susceptibility to mutation, and can act as sentinels indicating elevated mutation rates and increased risk of cancer. The probability of mutant fixation within the intestinal epithelium is dictated by a combination of stem cell dynamics and mutation rate. Here, we exploit this relationship to infer microsatellite mutation rates. First a sensitive, multiplexed, and quantitative method for detecting somatic changes in microsatellite length was developed that allowed the parallel detection of mutant [CA]n sequences from hundreds of low-input tissue samples at up to 14 loci. The method was applied to colonic crypts in Mus musculus, and enabled detection of mutant subclones down to 20% of the cellularity of the crypt (∼50 of 250 cells). By quantifying age-related increases in clone frequencies for multiple loci, microsatellite mutation rates in wild-type and Msh2-deficient epithelium were established. An average 388-fold increase in mutation per mitosis rate was observed in Msh2-deficient epithelium (2.4 × 10-2) compared to wild-type epithelium (6.2 × 10-5).


Brain songs framework used for discovering the relevant timescale of the human brain.
A key unresolved problem in neuroscience is to determine the relevant timescale for understanding spatiotemporal dynamics across the whole brain. While resting state fMRI reveals networks at an ultraslow timescale (below 0.1 Hz), other neuroimaging modalities such as MEG and EEG suggest that much faster timescales may be equally or more relevant for discovering spatiotemporal structure. Here, we introduce a novel way to generate whole-brain neural dynamical activity at the millisecond scale from fMRI signals. This method allows us to study the different timescales through binning the output of the model. These timescales can then be investigated using a method (poetically named brain songs) to extract the spacetime motifs at a given timescale. Using independent measures of entropy and hierarchy to characterize the richness of the dynamical repertoire, we show that both methods find a similar optimum at a timescale of around 200 ms in resting state and in task data.


Up-regulation of microRNA-503 by high glucose reduces the migration and proliferation but promotes the apoptosis of human umbilical vein endothelial cells by inhibiting the expression of insulin-like growth factor-1 receptor.
The present study is aimed to investigate the regulatory effect of microRNA (miRNA or miR)-503 on endothelial functions, as well as the mechanism by which high glucose leads to injury of endothelial cells. When reaching 80% confluency, human umbilical vein endothelial cells (HUVECs) were subjected to non-serum synchronization for 12 h, and medium of cells in high-glucose (HG) group was replaced by normal medium supplemented with 25 mmol/L D-glucose. HUVECs cultured in normal glucose (NG) medium were used as control. To overexpress miR-503, HUVECs were transfected with miR-503 mimics. To silence insulin-like growth factor-1 receptor (IGF-1R) mRNA, HUVECs were transfected with small interfering RNA (siRNA). To predict whether miR-503 targets IGF-1R, bioinformatics was performed. Quantitative Real-time polymerase chain reaction was used to determine miR-503 and IGF-1R mRNA expression, and Western blotting was employed to measure IGF-1R protein expression. Cell-Counting Kit 8 assay was used to determine HUVECs proliferation, while wound-healing assay was used to evaluate HUVECs migration. HUVECs apoptosis was investigated by measuring caspase 3 activity. Expression of IGF-1R in HUVECs in high glucose was decreased compared to that in normal glucose. miR-503 was predicted to target IGF-1R mRNA, and miR-503 expression in HUVECs in high glucose was higher than that in normal glucose. Overexpression of miR-503 inhibited the transcription and the translation of IGF-1R gene reducing migration, suppressed proliferation and promoted apoptosis. Transfection with IGF-1R siRNA decreased IGF-1R protein expression in HUVECs. Down-regulated IGF-1R expression reduced migration and proliferation, but promoted apoptosis of HUVECs. The present study demonstrates that miR-503 expression in HUVECs is elevated in high glucose condition. Also, miR-503 reduces migration and proliferation, but promotes apoptosis of HUVECs by inhibiting IGF-1R expression.


Rapid online-SPE-MS/MS method for ketoprofen determination in dermal interstitial fluid samples from rats obtained by microdialysis or open-flow microperfusion.
Pharmacokinetic studies of topical ketoprofen formulations using continuous sampling techniques such as microdialysis (MD) or open-flow microperfusion (OFM) require sensitive assays due to small sample volumes. A simple and easy online-SPE-MS/MS method for ketoprofen analysis was developed for both MD and OFM samples obtained from rat dermal tissue. The quantification range is 25-5000 ng/ml with a limit of detection of 3 ng/ml using only 10 microl sample volume. The method is characterized by a simple setup using a short polymeric SPE column (OASIS HLB) for desalting with 1.5 min run times in combination with a sensitive MS detection in negative ESI MRM mode. An easy sample workup procedure was used which enables high throughput analysis of a large number of samples for pharmacokinetic studies. In addition, a commercial available (fenoprofen) as well as an isotopically labelled (deuterated ketoprofen) standard were investigated as potential internal standards. The method was validated according to FDA guidelines for bioanalytical validation in terms of accuracy, intra-batch and inter-batch precision, linearity, matrix effect, recovery and stability for both internal standards. Accuracies were 98-113% (fenoprofen) and 95-108% (deuterated ketoprofen), intra-batch precision was 2-3% R.S.D. (fenoprofen) and 2-6% R.S.D. (deuterated ketoprofen), and inter-batch precision was 2-6% R.S.D. (fenoprofen) and 3-6% R.S.D. (deuterated ketoprofen) over the entire quantification range. The presented method was applied to dermal interstitial fluid samples obtained in a topical administration study of ketoprofen in rats.


HEY1 is expressed independent of NOTCH1 and is associated with poor prognosis in head and neck squamous cell carcinoma.
Notch signaling is frequently altered in head and neck squamous cell carcinoma (HNSCC). However, the nature and clinical implications of this dysregulation are not well understood. We previously described an association of transcriptionally active NOTCH1 Intracellular Domain (NICD1) immunohistochemical (IHC) expression pattern with high-risk pathologic characteristics. Here we further characterize Notch signaling in HNSCC. IHC expression patterns and clinicopathologic associations of Notch pathway molecules were evaluated among 78 tumors with known NOTCH1 mutation status. IHC was performed for JAG1, a NOTCH1 activating ligand, and HEY1, an NICD1 transcriptional target and Notch pathway activation marker. IHC pattern and H-score (% staining × intensity) were recorded and compared to clinicopathologic characteristics and survival. Survival was analyzed using Kaplan Meier method and Cox proportional hazards models (HR). JAG1 and NICD1 expression patterns were highly concordant among tumors without truncating NOTCH1 mutations (p < 0.001), but were dissimilar among tumors with truncating NOTCH1 mutations (p = 0.24). There was evidence for JAG1-independent NOTCH1 activation among seven tumors, all with wild-type NOTCH1. HEY1 expression was associated with neither JAG1 nor NICD1 expression, but was associated with NOTCH1 mutation status (p = 0.03). Twelve (16%) tumors expressed HEY1 but not NICD1. Higher HEY1 H-score was significantly associated with worse overall (adjusted hazard ratio [aHR] 2.0, 95% CI = 1.0-4.2) and disease-specific (aHR = 3.3, 95% CI = 1.4-7.9) survival, whereas JAG1 and NICD1 expression were not associated with survival. These findings suggest both NOTCH1-dependent and -independent HEY1 regulation, and imply a previously unrecognized prognostic role for HEY1 in HNSCC.


Association of methylphenidate-induced craving with changes in right striato-orbitofrontal metabolism in cocaine abusers: implications in addiction.
The authors have shown that decreases in dopamine D2 receptors in cocaine abusers were associated with decreased metabolism in the cingulate and prefrontal and orbitofrontal cortices. To assess whether increasing dopamine would reverse these metabolic decrements, they measured the effects of methylphenidate, a drug that increases dopamine, on brain glucose metabolism in 20 cocaine abusers. The subjects underwent two [18F]fluorodeoxyglucose positron emission tomography scans, one after two sequential placebo injections and one after two intravenous doses of methylphenidate. D2 receptors were measured with [11C]raclopride to evaluate their relation to methylphenidate-induced metabolic changes. Methylphenidate induced variable changes in brain metabolism: subjects with the higher D2 measures tended to increase metabolism, whereas those with the lower D2 measures tended to decrease metabolism. Methylphenidate's effects were significant for increases in metabolism in the superior cingulate, right thalamus, and cerebellum. Methylphenidate-induced changes in the right orbitofrontal cortex and right striatum were associated with craving, and those in the prefrontal cortex were associated with mood. Although methylphenidate increased metabolism in the superior cingulate, it only increased metabolism in orbitofrontal or prefrontal cortices in the subjects in whom it enhanced craving and mood, respectively. This indicates that dopamine enhancement is not sufficient per se to increase metabolism in these frontal regions. Activation of the right orbitofrontal cortex and right striatum (brain regions found to be abnormal in compulsive disorders) in the subjects reporting craving may be one of the mechanisms underlying compulsive drug administration in addicted persons. The predominant correlation of craving with right but not left brain regions suggests laterality of reinforcing and/or conditioned responses.


Evidence for New Light-Independent Pathways for Generation of the Endogenous Aryl Hydrocarbon Receptor Agonist FICZ.
Activation of the aryl hydrocarbon receptor (AhR), a conserved transcription factor best known as a target for highly toxic halogenated substances such as dioxin, under normal xenobiotic-free conditions is of considerable scientific interest. We have demonstrated previously that a photoproduct of tryptophan, 6-formylindolo[3,2-b]carbazole (FICZ), fulfills the criteria for an endogenous ligand for this receptor and proposed that this compound is the enigmatic mediator of the physiological functions of AhR. Here, we describe novel light-independent pathways by which FICZ can be formed. The oxidant H2O2 was shown to convert tryptophan to FICZ on its own in the absence of light. The enzymatic deamination of tryptamine yielded indole-3-acetaldehyde (I3A), which then rearranged to FICZ and its oxidation product, indolo[3,2-b]carbazole-6-carboxylic acid (CICZ). Indole-3-pyruvate (I3P) also produced I3A, FICZ, and CICZ. Malassezia yeast species, which constitute a part of the normal skin microbiota, produce a number of AhR activators from tryptophan. We identified both FICZ and CICZ among those products. Formation of FICZ from tryptophan or I3P produces a complex mixture of indole derivatives, some of which are CYP1A1 inhibitors. These can hinder the cellular clearance of FICZ and thereby increase its power as an AhR agonist. We present a general molecular mechanism involving dehydrogenations and oxidative coupling for the formation of FICZ in which I3A is the important precursor. In conclusion, our results suggest that FICZ is likely to be formed systemically.


Far field scattering pattern of differently structured butterfly scales.
The angular and spectral reflectance of single scales of five different butterfly species was measured and related to the scale anatomy. The scales of the pierids Pieris rapae and Delias nigrina scatter white light randomly, in close agreement with Lambert's cosine law, which can be well understood from the randomly organized beads on the scale crossribs. The reflectance of the iridescent blue scales of Morpho aega is determined by multilayer structures in the scale ridges, causing diffraction in approximately a plane. The purple scales in the dorsal wing tips of the male Colotis regina act similarly as the Morpho scale in the blue, due to multilayers in the ridges, but the scattering in the red occurs as in the Pieris scale, because the scales contain beads with pigment that does not absorb in the red wavelength range. The green-yellow scales of Urania fulgens backscatter light in a narrow spatial angle, because of a multilayer structure in the scale body.


Structural changes in mixed Col I/Col V collagen gels probed by SHG microscopy: implications for probing stromal alterations in human breast cancer.
Second Harmonic Generation (SHG) microscopy has been previously used to describe the morphology of collagen in the extracellular matrix (ECM) in different stages of invasion in breast cancer. Here this concept is extended by using SHG to provide quantitative discrimination of self-assembled collagen gels, consisting of mixtures of type I (Col I) and type V (Col V) isoforms which serve as models of changes in the ECM during invasion in vivo. To investigate if SHG is sensitive to changes due to Col V incorporation into Col I fibrils, gels were prepared with 0-20% Col V with the balance consisting of Col I. Using the metrics of SHG intensity, fiber length, emission directionality, and depth-dependent intensities, we found similar responses for gels comprised of 100% Col I, and 95% Col I/5% Col V, where these metrics were all significantly different from those of the 80% Col I/20% Col V gels. Specifically, the gels of lower Col V content produce brighter SHG, are characterized by longer fibers, and have a higher forward/backward emission ratio. These attributes are all consistent with more highly organized collagen fibrils/fibers and are in agreement with previous TEM characterization as well as predictions based on phase matching considerations. These results suggest that SHG can be developed to discriminate Col I/Col V composition in tissues to characterize and follow breast cancer invasion.


Evolution and function in serotonergic systems.
Serotonergic systems of invertebrate and vertebrate central nervous systems (CNS) are functionally similar in multiple characters. Serotonin (5-HT) neurons dispersed throughout the CNS of lophotrochozoan invertebrates (molluscs and leeches) are analogous to vertebrate 5-HT neurons concentrated in the raphe nuclei of mid- and hindbrain: they innervate specific central pattern generators and other circuits of the CNS, receive feedback from them, and support general behavioral arousal. In both groups 5-HT regulates excitatory gain of CNS circuitry and uses similarly diverse 5-HT receptors. Marked contrast, however, exists for roles of 5-HT in regulation of appetite. Where invertebrate 5-HT neurons promote an appetitive state, this role is supplanted in the vertebrates by a peptidergic network centered around orexins/hypocretins, to which the role of 5-HT in arousal is subordinate. In the vertebrates, 5-HT has appetite-suppressant properties. This is paralleled by differing complexities of mechanisms that bring about satiety. Lophotrozoans appear to rely on simple stretching of the gut, with no obvious feedback from true nutrient stores. In contrast, vertebrate appetite is regulated by hypothalamic sensitivity to hormonal signals reporting separately on the status of fat cells and digestive activity, and to blood glucose, in addition to gut stretch. The simple satiety mechanism of a mollusc can be used in value-based foraging decisions that integrate hunger state, taste, and experience (Gillette and others 2000). For vertebrates, where appetite and arousal are regulated by signals from long-lived nutrient stores, decisions can be based on resource need going far beyond simple gut content, enabling value estimation and risk assessment in the longer-term. Thus, connection of nutrient storage depots to CNS circuitry mediating appetite may supply critical substrate for evolving complexity in brain and behavior. This hypothesis may be tested in expanded comparative studies of 5-HT and peptidergic functions in appetite and arousal.


Perceived health status of women with knee osteoarthritis: a cross-sectional study of the relationships of age, body mass, pain and walking limitations.
Persons with knee osteoarthritis (OA) often experience considerable physical disability. Although some studies suggest women with this condition suffer more than men, few have attempted to characterize the magnitude and that impact of this condition specifically among women with moderate knee osteoarthritis as well as the relationships that exist between their perceived health status and well established physical, emotional and perceptual factors found in this disease. This exploratory study strove to better understand factors that underpin the perceived impact of the condition, and to describe the extent of pain and function among women with mild to moderate knee osteoarthritis, and how this impacts this condition. The records of 20 women with the condition who had undergone multiple tests using a standardized protocols and validated instruments were examined. The primary outcome measure was the perceived impact of the disease using the Arthritis Impact Measurement Scale. Secondary outcome measures included six minute walking distance, fastest walking velocity, self-reported pain, pain and functional self-efficacy, body mass, and depression. The variables were subjected to t-tests, and correlational analyses. Results demonstrated pain is the clinical factor most consistently impacting the disease experience, along with deficiencies in walking ability (p <0.05). Important mediating variables of ambulatory capacity were body mass and pain self-efficacy.


Is policy 'liberalization' associated with higher odds of adolescent cannabis use? A re-analysis of data from 38 countries.
Policy makers worldwide face the choice of whether to reform cannabis policy from the 'full prohibition' model. A paper by Shi et al. (2015) suggested that such 'liberalization' is significantly associated with higher odds of adolescent cannabis use. To test the validity and reliability of Shi et al's conclusion that the HBSC data show an association between policy 'liberalization' and increased likelihood of adolescent cannabis use. Replication and re-analysis of the same pooled data from three waves of the Health Behaviour in School-aged Children (HBSC) survey (2001/2, 2005/6 and 2009/10). This replicates - as far as possible - the coding and analytical strategy of the Shi et al article. The re-analysis makes some improvements by: excluding a variable ('number of siblings') for which many cases have missing data; including available data from the theoretically relevant case of Sweden for the latter two waves of the HBSC survey, which Shi et al omit; and including random slopes for gender between countries as well as random intercepts for countries in the mixed effects model, as the predictive effect of gender on cannabis use varies across countries. Shi et al's verbal summary of their findings is not supported by detailed interpretation of their own numerical results. Without making the suggested amendments, it is possible to find a statistically significant association between policy 'liberalization' and higher odds of some measures of adolescent cannabis use. But when these improvements are made, this association becomes statistically non-significant. Using a larger and more theoretically relevant sample of the HBSC respondents and an improved statistical model shows that the HBSC data do not reveal a statistically significant association between policy 'liberalization' and higher odds of adolescent cannabis use.


Osteogenic potential of mesenchymal stem cells on expanded polytetrafluoroethylene coated with both a poly-amino-Acid urethane copolymer and collagen.
We have developed an expanded polytetrafluoroethylene polymer (e-P) coated with both poly-amino-acid urethane copolymer and collagen (e-PPC) to be used for cell culture substrata. Rat mesenchymal stem cells (MSC) were cultured on the e-P and e-PPC polymers in the presence of dexamethasone. After 24 h, the MSC contacted well with the e-PPC surface and after 14 days, the MSC showed high levels of alkaline phosphatase activity, and calcium and bone-specific osteocalcin protein deposition. The MSC cultured on these polymers for 1 week were then implanted at rat subcutaneous sites and harvested after 4 weeks. Microcomputed tomography as well as histological analyses showed that any hard tissue could not be seen in implants of the MSC/e-P composites, whereas new bone formation could be detected in the MSC/e-PPC composites. These in vitro as well as in vivo results confirmed the importance of polymer surface to support the osteogenic differentiation, which resulted in new bone formation. The surface modification using poly-amino-acid urethane copolymer and collagen together with tissue engineering technology might facilitate bone anchoring to the polymers for dental and orthopedic applications.


The potential of Thelypteris palustris and Asparagus sprengeri in phytoremediation of arsenic contamination.
The potential of two plants, Thelypteris palustris (marsh fern) and Asparagus sprengeri (asparagus fern), for phytoremediation of arsenic contamination was evaluated. The plants were chosen for this study because of the discovery of the arsenic hyperaccumulating fern, Pteris vittata (Ma et al., 2001) and previous research indicating asparagus fern's ability to tolerate > 1200 ppm soil arsenic. Objectives were (1) to assess if selected plants are arsenic hyperaccumulators; and (2) to assess changes in the species of arsenic upon accumulation in selected plants. Greenhouse hydroponic experiments arsenic treatment levels were established by adding potassium arsenate to solution. All plants were placed into the hydroponic experiments while still potted in their growth media. Marsh fern and Asparagus fern can both accumulate arsenic. Marsh fern bioaccumulation factors (> 10) are in the range of known hyperaccumulator, Pteris vittata Therefore, Thelypteris palustris is may be a good candidate for remediation of arsenic soil contamination levels of < or = 500 microg/L arsenic. Total oxidation of As (III) to As (V) does not occur in asparagus fern. The asparagus fern is arsenic tolerant (bioaccumulation factors < 10), but is not considered a good potential phytoremediation candidate.


Toll-like Receptor 2 Signalling and the Lysosomal Machinery in Barrett's Esophagus.
Inflammation plays an important role in the development of esophageal adenocarcinoma and its metaplastic precursor lesion, Barrett's esophagus. Toll-like receptor (TLR) 2 signalling and lysosomal function have been linked to inflammation-associated carcinogenesis. We examined the expression of TLR2 in the esophagus and the effect of long-term TLR2 activation on morphological changes and expression of factors involved in lysosomal function in a Barrett's esophagus epithelium cell line. TLR2 expression in normal squamous esophagus, reflux esophagitis, Barrett's esophagus and esophageal adenocarcinoma biopsies was assessed with Q-RT-PCR, in situ hybridization and immunohistochemistry. Barrett's esophagus epithelium cells (BAR-T) were incubated with acid and bile salts in the presence or absence of the TLR2 agonist Pam3CSK4 for a period up to 4 weeks. Morphological changes were assessed with electron microscopy, while Q-RT-PCR was used to determine the expression of lysosomal enzymes (Cathepsin B and C) and factors involved in endocytosis (LAMP-1 and M6PR) and autophagy (LC3 and Rab7). TLR2 was expressed in normal squamous esophagus, reflux esophagitis, Barrett's esophagus but was most prominent in esophageal adenocarcinoma. Long-term TLR2 activation in acid and bile salts exposed BAR-T cells resulted in more and larger lysosomes, more mitochondria and increased expression of LAMP-1, M6PR, Cathepsin B and C when compared to BAR-T cells incubated with acid and bile salts but no TLR2 agonist. Factors associated with autophagy (LC3 and Rab7) expression remained largely unchanged. Activation of TLR2 in acid and bile salts exposed Barrett epithelium cells resulted in an increased number of mitochondria and lysosomes and increased expression of lysosomal enzymes and factors involved in endocytosis.


Predictors of skin and soft tissue infections in HIV-infected outpatients in the community-associated methicillin-resistant Staphylococcus aureus era.
Skin and soft tissue infections (SSTIs) are common in the era of community-associated methicillin-resistant Staphylococcus aureus (MRSA) among human immunodeficiency virus (HIV)-infected patients, but the risk factors are not well defined. We sought to elucidate the risk factors for SSTI occurrence in an HIV cohort. This investigation was a retrospective, single-center cohort study, carried out during the period 2005-2009. In this cohort of 511 HIV-infected individuals, 133 SSTIs occurred in 87 individuals over 1,228.6 person-years of follow-up, for an incidence of 108 SSTIs/1,000 person-years [95 % confidence interval (CI) 87-135]. The incidence declined significantly over time (p < 0.01). In a multivariable Cox regression, diabetes [hazard ratio (HR) 2.01; 95 % CI 1.04-3.89], psoriasis (HR 5.77; 95 % CI 1.86-17.9), lymphedema (HR 6.84; 95 % CI 2.59-18.1), intravenous catheter presence (HR 3.38; 95 % CI 1.00-11.5), and HIV viral load greater than 1,000 copies/mL (HR 2.13; 95 % CI 1.33-3.41) were most strongly associated with development of the first SSTI. Trends toward an association between SSTI risk and Medicaid insurance (HR 1.67; 95 % CI 0.98-2.83) and sexually transmitted disease during follow-up (HR 1.66; 0.99-2.78) were present. CD4+ count and trimethoprim-sulfamethoxazole use were not associated with SSTI risk. HIV-infected individuals are at high risk for SSTIs. In a primarily urban, African-American cohort, we found that a number of immunologic and demographic factors were associated with SSTI risk.


Exercise based rehabilitation for heart failure.
The prevalence of chronic heart failure is increasing, and increases with increasing age. Major symptoms include breathlessness and restricted activities of daily living due to reduced functional capacity, which in turn affects quality of life. Exercise training has been shown to be effective in patients with coronary heart disease and has been proposed as an intervention to improve exercise tolerance in patients with heart failure. To determine the effectiveness of exercise based interventions compared with usual medical care on the mortality, morbidity, exercise capacity and health related quality of life, of patients with heart failure. We searched the Cochrane Controlled Trials Register (The Cochrane Library Issue 2, 2001), MEDLINE (2000 to March 2001), EMBASE (1998 to March 2001), CINAHL (1984 to March 2001) and reference lists of articles. We also sought advice from experts. RCTs of exercise based interventions. The comparison group was usual medical care as defined by the study, or placebo. Adults of all ages with chronic heart failure. Only those studies with criteria for diagnosis of heart failure (based on clinical findings or objective indices) have been included. Studies were selected, and data were abstracted, independently by two reviewers. Authors were contacted where possible to obtain missing information. Twenty-nine studies met the inclusion criteria, with 1126 patients randomised. The majority of studies included both patients with primary and secondary heart failure, NYHA class II or III. None of the studies specifically examined the effect of exercise training on mortality and morbidity as most were of short duration. Exercise training significantly increased VO(2) max by (WMD random effects model) 2.16 ml/kg/min (95% CI 2.82 to 1.49), exercise duration increased by 2.38 minutes (95% CI 2.85 to 1.9), work capacity by 15.1 Watts (95% CI 17.7 to 12.6) and distance on the six minute walk by 40.9 metres (95% CI 64.7 to 17.1). Improvements in VO(2) max were greater for training programmes of greater intensity and duration. HRQoL improved in the seven of nine trials that measured this outcome. Exercise training improves exercise capacity and quality of life in patients mild to moderate heart failure in the short term. There is currently no information regarding the effect of exercise training on clinical outcomes. The findings are based on small-scale trials in patients who are unrepresentative of the total population of patients with heart failure. Other groups (more severe patients, the elderly, women) may also benefit. Large-scale pragmatic trials of exercise training of longer duration, recruiting a wider spectrum of patients are needed to address these issues.


Finding the right position: a three-year, single-center experience with the "self-locating" catheter.
Peritoneal dialysis (PD) catheter displacement is a major cause of dysfunction. Various catheter forms have been designed to improve outcome. One of them is based on the classical Tenckhoff catheter, but includes a small tungsten cylinder at the distal end to optimize location and to prevent dislocations. ♢ In this retrospective study, we analyzed the functionality and complication profile of the self-locating catheter implanted at our center from November 2005 to September 2008 in our PD program. ♢ Data from 27 self-locating catheters implanted in 25 patients (12 women) were obtained. Patients were followed for up to 30 months (median observation time: 13 months), resulting in a cumulative study period of 357 PD months. A total of 22 complications were identified, including 1 episode of peritonitis per 52 patient-months, 1 tunnel infection, and 1 exit-site infection. The "functional catheter failure incidence rate" was approximately 0.01 catheters per month on PD (or approximately 1 catheter loss per 100 PD months). The catheter survival was 93% at 1 year. ♢ The "self-locating" PD catheter demonstrates low rates of catheter migration and dislocations, and good catheter survival.


Prevalence and seasonality of tick-borne pathogens in questing Ixodes ricinus ticks from Luxembourg.
In Europe, ixodid ticks are important arthropod vectors of human and animal pathogens, but comprehensive studies of the prevalence of all relevant pathogens in Central Europe are scarce. As a result of ecological changes, the incidences of tick-borne infections are expected to increase. In this study, 1,394 nymphal and adult Ixodes ricinus ticks sampled monthly during the active season from 33 ecologically distinct collection sites throughout Luxembourg were screened for all human tick-borne pathogens relevant in Central Europe. Species were identified by sequence analysis of detection PCR amplicons. Mean infection rates of ticks were 11.3% for Borrelia burgdorferi sensu lato, 5.1% for Rickettsia sp., 2.7% for Babesia sp., and 1.9% for Anaplasma phagocytophilum. No tick was found to be infected with Coxiella sp., Francisella tularensis subsp., or Tick-borne encephalitis virus (TBEV). A total of 3.2% of ticks were infected with more than one pathogen species, including mixed Borrelia infections (1.5%). Seasonal variations of tick infection rates were observed for Borrelia, Babesia, and Anaplasma, possibly reflecting a behavioral adaptation strategy of questing ticks. A positive correlation between the grade of urbanization and Borrelia infection rate of ticks was observed, suggesting an established urban zoonotic cycle. We also found Hepatozoon canis (0.1%) and Bartonella henselae (0.3%), which so far have not been found in questing Ixodes ricinus ticks in Central Europe.


Assessment of response consistency and respective participant profiles in the Internet-based NutriNet-Santé Cohort.
Whereas the feasibility and effectiveness of Internet-based epidemiologic research have been established, methodological support for the quality of such data is still accumulating. We aimed to identify sociodemographic differences among members of a French cohort according to willingness to provide part of one's 15-digit national identification number (personal Social Security number (PSSN)) and to assess response consistency based on information reported on the sociodemographic questionnaire and that reflected in the PSSN. We studied 100,118 persons enrolled in an Internet-based prospective cohort study, the NutriNet-Santé Study, between 2009 and 2013. Persons aged 18 years or more who resided in France and had Internet access were eligible for enrollment. The sociodemographic profiles of participants with discordant data were compared against those of participants with concordant data via 2-sided polytomous logistic regression. In total, 84,442 participants (84.3%) provided the first 7 digits of their PSSN, and among them 5,141 (6.1%) had discordant data. Our multivariate analysis revealed differences by sex, age, education, and employment as regards response consistency patterns. The results support the quality of sociodemographic data obtained online from a large and diverse volunteer sample. The quantitative description of participant profiles according to response consistency patterns could inform future methodological work in e-epidemiology.


Effect of fluctuation of oxygenation and time spent in the target range on retinopathy of prematurity in extremely low birth weight infants.
The association between saturation of peripheral oxygenation (SpO2) fluctuation and severity of retinopathy of prematurity (ROP) is well elucidated in extremely low birth weight (ELBW) infants. Time spent in the Target range of SpO2 is also associated with the severity of ROP. In a prospective observational study, the SpO2 of all ELBW infants admitted to our unit were monitored for the first four weeks of life, and averaged every minute for analysis. The percent time spent at SpO2 <90%, 90-95%, and >95% and weekly SpO2 fluctuations [as SpO2 coefficient of variation (CoV)] were calculated. During the study period 21 infants had moderate to severe ROP and 35 infants served as controls. Infants with moderate to severe ROP were smaller and younger than their controls [676±124 grams vs. 796±148 grams (p < 0.001); and 24.0±1.0 weeks vs. 25.0±1.7 weeks (p < 0.001) respectively]. There were no significant differences in time spent in the 90-95% range between groups (p = 0.66). However there was a significant increase in weekly SpO2 CoV in infants with moderate to severe ROP vs. controls (p = 0.007). In ELBW infants, there was an association between SpO2 fluctuation during the first four weeks of life and severity of ROP, although, no association was established with time spent in the target range of SpO2.


Effects of semi-group housing and floor type on pododermatitis, spinal deformation and bone quality in rabbit does.
The most common housing system for reproduction rabbits, individual cage housing on a wire floor, is increasingly scrutinized because of its potential detrimental impact on animal welfare. We compared three types of housing: (1) individual cage housing on a wire floor (3952 cm2/doe, maximum roof height 63 cm, one 1000 cm2 plastic footrest/doe), (2) semi-group housing on a wire floor (5000 cm2/doe, roofless, one 1000 cm2 plastic footrest/doe) and (3) the same semi-group housing, but with a fully plastic slatted floor. In all housing systems, does had free access to an elevated platform. In the semi-group housing pens, four does were housed communally during 21 days of the reproduction cycle (to allow more space for locomotion and to increase opportunities for social contact), and individually during the other 21 days of the cycle (to minimize doe-doe and doe-kit aggression that peaks around kindling). In all, 24 Hycole does were included per system. The does entered the experiment at 203 days of age (after their first parity). The experiment consisted of four reproductive cycles, ending at 369 days of age. Pododermatitis was scored in cycles 1, 2 and 4. At the end of the 4th cycle the does were euthanized and X-rays were taken to assess spinal deformation. Tibia and femur length, width and cortical thickness were determined and bone strength was assessed using a shear test, as a measure of bone quality. Although severe pododermatitis was absent, the prevalence of plantar hyperkeratosis (hair loss and callus formation) at the end of the 4th cycle was much greater on the wire floor (65% and 68% for semi-group housing and individual cages, respectively) than on the plastic floor (5%, P<0.0001), even though the wire floors were equipped with a plastic footrest known to decrease hyperkeratosis. In contrast to our expectations, semi-group housing did not affect the prevalence of spinal deformations (P>0.10), but in line with our expectations bone quality was affected favourably by semi-group housing. The tibial cortex (and to a lesser extent the femoral cortex) was thicker in semi-group housing than in individual cages (1.45, 1.46 and 1.38 mm for semi-group housing on wire, semi-group housing on plastic and individual housing on wire, respectively, P=0.045). What this increase in cortical thickness means in terms of doe welfare requires further study, as it may reflect an increase in activity resulting either from increased space for locomotion, or from fleeing aggressive pen mates.


Photosynthetic efficiency and mesophyll conductance are unaffected in Arabidopsis thaliana aquaporin knock-out lines.
Improving photosynthetic efficiency is widely regarded as a major route to achieving much-needed yield gains in crop plants. In plants with C3 photosynthesis, increasing the diffusion conductance for CO2 transfer from substomatal cavity to chloroplast stroma (gm) could help to improve the efficiencies of CO2 assimilation and photosynthetic water use in parallel. The diffusion pathway from substomatal cavity to chloroplast traverses cell wall, plasma membrane, cytosol, chloroplast envelope membranes, and chloroplast stroma. Specific membrane intrinsic proteins of the aquaporin family can facilitate CO2 diffusion across membranes. Some of these aquaporins, such as PIP1;2 in Arabidopsis thaliana, have been suggested to exert control over gm and the magnitude of the CO2 assimilation flux, but the evidence for a direct physiological role of aquaporins in determining gm is limited. Here, we estimated gm with four different methods under a range of light intensities and CO2 concentrations in two previously characterized pip1;2 knock-out lines as well as pip1;3 and pip2;6 knock-out lines, which have not been previously evaluated for a role in gm. This study presents the most in-depth analysis of gm in Arabidopsis aquaporin knock-out mutants to date. Surprisingly, all methods failed to show any significant differences between the pip1;2, pip1;3, or pip2;6 mutants and the Col-0 control.


Synaptic current at the squid giant synapse.
Transmission in the giant synapse of squid was studied by measuring synaptic currents in the voltage-clamped postsynaptic giant axon. These currents varied linearly with the axon's membrane potential, and showed an intercept on the voltage axis at, or near, the sodium equilibrium potential. The intercept shifted in seawater containing less sodium by even more than the shift in the sodium equilibrium potential. It is concluded that the transmitter at this synapse causes a significant change in the sodium conductance only.