Clonorchis sinensis excretory-secretory products increase malignant characteristics of cholangiocarcinoma cells in three-dimensional co-culture with biliary ductal plates.
Clonorchis sinensis is a carcinogenic human liver fluke, prolonged infection which provokes chronic inflammation, epithelial hyperplasia, periductal fibrosis, and even cholangiocarcinoma (CCA). These effects are driven by direct physical damage caused by the worms, as well as chemical irritation from their excretory-secretory products (ESPs) in the bile duct and surrounding liver tissues. We investigated the C. sinensis ESP-mediated malignant features of CCA cells (HuCCT1) in a three-dimensional microfluidic culture model that mimics an in vitro tumor microenvironment. This system consisted of a type I collagen extracellular matrix, applied ESPs, GFP-labeled HuCCT1 cells and quiescent biliary ductal plates formed by normal cholangiocytes (H69 cells). HuCCT1 cells were attracted by a gradient of ESPs in a concentration-dependent manner and migrated in the direction of the ESPs. Meanwhile, single cell invasion by HuCCT1 cells increased independently of the direction of the ESP gradient. ESP treatment resulted in elevated secretion of interleukin-6 (IL-6) and transforming growth factor-beta1 (TGF-β1) by H69 cells and a cadherin switch (decrease in E-cadherin/increase in N-cadherin expression) in HuCCT1 cells, indicating an increase in epithelial-mesenchymal transition-like changes by HuCCT1 cells. Our findings suggest that C. sinensis ESPs promote the progression of CCA in a tumor microenvironment via the interaction between normal cholangiocytes and CCA cells. These observations broaden our understanding of the progression of CCA caused by liver fluke infection and suggest a new approach for the development of chemotherapeutic for this infectious cancer.


Weak outcome predictors of multimodal rehabilitation at one-year follow-up in patients with chronic pain-a practice based evidence study from two SQRP centres.
For patients with chronic pain, the heterogeneity of clinical presentations makes it difficult to identify patients who would benefit from multimodal rehabilitation programs (MMRP). Yet, there is limited knowledge regarding the predictors of MMRP's outcomes. This study identifies predictors of outcome of MMRPs at a 12-month follow-up (FU-12) based on data from the Swedish Quality Registry for Pain Rehabilitation (SQRP). Patients with chronic pain from two clinical departments in Sweden completed the SQRP questionnaires-background, pain characteristics, psychological symptoms, function, activity/participation, health and quality of life-on three occasions: 1) during their first visit; 2) immediately after the completion of their MMRP; and 3) 12 months after completing the MMRP (n = 227). During the FU-12, the patients also retrospectively reported their global impressions of any changes in their perception of pain and their ability to handle their life situation in general. Significant improvements were found for pain, psychological symptoms, activity/participation, health, and quality of life aspects with low/medium strong effects. A general pattern was observed from the analyses of the changes from baseline to FU-12; the largest improvements in outcomes were significantly associated with poor situations according to their respective baseline scores. Although significant regressors of the investigated outcomes were found, the significant predictors were weak and explained a minor part of the variation in outcomes (15-25%). At the FU-12, 53.6% of the patients reported that their pain had decreased and 80.1% reported that their life situation in general had improved. These improvements were associated with high education, low pain intensity, high health level, and work importance (only pain perception). The explained variations were low (9-11%). Representing patients in real-world clinical settings, this study confirmed systematic reviews that outcomes of MMRP are associated with broad positive effects. A mix of background and baseline variables influenced the outcomes investigated, but the explained variations in outcomes were low. There is still a need to develop standardized and relatively simple outcomes that can be used to evaluate MMRP in trials, in clinical evaluations at group level, and for individual patients.


Opportunities and challenges in school-based sex and sexual health education in Nepal.
This article identifies and addresses opportunities for and challenges to current school-based sex and sexual health education in Nepal. Key literature searches were conducted of electronic databases and relevant web-sites, furthermore personal contact with experts and the hand searching of key journals was included. The review of this literature generated the following challenges: Limitations to teaching including lack of life skill-based and human right-based approach, inappropriate teaching aid and reliance on conventional methods, existing policy and practice, parental/community support, and lack of research into and evaluation of sex education. Diverse methodology in teaching, implementation of peer education programme, partnership with parents, involvement of external agencies and health professionals, capacity building of teachers, access to support and service organisation, and research and evaluation in sex education have been suggested for improving the current practice of sex and sexual health education in Nepalese schools. Key words: Sex education, education, school, adolescence, Nepal.


The effect of perhexiline on myocardial protection during coronary artery surgery: a two-centre, randomized, double-blind, placebo-controlled trial.
Perhexiline is thought to modulate metabolism by inhibiting mitochondrial carnitine palmitoyltransferase-1, reducing fatty acid uptake and increasing carbohydrate utilization. This study assessed whether preoperative perhexiline improves markers of myocardial protection in patients undergoing coronary artery bypass graft surgery and analysed its effect on the myocardial metabolome. In a prospective, randomized, double-blind, placebo-controlled trial, patients at two centres were randomized to receive either oral perhexiline or placebo for at least 5 days prior to surgery. The primary outcome was a low cardiac output episode in the first 6 h. All pre-specified analyses were conducted according to the intention-to-treat principle with a statistical power of 90% to detect a relative risk of 0.5 and a conventional one-sided α-value of 0.025. A subset of pre-ischaemic left ventricular biopsies was analysed using mass spectrometry-based metabolomics. Over a 3-year period, 286 patients were randomized, received the intervention and were included in the analysis. The incidence rate of a low cardiac output episode in the perhexiline arm was 36.7% (51/139) vs 34.7% (51/147) in the control arm [odds ratio (OR) 0.92, 95% confidence interval (CI) 0.56-1.50, P = 0.74]. Perhexiline was associated with a reduction in the cardiac index at 6 h [difference in means 0.19, 95% CI 0.07-0.31, P = 0.001] and an increase in inotropic support in the first 12 h (OR 0.55, 95% CI 0.34-0.89, P = 0.015). There were no significant differences in myocardial injury with troponin-T or electrocardiogram, reoperation, renal dysfunction or length of stay. No difference in the preischaemic left ventricular metabolism was identified between groups on metabolomics analysis. Preoperative perhexiline does not improve myocardial protection in patients undergoing coronary surgery and in fact reduced perioperative cardiac output, increasing the need for inotropic support. Perhexiline has no significant effect on the mass spectrometry-visible polar myocardial metabolome in vivo in humans, supporting the suggestion that it acts via a pathway that is independent of myocardial carnitine palmitoyltransferase inhibition and may explain the lack of clinical benefit observed following surgery. NCT00845364.


Targeting mTOR dependency in pancreatic cancer.
Pancreatic cancer is a leading cause of cancer-related death in the Western world. Current chemotherapy regimens have modest survival benefit. Thus, novel, effective therapies are required for treatment of this disease. Activating KRAS mutation almost always drives pancreatic tumour initiation, however, deregulation of other potentially druggable pathways promotes tumour progression. PTEN loss leads to acceleration of Kras(G12D)-driven pancreatic ductal adenocarcinoma (PDAC) in mice and these tumours have high levels of mammalian target of rapamycin (mTOR) signalling. To test whether these KRAS PTEN pancreatic tumours show mTOR dependence, we compared response to mTOR inhibition in this model, to the response in another established model of pancreatic cancer, KRAS P53. We also assessed whether there was a subset of pancreatic cancer patients who may respond to mTOR inhibition. We found that tumours in KRAS PTEN mice exhibit a remarkable dependence on mTOR signalling. In these tumours, mTOR inhibition leads to proliferative arrest and even tumour regression. Further, we could measure response using clinically applicable positron emission tomography imaging. Importantly, pancreatic tumours driven by activated KRAS and mutant p53 did not respond to treatment. In human tumours, approximately 20% of cases demonstrated low PTEN expression and a gene expression signature that overlaps with murine KRAS PTEN tumours. KRAS PTEN tumours are uniquely responsive to mTOR inhibition. Targeted anti-mTOR therapies may offer clinical benefit in subsets of human PDAC selected based on genotype, that are dependent on mTOR signalling. Thus, the genetic signatures of human tumours could be used to direct pancreatic cancer treatment in the future.


Center-surround patterns emerge as optimal predictors for human saccade targets.
The human visual system is foveated, that is, outside the central visual field resolution and acuity drop rapidly. Nonetheless much of a visual scene is perceived after only a few saccadic eye movements, suggesting an effective strategy for selecting saccade targets. It has been known for some time that local image structure at saccade targets influences the selection process. However, the question of what the most relevant visual features are is still under debate. Here we show that center-surround patterns emerge as the optimal solution for predicting saccade targets from their local image structure. The resulting model, a one-layer feed-forward network, is surprisingly simple compared to previously suggested models which assume much more complex computations such as multi-scale processing and multiple feature channels. Nevertheless, our model is equally predictive. Furthermore, our findings are consistent with neurophysiological hardware in the superior colliculus. Bottom-up visual saliency may thus not be computed cortically as has been thought previously.


Does prolonged breast-feeding impair child growth? A critical review.
Recent epidemiologic evidence suggests that children in developing countries who terminate breast-feeding before their first birthday are less likely to be malnourished than those who are breast-fed for longer durations. This finding calls into question the advice women are given to breast-feed as long as possible. This review examines several studies that found a relationship between prolonged breast-feeding and malnutrition. Many studies have shown a negative association between prolonged breast-feeding and growth, but there is little reason to expect the association to be causal. Problems in study design and analysis, such as failure to control for confounding and reverse causality, leave the nature of the association largely unspecified.


Toxoplasma gondii in stranded marine mammals from the North Sea and Eastern Atlantic Ocean: Findings and diagnostic difficulties.
The occurrence of the zoonotic protozoan parasite Toxoplasma gondii in marine mammals remains a poorly understood phenomenon. In this study, samples from 589 marine mammal species and 34 European otters (Lutra lutra), stranded on the coasts of Scotland, Belgium, France, The Netherlands and Germany, were tested for the presence of T. gondii. Brain samples were analysed by polymerase chain reaction (PCR) for detection of parasite DNA. Blood and muscle fluid samples were tested for specific antibodies using a modified agglutination test (MAT), a commercial multi-species enzyme-linked immunosorbent assay (ELISA) and an immunofluorescence assay (IFA). Out of 193 animals tested by PCR, only two harbour porpoise (Phocoena phocoena) cerebrum samples, obtained from animals stranded on the Dutch coast, tested positive. The serological results showed a wide variation depending on the test used. Using a cut-off value of 1/40 dilution in MAT, 141 out of 292 animals (41%) were positive. Using IFA, 30 out of 244 tested samples (12%) were positive at a 1/50 dilution. The commercial ELISA yielded 7% positives with a cut-off of the sample-to-positive (S/P) ratio≥50; and 12% when the cut-off was set at S/P ratio≥20. The high number of positives in MAT may be an overestimation due to the high degree of haemolysis of the samples and/or the presence of lipids. The ELISA results could be an underestimation due to the use of a multispecies conjugate. Our results confirm the presence of T. gondii in marine mammals in The Netherlands and show exposure to the parasite in both the North Sea and the Eastern Atlantic Ocean. We also highlight the limitations of the tests used to diagnose T. gondii in stranded marine mammals.


Static polarizability and second hyperpolarizability of closed- and open-shell pi-conjugated polymers.
The static longitudinal linear polarizability (alpha) and second order hyperpolarizability (gamma) for neutral and charged, closed- and open-shell trans-polyacetylene (PA) chains C(2n)H(2n+2), C(2n-1)H(2n+1), C(2n-1)H(2n+1) (+), C(2n)H(2n+2) (+), and C(2n)H(2n+2) (2+) are systematically investigated and compared. The polarizabilities are calculated within the Pariser-Parr-Pople model, and the electron correlation effect is included through density matrix renormalization group. It turns out that for both alpha, and gamma, two neutral PA chains C(2n)H(2n+2) and C(2n-1)H(2n+1) give similar values, while both singly charged and doubly charged systems present significantly larger magnitude of alpha and gamma values than the two neutral chains. The two singly charged PA chains C(2n-1)H(2n+1) (+) and C(2n)H(2n+2) (+) give more apparent nonlinear optical responses than doubly charged case C(2n)H(2n+2) (2+) and both present negative second order hyperpolarizabilities for short to medium sized oligomers. The sign inversion of gamma values in singly charged PA molecules is anticipated to take place at the much longer length than ever observed due to the significant effects of electron correlation and geometry.


Visualization features of canalis sinuosus with cone beam computed tomography.
One of the key stages of evaluating an edentulous ridge prior to dental implant placement is the analysis of neighboring anatomical structures such as canalis sinuosus (CS) with cone-beam computed tomography (CBCT). CBCT visualization has its own features, such as the change of slice thickness. The aim of the study was to analyze CS prevalence in relation to the slice thickness and CS diameter according to CBCT scans. In total, 100 CBCT scans of 39 males and 61 females in the age span of 46 to 81 years were retrospectively studied. Terminal alveolar part of CS was analyzed in Ez3D2009 (Vatech) software on panoramic and cross-sectional views with 0.5 mm, 1 mm, 3 mm, and 10 mm slice thicknesses. The prevalence was documented with regard to the diameter and slice thickness and was statistically compared in age and gender groups. This CBCT study demonstrated 55.5% CS prevalence. There was no statistically significant difference in presence between age and gender groups regarding diameter as well as within different slice thickness. The study also showed the importance of the slice thickness choice for CS visualization: the best visualization was reached with 0.5 mm and 1 mm slice thicknesses. Visualization with 0.5 mm and 1 mm slice thicknesses was absolutely the same (55.5%). General CS prevalence with a 10 mm slice thickness (16.5%) was significantly lower (P < 0.05) than the prevalence with a 3 mm slice thickness (20.5%). The correlation between the slice thickness and a diameter more than 1.5 mm in size was not found. CBCT analysis showed that the highest CS prevalence was detected with the use of 0.5/1 mm slice thickness. As well, the higher CS diameter, the better is its visualization.


Risk of burnout in perioperative clinicians: a survey study and literature review.
Burnout can lead to health and psychologic problems and is apparently increasing in physicians and nurses. Previous studies have not evaluated all healthcare workers within a single work unit. This study evaluates the risk of burnout in all medical personnel in one perioperative unit. We developed an online survey that included demographics, a modified version of the Maslach Burnout Inventory-Human Services Survey, and the Social Support and Personal Coping Survey. Survey constructs (e.g., depersonalization and health) and a global score were calculated. Larger construct and global values were associated with higher risk of burnout. These were separately regressed on role, age, and sex. The global score was then regressed on each of the survey constructs. Of the 145 responses, 46.2% were physicians (22.8% residents), 43.4% were nurses or nurse anesthetists, and 10.3% were other personnel. After adjusting for sex and age, residents scored higher than other physicians on the following (expected change [95% confidence interval]): global score (1.12 [0.43-1.82]), emotional exhaustion (1.54 [0.44-2.60]), and depersonalization (1.09 [0.23-1.95]). Compared with nonphysicians, residents were 1 U or more higher on these items (P < 0.05 in all cases). Residents had higher health (1.49 [0.48-2.50]) and workload (1.23 [0.07-2.40]) values compared with physicians. Better health, personal support, and work satisfaction scores were related to decreased global scores (P < 0.05). Physicians (particularly residents) had the largest global burnout scores, implying increased risk of burnout. Improving overall health, increasing personal support, and improving work satisfaction may decrease burnout among perioperative team members.


Fanconi syndrome and lactic acidosis associated with stavudine and lamivudine therapy.
We report a case of the nucleoside reverse transcriptase inhibitors, stavudine and lamivudine inducing Fanconi syndrome in a patient. The presence of simultaneous lactic acidosis suggests mitochondrial toxicity within the proximal renal tubular cells as the likely pathogenesis. We recommend that both the above nucleosides be added to the list of antiretroviral drugs that can induce Fanconi syndrome and that patients on stavudine and lamivudine be monitored carefully for early signs of Fanconi syndrome.


Choosing and using climate-change scenarios for ecological-impact assessments and conservation decisions.
Increased concern over climate change is demonstrated by the many efforts to assess climate effects and develop adaptation strategies. Scientists, resource managers, and decision makers are increasingly expected to use climate information, but they struggle with its uncertainty. With the current proliferation of climate simulations and downscaling methods, scientifically credible strategies for selecting a subset for analysis and decision making are needed. Drawing on a rich literature in climate science and impact assessment and on experience working with natural resource scientists and decision makers, we devised guidelines for choosing climate-change scenarios for ecological impact assessment that recognize irreducible uncertainty in climate projections and address common misconceptions about this uncertainty. This approach involves identifying primary local climate drivers by climate sensitivity of the biological system of interest; determining appropriate sources of information for future changes in those drivers; considering how well processes controlling local climate are spatially resolved; and selecting scenarios based on considering observed emission trends, relative importance of natural climate variability, and risk tolerance and time horizon of the associated decision. The most appropriate scenarios for a particular analysis will not necessarily be the most appropriate for another due to differences in local climate drivers, biophysical linkages to climate, decision characteristics, and how well a model simulates the climate parameters and processes of interest. Given these complexities, we recommend interaction among climate scientists, natural and physical scientists, and decision makers throughout the process of choosing and using climate-change scenarios for ecological impact assessment. Selección y Uso de Escenarios de Cambio Climático para Estudios de Impacto Ecológico y Decisiones de Conservación.


Highly efficient near-infrared emission in Er3+ doped silica films containing size-tunable SnO2 nanocrystals.
Co-doping size-tunable SnO2 nanocrystals into Er(3+) ions embedded silica thin films produces an enhancement of Er-related near-infrared emission by three orders of magnitude. Selective PL and PLE measurements show that energy transfer process occurs between SnO2 nanocrystals and Er(3+) ions. Quantitative studies of PL decay lifetime and photoluminescence temperature-dependence demonstrate that both high energy transfer efficiency from SnO2 nanocrystals to Er(3+) ions and the partial incorporation of Er(3+) ions into SnO2 nanocrystals contribute to the near-infrared emission enhancement. All these results indicated that SnO2 nanocrystals with suitable size have great potentials in fabricating high-efficiency near-infrared luminous materials as sensitizers of Er(3+) ions.


Effect of a school-based intervention to promote healthy lifestyles in 7-11 year old children.
Physical inactivity is recognised as a public health concern within children and interventions to increase physical activity are needed. The purpose of this research was to evaluate the effect of a school-based healthy lifestyles intervention on physical activity, fruit and vegetable consumption, body composition, knowledge, and psychological variables. A non-randomised controlled study involving 8 primary schools (4 intervention, 4 control). Participants were 589 children aged 7-11 years. The intervention lasted 10 months and comprised a CD-rom learning and teaching resource for teachers; an interactive website for pupils, teachers and parents; two highlight physical activity events (1 mile school runs/walks); a local media campaign; and a summer activity wall planner and record. Primary outcome measures were objectively measured physical activity (pedometers and accelerometers) and fruit and vegetable consumption. Secondary outcomes included body mass index, waist circumference, estimated percent body fat, knowledge, psychological variables. Multi-level modelling was employed for the data analysis. Relative to children in control schools, those in intervention schools significantly increased their total time in moderate-to-vigorous physical activity (MVPA) (by 9 minutes/day vs a decrease of 10 minutes/day), their time in MVPA bouts lasting at least one minute (10 minutes/day increase vs no change) and increased daily steps (3059 steps per day increase vs 1527 steps per day increase). A similar pattern of results was seen in a subset of the least active participants at baseline. Older participants in intervention schools showed a significant slowing in the rate of increase in estimated percent body fat, BMI, and waist circumference. There were no differences between groups in fruit and vegetable intake. Extrinsic motivation decreased more in the intervention group. The intervention produced positive changes in physical activity levels and body composition. It appeared to have little or no effect on consumption of fruit and vegetables. Schools are a suitable setting for the promotion of healthy lifestyles although more work, particularly focussed on dietary change, is needed in a variety of schools and social settings.


Adenovirus E4orf4 protein reduces phosphorylation of c-Fos and E1A proteins while simultaneously reducing the level of AP-1.
Adenovirus E1A protein and cyclic AMP cooperate to induce transcription factor AP-1 and viral gene expression in mouse S49 cells. We report that a protein encoded within the viral E4 gene region acts to counterbalance the induction of AP-1 DNA-binding activity by E1A and cyclic AMP. Studies with mutant adenoviruses demonstrated that in the absence of E4orf4 protein, AP-1 DNA-binding activity is induced to substantially higher levels than in wild-type virus-infected cells. The induction is the result of increased production of JunB and c-Fos proteins. Hyperphosphorylated forms of c-Fos and E1A proteins accumulate in the absence of functional E4orf4 protein. We propose that the E4orf4 protein acts to inhibit the activity of a cellular kinase that phosphorylates both the E1A and c-Fos proteins. Phosphorylation-dependent alterations in the activity of c-Fos, E1A, or some unidentified protein might, then, lead to decreased synthesis of AP-1 components. This E4 function likely plays an important role in natural infections, since a mutant virus unable to express the E4orf4 protein is considerably more cytotoxic than the wild-type virus.


Age-dependent tolerance to an endogenous tumor-associated antigen.
Immunologic tolerance to endogenous antigens reduces antitumor responses. Gp70 is an endogenous tumor-associated antigen (TAA) of the BALB/c-derived colon carcinoma CT26. We found that expression of gp70 mRNA is detectable in tissues of mice 8 months of age and older. We showed that expression of gp70 establishes immunologic tolerance and affects antitumor immunity in a similarly age-dependent manner using gp70-deficient mice. We found that tumors grew in all gp70-sufficient mice, while approximately half of gp70-deficient mice controlled tumor growth with endogenous T-cell responses. Protection in gp70-deficient mice correlated with more robust gp70-specific CTL responses, and increased numbers and avidity of responding antigen-specific T cells after vaccination. We conclude that immunosurveillance may decline with age due to increased or de novo peripheral expression of endogenous TAAs.


Water- and Nitrogen-Use Efficiencies of Hemp (Cannabis sativa L.) Based on Whole-Canopy Measurements and Modeling.
Interest in hemp (Cannabis sativa L.) as a crop for the biobased economy is growing worldwide because hemp produces a high and valuable biomass while requiring low inputs. To understand the physiological basis of hemp's resource-use efficiency, canopy gas exchange was assessed using a chamber technique on canopies exposed to a range of nitrogen (N) and water levels. Since canopy transpiration and carbon assimilation were very sensitive to variations in microclimate among canopy chambers, observations were adjusted for microclimatic differences using a physiological canopy model, with leaf-level parameters estimated for hemp from our previous study. Canopy photosynthetic water-use efficiency (PWUEc), defined as the ratio of gross canopy photosynthesis to canopy transpiration, ranged from 4.0 mmol CO2 (mol H2O)-1 to 7.5 mmol CO2 (mol H2O)-1. Canopy photosynthetic nitrogen-use efficiency (PNUEc), the ratio of the gross canopy photosynthesis to canopy leaf-N content, ranged from 0.3 mol CO2 d-1 (g N)-1 to 0.7 mol CO2 d-1 (g N)-1. The effect of N-input levels on PWUEc and PNUEc was largely determined by the N effect on canopy size or leaf area index (LAI), whereas the effect of water-input levels differed between short- and long-term stresses. The effect of short-term water stress was reflected by stomatal regulation. The long-term stress increased leaf senescence, decreased LAI but retained total canopy N content; however, the increased average leaf-N could not compensate for the lost LAI, leading to a decreased PNUEc. Although hemp is known as a resource-use efficient crop, its final biomass yield and nitrogen use efficiency may be restricted by water limitation during growth. Our results also suggest that crop models should take stress-induced senescence into account in addition to stomatal effects if crops experience a prolonged water stress during growth.


Manipulation of adenosine kinase affects sleep regulation in mice.
Sleep and sleep intensity are enhanced by adenosine and its receptor agonists, whereas adenosine receptor antagonists induce wakefulness. Adenosine kinase (ADK) is the primary enzyme metabolizing adenosine in adult brain. To investigate whether adenosine metabolism or clearance affects sleep, we recorded sleep in mice with engineered mutations in Adk. Adk-tg mice overexpress a transgene encoding the cytoplasmic isoform of ADK in the brain but lack the nuclear isoform of the enzyme. Wild-type mice and Adk(+/-) mice that have a 50% reduction of the cytoplasmic and the nuclear isoforms of ADK served as controls. Adk-tg mice showed a remarkable reduction of EEG power in low frequencies in all vigilance states and in theta activity (6.25-11 Hz) in rapid eye movement (REM) sleep and waking. Adk-tg mice were awake 58 min more per day than wild-type mice and spent significantly less time in REM sleep (102 ± 3 vs 128 ± 3 min in wild type). After sleep deprivation, slow-wave activity (0.75-4 Hz), the intensity component of non-rapid eye movement sleep, increased significantly less in Adk-tg mice and their slow-wave energy was reduced. In contrast, the vigilance states and EEG spectra of Adk(+/-) and wild-type mice did not differ. Our data suggest that overexpression of the cytoplasmic isoform of ADK is sufficient to alter sleep physiology. ADK might orchestrate neurotransmitter pathways involved in the generation of EEG oscillations and regulation of sleep.


Responsiveness of the Smoking Cessation Quality of Life (SCQoL) questionnaire.
The Smoking Cessation Quality of Life (SCQoL) questionnaire was developed to quantify changes in self-reported functioning and well-being associated with the smoking cessation process and to facilitate comparisons among smokers, former smokers, and nonsmokers. The SCQoL includes 5 cessation-targeted scales and the 8 multi-item scales of the Medical Outcomes Study 36-Item Short-Form Health Survey. This study was conducted to assess the responsiveness of the SCQoL by analyzing associations between SCQoL scale scores and duration of smoking abstinence. The SCQoL was administered at a screening visit and 2 to 6 weeks after screening as part of a longitudinal study. Study participants included smokers who intended to quit smoking. Subjects were required to purchase nicotine inhalers and were prompted to quit smoking before follow-up. Based on self-reported duration of abstinence at follow-up, subjects were categorized as recidivists (0 days smoke free), short-term abstainers (1-7 days smoke free), or longer-term abstainers (>7 days smoke free). Kruskal-Wallis tests were used to compare changes in scale scores from screening to follow-up among the 3 groups. The internal consistency reliability of the 13 SCQoL scales ranged from 0.67 to 0.92. Subjects who maintained abstinence for longer periods experienced smaller declines in health-related quality of life between the screening and follow-up assessments. Differences among the 3 groups were driven primarily by differences between recidivists and longer-term abstainers. The results are consistent with research indicating that recidivists report a greater number of (and more severe) cessation-related symptoms than abstainers. The findings of this investigation provide evidence for the responsiveness of the SCQoL.


Optical resonances and two-sphere systems.
After a historical review of previous research into cooperative scattering, a theoretical investigation of the effects of interparticle coupling on morphology-dependent resonances of spheres is conducted. Bispheres composed of identical, slightly dissimilar, and very different monomers are considered. Calculations are presented of resonance spectra for selected orientations of the bispheres relative to the incident wave vector along with spectra that should prove useful in describing the scattering properties of a monodisperse ensemble of randomly oriented bispheres. (The bispheres in this dispersion may be constituted from dissimilar monomers.) Normalized source functions for regions inside and near the scatterers are also provided. Finally a numerical simulation of an interesting experiment is carried out in which a resonating aerosol passes through the focal volume of a relatively large, spherical microlens.


Microtubule organization by kinesin motors and microtubule crosslinking protein MAP65.
Microtubules are rigid, proteinaceous filaments required to organize and rearrange the interior of cells. They organize space by two mechanisms, including acting as the tracks for long-distance cargo transporters, such as kinesin-1, and by forming a network that supports the shape of the cell. The microtubule network is composed of microtubules and a bevy of associated proteins and enzymes that self-organize using non-equilibrium dynamic processes. In order to address the effects of self-organization of microtubules, we have utilized the filament-gliding assay with kinesin-1 motors driving microtubule motion. To further enhance the complexity of the system and determine if new patterns are formed, we added the microtubule crosslinking protein MAP65-1. MAP65-1 is a microtubule-associated protein from plants that crosslinks antiparallel microtubules, similar to mammalian PRC1 and fission yeast Ase1. We find that MAP65 can slow and halt the velocity of microtubules in gliding assays, but when pre-formed microtubule bundles are added to gliding assays, kinesin-1 motors can pull apart the bundles and reconstitute cell-like protrusions.


Estimates of genetic correlations between days to calving and reproductive and weight traits in Nelore cattle.
Data comprising 53,181 calving records were analyzed to estimate the genetic correlation between days to calving (DC), and days to first calving (DFC), and the following traits: scrotal circumference (SC), age at first calving (AFC), and weight adjusted for 550 d of age (W550) in a Nelore herd. (Co)variance components were estimated using the REML method fitting bivariate animal models. The fixed effects considered for DC were contemporary group, month of last calving, and age at breeding season (linear and quadratic effects). Contemporary groups were composed by herd, year, season, and management group at birth; herd and management group at weaning; herd, season, and management group at mating; and sex of calf and mating type (multiple sires, single sire, or AI). In DFC analysis, the same fixed effects were considered excluding the month of last calving. For DC, a repeatability animal model was applied. Noncalvers were not considered in analyses because an attempt to include them, attributing a penalty, did not improve the identification of genetic differences between animals. Heritability estimates ranged from 0.04 to 0.06 for DC, from 0.06 to 0.13 for DFC, from 0.42 to 0.44 for SC, from 0.06 to 0.08 for AFC, and was 0.30 for W550. The genetic correlation estimated between DC and SC was low and negative (-0.10), between DC and AFC was high and positive (0.76), and between DC and W550 was almost null (0.07). Similar results were found for genetic correlation estimates between DFC and SC (-0.14), AFC (0.94), and W550 (-0.02). The genetic correlation estimates indicate that the use of DC in the selection of beef cattle may promote favorable correlated responses to age at first mating and, consequently, higher gains in sexual precocity can be expected.


A randomized controlled trial of psychosocial interventions using the psychophysiological framework for Chinese breast cancer patients.
This study aimed to investigate the psychophysiological outcomes of different psychosocial interventions for breast cancer patients. Participants were randomly assigned into 3 intervention groups, namely, Body-Mind-Spirit (BMS), Supportive-Expressive (SE), and Social Support Self-Help (SS) groups; a no-intervention group was used as control. Salivary cortisol was used as the physiological stress marker. Distress level, mental adjustment, emotional control, and social support were measured. Data were collected at baseline, 4 month, and 8 month. Preliminary results indicated that BMS intervention produced the greatest and the most sustained effects. It enhanced positive social support, reduced psychological distress, emotional control, and negative mental adjustment. Total salivary cortisol was lowered after 8 months. Most participants in SE groups indicated the treatment helpful, but changes in psychophysiological outcomes were not statistically significant. Participants in SS groups seemed less likely to benefit from the intervention. The no intervention control group indicated a reduction in social support. These outcomes suggest that active professional intervention is more likely to yield therapeutic effects. In particular, psychosocial intervention attending to the spiritual dimension contributes to positive outcomes.


Avelumab Maintenance Therapy for Advanced or Metastatic Urothelial Carcinoma.
Platinum-based chemotherapy is standard-of-care first-line treatment for advanced urothelial carcinoma. However, progression-free survival and overall survival are limited by chemotherapy resistance. In a phase 3 trial, we randomly assigned patients with unresectable locally advanced or metastatic urothelial cancer who did not have disease progression with first-line chemotherapy (four to six cycles of gemcitabine plus cisplatin or carboplatin) to receive best supportive care with or without maintenance avelumab. The primary end point was overall survival, assessed among all patients who underwent randomization (overall population) and among those with tumors positive for programmed cell death ligand 1 (PD-L1). Secondary end points included progression-free survival and safety. Among all 700 patients who underwent randomization, the addition of maintenance avelumab to best supportive care significantly prolonged overall survival as compared with best supportive care alone (control). Overall survival at 1 year was 71.3% in the avelumab group and 58.4% in the control group (median overall survival, 21.4 months vs. 14.3 months; hazard ratio for death, 0.69; 95% confidence interval [CI], 0.56 to 0.86; P = 0.001). Avelumab also significantly prolonged overall survival in the PD-L1-positive population; overall survival at 1 year was 79.1% in the avelumab group and 60.4% in the control group (hazard ratio, 0.56; 95% CI, 0.40 to 0.79; P<0.001). The median progression-free survival was 3.7 months in the avelumab group and 2.0 months in the control group in the overall population (hazard ratio for disease progression or death, 0.62; 95% CI, 0.52 to 0.75) and 5.7 months and 2.1 months, respectively, in the PD-L1-positive population (hazard ratio, 0.56; 95% CI, 0.43 to 0.73). The incidence of adverse events from any cause was 98.0% in the avelumab group and 77.7% in the control group; the incidence of adverse events of grade 3 or higher was 47.4% and 25.2%, respectively. Maintenance avelumab plus best supportive care significantly prolonged overall survival, as compared with best supportive care alone, among patients with urothelial cancer who had disease that had not progressed with first-line chemotherapy. (Funded by Pfizer and Merck [Darmstadt, Germany]; JAVELIN Bladder 100 ClinicalTrials.gov number, NCT02603432.).


Achalasia as a complication of bulimia nervosa: A case report.
Oesophageal achalasia is a medical condition characterised by oesophageal aperistalsis, an increased resting pressure with partial or incomplete relaxation of the lower oesophageal sphincter. Bulimia nervosa (BN) is an eating disorder manifested by binge eating attacks followed by recurrent inappropriate compensatory behaviours, such as self-induced vomiting and excessive exercise. Dysphagia, regurgitation, vomiting, retrosternal pain, heartburn, weight loss, avoidance of eating, consumption of large amount of liquids and aberrant eating behaviours are symptoms of both achalasia and BN. Owing to these common signs and symptoms, oesophageal achalasia can be misdiagnosed as an eating disorder. In addition, oesophageal achalasia can occur as a complication of BN. It is often difficult to distinguish organic and psychological vomiting or comorbidity because of overlapping of the symptoms. We report the case of a patient who developed oesophageal achalasia following severe, repetitive vomiting as a complication of BN. We want to raise awareness regarding the development of a medical illness in the presence of a psychiatric disorder. Importantly, physicians should have a fundamental knowledge of these two diseases regarding their clinical patterns to differentially diagnose one or both disorders as quickly as possible.


Effect of Molecular Architecture of PDMAEMA-POEGMA Random and Block Copolymers on Their Adsorption on Regenerated and Anionic Nanocelluloses and Evidence of Interfacial Water Expulsion.
Block copolymers of poly(2-(dimethylamino)ethyl methacrylate) (PDMAEMA) and poly(oligo(ethylene glycol) methyl ether methacrylate) (POEGMA) with varying block sizes were synthesized by consecutive reversible addition-fragmentation chain transfer (RAFT) polymerization and then exposed to cellulose substrates with different anionic charge density. The extent and dynamics of quaternized PDMAEMA-b-POEGMA adsorption on regenerated cellulose, cellulose nanofibrils (CNF), and (2,2,6,6-tetramethylpiperidin-1-yl)oxyl (TEMPO)-oxidized cellulose nanofibrils (TOCNF) was determined by using electromechanical and optical techniques, namely, quartz crystal microbalance (QCM-D) and surface plasmon resonance (SPR), respectively. PDMAEMA-b-POEGMA equilibrium adsorption increased with the anionic charge of cellulose, an indication of electrostatic interactions. Such an observation was further confirmed by atomic force microscopy (AFM) and X-ray photoelectron spectroscopy (XPS). Depending on their architecture, adsorption on TOCNF of some of the PDMAEMA-b-POEGMA copolymers produced a significant reduction in QCM frequency, as expected from large mass uptake, while surprisingly, other copolymers induced the opposite effect. This latter, remarkable behavior was ascribed to coupled water expulsion from the interface upon charge neutralization of anionic surface sites with adsorbing cationic polymer segments. These observations were further investigated with SPR and QCM-D measurements using deuterium oxide solvent exchange to determine the amount of coupled water at the TOCNF-block copolymer interface. Finally, random copolymers with similar composition adsorbed to a larger extent compared to the respective block copolymers, revealing the effect of adsorbed loops and tails as well as hydration.


Scoring pairwise genomic sequence alignments.
The parameters by which alignments are scored can strongly affect sensitivity and specificity of alignment procedures. While appropriate parameter choices are well understood for protein alignments, much less is known for genomic DNA sequences. We describe a straightforward approach to scoring nucleotide substitutions in genomic sequence alignments, especially human-mouse comparisons. Scores are obtained from relative frequencies of aligned nucleotides observed in alignments of non-coding, non-repetitive genomic regions, and can be theoretically motivated through substitution models. Additional accuracy can be attained by down-weighting alignments characterized by low compositional complexity. We also describe an evaluation protocol that is relevant when alignments are intended to identify all and only the orthologous positions. One particular scoring matrix, called HOXD70, has proven to be generally effective for human-mouse comparisons, and has been used by the PipMaker server since July, 2000. We discuss but leave open the problem of effectively scoring regions of strongly biased nucleotide composition, such as low G + C content.


Drowning deaths in Sweden with emphasis on the presence of alcohol and drugs - a retrospective study, 1992-2009.
Drowning deaths constitute a significant proportion of unnatural deaths globally. In Sweden and other high-income countries, drowning deaths have decreased. This study investigates the epidemiology and current trends of unintentional, intentional, and undetermined drowning deaths with emphasis on the presence of alcohol and other drugs. During an 18-years period, 5,125 drowning deaths were autopsied in Sweden. Data on cases including toxicological analysis on alcohol, pharmaceutical drugs, and illicit drugs were obtained from the National Board of Forensic Medicine. During the study period, the annual incidence of drowning deaths in Sweden was 3.1/100,000 inhabitants and decreased on average by about 2% each year (p<0.001). The highest incidence was found among males and in middle/older age groups. The incidence increased 3% for each year of age. Children/adolescents (≤18 years) constituted 5% of all drowning deaths. Of all drowned females in the study, 55% (847/1,547) committed suicide, which was a significantly higher proportion compared with males (21%, 763/3,578) (p<0.001). In total, 38% (1,656/4,377) of tested drowned persons had alcohol in their blood and the mean concentration was 1.8 g/l. In the unintentional drowning group, intentional drowning group, and the undetermined group, the proportion of alcohol positive was 44%, 24%, and 45%, respectively. One or several psychoactive drugs were present in the blood in 40% (1,688 /4,181) of all tested persons and in 69% (965/1,394) of tested persons who died from suicidal drowning. The most common drug was benzodiazepines (21%, 891/4,181). Illicit drugs were detected in 10% (82/854) of tested persons. Presence of alcohol and drugs were frequent and may have contributed to the drowning deaths. The incidence of drowning deaths significantly decreased during the study period. Males and the middle/older age groups had a higher incidence compared to females and children. Suicidal drowning was common especially among women. Alcohol and drugs are significant contributors in drowning deaths in Sweden and should be considered as part of a comprehensive prevention program.


Delineation of the endocytic pathway of substance P and its seven-transmembrane domain NK1 receptor.
Many of the actions of the neuropeptide substance P (SP) that are mediated by the neurokinin 1 receptor (NK1-R) desensitize and resensitize, which may be associated with NK1-R endocytosis and recycling. We delineated this endocytic pathway in transfected cells by confocal microscopy using cyanine 3-SP and NK1-R antibodies. SP and the NK1-R were internalized into the same clathrin immunoreactive vesicles, and then sorted into different compartments. The NK1-R was colocalized with a marker of early endosomes, but not with markers of late endosomes or lysosomes. We quantified the NK1-R at the cell surface by incubating cells with an antibody to an extracellular epitope. After exposure to SP, there was a loss and subsequent recovery of surface NK1-R. The loss was prevented by hypertonic sucrose and potassium depletion, inhibitors of clathrin-mediated endocytosis. Recovery was independent of new protein synthesis because it was unaffected by cycloheximide. Recovery required endosomal acidification because it was prevented by an H(+)-ATPase inhibitor. The fate of internalized 125I-SP was examined by chromatography. SP was intact at the cell surface and in early endosomes, but slowly degraded in perinuclear vesicles. We conclude that SP induces clathrin-dependent internalization of the NK1-R. The SP/NK1-R complex dissociates in acidified endosomes. SP is degraded, whereas the NK1-R recycles to the cell surface.


Measurements of reactive ion etching process effect using long-period fiber gratings.
The paper presents for the first time a study of long-period fiber gratings (LPFGs) applied for the measurements of reactive ion etching (RIE) process effect in various places of a plasma reactor. For the purposes of the experiment a number of highly sensitive LPFGs working at the dispersion turning point was fabricated using electric arc discharges. We show that the LPFGs allow for monitoring of the phenomena taking place in the reactor, especially those resulting in reduction of the LPFG diameter. Results of the measurements supported by simulations have shown that etching rate significantly decreases with elevation of the sample up to 3.6 mm over the electrode in the reactor, and stays constant above this height.


Significant enhancement of broadband optical limiting behavior using off-resonant sub-wavelength coupled plasmonic waves.
We present the numerical investigation of the optical limiting behavior based on coupled nonlinear plasmonic waveguides. Exploiting the strong localization of the electromagnetic fields at metal-dielectric interfaces, significant enhancement of the nonlinear absorption was achieved. Two types of optical limiters (OLs), one based on the enhanced optical Kerr (OK) effect and the other based on the enhanced two-photon absorption (TPA), are proposed. Their transmission characteristics at off-resonant band of Au are investigated. The simulation results reveal that the linear transmittances in both cases are higher than 85%, and the limiting thresholds are 250 GW/cm(2) and 42.69 GW/cm(2) for the OK and TPA based OLs, respectively. As compared with the non-structured slab waveguides, the optical limiting thresholds are greatly reduced. Wideband operation over 200 nm was confirmed and TPA induced free carrier absorption (FCA) discussed.


A toolkit for ARB to integrate custom databases and externally built phylogenies.
Researchers are perpetually amassing biological sequence data. The computational approaches employed by ecologists for organizing this data (e.g. alignment, phylogeny, etc.) typically scale nonlinearly in execution time with the size of the dataset. This often serves as a bottleneck for processing experimental data since many molecular studies are characterized by massive datasets. To keep up with experimental data demands, ecologists are forced to choose between continually upgrading expensive in-house computer hardware or outsourcing the most demanding computations to the cloud. Outsourcing is attractive since it is the least expensive option, but does not necessarily allow direct user interaction with the data for exploratory analysis. Desktop analytical tools such as ARB are indispensable for this purpose, but they do not necessarily offer a convenient solution for the coordination and integration of datasets between local and outsourced destinations. Therefore, researchers are currently left with an undesirable tradeoff between computational throughput and analytical capability. To mitigate this tradeoff we introduce a software package to leverage the utility of the interactive exploratory tools offered by ARB with the computational throughput of cloud-based resources. Our pipeline serves as middleware between the desktop and the cloud allowing researchers to form local custom databases containing sequences and metadata from multiple resources and a method for linking data outsourced for computation back to the local database. A tutorial implementation of the toolkit is provided in the supporting information, S1 Tutorial. http://www.ece.drexel.edu/gailr/EESI/tutorial.php.


The Effect of Coatings on Cutting Force in Turning of C45 Steel.
This article focuses on the development of a mathematical model of a cutting force that is applicable for coated and uncoated cutting tool inserts and aims to enable more accurate calculation of the cutting force. Two common PVD coatings, AlTiN and TiAlCrN, were used. Firstly, a mathematical model of the cutting force based on the specific cutting force and cutting area is proposed. This mathematical model considers the cutting speed and coating correction factor as well as the real cutting edge geometry, i.e., it includes both the straight and rounded parts of the cutting edge. For this proposed model, material constants for C45 steel, which was machined with uncoated inserts, were obtained. Before determining an equation for a coating correction factor and implementing it into the model, experimental cutting force data for coated and uncoated inserts were compared using a paired t-test. The result was that the difference between them was statistically significant. Their percentage difference was found to be up to 4%. The correction factor equation that was obtained and implemented into the mathematical model was applied to compare the calculated and experimental data of the coated inserts, also using a paired t-test. The result was that the difference between them was insignificant. Moreover, their percentage difference was found to be up to 0.6%.


p38γ and p38δ: From Spectators to Key Physiological Players.
Although the physiological roles of p38γ and p38δ signalling pathways are largely unknown, new genetic and pharmacological tools are providing groundbreaking information on the function of these two stress-activated protein kinases. Recent studies show the importance of p38γ and p38δ in the regulation of processes as diverse as cytokine production, protein synthesis, exocytosis, cell migration, gene expression, and neuron activity, which have an acute impact on the development of pathologies related to inflammation, diabetes, neurodegeneration, and cancer. These recent breakthroughs are resolving some of the questions that have long been asked regarding the function of p38γ and p38δ in biology and pathology.


The Protective Effect of Polygonum cuspidatum (PCE) Aqueous Extract in a Dry Eye Model.
Dry eyes are caused by highly increased osmolarity of tear film, inflammation, and apoptosis of the ocular surface. In this study, we investigated the effect of Polygonum cuspidatum (PCE) aqueous extract in in vivo and in vitro dry eye models. Dry eye was induced by excision of the lacrimal gland and hyperosmotic media. In vivo, oral administration of PCE in exorbital lacrimal gland-excised rats recovered tear volume and Mucin4 (MUC4) expression by inhibiting corneal irregularity and expression of inflammatory cytokines. In vitro, hyperosmotic media induced human corneal epithelial cell (HCEC) cytotoxicity though increased inflammation, apoptosis, and oxidative stress. PCE treatment significantly inhibited expression of cyclooxygenase-2 and inflammatory cytokines (interleukin-6 and tumor necrosis factor-α), and activation of NF-κB p65 in hyperosmolar stress-induced HCECs. Hyperosmolarity-induced increase in Bcl-2-associated X protein (BAX) expression and activation of cleaved poly (ADP-ribose) polymerase and caspase 3 were attenuated in a concentration-dependent manner by PCE. PCE treatment restored anti-oxidative proteins such as heme oxygenase-1 (HO-1), superoxide dismutase-1 (SOD-1), and glutathione peroxidase (GPx) in hyperosmolar stress-induced HCECs. These data demonstrate that PCE prevents adverse changes in the ocular surface and tear fluid through inhibition of hyperosmolar stress-induced inflammation, apoptosis, and oxidation, suggesting that PCE may have the potential to preserve eye health.


Transformation of chlorpyrifos and chlorpyrifos-methyl in prairie pothole pore waters.
Non-point source pesticide pollution is a concern for wetlands in the prairie pothole region (PPR). Recent studies have demonstrated that reduced sulfur species (e.g., bisulfide and polysulfides) in PPR wetland pore waters directly undergo reactions with chloroacetanilide and dinitroaniline compounds. In this paper, the abiotic transformation of two organophosphate compounds, chlorpyrifos and chlorpyrifos-methyl, was studied in PPR wetland pore waters. Chlorpyrifos-methyl reacted significantly faster (up to 4 times) in pore water with reduced sulfur species relative to hydrolysis. No rate enhancement was observed in the transformation of chlorpyrifos in pore water with reduced sulfur species. The lack of reactivity was most likely caused by steric hindrance from the ethyl groups and partitioning to dissolved organic matter (DOM), thereby shielding chlorpyrifos from nucleophilic attack. Significant decreases in reaction rates were observed for chlorpyrifos in pore water with high concentrations of DOM. Rate enhancement due to other reactive species (e.g., organo-sulfur compounds) in pore water was minor for both compounds relative to the influence of bisulfide and DOM.


Reduced insulin-like growth factor I receptor and altered insulin receptor isoform mRNAs in normal mucosa predict colorectal adenoma risk.
Hyperinsulinemia resulting from obesity and insulin resistance is associated with increased risk of many cancers, but the biology underlying this risk is unclear. We hypothesized that increased mRNA levels of the insulin-like growth factor I receptor (IGFIR) versus the insulin receptor (IR) or elevated ratio of IR-A:IR-B isoforms in normal rectal mucosa would predict adenoma risk, particularly in individuals with high body mass index (BMI) or plasma insulin. Biopsies from normal rectal mucosa were obtained from consenting patients undergoing routine colonoscopy at University of North Carolina Hospitals (Chapel Hill, NC). Subjects with colorectal adenomas were classified as cases (n = 100) and were matched to adenoma-free controls (n = 98) based on age, sex, and BMI. IGFIR and IR mRNA levels were assessed by qRT-PCR, and IR-A:IR-B mRNA ratios by standard PCR. Plasma insulin and crypt apoptosis were measured by ELISA and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL), respectively. Logistic regression models examined relationships between receptor mRNAs, BMI, plasma insulin, and adenoma risk. Unexpectedly, cases were significantly more likely to have lower IGFIR mRNA levels than controls. No overall differences in total IR mRNA or IR-A:IR-B ratios were observed between cases and controls. Interestingly, in patients with high plasma insulin, increased IR-A:IR-B ratio was associated with increased likelihood of having adenomas. Our work shows novel findings that reduced IGFIR mRNA and, during high plasma insulin, increased IR-A:IR-B ratios in normal rectal mucosa are associated with colorectal adenoma risk. Our work provides evidence supporting a link between IGFIR and IR isoform expression levels and colorectal adenoma risk.


A mouse model for alpha-methylacyl-CoA racemase deficiency: adjustment of bile acid synthesis and intolerance to dietary methyl-branched lipids.
alpha-Methylacyl-CoA racemase (Amacr) deficiency in humans leads to sensory motor neuronal and liver abnormalities. The disorder is recessively inherited and caused by mutations in the AMACR gene, which encodes Amacr, an enzyme presumed to be essential for bile acid synthesis and to participate in the degradation of methyl-branched fatty acids. To generate a model to study the pathophysiology in Amacr deficiency we inactivated the mouse Amacr gene. As per human Amacr deficiency, the Amacr(-/-) mice showed accumulation (44-fold) of C27 bile acid precursors and decreased (over 50%) primary (C24) bile acids in bile, serum and liver, however the Amacr(-/-) mice were clinically symptomless. Real-time quantitative PCR analysis showed that, among other responses, the level of mRNA for peroxisomal multifunctional enzyme type 1 (pMFE-1) was increased 3-fold in Amacr(-/-) mice. This enzyme can be placed, together with CYP3A11 and CYP46A1, to make an Amacr-independent pathway for the generation of C24 bile acids. Exposure of Amacr(-/-) mice to a diet supplemented with phytol, a source for branched-chain fatty acids, triggered the development of a disease state with liver manifestations, redefining the physiological significance of Amacr. Amacr is indispensable for the detoxification of dietary methyl-branched lipids and, although it contributes normally to bile acid synthesis from cholesterol, the putative pMFE-1-mediated cholesterol degradation can provide for generation of bile acids, allowing survival without Amacr. Based upon our mouse model, we propose elimination of phytol from the diet of patients suffering from Amacr deficiency.


Factors That Influence the Success of Contact Lens Fitting in Presbyopes: A Multicentric Survey.
To evaluate the key factors behind successful and unsuccessful wear of contact lenses (CLs) for patients with presbyopia. A multicenter survey was conducted using a questionnaire, in eight CL centers, among two groups of presbyopes: successful wearers (SWs), who were presbyopic and wearing CLs successfully, and unsuccessful wearers (UWs), who tried unsuccessfully to wear CLs to manage their presbyopia and had stopped wearing CLs. A total of 237 completed questionnaires were returned; 178 from SWs and 59 from UWs. Successful wearers used CLs 5.8±1.5 days a week and additional reading spectacles were never used by half of the SWs. Among SWs, the most important reason to continue wearing CLs was convenience (61%), whereas among UWs, the most important reason to discontinue wearing CLs was poor vision (80%). A logistic regression analysis showed that the absence of astigmatism (P<0.05), a higher subjective satisfaction (P<0.05), a better subjective perceived vision at distance (P<0.01), and a lower subjective loss of visual contrast (P<0.05) were able to predict the success of CL use for presbyopia. This study explores the predictors for determining successful or unsuccessful CL wear in presbyopes. Among ophthalmic, demographic, lifestyle, and subjective variables, the latter seems to be of greater importance in determining the success of CL wear. Therefore, it is necessary that subjective variables are taken into account by the CL practitioner when approaching presbyopic CL fitting in practice.


Controlled intracellular generation of reactive oxygen species in human mesenchymal stem cells using porphyrin conjugated nanoparticles.
Nanoparticles capable of generating controlled amounts of intracellular reactive oxygen species (ROS), that advance the study of oxidative stress and cellular communication, were synthesized by functionalizing polyacrylamide nanoparticles with zinc(II) porphyrin photosensitisers. Controlled ROS production was demonstrated in human mesenchymal stem cells (hMSCs) through (1) production of nanoparticles functionalized with varying percentages of Zn(II) porphyrin and (2) modulating the number of doses of excitation light to internalized nanoparticles. hMSCs challenged with nanoparticles functionalized with increasing percentages of Zn(II) porphyrin and high numbers of irradiations of excitation light were found to generate greater amounts of ROS. A novel dye, which is transformed into fluorescent 7-hydroxy-4-trifluoromethyl-coumarin in the presence of hydrogen peroxide, provided an indirect indicator for cumulative ROS production. The mitochondrial membrane potential was monitored to investigate the destructive effect of increased intracellular ROS production. Flow cytometric analysis of nanoparticle treated hMSCs suggested irradiation with excitation light signalled controlled apoptotic cell death, rather than uncontrolled necrotic cell death. Increased intracellular ROS production did not induce phenotypic changes in hMSC subcultures.


Alterations of sorbin and SH3 domain containing 3 (SORBS3) in human skeletal muscle following Roux-en-Y gastric bypass surgery.
Obesity is a disease that is caused by genetic and environmental factors. However, epigenetic mechanisms of obesity are less well known. DNA methylation provides a mechanism whereby environmental factors can influence gene transcription. The aim of our study was to investigate skeletal muscle DNA methylation of sorbin and SH3 domain containing 3 (SORBS3) with weight loss induced by Roux-en-Y gastric bypass (RYGB). Previously, we had shown increased methylation (5.0 to 24.4%) and decreased gene expression (fold change - 1.9) of SORBS3 with obesity (BMI > 30 kg/m2) compared to lean controls. In the present study, basal muscle biopsies were obtained from seven morbidly obese (BMI > 40 kg/m2) female subjects pre- and 3 months post-RYGB surgery, in combination with euglycemic-hyperinsulinemic clamps to assess insulin sensitivity. We identified 30 significantly altered promoter and untranslated region methylation sites in SORBS3 using reduced representation bisulfite sequencing (RRBS). Twenty-nine of these sites were decreased (- 5.6 to - 24.2%) post-RYGB compared to pre-RYGB. We confirmed the methylation in 2 (Chr.8:22,423,690 and Chr.8:22,423,702) of the 29 decreased SORBS3 sites using pyrosequencing. This decreased methylation was associated with an increase in SORBS3 gene expression (fold change + 1.7) post-surgery. In addition, we demonstrated that SORBS3 promoter methylation in vitro significantly alters reporter gene expression (P < 0.0001). Two of the SORBS3 methylation sites (Chr.8:22,423,111 and Chr.8:22,423,205) were strongly correlated with fasting plasma glucose levels (r = 0.9, P = 0.00009 and r = 0.8, P = 0.0010). Changes in SORBS3 gene expression post-surgery were correlated with obesity measures and fasting insulin levels (r = 0.5 to 0.8; P < 0.05). These results demonstrate that SORBS3 methylation and gene expression are altered in obesity and restored to normal levels through weight loss induced by RYGB surgery.


Characteristics and long-term outcome of echocardiographic super-responders to cardiac resynchronisation therapy: 'real world' experience from a single tertiary care centre.
The individual benefit from cardiac resynchronisation therapy (CRT) varies largely among patients. To compare different definitions of echocardiographic super-response to CRT regarding their ability to predict the incidence of adverse events. Three definitions of super-response to CRT were evaluated in 110 consecutive patients with CRT implantation: (1) an absolute increase in ejection fraction of ≥ 10%; (2) a decrease in left ventricular end-systolic volume of ≥ 30%; and (3) a decrease in left ventricular end-diastolic volume of ≥ 20%. The primary endpoint was a combination of time to death, heart transplantation, ventricular assist device implantation and hospitalisation for heart failure. Secondary endpoints included time to first appropriate implantable cardioverter defibrillator (ICD) discharge during follow-up. All three definitions of super-response were highly predictive of a reduced risk for reaching the primary combined endpoint (3-year estimators: 64% ± 7% vs 82% ± 7% for ejection fraction ≥ 10%; 63% ± 8% vs 92% ± 5% for end-systolic volume ≥ 30%; and 62% ± 8% vs 94% ± 4% for end-diastolic volume ≥ 20%; all p<0.001). In all three analyses, super-responders had a significantly shorter time from diagnosis of heart failure until the time point of CRT implantation. However, even super-responders, independent of the definition, did experience appropriate ICD discharges during follow-up. All three definitions of super-response are highly predictive for a favourable outcome after CRT. However, even patients with pronounced reverse left ventricular remodelling experience appropriate ICD discharges during follow-up.


Action of vasopressin on the permeability of mesentery.
Contemporaneous movements of Rb86 cations and of P(32) orthophosphate across isolated rabbit mesentery display kinetic patterns that are generally associated with passive diffusion. Vasopressin at a concentration of 100 milliunit/ml produced a significant increase in the permeability constant for P(32) and at the same time a significant decrease in the permeability to Rb(86). At lower hormone concentrations (0.1 milliunit/ml) the P32 response was less marked but still significant (P < .01), while the Rb(86) effect was not (P > .05). Hyaluronidase did not mimic these actions of vasopressin.


A systematic review of intravaginal testosterone for the treatment of vulvovaginal atrophy.
Intravaginal testosterone has emerged as a potential treatment for vulvovaginal atrophy (VVA) in women, in general, and women taking an aromatase inhibitor (AI). A systematic review of the literature was undertaken to determine whether available clinical trial data support efficacy and safety of intravaginal testosterone for the treatment of VVA. Scopus, MEDLINE, EMBASE, and the Cochrane Library databases were systematically searched on July 26, 2017, for human studies published in English of clinical trials of intravaginal testosterone. Six separate clinical trials were identified that ranged in size from 10 to 80 participants, with either single dose, or durations of 4 to 12 weeks. Only one study incorporated a double-blind design. Three studies were of women taking an AI.Taken together, the studies suggest that intravaginal testosterone may lower vaginal pH, increase the proportion of vaginal lactobacilli, and possibly improve the vaginal maturation index. The lack of a placebo treatment in four studies, and failure to adjust for baseline differences, resulted in uncertainty of the effect on sexual function. Safety remains uncertain because of the small number of women exposed, short study durations, and inconsistent and incomplete outcome reporting for sex steroid levels. Adequately powered double-blind, placebo-controlled clinical trials of intravaginal testosterone therapy are needed to establish both efficacy and safety.


TGF beta1 and related-Smads contribute to pulmonary metastasis of hepatocellular carcinoma in mice model.
Recent studies indicate that Transforming Growth Factor beta (TGF β) correlated with pulmonary metastasis of cancers. However, the correlation between TGF β and pulmonary metastasis of hepatocellular carcinoma (HCC) is till unknown. We detected the in vitro and in vivo expression levels of TGF β1/Smads by Real-time PCR and Western blot in MHCC97-H and MHCC97-L cell lines, which are HCC cell lines and have higher and lower pulmonary metastatic potential respectively. TGF β1 mRNA level in MHCC97-L tumors were higher than that in MHCC97-H tumors, (2.81±1.61 vs. 1.24±0.96, P=0.002), TGF β1 protein level in MHCC97-L tumors were also higher than that in MHCC97-H tumors (1.37±0.95 vs. 0.32±0.22, P<0.001). In addition, the TGF β1 mRNA level positively correlated with pulmonary metastasis, and the relations between TGF β1 and Smads were also found (R2=0.12 and 0.40, respectively). Our results suggest that TGF β/ Smads promote pulmonary metastasis of HCC.


High HIV risk-taking among young gay men.
Previous research has shown younger age to be correlated with greater HIV sexual risk-taking among gay men. The purpose of this study was to identify variables associated with HIV risk-taking among younger gay men. Ninety-nine gay men aged 18-25 in three medium-sized West Coast communities completed self-report questionnaires regarding HIV-related behaviors and attitudes. Of the respondents, 43% reported having engaged in unprotected anal intercourse during the previous 6 months. Men who engaged in unprotected anal intercourse reported greater enjoyment of unprotected anal intercourse, perceived less risk of unprotected anal intercourse, labeled themselves as more at risk for AIDS, reported poorer communication skills with sexual partners, and were more likely to have a boyfriend/lover than men who had not engaged in high-risk sex. In addition, respondents perceived the likelihood of acquiring HIV from unprotected anal intercourse with young gay men to be significantly lower than with older gay men. These findings highlight the need for HIV risk-reduction interventions designed specifically for young gay men and identify critical areas to be targeted in such interventions.


Electro-optic lightning detector.
The design, alignment, calibration, and field deployment of a solid-state lightning detector is described. The primary sensing component of the detector is a potassium dihydrogen phosphate electro-optic crystal that is attached in series to a flat-plate aluminum antenna; the antenna is exposed to the ambient thundercloud electric field. A semiconductor laser diode (lambda = 685 nm), polarizing optics, and the crystal are arranged in a Pockels cell configuration. Lightning-caused electric field changes are related to small changes in the transmission of laser light through the optical cell. Several hundred lightning electric field change excursions were recorded during five thunderstorms that occurred in the summer of 1998 at the NASA Marshall Space Flight Center in northern Alabama.


Multivalency effects on Pseudomonas aeruginosa biofilm inhibition and dispersal by glycopeptide dendrimers targeting lectin LecA.
The galactose specific lectin LecA partly mediates the formation of antibiotic resistant biofilms by Pseudomonas aeruginosa, an opportunistic pathogen causing lethal airways infections in immunocompromised and cystic fibrosis patients, suggesting that preventing LecA binding to natural saccharides might provide new opportunities for treatment. Here 8-fold (G3) and 16-fold (G4) galactosylated analogs of GalAG2, a tetravalent G2 glycopeptide dendrimer LecA ligand and P. aeruginosa biofilm inhibitor, were obtained by convergent chloroacetyl thioether (ClAc) ligation between 4-fold or 8-fold chloroacetylated dendrimer cores and digalactosylated dendritic arms. Hemagglutination inhibition, isothermal titration calorimetry and biofilm inhibition assays showed that G3 dendrimers bind LecA slightly better than their parent G2 dendrimers and induce complete biofilm inhibition and dispersal of P. aeruginosa biofilms, while G4 dendrimers show reduced binding and no biofilm inhibition. A binding model accounting for the observed saturation of glycopeptide dendrimer galactosyl groups and LecA binding sites is proposed based on the crystal structure of a G3 dendrimer LecA complex.


Orthopaedic Surgery Pediatric Sports Medicine: Characterizing Practice Patterns and Subspecialization.
Pediatric sports medicine is a new and rapidly growing subspecialty within orthopaedic surgery. However, there is very limited literature on the practice of pediatric sports medicine in North America. Therefore, the purpose of this study was to evaluate and describe the current practice patterns of orthopaedic surgeons specializing in pediatric sports medicine. An online survey was distributed to orthopaedic surgeons specializing in pediatric sports medicine through the Pediatric Research in Sports Medicine Society. The purpose of the survey was to characterize (1) surgeon demographics, (2) the breakdown of different joint specialization, and (3) the specific procedures for joints that the surgeons specialize in. Responses from 55 orthopaedic surgeons were collected and analyzed. Most respondents considered pediatric sports medicine as the primary focus of their practice (89.1%, n=49/55). The number of fellowships completed was almost evenly split between either a single fellowship (52.7%, n=29/55) or 2 or more (47.3%, n=26/55). The most common combination of fellowships was pediatric orthopaedics and adult sports medicine (32.7%, n=18/55). Most survey respondents had been in practice for <10 years (69.0%, n=38/55) and were affiliated with an academic center (61.8%, n=34/55). On average, 77.5% of the patients treated were <18 years old. The knee joint was the most specialized joint, with 98.2% (n=54/55) respondents reporting that the knee joint constituted ≥25% of their practice. The knee joint constituted a mean of 52.1% of the respondents' overall practice, followed by the shoulder (15.2%), hip (13.9%), ankle (7.5%), elbow (7.1%), and wrist (4.2%). Pediatric sports medicine practices are variable and have distinct practice patterns in pediatric, orthopaedic, and adult sports practices. In the current study, most surgeons are less than 10 years into practice, affiliated with academic centers, and have typically completed either 1 or 2 fellowships after residency. Surgeons were most commonly specialized in the knee joint and cared for patients <18 years old. Level of evidence IV.


Comparison of the surgical outcomes in patients with synchronous versus metachronous multiple hepatocellular carcinoma.
Multiplicity is one of the characteristics of hepatocellular carcinoma (HCC), and patients with multiple HCC (≤ 3 nodules) are recommended as candidates for liver resection. To confirm the validity of resecting multiple HCC, we compared the surgical outcomes in patients with synchronous and metachronous multiple HCC. Patients who underwent resection for multiple HCC (2 or 3 nodules) were classified into the "synchronous multiple HCC" group, while those undergoing resection for solitary HCC and repeated resection for 1 or 2 recurrent nodules within 2 years after initial operation were classified into the "metachronous multiple HCC" group. After one-to-one matching, longer operation time and more bleeding were seen in the synchronous multiple HCC group (n = 98) than those in the metachronous multiple HCC group (n = 98); however, the complication rates were not different between the two groups. The median overall survival times were 4.0 years (95% CI, 3.0-5.9) and 5.9 years (4.0-NA) for the synchronous and metachronous multiple HCC (after second operation) groups, respectively (P = 0.041). The recurrence-free survival times were shorter in the synchronous multiple HCC group than in the metachronous multiple HCC group (median, 1.5 years [95% CI, 0.9-1.8] versus 1.8 years, [1.3-2.2]) (P = 0.039). On multivariate analysis, independent factors for overall survivals in the synchronous multiple HCC group were older age, cirrhosis, larger tumor, and tumor thrombus. Taken together, resection of metachronous multiple HCC still has good therapeutic effect, even better than synchronous multiple HCC, so resection is suggested for metachronous multiple HCC.


Insight into the relationship between the cell culture model, cell trafficking and siRNA silencing efficiency.
Despite research efforts, cell uptake processes determining siRNA silencing efficiency remain unclear. Here, we examine the relationship between in vitro cell culture models, cellular trafficking and siRNA silencing efficiency to provide a mechanistic insight on siRNA delivery system design. Model siRNA-polyplexes, based on chitosan as a 'classical' condensing agent, were applied to a panel of lung epithelial cell lines, H1299, A549 and Calu-3 and cell internalization levels, trafficking pathways and gene silencing assessed on exposure to pharmacological inhibitors. The data reveal striking differences in the internalization behaviour and gene silencing efficiency in the tested cell lines, despite their common lung epithelial origins. The model system's silencing was lower where clathrin internalization pathway predominated in Calu-3, relative to silencing in H1299 cells where a non-clathrin internalization appears dominant. Increased silencing on endosomal disruption was apparent in Calu-3 cells, but absent when cellular internalization was not predominantly clathrin-mediated in A549 cells. This highlights that identifying cell trafficking pathways before incorporation of functional components to siRNA delivery systems (e.g. endosomolytic compounds) is crucial. The study hence stresses the importance of selection of appropriate cell culture model, relevant to in vivo target, to assess the gene silencing efficiency and decide which functionalities the 'stratified siRNA silencing vector' requires.


AAA+ chaperones and acyldepsipeptides activate the ClpP protease via conformational control.
The Clp protease complex degrades a multitude of substrates, which are engaged by a AAA+ chaperone such as ClpX and subsequently digested by the dynamic, barrel-shaped ClpP protease. Acyldepsipeptides (ADEPs) are natural product-derived antibiotics that activate ClpP for chaperone-independent protein digestion. Here we show that both protein and small-molecule activators of ClpP allosterically control the ClpP barrel conformation. We dissect the catalytic mechanism with chemical probes and show that ADEP in addition to opening the axial pore directly stimulates ClpP activity through cooperative binding. ClpP activation thus reaches beyond active site accessibility and also involves conformational control of the catalytic residues. Moreover, we demonstrate that substoichiometric amounts of ADEP potently prevent binding of ClpX to ClpP and, at the same time, partially inhibit ClpP through conformational perturbance. Collectively, our results establish the hydrophobic binding pocket as a major conformational regulatory site with implications for both ClpXP proteolysis and ADEP-based anti-bacterial activity.


Effect of Cephalexin Plus Trimethoprim-Sulfamethoxazole vs Cephalexin Alone on Clinical Cure of Uncomplicated Cellulitis: A Randomized Clinical Trial.
Emergency department visits for skin infections in the United States have increased with the emergence of methicillin-resistant Staphylococcus aureus (MRSA). For cellulitis without purulent drainage, β-hemolytic streptococci are presumed to be the predominant pathogens. It is unknown if antimicrobial regimens possessing in vitro MRSA activity provide improved outcomes compared with treatments lacking MRSA activity. To determine whether cephalexin plus trimethoprim-sulfamethoxazole yields a higher clinical cure rate of uncomplicated cellulitis than cephalexin alone. Multicenter, double-blind, randomized superiority trial in 5 US emergency departments among outpatients older than 12 years with cellulitis and no wound, purulent drainage, or abscess enrolled from April 2009 through June 2012. All participants had soft tissue ultrasound performed at the time of enrollment to exclude abscess. Final follow-up was August 2012. Cephalexin, 500 mg 4 times daily, plus trimethoprim-sulfamethoxazole, 320 mg/1600 mg twice daily, for 7 days (n = 248 participants) or cephalexin plus placebo for 7 days (n = 248 participants). The primary outcome determined a priori in the per-protocol group was clinical cure, defined as absence of these clinical failure criteria at follow-up visits: fever; increase in erythema (>25%), swelling, or tenderness (days 3-4); no decrease in erythema, swelling, or tenderness (days 8-10); and more than minimal erythema, swelling, or tenderness (days 14-21). A clinically significant difference was defined as greater than 10%. Among 500 randomized participants, 496 (99%) were included in the modified intention-to-treat analysis and 411 (82.2%) in the per-protocol analysis (median age, 40 years [range, 15-78 years]; 58.4% male; 10.9% had diabetes). Median length and width of erythema were 13.0 cm and 10.0 cm. In the per-protocol population, clinical cure occurred in 182 (83.5%) of 218 participants in the cephalexin plus trimethoprim-sulfamethoxazole group vs 165 (85.5%) of 193 in the cephalexin group (difference, -2.0%; 95% CI, -9.7% to 5.7%; P = .50). In the modified intention-to-treat population, clinical cure occurred in 189 (76.2%) of 248 participants in the cephalexin plus trimethoprim-sulfamethoxazole group vs 171 (69.0%) of 248 in the cephalexin group (difference, 7.3%; 95% CI, -1.0% to 15.5%; P = .07). Between-group adverse event rates and secondary outcomes through 7 to 9 weeks, including overnight hospitalization, recurrent skin infections, and similar infection in household contacts, did not differ significantly. Among patients with uncomplicated cellulitis, the use of cephalexin plus trimethoprim-sulfamethoxazole compared to cephalexin alone did not result in higher rates of clinical resolution of cellulitis in the per-protocol analysis. However, because imprecision around the findings in the modified intention-to-treat analysis included a clinically important difference favoring cephalexin plus trimethoprim-sulfamethoxazole, further research may be needed. clinicaltrials.gov Identifier: NCT00729937.


Childhood Tuberculosis in a Sub-Saharan Tertiary Facility: Epidemiology and Factors Associated with Treatment Outcome.
Childhood tuberculosis (TB) is a diagnostic challenge in developing countries, and patient outcome can be influenced by certain factors. We report the disease course, clinical profile and factors associated with treatment outcome in a tertiary facility of Kinshasa. Documentary and analytical studies were conducted using clinical and exploratory data for children aged up to 15 years who were admitted to the University Clinics of Kinshasa for TB. Data are presented as frequencies and averages, and binary and logistic regression analyses were performed. Of 283 children with TB, 82 (29.0%) had smear-negative TB, 40 (14.1%) had smear-positive TB, 159 (56.1%) had extra-pulmonary TB (EPTB), 2 (0.7%) had multidrug-resistant TB (MDR-TB), 167 (59.0%) completed treatment, 30 (10.6%) were cured, 7 (2.5%) failed treatment, 4 (1.4%) died, 55 (19.4%) were transferred to health centers nearest their home, and 20 (7.0%) were defaulters. In the binary analysis, reported TB contacts (p = 0.048), type of TB (p = 0.000), HIV status (p = 0.050), Ziehl-Nielsen test result (p = 0.000), Lowenstein culture (p = 0.004) and chest X-ray (p = 0.057) were associated with outcome. In the logistic regression, none of these factors was a significant predictor of outcome. Tertiary level care facilities must improve the diagnosis and care of patients with childhood TB, which justifies the development of alternative diagnostic techniques and the assessment of other factors that potentially affect outcome.


HELP LDL apheresis reduces plasma pentraxin 3 in familial hypercholesterolemia.
Pentraxin 3 (PTX3), a key component of the humoral arm of innate immunity, is secreted by vascular cells in response to injury, possibly aiming at tuning arterial activation associated with vascular damage. Severe hypercholesterolemia as in familial hypercholesterolemia (FH) promotes vascular inflammation and atherosclerosis; low-density lipoprotein (LDL) apheresis is currently the treatment of choice to reduce plasma lipids in FH. HELP LDL apheresis affects pro- and antiinflammatory biomarkers, however its effects on PTX3 levels are unknown. We assessed the impact of FH and of LDL removal by HELP apheresis on PTX3. Plasma lipids, PTX3, and CRP were measured in 19 patients with FH undergoing chronic HELP LDL apheresis before and after treatment and in 20 control subjects. In the patients assessment of inflammation and oxidative stress markers included also plasma TNFα, fibrinogen and TBARS. At baseline, FH patients had higher (p = 0.0002) plasma PTX3 than matched control subjects. In FH PTX3 correlated positively (p≤0.05) with age, gender and CRP and negatively (p = 0.01) with HELP LDL apheresis vintage. The latter association was confirmed after correction for age, gender and CRP. HELP LDL apheresis acutely reduced (p≤0.04) plasma PTX3, CRP, fibrinogen, TBARS and lipids, but not TNFα. No association was observed between mean decrease in PTX3 and in LDL cholesterol. PTX3 paralleled lipids, oxidative stress and inflammation markers in time-course study. FH is associated with increased plasma PTX3, which is acutely reduced by HELP LDL apheresis independently of LDL cholesterol, as reflected by the lack of association between change in PTX3 and in LDL levels. These results, together with the finding of a negative relationship between PTX3 and duration of treatment suggest that HELP LDL apheresis may influence both acutely and chronically cardiovascular outcomes in FH by modulating PTX3.


Stimulation of gross chromosomal rearrangements by the human CEB1 and CEB25 minisatellites in Saccharomyces cerevisiae depends on G-quadruplexes or Cdc13.
Genomes contain tandem repeats that are at risk of internal rearrangements and a threat to genome integrity. Here, we investigated the behavior of the human subtelomeric minisatellites HRAS1, CEB1, and CEB25 in Saccharomyces cerevisiae. In mitotically growing wild-type cells, these GC-rich tandem arrays stimulate the rate of gross chromosomal rearrangements (GCR) by 20, 1,620, and 276,000-fold, respectively. In the absence of the Pif1 helicase, known to inhibit GCR by telomere addition and to unwind G-quadruplexes, the GCR rate is further increased in the presence of CEB1, by 385-fold compared to the pif1Δ control strain. The behavior of CEB1 is strongly dependent on its capacity to form G-quadruplexes, since the treatment of WT cells with the Phen-DC(3) G-quadruplex ligand has a 52-fold stimulating effect while the mutation of the G-quadruplex-forming motif reduced the GCR rate 30-fold in WT and 100-fold in pif1Δ cells. The GCR events are telomere additions within CEB1. Differently, the extreme stimulation of CEB25 GCR depends on its affinity for Cdc13, which binds the TG-rich ssDNA telomere overhang. This property confers a biased orientation-dependent behavior to CEB25, while CEB1 and HRAS1 increase GCR similarly in either orientation. Furthermore, we analyzed the minisatellites' distribution in the human genome and discuss their potential role to trigger subtelomeric rearrangements.


Cost-effectiveness of Roux-en-Y gastric bypass in type 2 diabetes patients.
To assess the cost-effectiveness of Roux-en-Y gastric bypass for treating type 2 diabetes mellitus (T2DM) in the United States compared with standard medical management, using clinical data from a prospective observational study conducted at an academic medical center. Our study used a predictive health economic model (the CORE Diabetes Model) to project the long-term costs and clinical effectiveness of Roux-en-Y gastric bypass as a treatment for T2DM using the prospective observational study as the basis for our clinical effectiveness assumptions. The CORE Diabetes Model used Monte Carlo simulation with tracker variables to estimate the lifetime costs and clinical outcomes of Roux-en-Y gastric bypass compared with standard medical management of obese T2DM patients. Sensitivity analyses were performed on key clinical assumptions, discount rates, and shorter time horizons. The base-case scenario yielded an incremental cost-effectiveness ratio (ICER) of $21,973 per quality-adjusted life-year (QALY) gained. In sensitivity analyses, shortening the time horizon to 5 and 10 years and excluding the negative impact of increased body mass index on the patient's quality of life had the greatest adverse impact on the ICERs (ie, higher cost per QALY). Under base-case assumptions, Roux-en-Y gastric bypass is cost-effective in the treatment of T2DM in the United States with an ICER below $50,000 per QALY gained. Sensitivity analyses indicated that bariatric surgery is not cost-effective over shorter time horizons, or if the negative quality-of-life impact of increased body mass index is ignored.


Blood-brain barrier disruption and intra-arterial methotrexate-based therapy for newly diagnosed primary CNS lymphoma: a multi-institutional experience.
PURPOSE Primary CNS lymphoma (PCNSL) is confined to the CNS and/or the eyes at presentation and is usually initially treated with intravenous methotrexate-based chemotherapy and whole-brain radiotherapy (WBRT). However, the intact blood-brain barrier (BBB) can limit diffusion of methotrexate into brain and tumor. With BBB disruption (BBBD), enhanced drug delivery to the tumor can be achieved. PATIENTS AND METHODS This report summarizes the multi-institutional experience of 149 newly diagnosed (with no prior WBRT) patients with PCNSL treated with osmotic BBBD and intra-arterial (IA) methotrexate at four institutions from 1982 to 2005. In this series, 47.6% of patients were age > or = 60 years, and 42.3% had Karnofsky performance score (KPS) less than 70 at diagnosis. Results The overall response rate was 81.9% (57.8% complete; 24.2% partial). Median overall survival (OS) was 3.1 years (25% estimated survival at 8.5 years). Median progression-free survival (PFS) was 1.8 years, with 5-year PFS of 31% and 7-year PFS of 25%. In low-risk patients (age < 60 years and KPS > or = 70), median OS was approximately 14 years, with a plateau after approximately 8 years. Procedures were generally well tolerated; focal seizures (9.2%) were the most frequent side effect and lacked long-term sequelae. CONCLUSION This large series of patients treated over a 23-year period demonstrates that BBBD/IA methotrexate-based chemotherapy results in successful and durable tumor control and outcomes that are comparable or superior to other PCNSL treatment regimens.


Simultaneous silencing of isoamylases ISA1, ISA2 and ISA3 by multi-target RNAi in potato tubers leads to decreased starch content and an early sprouting phenotype.
Isoamylases hydrolyse (1-6)-alpha-D-glucosidic linkages in starch and are involved in both starch granule formation and starch degradation. In plants, three isoamylase isoforms with distinct functions in starch synthesis (ISA1 and ISA2) and degradation (ISA3) have been described. Here, we created transgenic potato plants with simultaneously decreased expression of all three isoamylases using a chimeric RNAi construct targeting all three isoforms. Constitutive expression of the hairpin RNA using the 35S CaMV promoter resulted in efficient silencing of all three isoforms in leaves, growing tubers, and sprouting tubers. Neither plant growth nor tuber yield was effected in isoamylase-deficient potato lines. Interestingly, starch metabolism was found to be impaired in a tissue-specific manner. While leaf starch content was unaffected, tuber starch was significantly reduced. The reduction in tuber starch content in the transgenic plants was accompanied by a decrease in starch granules size, an increased sucrose content and decreased hexose levels. Despite the effects on granule size, only little changes in chain length composition of soluble and insoluble glucose polymers were detected. The transgenic tubers displayed an early sprouting phenotype that was accompanied by an increased level of sucrose in parenchyma cells below the outgrowing bud. Since high sucrose levels promote sprouting, we propose that the increased number of small starch granules may cause an accelerated turnover of glucan chains and hence a more rapid synthesis of sucrose. This observation links alterations in starch structure/degradation with developmental processes like meristem activation and sprout outgrowth in potato tubers.


Holding thermal receipt paper and eating food after using hand sanitizer results in high serum bioactive and urine total levels of bisphenol A (BPA).
Bisphenol A (BPA) is an endocrine disrupting environmental contaminant used in a wide variety of products, and BPA metabolites are found in almost everyone's urine, suggesting widespread exposure from multiple sources. Regulatory agencies estimate that virtually all BPA exposure is from food and beverage packaging. However, free BPA is applied to the outer layer of thermal receipt paper present in very high (∼20 mg BPA/g paper) quantities as a print developer. Not taken into account when considering thermal paper as a source of BPA exposure is that some commonly used hand sanitizers, as well as other skin care products, contain mixtures of dermal penetration enhancing chemicals that can increase by up to 100 fold the dermal absorption of lipophilic compounds such as BPA. We found that when men and women held thermal receipt paper immediately after using a hand sanitizer with penetration enhancing chemicals, significant free BPA was transferred to their hands and then to French fries that were eaten, and the combination of dermal and oral BPA absorption led to a rapid and dramatic average maximum increase (Cmax) in unconjugated (bioactive) BPA of ∼7 ng/mL in serum and ∼20 µg total BPA/g creatinine in urine within 90 min. The default method used by regulatory agencies to test for hazards posed by chemicals is intra-gastric gavage. For BPA this approach results in less than 1% of the administered dose being bioavailable in blood. It also ignores dermal absorption as well as sublingual absorption in the mouth that both bypass first-pass liver metabolism. The elevated levels of BPA that we observed due to holding thermal paper after using a product containing dermal penetration enhancing chemicals have been related to an increased risk for a wide range of developmental abnormalities as well as diseases in adults.


Texas needs physicians trained in public health: a new 4-year integrated MD/MPH degree program.
Traditionally, medicine and public health have not worked as synergistic disciplines because they are based on fundamentally different models. However, a number of very recent imperatives emphasize the need for dual training in these fields to address major public health problems facing society as well as the documented and forecasted workforce shortages. In response to this need, two University of Texas institutions based in San Antonio, Texas, partnered in 2007 to offer a dual 4-year Doctor of Medicine/Master of Public health (MD/MPH) degree program, one of a handful in the nation. Approximately 65 students (or 10% of three consecutive medical school classes) are currently enrolled. The dual-degree program meets the requirements of both degree programs while giving shared MPH credit for relevant courses taken in the medical curriculum and medical school credit for some courses in the public health curriculum. However, 75% of the MPH coursework originates at the School of Public Health. Initial results from focus groups conducted after the first year showed a high degree of student satisfaction, with frequent comments that the program was broadening their perspective on medicine and influencing their career and life goals. A dual MD/MPH degree is an important option for all medical students as a means of addressing pressing health issues in our society through combined training in medicine and the broader areas of prevention and population health. The four-year MD/MPH program, while posing challenges for faculty and students, attracts community- and prevention-minded medical students, reduces training costs (housing/living costs and lost time and wages before entering residency), and allows students to progress with the rest of their class.