Viral phylogeny in court: the unusual case of the Valencian anesthetist.
A large and complex outbreak of hepatitis C virus in Valencia, Spain that began 25 years ago led to the prosecution and conviction of an anesthetist who was accused of infecting hundreds of his patients. Evolutionary analyses of viral gene sequences were presented as evidence in the trial, and these are now described in detail by González-Candelas and colleagues in a paper published in BMC Biology. Their study illustrates the challenges and opportunities that arise from the use of phylogenetic inference in criminal trials concerning virus transmission.


HSP90 inhibitors disrupt a transient HSP90-HSF1 interaction and identify a noncanonical model of HSP90-mediated HSF1 regulation.
Heat shock factor 1 (HSF1) initiates a broad transcriptional response to proteotoxic stress while also mediating a cancer-specific transcriptional program. HSF1 is thought to be regulated by molecular chaperones, including Heat Shock Protein 90 (HSP90). HSP90 is proposed to sequester HSF1 in unstressed cells, but visualization of this interaction in vivo requires protein crosslinking. In this report, we show that HSP90 binding to HSF1 depends on HSP90 conformation and is only readily visualized for the ATP-dependent, N-domain dimerized chaperone, a conformation only rarely sampled by mammalian HSP90. We have used this mutationally fixed conformation to map HSP90 binding sites on HSF1. Further, we show that ATP-competitive, N-domain targeted HSP90 inhibitors disrupt this interaction, resulting in the increased duration of HSF1 occupancy of the hsp70 promoter and significant prolongation of both the constitutive and heat-induced HSF1 transcriptional activity. While our data do not support a role for HSP90 in sequestering HSF1 monomers to suppress HSF1 transcriptional activity, our findings do identify a noncanonical role for HSP90 in providing dynamic modulation of HSF1 activity by participating in removal of HSF1 trimers from heat shock elements in DNA, thus terminating the heat shock response.


Overexpression of hepatocyte growth factor receptor in scleroderma dermal fibroblasts is caused by autocrine transforming growth factor β signaling.
Cutaneous fibrosis seen in systemic sclerosis (SSc) is caused by fibroblast activation and abnormal collagen accumulation due to 'autocrine transforming growth factor (TGF)-β/Smad signaling'. Hepatocyte growth factor (HGF) may have therapeutic value against SSc, because of its inducible effect on the expression of matrix metalloproteinase (MMP)-1. Previous studies indicated SSc dermal fibroblasts overexpress HGF receptor c-met, which suggest specific and effective induction of MMP-1 in SSc fibroblasts caused by HGF treatment. However, the exact mechanism of c-met overexpression in SSc cells was hardly investigated. We hypothesized that such c-met overexpression is also caused by autocrine TGF-β/Smad signaling. Expression of c-met protein in cultured SSc dermal fibroblasts was significantly up-regulated compared with that in normal fibroblasts. Ectopic TGF-β stimulation induced c-met synthesis in normal fibroblasts, while a TGF-β knockdown normalized the up-regulated c-met levels in SSc fibroblasts. Furthermore, we found the c-met promoter contains a putative binding site for Smads, and the binding activity of Smad2/3 to the c-met promoter was constitutively up-regulated in SSc fibroblasts as well as in normal fibroblasts treated with exogenous TGF-β1. Taken together, c-met may be overexpressed due to autocrine TGF-β/Smad signaling in SSc. Considering that HGF has an antifibrotic effect, such c-met overexpression in SSc fibroblasts may be a negative feedback against cutaneous fibrosis. Clarifying the mechanisms of c-met overexpression and controlling the HGF/c-met pathway may lead to a new therapeutic approach for this disease.


Programme factors that influence completion of residential treatment.
The aim of this study was to predict retention in residential rehabilitation (RR) services for drug users, focusing on service provider factors. A national postal survey of RR services in England and Wales was carried out and information was obtained from 57 of 87 services identified (65.5%). Service managers were asked to complete a questionnaire asking about treatment philosophy, treatments provided, staff characteristics and staffing levels, as well as overall service size and funding. Services also provided information on the number of clients admitted and the number who had completed, dropped out and been asked to leave in the past year. Completion rates varied widely, from 3% to 92%, with an average of 48%. Higher completion rates were associated with lower counsellor caseloads, fewer beds, single rooms, shorter scheduled treatment durations, higher fees per client and provision of what could be termed a balanced treatment programme containing adequate amounts of individual counselling and programme-free time, and with only moderate demands for domestic duties. Programmes with more drug than alcohol users had lower completion rates, but the proportion of dual diagnosis or criminal justice referred clients did not appear to affect retention. Completion rates varied as a function of a number of service factors that are amenable to manipulation. To retain clients successfully, programmes should not be too large and should have adequate levels of therapeutic staff, a well-developed treatment schedule which is not too demanding for the client in terms of duties or overall time spent in structured activities, and which incorporates sufficient levels of individual counselling. [Meier PS, Best D. Programme factors that influence completion of residential treatment. Drug Alcohol Rev2006;25:349 - 355].


Effects of a light touch on fixed or mobile supports on gait parameters in visually restricted young adults.
We investigated the effects of lightly touching fixed and mobile supports on gait parameters and center of mass oscillations in visually restricted young adults. Fourteen healthy male and female adults of mean=23.6 years (SD=1.6 years). Twelve walking conditions were completed on the GAITRite measurement system, resulting from crossing 3 conditions of visual restriction (no restriction, partial restriction, blindfolded) and 4 conditions of haptic supplementation (no supplementation, with a cane used as "light touch", with a cane sliding on the ground, while touching a soft elastic handrail). Gait speed, stance time, step length and the basis of support were measured. Accelerations of center of mass were also recorded through an accelerometer and the displacements of center of mass were analyzed. Results showed that visual restriction decreased in gait speed, reduced step length, while it increased the base of support and the amplitude of CoM displacements. In the full restriction condition, haptic supplementation provided by the use of the classic cane improved normalized stance time (%). In addition, in the no vision condition, both the classic cane and the soft handrail increased step length and reduced medio-lateral oscillations of the CoM. These results suggest that in visually restricted healthy adults, lightly touching a fixed (soft handrail) or a mobile (classic cane) support contributes to adaptation of gait parameters and postural control during locomotion.


Patient and Provider Reported Reasons for Lost to Follow Up in MDRTB Treatment: A Qualitative Study from a Drug Resistant TB Centre in India.
Multidrug-resistant Tuberculosis (MDR TB) is emerging public health concern globally. Lost to follow-up (LTFU) is one of the key challenge in MDRTB treatment. In 2013, 18% of MDR TB patients were reported LTFU in India. A qualitative study was conducted to obtain better understanding of both patient and provider related factors for LTFU among MDR TB treatment. Qualitative semi-structured personal interviews were conducted with 20 MDRTB patients reported as LTFU and 10 treatment providers in seven districts linked to Nagpur Drug resistant TB Centre (DRTBC) during August 2012-February 2013. Interviews were transcribed and inductive content analysis was performed to derive emergent themes. We found multiple factors influencing MDR TB treatment adherence. Barriers to treatment adherence included drug side effects, a perceived lack of provider support, patient financial constraints, conflicts with the timing of treatment services, alcoholism and social stigma. Patient adherence to treatment is multi-factorial and involves individual patient factors, provider factors, and community factors. Addressing issue of LTFU during MDRTB treatment requires enhanced efforts towards resolving medical problems like adverse drug effects, developing short duration treatment regimens, reducing pill burden, motivational counselling, flexible timings for DOT services, social, family support for patients & improving awareness about disease.


Aptasensors for rapid detection of Escherichia coli O157:H7 and Salmonella typhimurium.
Herein we reported the development of aptamer-based biosensors (aptasensors) based on label-free aptamers and gold nanoparticles (AuNPs) for detection of Escherichia coli (E. coli) O157:H7 and Salmonella typhimurium. Target bacteria binding aptamers are adsorbed on the surface of unmodified AuNPs to capture target bacteria, and the detection was accomplished by target bacteria-induced aggregation of the aptasensor which is associated as red-to-purple color change upon high-salt conditions. By employing anti-E. coli O157:H7 aptamer and anti-S. typhimurium aptamer, we developed a convenient and rapid approach that could selectively detect bacteria without specialized instrumentation and pretreatment steps such as cell lysis. The aptasensor could detect as low as 105colony-forming units (CFU)/ml target bacteria within 20 min or less and its specificity was 100%. This novel method has a great potential application in rapid detection of bacteria in the near future.


Economic implications of type 2 diabetes management.
Diabetes mellitus (DM) is a complex and progressive disease associated with significant morbidity and mortality. Both the prevalence and economic burden associated with DM have been steadily increasing and are projected to continue to increase. A series of cost reports published by the American Diabetes Association (ADA) between 1997 and 2012 have suggested that direct medical costs are the primary cost driver in DM. Direct costs have increased over 5-fold during this time frame, from $44 billion to $176 billion annually, likely due to both the increased prevalence of DM and increased costs of providing DM-related care. Trends in anti-diabetic medication usage as reported by Centers for Disease Control data from 1997 to 2011 have found that the proportion of diagnosed patients treated for DM has remained fairly stable. However, there has been an increase in the use of oral agents alone, a decrease in patients on insulin only, and a relatively small increase in the percentage of patients using both insulin and oral agents, suggesting stagnation regarding the uptake of insulin use. In parallel, the ADA cost and utilization data suggest that use of medications to treat DM-related comorbidities and complications has risen, which contributes to the risk of medication errors, avoidable adverse drug events, and noncompliance. These data highlight a few of the challenges involved in managing a population of patients with DM. A comprehensive assessment of unmet needs for the population with DM is essential for effective population management. An approach that strives to identify and reduce barriers to patients receiving optimal care and being active participants in their DM management will facilitate systemwide efforts to improve DM outcomes.


Physician Attitudes and Perceptions of Complementary and Alternative Medicine (CAM): A Multicentre Italian Study.
Purpose: Complementary and Alternative Medicine (CAM) interventions are widely used by patients with chronic disorders, including cancer, and may interact with cancer treatment. Physicians are often unaware of this, probably due to poor patient-physician communication on CAM. The purpose of this study was to evaluate physicians' knowledge, attitudes and practice patterns regarding CAM in a survey conducted in Italy. Methods: A questionnaire was administered to 438 physicians (11 Italian hospitals) who predominantly treat patients with chronic disease, to collect personal and professional data and information on attitudes toward CAM and its possible role in Conventional Medicine (CM). Results: Of the 438 participants, most were specialists in oncology (18%), internal medicine (17%), surgery (15%), and radiotherapy (11%). Most worked at university (44%) or research hospitals (31%). Forty-two percent of participants believed that CAM could have an integrative role within CM. Oncologists were the physicians who were best informed on CAM (58%). Physicians working at research institutes or university hospitals had a greater knowledge of CAM than those employed at general hospitals (p < 0.0001), and those who were also involved in research activity had a greater knowledge of CAM than those who were not (p < 0.003). Length of work experience was significantly related to CAM knowledge. Moreover, 55% of participants suggest CAM interventions to their patients and 44% discuss CAM with them. The best-known interventions were acupuncture, Aloe vera and high-dose vitamin C. Conclusion: CAM use by patients with chronic disease and/or cancer has become a topical issue for the scientific community and for physicians. Knowing the reasons that prompt these patients to use CAM and guiding them in their decisions would improve treatment and outcomes and also benefit healthcare systems. Our findings contribute to a greater understanding of CAM knowledge, attitudes, and practice among Italian physicians. Further research is needed to identify the more effective CAM treatments and to work toward an integrated healthcare model.


Molecular analysis of 250 patients with autosomal recessive congenital ichthyosis: evidence for mutation hotspots in ALOXE3 and allelic heterogeneity in ALOX12B.
In recent years several new genes for autosomal recessive congenital ichthyosis (ARCI) have been identified. However, little is known about the molecular epidemiology and pathophysiology of this genetically and clinically heterogeneous group of severe disorders of keratinization. ARCI is characterized by intense scaling of the whole integument often associated with erythema. We and others have shown that mutations in ALOX12B and ALOXE3, coding for the lipoxygenases 12R-LOX and eLOX-3 predominantly synthesized in the epidermis, can underlie this rare condition. Here we have surveyed a large group of 250 patients with ARCI for mutations in these two genes. We have identified 11 different previously unreported mutations in ALOX12B and ALOXE3 in 21 ARCI patients from 19 unrelated families and demonstrated that mutations in the two genes are the second most common cause for ARCI in this cohort of patients. Examination of the molecular data revealed allelic heterogeneity for ALOX12B and two mutational hotspots in ALOXE3. Functional analysis of all missense mutations and a splice site mutation demonstrated that complete loss of function of the enzymes underlies the phenotype. Our findings further establish the pivotal role of the 12-lipoxygenase pathway during epidermal differentiation.


Assessing articulatory speed performance as a potential factor of slowed speech in older adults.
PURPOSE To improve our understanding about the underlying factors of aging-related speaking rate decline, the authors sought to determine if lip and jaw speeds are physiologically constrained in older adults. METHOD Thirty-six females-10 young adults (ages 22-27 years), 9 middle-aged adults (ages 45-55 years), 10 young-old adults (65-74 years), and 7 very old adults (ages 87-95 years)-completed metronome-paced syllable repetitions while moving the lower lip or jaw to a fixed target with each repetition. Metronome paces incrementally increased from 1.4 Hz to 6.7 Hz. Lip and jaw movements were tracked using a 3-dimensional motion capture system. RESULTS Older adults' maximum percent increase in lip and jaw peak speed was comparable to or tended to be even greater than that of middle-aged and young adults. By contrast, lip and jaw stiffness, indexed by peak speed-displacement ratios, tended to decrease with age during fast and very fast repetition rates and were associated with mildly prolonged movement durations. CONCLUSIONS The findings suggest that lip and jaw speeds are not constrained in older adults. The trend of reduced stiffness during fast rates, however, suggests that fine-force regulation becomes difficult for older adults. Thus, older adults may implement reduced habitual speaking rates as a behavioral strategy to compensate for diminished articulatory control.


Pathogenetics of alveolar capillary dysplasia with misalignment of pulmonary veins.
Alveolar capillary dysplasia with misalignment of pulmonary veins (ACDMPV) is a lethal lung developmental disorder caused by heterozygous point mutations or genomic deletion copy-number variants (CNVs) of FOXF1 or its upstream enhancer involving fetal lung-expressed long noncoding RNA genes LINC01081 and LINC01082. Using custom-designed array comparative genomic hybridization, Sanger sequencing, whole exome sequencing (WES), and bioinformatic analyses, we studied 22 new unrelated families (20 postnatal and two prenatal) with clinically diagnosed ACDMPV. We describe novel deletion CNVs at the FOXF1 locus in 13 unrelated ACDMPV patients. Together with the previously reported cases, all 31 genomic deletions in 16q24.1, pathogenic for ACDMPV, for which parental origin was determined, arose de novo with 30 of them occurring on the maternally inherited chromosome 16, strongly implicating genomic imprinting of the FOXF1 locus in human lungs. Surprisingly, we have also identified four ACDMPV families with the pathogenic variants in the FOXF1 locus that arose on paternal chromosome 16. Interestingly, a combination of the severe cardiac defects, including hypoplastic left heart, and single umbilical artery were observed only in children with deletion CNVs involving FOXF1 and its upstream enhancer. Our data demonstrate that genomic imprinting at 16q24.1 plays an important role in variable ACDMPV manifestation likely through long-range regulation of FOXF1 expression, and may be also responsible for key phenotypic features of maternal uniparental disomy 16. Moreover, in one family, WES revealed a de novo missense variant in ESRP1, potentially implicating FGF signaling in the etiology of ACDMPV.


Evanescent-wave-induced frequency shift for optical superresolution imaging.
We propose a method to enhance the resolution and break the diffraction limit. The superresolution imaging is realized by incorporating total internal reflection (TIR) illumination with a passive spatial frequency shift mechanism. Meanwhile, TIR supplies a surface field with a limited penetration depth, which demonstrates that the axial resolution can be improved simultaneously. The superresolution capability is confirmed both theoretically and experimentally. Compared with microfiber-based former work, this idea possesses promising merits, providing a wider viewing field and a simpler configuration for variable illumination orientations, thereby implying abundant application potentials.


Suicidal behavior in adolescents with post-traumatic stress disorder.
Recently, the prevalence of post-traumatic stress disorder (PTSD) in adolescence is higher than the prevalence of PTSD in adult populations. PTSD and suicidality are often found in populations of adolescents presenting with other emotional disorders (particularly mood disorders), traumatic grief, childhood abuse, and/or a family or peer history of suicide. The reasons and developments of the association between PTSD and suicidality in adolescence, however, remain unclear. Core psychobiological changes contributing to PTSD affect emotion, arousal, perception of the self and the world, irritability, impulsivity, anger, aggression and depression. There is evidence that the aforementioned factors, as well as alcohol and other drug use may act to moderate the influence of stressful life events and lead to eventual suicidality. Both PTSD and suicidality in adolescents have also been hypothesized to be a result of exposure to violence and negative coping styles. There are many treatment challenges for these populations, yet the most promising prevention and treatments include suicide risk screenings, suicide education, Dialectical Behavioral Therapy, addressing associated coping mechanisms and prescribing anti-depressant and anti-anxiety medications. However, when prescribing medications, physicians do need to be careful to consider the weaknesses and strengths of each of the pharmacological options as they apply to adolescents presenting with PTSD and suicidality.


Studying molecular motor-based cargo transport: what is real and what is noise?
Noise is a major problem in analyzing tracking data of cargos moved by molecular motors. We use Bayesian statistics to incorporate what is known about the noise in parsing the trajectory of a cargo into a series of constant velocity segments. Tracks with just noise and no underlying motion are fit with constant velocity segments to produce a calibration curve of fit quality versus average segment duration. Fits to tracks of moving cargos are compared to the calibration curves with similar noise. The fit with the optimum number of constant velocity states has the least number of segments needed to match the fit quality of the calibration curve. We have tested this approach using tracks with known underlying motion generated by computer simulations and with a specially designed in vitro experiment. We present the results of using this parsing approach to analyze transport of lipid droplets in Drosophila embryos.


Microsatellite instability and expression of hMLH1 and hMSH2 proteins in ovarian endometrioid cancer.
Microsatellite instability and loss of heterozygosity has been implicated in ovarian carcinogenesis. The reported frequency of microsatellite instability in human ovarian cancer varies significantly owing to the use of heterogeneous tumor histotypes and various microsatellite markers in different laboratories. In this study, we determined the frequency of microsatellite instability in 74 ovarian endometrioid carcinomas using four microsatellite markers (BAT25, BAT26, D5S346, D17S250), and examined hMLH1 and hMSH2 protein expression. In all, 20% of the tumors were microsatellite instability high (two or more markers showing instability) and 12% were microsatellite instability low (one marker showed instability). Loss of hMLH1 and/or hMSH2 expression was found in nine of 15 microsatellite instability-high tumors. The microsatellite instability-high phenotype tended to occur more frequently in low-grade tumors (P=0.053), but did not correlate with clinical stage. Totally, 38% of cases also displayed loss of heterozygosity at D17S250; this loss of heterozygosity was associated with high clinical stage (P=0.097). Our results indicate that both microsatellite and loss of heterozygosity at D17S250 are involved in the development of ovarian endometrioid carcinoma.


The influence of sentential position on noun phrase structure priming.
This article explores the interaction between global sentence context and local syntactic decision making. Specifically, four noun phrase (NP) structural priming experiments investigated whether the position of an NP within a sentence increased speakers' tendency to repeat primed structure. We crossed the position of the NP with the structure of the NP, such that NPs could be sentence initial or final in prime sentences. We further manipulated whether the to-be-modified target NP was sentence initial or final. Structural persistence effects were consistently observed, but there was no influence of parallel position. Rather, sentence-initial NP primes had a stronger influence on subsequent syntactic decisions than sentence-final primes, suggesting a primacy effect. Sentence-initial target NPs contributed to this primacy effect, while sentence-final target NPs did not. We argue that this primacy effect arises as the result of greater processing demands and resources for early than for late sentence constituents as well as deeper encoding and more focused attention when processing the beginnings of sentences.


DAF-16-dependent and independent expression targets of DAF-2 insulin receptor-like pathway in Caenorhabditis elegans include FKBPs.
The daf-2 insulin-like receptor pathway regulates development and life-span in Caenorhabditis elegans. Reduced DAF-2 signaling leads to changes in downstream targets via the daf-16 gene, a fork-head transcription factor which is regulated by DAF-2, and results in extended life-span. Here, we describe the first identification of genes whose expression is controlled by the DAF-2 signaling cascade. dao-1, dao-2, dao-3, dao-4, dao-8 and dao-9 are down-regulated in daf-2 mutant adults compared to wild-type adults, whereas dao-5, dao-6 and dao-7 are up-regulated. The latter genes are negatively regulated by DAF-2 signaling and positively regulated by DAF-16. Positive regulation by DAF-2 on dao-1, dao-4 and dao-8 was mediated by DAF-16, whereas daf-16 mediates only part of DAF-2 signaling for dao-2 and dao-9. Regulation by DAF-2 is most likely DAF-16 independent for dao-3 and hsp-90. RNA levels of dao-5 and dao-6 showed elevated expression in daf-2 adults, as well as being strongly expressed in dauer larvae. In contrast, hsp-90 transcript levels are low in daf-2 mutant adults though they are enriched in dauer larvae, indicating overlapping but not identical mechanisms of efficient life maintenance in stress-resistant dauer larvae and long-lived daf-2 mutant adults. dao-1, dao-8 and dao-9 are homologs of the FK506 binding proteins that interact with the mammalian insulin pathway. dao-3 encodes a putative methylenetetrahydrofolate dehydrogenase. DAO-5 shows 33 % identity with human nucleolar phosphoprotein P130. dao-7 is similar to the mammalian ZFP36 protein. Distinct regulatory patterns of dao genes implicate their diverse positions within the signaling network of DAF-2 pathway, and suggest they have unique contributions to development, metabolism and longevity.


Impact of Social Isolation on the Behavioral, Functional Profiles, and Hippocampal Atrophy Asymmetry in Dementia in Times of Coronavirus Pandemic (COVID-19): A Translational Neuroscience Approach.
The impact of COVID-19 on the elderly is devastating, and nursing homes are struggling to provide the best care to the most fragile. The urgency and severity of the pandemic forces the use of segregation in restricted areas and confinement in individual rooms as desperate strategies to avoid the spread of disease and the worst-case scenario of becoming a deadly trap. The conceptualization of the post-COVID-19 era implies strong efforts to redesign all living conditions, care/rehabilitation interventions, and management of loneliness forced by social distance measures. Recently, a study of gender differences in COVID-19 found that men are more likely to suffer more severe effects of the disease and are over twice as likely to die. It is well-known that dementia is associated with increased mortality, and males have worse survival and deranged neuro-immuno-endocrine systems than females. The present study examines the impact of long-term isolation in male 3xTg-AD mice modeling advanced stages of Alzheimer's disease (AD) and as compared to age-matched counterparts with normal aging. We used a battery of ethological and unconditioned tests resembling several areas in nursing homes. The main findings refer to an exacerbated (two-fold increase) hyperactivity and emergence of bizarre behaviors in isolated 3xTg-AD mice, worrisome results since agitation is a challenge in the clinical management of dementia and an important cause of caregiver burden. This increase was consistently shown in gross (activity in most of the tests) and fine (thermoregulatory nesting) motor functions. Isolated animals also exhibited re-structured anxiety-like patterns and coping-with-stress strategies. Bodyweight and kidney weight loss were found in AD-phenotypes and increased by isolation. Spleen weight loss was isolation dependent. Hippocampal tau pathology was not modified, but asymmetric atrophy of the hippocampus, recently described in human patients with dementia and modeled here for the first time in an animal model of AD, was found to increase with isolation. Overall, the results show awareness of the impact of isolation in elderly patients with dementia, offering some guidance from translational neuroscience in these times of coronavirus and post-COVID-19 pandemic. They also highlight the relevance of personalized-based interventions tailored to the heterogeneous and complex clinical profile of the individuals with dementia and to consider the implications on caregiver burden.


Adaptive paternal effects? Experimental evidence that the paternal environment affects offspring performance.
The ability of females to adaptively influence offspring phenotype via maternal effects is widely acknowledged, but corresponding nongenetic paternal effects remain unexplored. Males can adjust sperm phenotype in response to local conditions, but the transgenerational consequences of this plasticity are unknown. We manipulated paternal density of a broadcast spawner (Styela plicata, a solitary ascidean) using methods shown previously to alter sperm phenotype in the field, then conducted in vitro fertilizations that excluded maternal effects and estimated offspring performance under natural conditions. Offspring sired by males from low-density experimental populations developed faster and had a higher hatching success than offspring sired by males living in high densities. In the field, offspring survived relatively better when their environment matched their father's, raising the possibility that fathers can adaptively influence the phenotype of their offspring according to local conditions. As the only difference between offspring is whether they were artificially fertilized by sperm from males kept in high- vs. low-density cages, we can unequivocally attribute any differences in offspring performance to an environmentally induced paternal effect. Males of many species manipulate the phenotype of their sperm in response to sperm competition: our results show this plasticity can influence offspring fitness, potentially in adaptive ways, raising the possibility that adaptive nongenetic paternal effects may be more common than previously thought.


The burden of influenza complications in different high-risk groups: a targeted literature review.
The objective was to review the published literature on seasonal influenza to assess the differences between complications and mortality rates for those adults at high risk of influenza complications, including the resource use of those hospitalized with influenza complications. A targeted literature review was performed using electronic database keyword searches, specific inclusion criteria, quality rating of the reviewed full-text articles and abstraction of data to present published evidence on the incidence, complication rates and health service use associated with clinical influenza in different adult high-risk groups including those who are aged 65 years and older or those with different chronic underlying medical conditions. Key findings for incidence rates of clinical influenza were that incidence rates are similar among people with chronic cardiovascular or respiratory comorbidity, and may be higher in those with allogeneic stem cell transplants compared to those with autologous transplants. Rates of hospitalization and/or pneumonia or lower respiratory tract infection for those with chronic conditions or those who are immunocompromised are substantially higher than those in people over age 65 but without additional high-risk factors. A person who is hospitalized and has a laboratory-confirmed influenza diagnosis has a probability of intensive care unit admission of between 11.8-28.6% and of death of between 2.9-14.3%. These findings indicate that although the burden of influenza varied across high-risk groups, it also varied widely across studies within a single high-risk group. A key finding was that those over 65 years of age but without additional high-risk factors had a low risk of influenza complications. A limitation of the review is that most of the studies of hospitalized patients did not present outcomes data separately by high-risk group and only limited data were identified on rates of hospitalization or lower respiratory tract infection for most high-risk groups. Information about influenza complication rates and resource use, including influenza vaccines, chemoprophylaxis and/or treatment strategies for different high-risk groups, is needed to evaluate new interventions.


The struggle for web eQuality by persons with cognitive disabilities.
This article is based on the book eQuality: The Struggle for Web Accessibility by Persons with Cognitive Disabilities (2014, Cambridge University Press). It contends that the rights of individuals with cognitive disabilities to equal access to web content are not only protected under law, but may also be implemented and supported by current user-based, semantic and cloud technologies. Consistent with the Americans with Disabilities Act and the U.N. Convention on the Rights of Persons with Disabilities, web content equality is defined through functional, rather than disability-specific, approaches and techniques to enable personalization and customized usage across online functions. Legal challenges brought forward by individuals with cognitive and other disabilities illustrate the barriers still faced by individuals with disabilities to web equality as well as some of the solutions to and outcomes of these challenges. In closing, a view for the full and equal enjoyment of web content, which considers technology, financial benefits, and the role of advocacy and regulations, is discussed.


Alterations in the expression of colony-stimulating factor-1 and its receptor during an acute graft-vs-host reaction in mice.
During the course of an acute graft-vs-host reaction in the mouse we observed a progressive increase in the concentration of CSF-1 in serum and liver, peaking at day 14. In contrast, there was a progressive decrease in the splenic CSF-1 concentration. In vivo studies of 125I-CSF-1 uptake and degradation and in vitro studies of 125I-CSF-1 binding by splenic cells demonstrated that within 24 h of the reaction the number of CSF-1 receptor+ cells had increased by 2-fold and their capacity to express the CSF-1 receptor by approximately 3-fold, resulting in a approximately 2.5-fold increase in the splenic clearance of CSF-1 from the circulation. Inasmuch as at 24 h, serum CSF-1 was not significantly altered, these results are suggestive of an increased rate of release of CSF-1 into the circulation early in the response. The splenic CSF-1-receptor bearing cells were in a Mac-1+ fraction that is consistent with a role for CSF-1 in the generation of host-derived splenic macrophages in acute graft-vs-host reaction.


Are driving and overtaking on right curves more dangerous than on left curves?
It is well known that crashes on horizontal curves are a cause for concern in all countries due to the frequency and severity of crashes at curves compared to road tangents. A recent study of crashes in western Sweden reported a higher rate of crashes in right curves than left curves. To further understand this result, this paper reports the results of novel analyses of the responses of vehicles and drivers during negotiating and overtaking maneuvers on curves for right hand traffic. The overall objectives of the study were to find road parameters for curves that affect vehicle dynamic responses, to analyze these responses during overtaking maneuvers on curves, and to link the results with driver behavior for different curve directions. The studied road features were speed, super-elevation, radius and friction including their interactions, while the analyzed vehicle dynamic factors were lateral acceleration and yaw angular velocity. A simulation program, PC-Crash, has been used to simulate road parameters and vehicle response interaction in curves. Overtaking maneuvers have been simulated for all road feature combinations in a total of 108 runs. Analysis of variances (ANOVA) was performed, using two sided randomized block design, to find differences in vehicle responses for the curve parameters. To study driver response, a field test using an instrumented vehicle and 32 participants was reviewed as it contained longitudinal speed and acceleration data for analysis. The simulation results showed that road features affect overtaking performance in right and left curves differently. Overtaking on right curves was sensitive to radius and the interaction of radius with road condition; while overtaking on left curves was more sensitive to super-elevation. Comparisons of lateral acceleration and yaw angular velocity during these maneuvers showed different vehicle response configurations depending on curve direction and maneuver path. The field test experiments also showed that drivers behave differently depending on the curve direction where both speed and acceleration were higher on right than left curves. The implication of this study is that curve direction should be taken into consideration to a greater extent when designing and redesigning curves. It appears that the driver and the vehicle are influenced by different infrastructure factors depending on the curve direction. In addition, the results suggest that the vehicle dynamics response alone cannot explain the higher crash risk in right curves. Further studies of the links between driver, vehicle, and highway characteristics are needed, such as naturalistic driving studies, to identify the key safety indicators for highway safety.


Adaptive vaccination strategies to mitigate pandemic influenza: Mexico as a case study.
We explore vaccination strategies against pandemic influenza in Mexico using an age-structured transmission model calibrated against local epidemiological data from the Spring 2009 A(H1N1) pandemic. In the context of limited vaccine supplies, we evaluate age-targeted allocation strategies that either prioritize youngest children and persons over 65 years of age, as for seasonal influenza, or adaptively prioritize age groups based on the age patterns of hospitalization and death monitored in real-time during the early stages of the pandemic. Overall the adaptive vaccination strategy outperformed the seasonal influenza vaccination allocation strategy for a wide range of disease and vaccine coverage parameters. This modeling approach could inform policies for Mexico and other countries with similar demographic features and vaccine resources issues, with regard to the mitigation of the S-OIV pandemic. We also discuss logistical issues associated with the implementation of adaptive vaccination strategies in the context of past and future influenza pandemics.


Adsorption and binding dynamics of graphene-supported phospholipid membranes using the QCM-D technique.
We report on the adsorption dynamics of phospholipid membranes on graphene-coated substrates using the quartz crystal microbalance with dissipation monitoring (QCM-D) technique. We compare the lipid vesicle interaction and membrane formation on gold and silicon dioxide QCM crystal surfaces with their graphene oxide (GO) and reduced (r)GO coated counterparts, and report on the different lipid structures obtained. We establish graphene derivative coatings as support surfaces with tuneable hydrophobicity for the formation of controllable lipid structures. One structure of interest formed is lipid monolayer membranes which were formed on rGO, which are otherwise challenging to produce. We also demonstrate and monitor biotin-avidin binding on such a membrane, which will then serve as a platform for a wide range of biosensing applications. The QCM-D technique could be extended to both fundamental studies and applications of other covalent and non-covalent interactions in 2-dimensional materials.


Axenic cultivation of Entamoeba hitolytica.
Entamoeba histolytica (National Institutes of Health strain 200) has been maintained in axenic culture through 50 transfers over a period of 6 months. The medium used is diphasic and contains a cell-free extract of chick embryo which is essential for growth. Inocula of 50,000 amoebas yield 150,000 to 200,000 organisms in 72 hours of incubation at 35 degrees C. An extract prepared from boiled embryos supports growth equally well.


Hypothermia and complications in postbariatric body contouring.
Postbariatric body contouring represents a rapidly growing field. With long operative times and high rates of minor complications, evidence-based guidelines for operative management are needed. Data analyzing the impact of perioperative factors on patient outcomes are currently limited. Patients who lost 50 pounds or more and underwent body contouring were enrolled in an institutional review board-approved prospective registry over 4 years. All cases were performed by the senior author at two community hospitals and two academic hospitals. Hypothermia was defined as a minimum temperature of 35°C or lower. Three hundred eight patients (272 women and 36 men) were analyzed. The average operative time was 4.7 hours, and 71.4 percent of cases were performed in an academic hospital. The average minimum operative temperature was 35.6±0.63°C (range, 34.0 to 38.0°C), and the average maximum operative temperature was 36.5±0.75°C (range, 34.8 to 39.0°C). On multivariate analysis, a lower minimum temperature was associated with both seroma (p=0.003; odds ratio, 3.1 per 1°C decrease) and transfusion (p=0.005; odds ratio, 2.4 per 1°C decrease). Operative hypothermia was present in one-fifth of cases. Lower intraoperative temperature was associated with an increased risk of seroma formation, blood loss, and the need for transfusion. Despite this, there were few major complications in lengthy cases involving substantial blood loss. Maintaining normothermia is a critical component of perioperative management. The authors have adopted a protocol involving patient prewarming, a higher operating room temperature, and the routine use of warmed fluids. Risk, II.


Peer Review Bias: A Critical Review.
Various types of bias and confounding have been described in the biomedical literature that can affect a study before, during, or after the intervention has been delivered. The peer review process can also introduce bias. A compelling ethical and moral rationale necessitates improving the peer review process. A double-blind peer review system is supported on equipoise and fair-play principles. Triple- and quadruple-blind systems have also been described but are not commonly used. The open peer review system introduces "Skin in the Game" heuristic principles for both authors and reviewers and has a small favorable effect on the quality of published reports. In this exposition, we present, on the basis of a comprehensive literature search of PubMed from its inception until October 20, 2017, various possible mechanisms by which the peer review process can distort research results, and we discuss the evidence supporting different strategies that may mitigate this bias. It is time to improve the quality, transparency, and accountability of the peer review system.


Confined Water in Layered Silicates: The Origin of Anomalous Thermal Expansion Behavior in Calcium-Silicate-Hydrates.
Water, under conditions of nanoscale confinement, exhibits anomalous dynamics, and enhanced thermal deformations, which may be further enhanced when such water is in contact with hydrophilic surfaces. Such heightened thermal deformations of water could control the volume stability of hydrated materials containing nanoconfined structural water. Understanding and predicting the thermal deformation coefficient (TDC, often referred to as the CTE, coefficient of thermal expansion), which represents volume changes induced in materials under conditions of changing temperature, is of critical importance for hydrated solids including: hydrogels, biological tissues, and calcium silicate hydrates, as changes in their volume can result in stress development, and cracking. By pioneering atomistic simulations, we examine the physical origin of thermal expansion in calcium-silicate-hydrates (C-S-H), the binding agent in concrete that is formed by the reaction of cement with water. We report that the TDC of C-S-H shows a sudden increase when the CaO/SiO2 (molar ratio; abbreviated as Ca/Si) exceeds 1.5. This anomalous behavior arises from a notable increase in the confinement of water contained in the C-S-H's nanostructure. We identify that confinement is dictated by the topology of the C-S-H's atomic network. Taken together, the results suggest that thermal deformations of hydrated silicates can be altered by inducing compositional changes, which in turn alter the atomic topology and the resultant volume stability of the solids.


Transmembrane Pickets Connect Cyto- and Pericellular Skeletons Forming Barriers to Receptor Engagement.
Phagocytic receptors must diffuse laterally to become activated upon clustering by multivalent targets. Receptor diffusion, however, can be obstructed by transmembrane proteins ("pickets") that are immobilized by interacting with the cortical cytoskeleton. The molecular identity of these pickets and their role in phagocytosis have not been defined. We used single-molecule tracking to study the interaction between Fcγ receptors and CD44, an abundant transmembrane protein capable of indirect association with F-actin, hence likely to serve as a picket. CD44 tethers reversibly to formin-induced actin filaments, curtailing receptor diffusion. Such linear filaments predominate in the trailing end of polarized macrophages, where receptor mobility was minimal. Conversely, receptors were most mobile at the leading edge, where Arp2/3-driven actin branching predominates. CD44 binds hyaluronan, anchoring a pericellular coat that also limits receptor displacement and obstructs access to phagocytic targets. Force must be applied to traverse the pericellular barrier, enabling receptors to engage their targets.


Characterization of repetitive DNA landscape in wheat homeologous group 4 chromosomes.
The number and complexity of repetitive elements varies between species, being in general most represented in those with larger genomes. Combining the flow-sorted chromosome arms approach to genome analysis with second generation DNA sequencing technologies provides a unique opportunity to study the repetitive portion of each chromosome, enabling comparisons among them. Additionally, different sequencing approaches may produce different depth of insight to repeatome content and structure. In this work we analyze and characterize the repetitive sequences of Triticum aestivum cv. Chinese Spring homeologous group 4 chromosome arms, obtained through Roche 454 and Illumina sequencing technologies, hereinafter marked by subscripts 454 and I, respectively. Repetitive sequences were identified with the RepeatMasker software using the interspersed repeat database mips-REdat_v9.0p. The input sequences consisted of our 4DS454 and 4DL454 scaffolds and 4ASI, 4ALI, 4BSI, 4BLI, 4DSI and 4DLI contigs, downloaded from the International Wheat Genome Sequencing Consortium (IWGSC). Repetitive sequences content varied from 55% to 63% for all chromosome arm assemblies except for 4DLI, in which the repeat content was 38%. Transposable elements, small RNA, satellites, simple repeats and low complexity sequences were analyzed. SSR frequency was found one per 24 to 27 kb for all chromosome assemblies except 4DLI, where it was three times higher. Dinucleotides and trinucleotides were the most abundant SSR repeat units. (GA)n/(TC)n was the most abundant SSR except for 4DLI where the most frequently identified SSR was (CCG/CGG)n. Retrotransposons followed by DNA transposons were the most highly represented sequence repeats, mainly composed of CACTA/En-Spm and Gypsy superfamilies, respectively. This whole chromosome sequence analysis allowed identification of three new LTR retrotransposon families belonging to the Copia superfamily, one belonging to the Gypsy superfamily and two TRIM retrotransposon families. Their physical distribution in wheat genome was analyzed by fluorescent in situ hybridization (FISH) and one of them, the Carmen retrotransposon, was found specific for centromeric regions of all wheat chromosomes. The presented work is the first deep report of wheat repetitive sequences analyzed at the chromosome arm level, revealing the first insight into the repeatome of T. aestivum chromosomes of homeologous group 4.


Post-licence driver education for the prevention of road traffic crashes.
Worldwide, each year over a million people are killed and some ten million people are permanently disabled in road traffic crashes. Post-licence driver education is used by many as a strategy to reduce traffic crashes. However, the effectiveness of post-licence driver education has yet to be ascertained. To quantify the effectiveness of post-licence driver education in reducing road traffic crashes. We searched the following electronic databases: the Cochrane Injuries Group's Specialised Register, Cochrane Central Register of Controlled Trials, MEDLINE, EMBASE, TRANSPORT (NTIS, TRIS, TRANSDOC, IRRD), Road Res (ARRB), ATRI, National Research Register, PsycInfo, ERIC, SPECTR, Zetoc, SIGLE, Science (and Social Science) Citation Index. We searched the Internet, checked reference lists of relevant papers and contacted appropriate organisations. The search was not restricted by language or publication status. Randomised controlled trials comparing post-licence driver education versus no education, or one form of post-licence driver education versus another. Two reviewers independently screened search results, extracted data and assessed methodological trial quality. We found 24 trials of driver education, 23 conducted in the USA and one in Sweden. Twenty trials studied remedial driver education. The methodological quality of the trials was poor and three reported data unsuitable for meta-analysis. Nineteen trials reported traffic offences: pooled relative risk (RR) = 0.96, 95% confidence interval (95% CI) = 0.94, 0.98); trial heterogeneity was significant (p=<0.00001). Fifteen trials reported traffic crashes: pooled RR = 0.98 (95% CI 0.96, 1.01), trial heterogeneity was not significant (p=0.75). Four trials reported injury crashes: pooled RR = 1.12 (95% CI 0.88, 1.41), trial heterogeneity was significant (p=<0.00001). No one form of education (correspondence, group or individual) was found to be substantially more effective than another, nor was a significant difference found between advanced driver education and remedial driver education. Funnel plots indicated the presence of publication bias affecting the traffic offence and crash outcomes. This systematic review provides no evidence that post-licence driver education is effective in preventing road traffic injuries or crashes. Although the results are compatible with a small reduction in the occurrence of traffic offences, this may be due to selection biases or bias in the included trials. Because of the large number of participants included in the meta-analysis (close to 300,000 for some outcomes) we can exclude, with reasonable precision, the possibility of even modest benefits.


Diagnosis and clinical manifestations of bacterial vaginosis.
Among 640 randomly selected women who were attending a sexually transmitted disease clinic and did not have trichomoniasis, 33% had bacterial vaginosis as defined by a composite of four clinical criteria: (1) Vaginal discharge was homogeneous; (2) vaginal discharge had a pH greater than or equal to 4.7; (3) vaginal discharge had an amine-like odor when mixed with 10% potassium hydroxide; (4) vaginal discharge contained clue cells representing greater than or equal to 20% of vaginal epithelial cells. Previously published Gram stain criteria for bacterial vaginosis correlated better than results of semiquantitative cultures for Gardnerella vaginalis with presence or absence of clue cells and with composite clinical criteria. Of 293 women with bacterial vaginosis by Gram stain criteria, 65% had symptoms of increased vaginal discharge and/or vaginal malodor, while 74% had signs of characteristic homogeneous vaginal discharge or amine-like odor. Elevated vaginal pH was the least specific and amine-like odor the least sensitive sign of bacterial vaginosis. Gram stain criteria for bacterial vaginosis were not associated with the concentrations of endocervical or vaginal inflammatory cells but were significantly associated with a clinical diagnosis of pelvic inflammatory disease. After adjusting for coinfection, sexual behavior, and other variables, bacterial vaginosis remained associated with adnexal tenderness (odds ratio = 9.2, p = 0.04). Bacterial vaginosis, previously implicated as a risk factor for obstetric infections, may be a risk factor for pelvic inflammatory disease.


Antifungal susceptibility testing in Aspergillus spp. according to EUCAST methodology.
The availability of new antifungal agents has multiplied the demand for in vitro antifungal susceptibility testing for Aspergillus spp. The European Committee on Antimicrobial Susceptibility Testing (EUCAST) has charged its Antifungal Susceptibility Testing Subcommittee (AFST-EUCAST) with the preparation of new guidelines for in vitro susceptibility testing of antifungals against Aspergillus spp (EUCAST-AST-ASPERGILLUS). This committee has modified the reference method for broth dilution antifungal susceptibility testing of filamentous fungi (M38-A) as follows: (i) RPMI 1640 supplemented with 2% glucose (RPMI 2%G) as assay medium, (ii) inoculum preparation by conidium counting in a haemocytometer and (iii) an inoculum size of 2×105-5×105 CFU/ml. The incubation time is about 48 h at 35°C and MIC is read visually. The MIC value is a no-growth visual endpoint. The standard method described herein is intended to provide a valid and economic method for testing the susceptibility to antifungal agents of Aspergillus spp., to identify resistance, and to facilitate an acceptable degree of conformity, e.g. agreement within specified ranges and between laboratories in measuring the susceptibility.


Normative steps/day and peak cadence values for united states children and adolescents: National Health and Nutrition Examination Survey 2005-2006.
To provide sex-and-age specific normative values for children and adolescents' accelerometer-determined steps/day, and peak 60-minute cadence adjusted to a pedometer-based scale. The analysis sample was 2610 children and adolescents (1329 girls) from the 2005-2006 National Health and Nutrition Examination Survey. Accelerometer data were adjusted by eliminating steps counted when activity counts/min <500. Peak 60-minute cadence represented the 60 highest minutes of accumulated steps, averaged over monitored days. Normative data included quintile-defined categories of adjusted steps/day and peak 60-minute cadence for 7 age groups (6-7, 8-9, 10-11, 12-13, 14-15, 16-17, and 18-19 years). LMSChartmakerPro produced 10 age-group-specific smoothed curves (5 for each sex) showing the 5th, 25th, 50th, 75th, and 95th percentiles, respectively, for steps/day and peak 60-minute cadence. Steps/day was inversely associated with age in both boys and girls. The age-associated reduction was apparent in only small decrements for boys; the girl's reduction was steeper. Boys appeared to maintain or increase their peak 60-minute cadence with increased age between 8 and 15 years of age, with a reduction apparent over the last 2 age groups investigated. The peak 60-minute cadence was more variable for girls; a similar sharp reduction (3-6 steps/min) in tandem with the steps/day was apparent between 10- to 11-year-old girls and 12- to 13-year-old girls. We provided detailed information and normative data pertaining to steps/d and peak 60-minute cadence in US children and adolescents. Like well-known body mass index growth curves, these data may be useful for scientists and clinical practitioners.


Weight-Based Guide Overestimates Endotracheal Tube Tip Position in Extremely Preterm Infants.
To evaluate the association between endotracheal tube (ETT) tip position and adverse pulmonary outcomes using chest X-ray (CXR) in extremely preterm infants in whom ETT insertion length was estimated using weight + 6 guide (adding 6 cms to the infant's weight in kg). CXRs of 85 infants performed in the first week were reviewed for right-sided atelectasis, air leaks, and uneven lung inflation. The first CXR was later reviewed to document the ETT tip. Regression analysis was performed to find the association between ETT tip position and adverse outcome after adjusting for other confounders. Forty (46%) infants had ETT tip placement between the first and second thoracic vertebrae (optimal position) compared with 45 (53%) who had the ETT tip placement outside this range (suboptimal position). Infants with suboptimal ETT were ventilated for a longer period (6.1 vs. 15.9 days; p = 0.004). The odds of adverse outcomes was 11.6 (95% confidence interval: 3.03, 44.1) times higher among infants who did not have ETT at the optimal position compared with infants who had ETT at the optimal position. Weight + 6 guide is not recommended to estimate ETT insertion length in extremely preterm infants. Gestation-based guide may be more appropriate to estimate ETT insertion length in this group of infants.


In-hospital outcome in patients presenting with acute coronary syndrome with left main coronary artery disease: A report from Japanese prospective multicenter percutaneous coronary intervention registry.
Patients presenting with acute coronary syndrome (ACS) from left main (LM) disease are at a high risk for mortality despite recent advancement in devices and techniques during percutaneous coronary interventions (PCI). We aimed to evaluate patient characteristics, clinical presentations, and key clinical characteristics associated with adverse in-hospital outcomes among ACS patients undergoing LM-PCI. We retrospectively identified 280 LM-ACS patients (3.7 %) from 7608 ACS patients in the prospective multicenter Japan Cardiovascular Database-Keio Inter-Hospital Cardiovascular Studies registry from March 2009 to May 2016 and divided them into those with/without PCI/coronary artery bypass grafting. We compared baseline demographics, coronary lesion characteristics, PCI details, and short-term outcomes, including in-hospital mortality and periprocedural complications, between the two groups. Among LM-ACS patients, 38.6 % presented with ST elevation myocardial infarction, 29.6 % with cardiogenic shock (CS), and 15.4 % with cardiac arrest. The observed in-hospital mortality rate was 18.9 % with presence of CS [odds ratio (OR): 10.16, 95 % confidence interval (CI): 4.51-22.91, p<0.001] and absence of prior revascularization (de novo patients; OR: 4.31, 95 % CI: 1.43-12.94, p=0.009) was independently associated with higher incidence of in-hospital mortality. Notably, the observed mortality rate was substantially higher among de novo patients than the predicted mortality rate with a contemporary risk model (observed: 25.1 %; predicted: 11.6 %). Prior revascularization act as a protective factor among LM-ACS patients in the contemporary era of PCI. Further studies to detect those at higher risk for LM coronary lesion progression are needed to fully implement these findings into clinical practice.


Dissecting direct reprogramming through integrative genomic analysis.
Somatic cells can be reprogrammed to a pluripotent state through the ectopic expression of defined transcription factors. Understanding the mechanism and kinetics of this transformation may shed light on the nature of developmental potency and suggest strategies with improved efficiency or safety. Here we report an integrative genomic analysis of reprogramming of mouse fibroblasts and B lymphocytes. Lineage-committed cells show a complex response to the ectopic expression involving induction of genes downstream of individual reprogramming factors. Fully reprogrammed cells show gene expression and epigenetic states that are highly similar to embryonic stem cells. In contrast, stable partially reprogrammed cell lines show reactivation of a distinctive subset of stem-cell-related genes, incomplete repression of lineage-specifying transcription factors, and DNA hypermethylation at pluripotency-related loci. These observations suggest that some cells may become trapped in partially reprogrammed states owing to incomplete repression of transcription factors, and that DNA de-methylation is an inefficient step in the transition to pluripotency. We demonstrate that RNA inhibition of transcription factors can facilitate reprogramming, and that treatment with DNA methyltransferase inhibitors can improve the overall efficiency of the reprogramming process.


Guidelines for the standardized collection of blood-based biomarkers in psychiatry: Steps for laboratory validity - a consensus of the Biomarkers Task Force from the WFSBP.
Recently, there has been a major shift in the field of psychiatry towards the exploration of complex relationships between blood-based biomarkers and the pathophysiology of psychiatric and neuropsychiatric disorders. However, issues with study reproducibility, validity and reliability have hindered progress towards the identification of clinically relevant biomarkers for psychiatry. The achievement of laboratory validity is a crucial first step for the posterior development of clinical validity. There is evidence that the variability observed in blood-based research studies may be minimised with the implementation of standardised pre-analytical methods and uniform clinical protocols (i.e., pre-venipuncture). It has been documented that errors made in the pre-analytical phase account for 46-68.2% of laboratory testing errors. Thus, standardising clinical assessment, ethical procedures and pre-analytical phase of clinical research is essential for the reproducibility, validity and reliability of blood marker assessment, and reducing the risk of invalid test results. Various other areas of research have already moved towards guidelines for the standardised collection of blood-based biomarkers. Here we aim to provide a set of guidelines that we believe would improve biomarker research: (1) pre-venipuncture information and documentation, (2) ethics of participant consent and (3) pre-analytical methods. Ultimately, we hope this will assist study planning and will improve data comparison across studies allowing for the discovery of biomarkers in psychiatry with both laboratorial and clinical validity.


Development and face validation of ultrasound-guided renal biopsy virtual trainer.
The overall prevalence of chronic kidney disease in the general population is ∼14% with more than 661,000 Americans having a kidney failure. Ultrasound (US)-guided renal biopsy is a critically important tool in the evaluation and management of renal pathologies. This Letter presents KBVTrainer, a virtual simulator that the authors developed to train clinicians to improve procedural skill competence in US-guided renal biopsy. The simulator was built using low-cost hardware components and open source software libraries. They conducted a face validation study with five experts who were either adult/pediatric nephrologists or interventional/diagnostic radiologists. The trainer was rated very highly (>4.4) for the usefulness of the real US images (highest at 4.8), potential usefulness of the trainer in training for needle visualization, tracking, steadiness and hand-eye coordination, and overall promise of the trainer to be useful for training US-guided needle biopsies. The lowest score of 2.4 was received for the look and feel of the US probe and needle compared to clinical practice. The force feedback received a moderate score of 3.0. The clinical experts provided abundant verbal and written subjective feedback and were highly enthusiastic about using the trainer as a valuable tool for future trainees.


Transforming growth factor-β2 increases extracellular matrix proteins in optic nerve head cells via activation of the Smad signaling pathway.
Transforming growth factor-β2 (TGF-β2) is associated with glaucomatous neuropathy, primarily via the increased synthesis and secretion of extracellular matrix (ECM) proteins and remodeling of the optic nerve head (ONH). Here, we investigated the signaling pathways used by TGF-β2 to stimulate ECM expression by ONH astrocytes and lamina cribrosa (LC) cells. TGF-β2 localization and secretion was examined in human donor tissues and ONH astrocytes and LC cells. To examine TGF-β2 signaling, ONH astrocytes and LC cells were treated with recombinant TGF-β2, and phosphorylation of Smad and non-Smad signaling proteins were examined by western blot analyses and immunostaining. TGF-β2 is significantly increased in the LC region of the ONH in glaucomatous eyes compared to age-matched normal eyes (n=4, p<0.0013). ONH astrocytes and LC cells secrete TGF-β2, indicating that these cells may be an in vivo source of TGF-β2 in the human ONH. In addition, treatment of ONH astrocytes and LC cells with exogenous TGF-β2 increased ECM protein synthesis and secretion. With respect to TGF-β2 signaling, recombinant TGF-β2 induced phosphorylation of canonical signaling proteins Smad2/3 but did not alter phosphorylation of non-canonical signaling proteins extracellular signal-regulated kinases (ERK)1/2, p38, and c-Jun N-terminal kinases (JNK)1/2 proteins in ONH astrocytes and LC cells. Exogenous TGF-β2 increased co-localization of pSmad2/3 with Co-Smad4 in the nucleus of ONH astrocytes and LC cells further indicating activation of the canonical Smad signaling pathway. Furthermore, inhibition of TGF-β I receptor activity by SB431542 or inhibition of Smad3 phosphorylation by SIS3 blocked TGF-β2 stimulated ECM expression as well as activation of downstream canonical pathway signaling molecules. Knockdown of either Smad2 or Smad3 via small interfering RNA (siRNA) reduced TGF-β2 stimulated ECM proteins in ONH astrocytes and LC cells. These studies indicate that TGF-β2 utilizes the canonical Smad signaling pathway to stimulate ECM synthesis in human ONH cells. Our studies also indicate that pSmad2/3 is required for TGF-β2 stimulation of ECM remodeling.


The Global Diffusion of Law: Transnational Crime and the Case of Human Trafficking.
The past few decades have seen the proliferation of new laws criminalizing certain transnational activities, from money laundering to corruption; from insider trading to trafficking in weapons and drugs. Human trafficking is one example. We argue criminalization of trafficking in persons has diffused in large part because of the way the issue has been framed: primarily as a problem of organized crime rather than predominantly an egregious human rights abuse. Framing human trafficking as an organized crime practice empowers states to confront cross border human movements viewed as potentially threatening. We show that the diffusion of criminalization is explained by road networks that reflect potential vulnerabilities to the diversion of transnational crime. We interpret our results as evidence of the importance of context and issue framing, which in turn affects perceptions of vulnerability to neighbors' policy choices. In doing so, we unify diffusion studies of liberalization with the spread of prohibition regimes to explain the globalization of aspects of criminal law.


Short telomeres limit tumor progression in vivo by inducing senescence.
Telomere maintenance is critical for cancer progression. To examine mechanisms of tumor suppression induced by short telomeres, we crossed mice deficient for the RNA component of telomerase, mTR(-/-), with Emu-myc transgenic mice, an established model of Burkitt's lymphoma. Short telomeres suppressed tumor formation in Emu-myc transgenic animals. Expression of Bcl2 blocked apoptosis in tumor cells, but surprisingly, mice with short telomeres were still resistant to tumor formation. Staining for markers of cellular senescence showed that pretumor cells induced senescence in response to short telomeres. Loss of p53 abrogated the short telomere response. This study provides in vivo evidence for the existence of a p53-mediated senescence mechanism in response to short telomeres that suppresses tumorigenesis.


Structured-Light Based 3D Reconstruction System for Cultural Relic Packaging.
The non-contact three-dimensional measurement and reconstruction techniques have played a significant role in the packaging and transportation of precious cultural relics. This paper develops a structured light based three-dimensional measurement system, with a low-cost for cultural relics packaging. The structured light based system performs rapid measurements and generates 3D point cloud data, which is then denoised, registered and merged to achieve accurate 3D reconstruction for cultural relics. The multi-frequency heterodyne method and the method in this paper are compared. It is shown that the relative accuracy of the proposed low-cost system can reach a level of 1/1000. The high efficiency of the system is demonstrated through experimental results.


Uveal Melanoma Associated With Myotonic Dystrophy: A Report of 6 Cases.
Patients with myotonic dystrophy (MD) have an increased risk of malignancy including uveal melanoma. This case series further explores the association between these 2 diseases. To describe a cohort of patients with uveal melanoma associated with MD, including a case of iris melanoma, and MD-associated uveal melanoma in relatives. Retrospective case series at 3 tertiary referral centers (Wills Eye Hospital, Philadelphia, Pennsylvania; Mayo Clinic, Rochester, Minnesota; and Moorfields Eye Hospital, London, England), between January 1, 2000, and August 31, 2017. The study included 6 patients with MD and uveal melanoma. Melanoma response to treatment and development of metastatic disease. There were 6 patients, 4 men and 2 women, with MD and uveal melanoma. The mean patient age at melanoma diagnosis was 47 years (median, 43 years; range, 30-67 years), and the tumor involved the choroid in 5 patients (83%) and iris in 1 patient (17%). The diagnosis of MD was known since young adulthood in 2 patients (33%) and was discovered in adulthood in 4 patients (67%). The main clinical features of MD included muscle weakness (n = 5; 83%), myotonia (n = 4; 67%), polychromatic cataract (n = 4; 67%), complications with general anesthesia (n = 4; 67%), myalgia (n = 3; 50%), cardiac arrhythmia (n = 2; 33%), and frontal baldness (n = 2; 33%). Genetic testing revealed MD type 1 (4 of 4 tested patients), and 2 patients demonstrated positive family history of MD with classic clinical features and preferred no testing. Melanoma treatment included plaque radiotherapy (n = 4; 67%), photodynamic therapy (n = 1; 17%), and declined treatment (n = 1; 17%). At follow-up of 6, 6, 41, 42, and 87 months (5 patients), findings included melanoma regression (4 of 5 tumors), melanoma recurrence (1 of 5 tumors), and no metastatic disease (5 of 5 patients). Six adult patients with MD demonstrated uveal melanoma involving the choroid or iris, emphasizing the association between these 2 diseases. Further research seems warranted to explore the pathogenesis of uveal melanoma in MD. These findings support the consideration of ophthalmic examination for uveal melanoma in patients with MD.


A robotic approach to 4D real-time tumor tracking for radiotherapy.
Respiratory and cardiac motions induce displacement and deformation of the tumor volumes in various internal organs. To accommodate this undesired movement and other errors, physicians incorporate a large margin around the tumor to delineate the planning target volume, so that the clinical target volume receives the prescribed radiation dose under any scenario. Consequently, a large volume of healthy tissue is irradiated and sometimes it is difficult to spare critical organs adjacent to the tumor. In this study we have proposed a novel approach to the 4D active tracking and dynamic delivery incorporating the tumor motion prediction technique. This method has been applied to the two commercially available robotic treatment couches. The proposed algorithm can predict the tumor position and the robotic systems are able to continuously track the tumor during radiation dose delivery. Therefore a precise dose is given to a moving target while the dose to the nearby critical organs is reduced to improve the patient treatment outcome. The efficacy of the proposed method has been investigated by extensive computer simulation. The tumor tracking method is simulated for two couches: HexaPOD robotic couch and ELEKTA Precise Table. The comparison results have been presented in this paper. In order to assess the clinical significance, dosimetric effects of the proposed method have been analyzed.


Optimal Population and Exhaustible Resource Constraints.
A large literature considers the optimal size and growth rate of the human population, trading off the utility value of additional people with the costs of a larger population. In this literature, an important parameter is the social weight placed on population size; a standard result is that a planner with a larger weight on population chooses larger population levels and growth rates. We demonstrate that this result is conditionally overturned when an exhaustible resource constraint is introduced: if the discount rate is small enough, the optimal population today decreases with the welfare weight on population size. That is, a more total-utilitarian social planner could prefer a smaller population today than a more average-utilitarian social planner. We also present a numerical illustration applied to the case of climate change, where we show that under plausible real-world parameter values, our result matters for the direction and magnitude of optimal population policy.


Blooms of single bacterial species in a coastal lagoon of the southwestern Atlantic Ocean.
We investigated seasonal differences in community structure and activity (leucine incorporation) of the planktonic bacterial assemblage in the freshwater and brackish-water zones of a shallow coastal lagoon of the southwestern Atlantic Ocean. Alphaproteobacteria formed the dominant microbial group in both zones throughout the sampling period. After an intrusion of marine water, members of the SAR11 lineage became abundant in the brackish-water zone. These bacteria were apparently distributed over the lagoon during the following months until they constituted almost 30% of all prokaryotic cells at both sampling sites. At the first sampling date (March 2003) a single alphaproteobacterial species unrelated to SAR11, Sphingomonas echinoides, dominated the microbial assemblages in both zones of the lagoon concomitantly with a bloom of filamentous cyanobacteria. Pronounced maxima of leucine incorporation were observed once in each zone of the lagoon. In the freshwater zone, this highly active microbial assemblage was a mix of the typical bacteria lineages expected in aquatic systems. By contrast, a single bacterial genotype with >99% similarity to the facultative pathogen gammaproteobacterial species Stenotrophomonas maltophilia formed >90% of the bacterial assemblage (>10(7) cell ml(-1)) in the brackish-water zone at the time point of highest bacterial leucine incorporation. Moreover, these bacteria were equally dominant, albeit less active, in the freshwater zone. Thus, the pelagic zone of the studied lagoon harbored repeated short-term blooms of single bacterial species. This finding may have consequences for environmental protection.


Induction of antigen-positive cell death by the expression of perforin, but not DTa, from a DNA vaccine enhances the immune response.
The failure of traditional protein-based vaccines to prevent infection by viruses such as HIV or hepatitis C highlights the need for novel vaccine strategies. DNA vaccines have shown promise in small animal models, and are effective at generating anti-viral T cell-mediated immune responses; however, they have proved to be poorly immunogenic in clinical trials. We propose that the induction of necrosis will enhance the immune response to vaccine antigens encoded by DNA vaccines, as necrotic cells are known to release a range of intracellular factors that lead to dendritic cell (DC) activation and enhanced cross-presentation of antigen. Here we provide evidence that induction of cell death in DNA vaccine-targeted cells provides an adjuvant effect following intradermal vaccination of mice; however, this enhancement of the immune response is dependent on both the mechanism and timing of cell death after antigen expression. We report that a DNA vaccine encoding the cytolytic protein, perforin, resulted in DC activation, enhanced broad and multifunctional CD8 T-cell responses to the HIV-1 antigen GAG and reduced viral load following challenge with a chimeric virus, EcoHIV, compared with the canonical GAG DNA vaccine. This effect was not observed for a DNA vaccine encoding an apoptosis-inducing toxin, DTa, or when the level of perforin expression was increased to induce cell death sooner after vaccination. Thus, inducing lytic cell death following a threshold level of expression of a viral antigen can improve the immunogenicity of DNA vaccines, whereas apoptotic cell death has an inhibitory effect on the immune response.


Child and parent attributions in chronic pediatric conditions: phenylketonuria (PKU) as an exemplar.
Attribution theory, self-regulation, self-handicapping and sick role theories all suggest that children with chronic disease may be held to different standards. This study assesses child and parent attributions in pediatric chronic health conditions and addresses how attributional style may be related to treatment adherence. Four different vignettes were utilized to compare the attributional style of children with phenylketonuria (PKU) and parents of children with PKU to two comparison groups: children with other chronic medical conditions and medically healthy children. In addition, the relationship between metabolic control and attribution ratings was assessed in the PKU sample. Parents of children with medical conditions provided attributions of less child control and more stability as well as more positive affective reactions when compared to parents of healthy children. Children without a medical condition viewed childhood problems as less controllable and more stable. Mothers were more disturbed affectively by behavioral dysregulation and fathers were more disturbed by academic difficulties. In the PKU sample, the higher the phenylalanine level in the child, the more likely the parent and child alike were to attribute childhood problems to external loci of control. Attributional styles appear not to be diagnosis-specific in pediatric populations. Attributional style may be an important variable to consider when targeting treatment adherence.


The use of arthrography in pediatric orthopaedic surgery.
Surgery near pediatric joints can be challenging because it is difficult to visualize vital articular structures. Assessment of underlying pathology is also challenging because the joint structures have not yet ossified. Arthrography is a useful tool that is quick and minimally invasive and allows adequate visualization of joint anatomy during surgery, which aids intraoperative decision making. In pediatric patients, arthrography is most useful for visualization of the elbow, knee, and hip joints. This tool can help the surgeon to refine the diagnosis and management of pediatric fractures and aids in surgical assessment during joint and limb reconstruction. Arthrography adds minimal time to surgery and carries a low risk of complications; it should be part of the armamentarium of any surgeon who treats pediatric orthopaedic patients.


A blinded randomized assessment of laser Doppler flowmetry efficacy in standardizing outcome from intraluminal filament MCAO in the rat.
Laser Doppler flowmetry (LDF) is widely used for estimating cerebral blood flow changes during intraluminal middle cerebral artery occlusion (MCAO). No investigation has systematically examined LDF efficacy in standardizing outcome. We examined MCAO histologic and behavioral outcome as a function of LDF measurement. Rats were subjected to 90min MCAO by 4 surgeons having different levels of MCAO surgical experience. LDF was measured in all rats during ischemia. By random assignment, LDF values were (Assisted) or were not (Blinded) made available to each surgeon during MCAO (n=12-17 per group). Neurologic and histologic outcomes were measured 7 days post-MCAO. A second study examined LDF effects on 1-day post-MCAO outcome. Pooled across surgeons, intra-ischemic %LDF change (P=0.12), neurologic scores (Assisted vs. Blinded=14±6 vs. 13±7, P=0.61, mean±standard deviation) and cerebral infarct volume (162±63mm(3)vs. 143±86mm(3), P=0.24) were not different between groups. Only for one surgeon (novice) did LDF use alter infarct volume (145±28mm(3)vs. 98±61mm(3), P=0.03). LDF use decreased infarct volume coefficient of variation (COV) by 35% (P=0.02), but had no effect on neurologic score COV. We compared intraluminal MCAO outcome as a function of LDF use. LDF measurement altered neither neurologic nor histologic MCAO outcome. LDF did not decrease neurologic deficit COV, but did decrease infarct volume COV. LDF may allow use of fewer animals if infarct volume is the primary dependent variable, but is unlikely to impact requisite sample sizes if neurologic function is of primary interest.


Spiroplasma spp. from transmissible spongiform encephalopathy brains or ticks induce spongiform encephalopathy in ruminants.
Spiroplasma, small motile wall-less bacteria, are linked by molecular and serological studies to the transmissible spongiform encephalopathies (TSEs), which include scrapie in sheep, chronic wasting disease (CWD) in deer and Creutzfeldt-Jakob disease in humans. In this study, two experiments were undertaken to determine the role of spiroplasma in the pathogenesis of TSE. In experiment 1, Spiroplasma mirum, a rabbit tick isolate that had previously been shown to experimentally induce spongiform encephalopathy in rodents, was inoculated intracranially (IC) into ruminants. S. mirum-inoculated deer manifested clinical signs of TSE after 1.5 to 5.5 months incubation. The deer, as well as sheep and goats, inoculated with S. mirum developed spongiform encephalopathy in a dose-dependent manner. In experiment 2, spiroplasma closely related to S. mirum were isolated from TSE-affected brains via passage in embryonated eggs, and propagated in cell-free M1D media. Spiroplasma spp. isolates from scrapie-affected sheep brain and from CWD-affected deer brain inoculated IC into sheep and goats induced spongiform encephalopathy closely resembling natural TSE in these animals. These data show spiroplasma to be consistently associated with TSE, and able experimentally to cause TSE in ruminant animal models, therein questioning the validity of studies that have concluded the prion, a miss-folded protease-resistant protein that builds up in TSE brains during the course of the disease, to be the sole causal agent. The spiroplasma infection models reported here will be important for investigating factors involved in the pathogenesis of TSE since ruminants are the natural hosts.


Autoimmune uveitis: study of treatment therapies.
Experimental autoimmune uveitis is an organ-specific T-cell mediated autoimmune disease characterized by inflammation and consequent destruction of the neural retina and adjacent tissues. Inflammation in experimental autoimmune uveitis may be induced in rodents by immunization with retinal antigens, such as interphotoreceptor retinoid-binding protein. We present a review of experimental studies that correlate primary immunobiological functions with this chronic disease and the possible use of molecules for the treatment of autoimmune uveitis.


African Haplogroup L mtDNA sequences show violations of clock-like evolution.
A set of 96 complete mtDNA sequences that belong to the three major African haplogroups (L1, L2, and L3) was analyzed to determine if mtDNA has evolved as a molecular clock. Likelihood ratio tests (LRTs) were carried out with each of the haplogroups and with combined haplogroup sequence sets. Evolution has not been clock-like, neither for the coding region nor for the control region, in combined sets of African haplogroup L mtDNA sequences. In tests of individual haplogroups, L2 mtDNAs showed violations of a molecular clock under all conditions and in both the control and coding regions. In contrast, haplogroup L1 and L3 sequences, both for the coding and control regions, show clock-like evolution. In clock tests of individual L2 subclades, the L2a sequences showed a marked violation of clock-like evolution within the coding region. In addition, the L2a and L2c branch lengths of both the coding and control regions were shorter relative to those of the L2b and L2d sequences, a result that indicates lower levels of sequence divergence. Reduced median network analyses of the L2a sequences indicated the occurrence of marked homoplasy at multiple sites in the control region. After exclusion of the L2a and L2c sequences, African mtDNA coding region evolution has not significantly departed from a molecular clock, despite the results of neutrality tests that indicate the mitochondrial coding region has evolved under nonneutral conditions. In contrast, control region evolution is clock-like only at the haplogroup level, and it thus appears to have evolved essentially independently from the coding region. The results of the clock tests, the network analyses, and the branch length comparisons all caution against the use of simple mtDNA clocks.


Non-dopaminergic treatments for motor control in Parkinson's disease.
The pathological processes underlying Parkinson's disease (PD) involve more than dopamine cell loss within the midbrain. These non-dopaminergic neurotransmitters include noradrenergic, serotonergic, glutamatergic, and cholinergic systems within cortical, brainstem and basal ganglia regions. Several non-dopaminergic treatments are now in clinical use to treat motor symptoms of PD, or are being evaluated as potential therapies. Agents for symptomatic monotherapy and as adjunct to dopaminergic therapies for motor symptoms include adenosine A2A antagonists and the mixed monoamine-B inhibitor (MAO-BI) and glutamate release agent safinamide. The largest area of potential use for non-dopaminergic drugs is as add-on therapy for motor fluctuations. Thus adenosine A2A antagonists, safinamide, and the antiepileptic agent zonisamide can extend the duration of action of levodopa. To reduce levodopa-induced dyskinesia, drugs that target overactive glutamatergic neurotransmission can be used, and include the non-selective N-methyl D-aspartate antagonist amantadine. More recently, selective metabotropic glutamate receptor (mGluR₅) antagonists are being evaluated in phase II randomized controlled trials. Serotonergic agents acting as 5-HT2A/2C antagonists, such as the atypical antipsychotic clozapine, may also reduce dyskinesia. 5-HT1A agonists theoretically can reduce dyskinesia, but in practice, may also worsen PD motor symptoms, and so clinical applicability has not yet been shown. Noradrenergic α2A antagonism using fipamezole can potentially reduce dyskinesia. Several non-dopaminergic agents have also been investigated to reduce non-levodopa-responsive motor symptoms such as gait and tremor. Thus the cholinesterase inhibitor donepezil showed mild benefit in gait, while the predominantly noradrenergic re-uptake inhibitor methylphenidate had conflicting results in advanced PD subjects. Tremor in PD may respond to muscarinic M4 cholinergic antagonists (anticholinergics), but tolerability is often poor. Alternatives include β-adrenergic antagonists such as propranolol. Other options include 5-HT2A antagonists, and drugs that have mixed binding properties involving serotonin and acetylcholine, such as clozapine and the antidepressant mirtazapine, can be effective in reducing PD tremor. Many other non-dopaminergic agents are in preclinical and phase I/II early stages of study, and the reader is directed to recent reviews. While levodopa remains the most effective agent to treat motor symptoms in PD, the overall approach to using non-dopaminergic drugs in PD is to reduce reliance on levodopa and to target non-levodopa-responsive symptoms.


Quality of reporting statistics in two Indian pharmacology journals.
To evaluate the reporting of the statistical methods in articles published in two Indian pharmacology journals. All original articles published since 2002 were downloaded from the journals' (Indian Journal of Pharmacology (IJP) and Indian Journal of Physiology and Pharmacology (IJPP)) website. These articles were evaluated on the basis of appropriateness of descriptive statistics and inferential statistics. Descriptive statistics was evaluated on the basis of reporting of method of description and central tendencies. Inferential statistics was evaluated on the basis of fulfilling of assumption of statistical methods and appropriateness of statistical tests. Values are described as frequencies, percentage, and 95% confidence interval (CI) around the percentages. Inappropriate descriptive statistics was observed in 150 (78.1%, 95% CI 71.7-83.3%) articles. Most common reason for this inappropriate descriptive statistics was use of mean ± SEM at the place of "mean (SD)" or "mean ± SD." Most common statistical method used was one-way ANOVA (58.4%). Information regarding checking of assumption of statistical test was mentioned in only two articles. Inappropriate statistical test was observed in 61 (31.7%, 95% CI 25.6-38.6%) articles. Most common reason for inappropriate statistical test was the use of two group test for three or more groups. Articles published in two Indian pharmacology journals are not devoid of statistical errors.


Long-term effects of artificial nighttime lighting and trophic complexity on plant biomass and foliar carbon and nitrogen in a grassland community.
The introduction of artificial nighttime lighting due to human settlements and transport networks is increasingly altering the timing, intensity, and spectra of natural light regimes worldwide. Much of the research on the impacts of nighttime light pollution on organisms has focused on animal species. Little is known about the impacts of daylength extension due to outdoor lighting technologies on wild plant communities, despite the fact that plant growth and development are under photoperiodic control. In a five-year field experiment, artificial ecosystems ("mesocosms") of grassland communities both alone or in combination with invertebrate herbivores and predators were exposed to light treatments that simulated street lighting technologies (low-pressure sodium, and light-emitting diode [LED]-based white lighting), at ground-level illuminance. Most of the plant species in the mesocosms did not exhibit changes in biomass accumulation after 5 years of exposure to the light treatments. However, the white LED treatment had a significant negative effect on biomass production in the herbaceous species Lotus pedunculatus. Likewise, the interaction between the white LED treatment and the presence of herbivores significantly reduced the mean shoot/root ratio of the grass species Holcus lanatus. Artificial nighttime lighting had no effect on the foliar carbon or nitrogen in most of the grassland species. Nevertheless, the white LED treatment significantly increased the leaf nitrogen content in Lotus corniculatus in the presence of herbivores. Long-term exposure to artificial light at night had no general effects on plant biomass responses in experimental grassland communities. However, species-specific and negative effects of cool white LED lighting at ground-level illuminance on biomass production and allocation in mixed plant communities are suggested by our findings. Further studies on the impacts of light pollution on biomass accumulation in plant communities are required as these effects could be mediated by different factors, including herbivory, competition, and soil nutrient availability.


MHC2TA promoter polymorphism (-168*G/A, rs3087456) is not associated with susceptibility to rheumatoid arthritis in British Caucasian rheumatoid arthritis patients.
To investigate the association of a single-nucleotide polymorphism (SNP) in the promoter region of MHC2TA gene (-168*G/A, rs3087456), which has previously been described in a Swedish rheumatoid arthritis (RA) cohort, in British Caucasian RA patients. We genotyped 733 RA patients and 613 healthy controls for MHC2TA -168*G/A SNP by amplification-refractory mutation system (ARMS). Data were analysed using SPSS version 13.0 software and the chi-square test was applied where appropriate. The MHC2TA -168*G/A SNP was not associated with increased susceptibility to RA in our patients. Stratifying the patients according to the presence or absence of rheumatoid factor (RF) showed the SNP to be more common in RF negative patients, but this did not reach statistical significance. We did not confirm the previously reported association of this MHC2TA polymorphism with RA in our UK population despite its ethnic similarities with the Swedish population in which it was first described.


Determinants of surface membrane maturation during the cercarial-schistosomula transformation of Schistosoma mansoni.
The transformation of free-living cercariae to parasitic schistosomula of Schistosoma mansoni is accompanied by the formation of a multilaminate surface membrane which binds concanavalin A (Con A). This transition normally occurs with penetration of intact skin of the mammalian host, but can be accomplished by vortexing and incubating organisms at 37 degrees C in complex tissue culture medium (e.g., Earle's lactalbumin). We determined the minimal conditions that initiate surface maturation by monitoring surface changes by transmission electron microscopy and surface fluorescein-conjugated Con A labeling. Cercariae incubated in water for 2 hr at 37 degrees C retained their trilaminate surface and were non-fluorescent. After incubation in 116 mM NaCl for 3 hr at 23 degrees C, no organisms were covered by intense surface fluorescence; increasing the temperature to 37 degrees C resulted in 50% of the parasites becoming fluorescent. The addition of 26 mM NaHCO3 (pH 7.4) to parasites incubated in NaCl at 37 degrees C resulted in 98% of the organisms binding Con A. No significant difference in surface Con A binding was found between organisms incubated with the chelators EGTA or EDTA and controls. Mechanical stimulus by itself was not effective in stimulating surface maturation. The conditions that initiated surface maturation were elevation of temperature to 37 degrees C and NaHCO3-containing medium.


Stigma, sexual risk and desire for HIV tests in Ghana.
Although various HIV prevention and treatment programs have been implemented in Ghana since 2003, desire for and uptake of HIV testing is still low, owing largely to HIV-related stigma. This study examined the effect of stigma on desire for HIV testing, while controlling for socio-demographic and other influences. The study used data from the 2003 Ghana Demographic and Health Survey to regress desire for an HIV test on levels of stigma while controlling for selected socio-demographic, socio-cognitive and socio-behavioural covariates. The study revealed significant associations between several socio-demographic and socio-cognitive variables and the desire for an HIV test. For example, both male (adjusted odds ratio (AOR) = 0.41; 95% confidence interval (CI) = 0.24-0.72) and female (AOR = 0.62; 95% CI = 0.41-0.93) respondents in the wealthiest households, and those who know someone infected with HIV (AOR = 0.65; 95% CI = 0.43-1.00) have lower odds of desiring an HIV test. The study showed a significant but negative interaction between risky sexual behaviours and community stigma (AOR = 0.44; 95% CI = 0.19-0.67), indicating that the positive effect of risky sex on HIV testing is attenuated among females who live in communities with high levels of stigma. Since community-level education and risk reduction programs have demonstrable influences on reducing HIV stigma, it is imperative that the Ghana government's ongoing anti-stigma campaigns and other HIV prevention programs recognise the role of community stigma in influencing HIV testing decisions, especially in the context of risky sexual behaviours.


Acceptability and feasibility of magnetic femoral nerve stimulation in older, functionally impaired patients.
Magnetic femoral nerve stimulation to test muscle function has been largely unexplored in older people. We assessed acceptability, feasibility, along with reproducibility and correlation with other physical function measures. Study 1 recruited older people with sarcopenia. Stimulation was performed at baseline and 2 weeks along with six minute walk (6MW), maximum voluntary quadriceps contraction, short physical performance battery and grip strength. Acceptability was measured using visual analog scales. Study 2 used baseline data from a trial of older people. We correlated stimulation results with 6MW, maximal voluntary contraction and muscle mass. Maximum quadriceps twitch tension was measured in both studies, evoked using biphasic magnetic stimulation of the femoral nerve. In study 1 (n = 12), magnetic stimulation was well tolerated with mean discomfort rating of 9% (range 0-40%) on a visual analog scale. Reproducibility was poor (intraclass correlation coefficient 0.06; p = 0.44). Study 2 (n = 64) showed only weak to moderate correlations for maximum quadriceps twitch tension with other measures of physical function (6 minute walk test r = 0.24, p = 0.06; maximal voluntary contraction r = 0.26; p = 0.04). We conclude that magnetic femoral nerve stimulation is acceptable and feasible but poorly reproducible in older, functionally impaired people.