Expert consensus statement on the science of HIV in the context of criminal law.
Globally, prosecutions for non-disclosure, exposure or transmission of HIV frequently relate to sexual activity, biting, or spitting. This includes instances in which no harm was intended, HIV transmission did not occur, and HIV transmission was extremely unlikely or not possible. This suggests prosecutions are not always guided by the best available scientific and medical evidence. Twenty scientists from regions across the world developed this Expert Consensus Statement to address the use of HIV science by the criminal justice system. A detailed analysis of the best available scientific and medical research data on HIV transmission, treatment effectiveness and forensic phylogenetic evidence was performed and described so it may be better understood in criminal law contexts. Description of the possibility of HIV transmission was limited to acts most often at issue in criminal cases. The possibility of HIV transmission during a single, specific act was positioned along a continuum of risk, noting that the possibility of HIV transmission varies according to a range of intersecting factors including viral load, condom use, and other risk reduction practices. Current evidence suggests the possibility of HIV transmission during a single episode of sex, biting or spitting ranges from no possibility to low possibility. Further research considered the positive health impact of modern antiretroviral therapies that have improved the life expectancy of most people living with HIV to a point similar to their HIV-negative counterparts, transforming HIV infection into a chronic, manageable health condition. Lastly, consideration of the use of scientific evidence in court found that phylogenetic analysis alone cannot prove beyond reasonable doubt that one person infected another although it can be used to exonerate a defendant. The application of up-to-date scientific evidence in criminal cases has the potential to limit unjust prosecutions and convictions. The authors recommend that caution be exercised when considering prosecution, and encourage governments and those working in legal and judicial systems to pay close attention to the significant advances in HIV science that have occurred over the last three decades to ensure current scientific knowledge informs application of the law in cases related to HIV.


High-Deductible Health Plans: Implications for Substance Use Treatment.
To examine whether high-deductible health plans are related to patient complexity, health services use, and medical care costs among substance use treatment patients. Electronic health record data from Kaiser Permanente Northern California; 2007-2011. Retrospective analysis of electronic health record data of substance use treatment patients (N = 31,001). We examined relationship of patient demographics, health comorbidities, and services use and cost to deductible level: none, low ($1-$999), and high (≥$1,000). Demographic, membership, diagnostic, and utilization data were merged with cost data. Utilization and costs were summarized into 6-month intervals. Generalized estimation methods for repeated measures with logistic, Poisson, and linear regression were used. Substance use patients with deductible plans were younger and had less comorbidity than those without deductibles. Patients with high deductibles had lower emergency room and hospital use 12- to 6-month pretreatment, but rates became similar to other groups in the 6 months immediately prior to treatment; treatment costs were similar. Immediately prior to entering treatment, substance use patients with and without high deductibles have similar patterns of health services utilization. We discuss implications for health policy and treatment, particularly in an era of health reform.


The learners' perspective on internal medicine ward rounds: a cross-sectional study.
Ward rounds form an integral part of Internal Medicine teaching. This study aimed to determine the trainees' opinions regarding various aspects of their ward rounds, including how well they cover their learning needs, how they would like the rounds to be conducted, and differences of opinion between medical students and postgraduates. A cross-sectional study was conducted on a total of 134 trainees in Internal Medicine, comprising medical students, interns, residents and fellows, who were asked to fill in a structured, self-designed questionnaire. Most of the responses required a rating on a scale of 1-5 (1 being highly unsatisfactory and 5 being highly satisfactory). Teaching of clinical skills and bedside teaching received the lowest overall mean score (Mean +/- SD 2.48 +/- 1.02 and 2.49 +/- 1.12 respectively). They were rated much lower by postgraduates as compared to students (p < 0.001). All respondents felt that management of patients was the aspect best covered by the current ward rounds (Mean +/- SD 3.71 +/- 0.72). For their desired ward rounds, management of patients received the highest score (Mean +/- SD 4.64 +/- 0.55), followed by bedside examinations (Mean +/- SD 4.60 +/- 0.61) and clinical skills teaching (Mean +/- SD 4.50 +/- 0.68). The postgraduates desired a lot more focus on communication skills, counselling and medical ethics as compared to students, whose primary focus was teaching of bedside examination and management. A majority of the respondents (87%) preferred bedside rounds over conference room rounds. Even though the duration of rounds was found to be adequate, a majority of the trainees (68%) felt there was a lack of individual attention during ward rounds. This study highlights important areas where ward rounds need improvement in order to maximize their benefit to the learners. There is a need to modify the current state of ward rounds in order to address the needs and expectations of trainees.


Overall Tumor Burden Dictates Outcomes for Patients Undergoing Resection of Multinodular Hepatocellular Carcinoma Beyond the Milan Criteria.
The objective of the current study was to define surgical outcomes after resection of multinodular hepatocellular carcinoma (HCC) beyond the Milan criteria, and develop a prediction tool to identify which patients likely benefit the most from resection. Liver resection for multinodular HCC, especially beyond the Milan criteria, remains controversial. Rigorous selection of the best candidates for resection is essential to achieve optimal outcomes after liver resection of advanced tumors. Patients who underwent resection for HCC between 2000 and 2017 were identified from an international multi-institutional database. Patients were categorized according to Milan criteria status. Pre- and postoperative overall survival (OS) prediction models that included HCC tumor burden score (TBS) among patients with multinodular HCC beyond Milan criteria were developed and validated. Among 1037 patients who underwent resection for HCC, 164 (15.8%) had multinodular HCC beyond the Milan criteria. Among patients with multinodular HCC, 25 (15.2%) patients experienced a serious complication and 90-day mortality was 3.7% (n = 6). Five-year OS after resection of multinodular HCC beyond Milan criteria was 52.8%. A preoperative TBS-based model (5-year OS: low-risk, 73.7% vs intermediate-risk, 45.1% vs high-risk, 13.1%), and postoperative TBS-based model (5-year OS: low-risk, 80.1% vs intermediate-risk, 37.2% vs high-risk, not reached) categorized patients into distinct prognostic groups relative to long-term prognosis (both P < 0.001). Pre- and postoperative models could accurately stratify OS in an external validation cohort (5-year OS; low vs medium vs high risk; pre: 66.3% vs 25.2% vs not reached, P = 0.012; post: 61.4% vs 42.5% vs not reached, P = 0.045) Predictive accuracy of the pre- and postoperative models was good in the training (c-index; pre: 0.68; post: 0.71), internal validation (n = 2000 resamples) (c-index, pre: 0.70; post: 0.72) and external validation (c-index, pre: 0.67; post 0.68) datasets. TBS alone could stratify patients relative to 5-year OS after resection of multinodular HCC beyond Milan criteria (c-index: 0.65; 5-year OS; low TBS: 70.2% vs medium TBS: 54.7% vs high TBS: 16.7%; P < 0.001). The vast majority of patients with low and intermediate TBS were deemed low or medium risk based on both the preoperative (98.4%) and postoperative risk scores (95.3%). Prognosis of patients with multinodular HCC was largely dependent on overall tumor burden. Liver resection should be considered among patients with multinodular HCC beyond the Milan criteria who have a low- or intermediate-TBS.


Lyssavirus matrix protein cooperates with phosphoprotein to modulate the Jak-Stat pathway.
Phosphoprotein (P) and matrix protein (M) cooperate to undermine the immune response to rabies virus (RABV) infections. While P is involved in the modulation of the Jak-Stat pathway through the cytoplasmic retention of interferon (IFN)-activated STAT1 (pSTAT1), M interacts with the RelAp43-p105-ABIN2-TPL2 complex, to efficiently inhibit the nuclear factor-κB (NF-κB) pathway. Using transfections, protein-complementation assays, reverse genetics and DNA ChIP, we identified a role of M protein in the control of Jak-Stat signaling pathway, in synergy with the P protein. In unstimulated cells, both M and P proteins were found to interact with JAK1. Upon type-I IFN stimulation, the M switches toward pSTAT1 interaction, which results in an enhanced capacity of P protein to interact with pSTAT1 and restrain it in the cytoplasm. Furthermore, the role for M-protein positions 77, 100, 104 and 110 was also demonstrated in interaction with both JAK1 and pY-STAT1, and confirmed in vivo. Together, these data indicate that M protein cooperates with P protein to restrain in parallel, and sequentially, NF-κB and Jak-Stat pathways.


Nanopore sequencing and full genome de novo assembly of human cytomegalovirus TB40/E reveals clonal diversity and structural variations.
Human cytomegalovirus (HCMV) has a double-stranded DNA genome of approximately 235 Kbp that is structurally complex including extended GC-rich repeated regions. Genomic recombination events are frequent in HCMV cultures but have also been observed in vivo. Thus, the assembly of HCMV whole genomes from technologies producing shorter than 500 bp sequences is technically challenging. Here we improved the reconstruction of HCMV full genomes by means of a hybrid, de novo genome-assembly bioinformatics pipeline upon data generated from the recently released MinION MkI B sequencer from Oxford Nanopore Technologies. The MinION run of the HCMV (strain TB40/E) library resulted in ~ 47,000 reads from a single R9 flowcell and in ~ 100× average read depth across the virus genome. We developed a novel, self-correcting bioinformatics algorithm to assemble the pooled HCMV genomes in three stages. In the first stage of the bioinformatics algorithm, long contigs (N50 = 21,892) of lower accuracy were reconstructed. In the second stage, short contigs (N50 = 5686) of higher accuracy were assembled, while in the final stage the high quality contigs served as template for the correction of the longer contigs resulting in a high-accuracy, full genome assembly (N50 = 41,056). We were able to reconstruct a single representative haplotype without employing any scaffolding steps. The majority (98.8%) of the genomic features from the reference strain were accurately annotated on this full genome construct. Our method also allowed the detection of multiple alternative sub-genomic fragments and non-canonical structures suggesting rearrangement events between the unique (UL /US) and the repeated (T/IRL/S) genomic regions. Third generation high-throughput sequencing technologies can accurately reconstruct full-length HCMV genomes including their low-complexity and highly repetitive regions. Full-length HCMV genomes could prove crucial in understanding the genetic determinants and viral evolution underpinning drug resistance, virulence and pathogenesis.


Unravelling selection signatures in a single dog breed suggests recent selection for morphological and behavioral traits.
Strong selection has resulted in substantial morphological and behavioral diversity across modern dog breeds, which makes dogs interesting model animals to study the underlying genetic architecture of these traits. However, results from between-breed analyses may confound selection signatures for behavior and morphological features that were coselected during breed development. In this study, we assess population genetic differences in a unique resource of dogs of the same breed but with systematic behavioral selection in only one population. We exploit these different breeding backgrounds to identify signatures of recent selection. Selection signatures within populations were found on chromosomes 4 and 19, with the strongest signals in behavior-related genes. Regions showing strong signals of divergent selection were located on chromosomes 1, 24, and 32, and include candidate genes for both physical features and behavior. Some of the selection signatures appear to be driven by loci associated with coat color (Chr 24; ASIP) and length (Chr 32; FGF5), while others showed evidence of association with behavior. Our findings suggest that signatures of selection within dog breeds have been driven by selection for morphology and behavior. Furthermore, we demonstrate that combining selection scans with association analyses is effective for dissecting the traits under selection.


Hypoxia-responsive micelles self-assembled from amphiphilic block copolymers for the controlled release of anticancer drugs.
Amphiphilic block copolymers poly(ethylene glycol)-block-poly(methacrylic acid-co-2-nitroimidazole methacrylate) (PEG-b-P(MAA-co-NIMA)) were synthesized by the combination of atom transfer radical polymerization (ATRP), hydrolysis and EDC reactions. These copolymers could self-assemble into spherical micelles in water. 2-Nitroimidazole (NI) groups presented hypoxia-responsive properties under hypoxia conditions. The hydrophobic NI groups could be converted into hydrophilic aminoimidazole (AI) groups, which would lead to the expansion of micelles. Moreover, the content of NI groups in the copolymers would affect the hydrophilic-hydrophobic balance and therefore influence the self-assembly behaviour of the copolymer and the morphologies of the micelles. The copolymer micelles were used as a drug delivery system for controlled release of anticancer drug doxorubicin (DOX). The in vitro cytotoxicity investigation revealed that the DOX-loaded micelles showed higher toxicity to hypoxic cells than to normoxic cells. As a result, the block copolymers are expected to be used as an intelligent carrier for hydrophobic drugs to treat hypoxia-associated diseases.


Feasibility and Preliminary Efficacy of a 10-Week Resistance and Aerobic Exercise Intervention During Neoadjuvant Chemoradiation Treatment in Rectal Cancer Patients.
Neoadjuvant chemoradiation treatment (CRT) in rectal cancer patients is associated with a reduction in physical capacity, lean mass and increased fatigue. As a countermeasure to these treatment-related adverse effects, we examined the feasibility and preliminary efficacy of a 10-week exercise program during CRT. Ten rectal cancer patients (7 men, aged 27-70 years, body mass index = 26.4 ± 3.8 kg/m2) receiving CRT undertook supervised resistance and aerobic exercise twice weekly. Assessments were undertaken pre- and post-intervention for upper and lower body muscle strength by 1-RM, muscle endurance, physical performance tests, body composition by dual X-ray absorptiometry, quality of life, and fatigue. There was a significant loss in appendicular skeletal muscle (-1.1 kg, P = .012), and fat mass (-0.8 kg, P = .029) following CRT. Despite the loss in skeletal muscle, leg press ( P = .030) and leg extension ( P = .046) strength improved by 27.2% and 22.7%, respectively, and leg press endurance by 76.7% ( P = .007). Changes in strength were accompanied by improved performance ( P < .05) in 6-m fast walking speed (6.9%) and dynamic balance as determined by the 6-m backwards walk (15.5%). There was minimal change in quality of life and fatigue, and no adverse events related to training. Exercise during neoadjuvant CRT appears to be feasible and well tolerated in rectal cancer patients and may enhance physical function while minimizing adverse changes in body composition and cancer-related fatigue. These initial findings need to be confirmed in randomized controlled trials.


Clarification of Photorespiratory Processes and the Role of Malic Enzyme in Diatoms.
Evidence suggests that diatom photorespiratory metabolism is distinct from other photosynthetic eukaryotes in that there may be at least two routes for the metabolism of the photorespiratory metabolite glycolate. One occurs primarily in the mitochondria and is similar to the C2 photorespiratory pathway, and the other processes glycolate through the peroxisomal glyoxylate cycle. Genomic analysis has identified the presence of key genes required for glycolate oxidation, the glyoxylate cycle, and malate metabolism, however, predictions of intracellular localization can be ambiguous and require verification. This knowledge gap leads to uncertainties surrounding how these individual pathways operate, either together or independently, to process photorespiratory intermediates under different environmental conditions. Here, we combine in silico sequence analysis, in vivo protein localization techniques and gene expression patterns to investigate key enzymes potentially involved in photorespiratory metabolism in the model diatom Thalassiosira pseudonana. We demonstrate the peroxisomal localization of isocitrate lyase and the mitochondrial localization of malic enzyme and a glycolate oxidase. Based on these analyses, we propose an updated model for photorespiratory metabolism in T. pseudonana, as well as a mechanism by which C2 photorespiratory metabolism and its associated pathways may operate during silicon starvation and growth arrest.


Environmental impacts of milk powder and butter manufactured in the Republic of Ireland.
The abolition of the milk quota system that was in place in Europe was abolished in 2015, which instigated an immediate increase in milk production in many European countries. This increase will aid in addressing the world's ever growing demand for food, but will incur increased stresses on the environmental impact and sustainability of the dairy industry. In this study, an environmental life cycle assessment was performed in order to estimate the environmental impacts associated with the manufacture of milk powder and butter in the Republic of Ireland. A farm gate to processing factory gate analysis, which includes raw milk transportation, processing into each product and packaging, is assessed in this study. Operational data was obtained from 5 dairy processing factories that produce milk powder (4 of which also produce butter). Results for each environmental impact category are presented per kilogram of product. Energy consumption (raw milk transportation and on-site electrical and thermal energy usage) contributes, on average, 89% and 78% of the total global warming potential, for milk powder and butter respectively, for the life cycle stages assessed. Similarly, energy consumption contributes, on average, 86% and 96% of the total terrestrial acidification potential for milk powder and butter respectively, for these life cycle stages. Emissions associated with wastewater treatment contribute approximately 10% and 40% to the total freshwater eutrophication potential and marine eutrophication potential, respectively, for both milk powder and butter production. In addition, packaging materials also has a significant contribution to these environmental impact categories for butter production. Results were also presented for three milk powder products being manufactured by the factories surveyed: skim milk powder, whole milk powder and full fat milk powder. The analysis presented in this paper helps to identify opportunities to reduce the environmental impacts associated with post-farm processing of milk powder and butter.


Influenza human monoclonal antibody 1F1 interacts with three major antigenic sites and residues mediating human receptor specificity in H1N1 viruses.
Most monoclonal antibodies (mAbs) to the influenza A virus hemagglutinin (HA) head domain exhibit very limited breadth of inhibitory activity due to antigenic drift in field strains. However, mAb 1F1, isolated from a 1918 influenza pandemic survivor, inhibits select human H1 viruses (1918, 1943, 1947, and 1977 isolates). The crystal structure of 1F1 in complex with the 1918 HA shows that 1F1 contacts residues that are classically defined as belonging to three distinct antigenic sites, Sa, Sb and Ca(2). The 1F1 heavy chain also reaches into the receptor binding site (RBS) and interacts with residues that contact sialoglycan receptors and determine HA receptor specificity. The 1F1 epitope is remarkably similar to the previously described murine HC63 H3 epitope, despite significant sequence differences between H1 and H3 HAs. Both antibodies potently inhibit receptor binding, but only HC63 can block the pH-induced conformational changes in HA that drive membrane fusion. Contacts within the RBS suggested that 1F1 may be sensitive to changes that alter HA receptor binding activity. Affinity assays confirmed that sequence changes that switch the HA to avian receptor specificity affect binding of 1F1 and a mAb possessing a closely related heavy chain, 1I20. To characterize 1F1 cross-reactivity, additional escape mutant selection and site-directed mutagenesis were performed. Residues 190 and 227 in the 1F1 epitope were found to be critical for 1F1 reactivity towards 1918, 1943 and 1977 HAs, as well as for 1I20 reactivity towards the 1918 HA. Therefore, 1F1 heavy-chain interactions with conserved RBS residues likely contribute to its ability to inhibit divergent HAs.


Nanoscale materials for hyperthermal theranostics.
Recently, the use of nanoscale materials has attracted considerable attention with the aim of designing personalized therapeutic approaches that can enhance both spatial and temporal control over drug release, permeability, and uptake. Potential benefits to patients include the reduction of overall drug dosages, enabling the parallel delivery of different pharmaceuticals, and the possibility of enabling additional functionalities such as hyperthermia or deep-tissue imaging (LIF, PET, etc.) that complement and extend the efficacy of traditional chemotherapy and surgery. This mini-review is focused on an emerging class of nanometer-scale materials that can be used both to heat malignant tissue to reduce angiogenesis and DNA-repair while simultaneously offering complementary imaging capabilities based on radioemission, optical fluorescence, magnetic resonance, and photoacoustic methods.


Immediate and delayed effects of growth conditions on ageing parameters in nestling zebra finches.
Conditions experienced during development and growth are of crucial importance as they can have a significant influence on the optimisation of life histories. Indeed, the ability of an organism to grow fast and achieve a large body size often confers short- and long-term fitness benefits. However, there is good evidence that organisms do not grow at their maximal rates as growth rates seem to have potential costs on subsequent lifespan. There are several potential proximate causes of such a reduced lifespan. Among them, one emerging hypothesis is that growth impacts adult survival and/or longevity through a shared, end point, ageing mechanism: telomere erosion. In this study, we manipulated brood size in order to investigate whether rapid growth (chicks in reduced broods) is effectively done at the cost of a short- (end of growth) and long-term (at adulthood) increase of oxidative damage and telomere loss. Contrary to what we expected, chicks from the enlarged broods displayed more oxidative damage and had shorter telomeres at the end of the growth period and at adulthood. Our study extends the understanding of the proximate mechanisms involved in the trade-off between growth and ageing. It highlights that adverse environmental conditions during growth can come at a cost via transient increased oxidative stress and pervasive eroded telomeres. Indeed, it suggests that telomeres are not only controlled by intrinsic growth rates per se but also may be under the control of some extrinsic environmental factors, which could complicate our understanding of the growth-ageing interaction.


Adaptive color polymorphism and unusually high local genetic diversity in the side-blotched lizard, Uta stansburiana.
Recently, studies of adaptive color variation have become popular as models for examining the genetics of natural selection. We examined color pattern polymorphism and genetic variation in a population of side-blotched lizards (Uta stansburiana) that is found in habitats with both dark (lava) and light colored (granite) substrates. We conducted a limited experiment for adult phenotypic plasticity in laboratory conditions. We recorded both substrate and lizard color patterns in the field to determine whether lizards tended to match their substrate. Finally we examined genetic variation in a gene (melanocortin 1 receptor) that has been shown to affect lizard color in other species and in a presumably neutral gene (mitochondrial cytochrome b). Populations were sampled in the immediate area of the lava flows as well as from a more distant site to examine the role of population structure. Our captive Uta did not change color to match their background. We show that side-blotched lizards tend to match the substrate on which it was caught in the field and that variation in the melanocortin 1 receptor gene does not correlate well with color pattern in this population. Perhaps the most remarkable result is that this population of side-blotched lizards shows extremely high levels of variation at both genetic markers, in the sense of allele numbers, with relatively low levels of between-allele sequence variation. Genetic variation across this small region was as great or greater than that seen in samples of pelagic fish species collected worldwide. Statistical analysis of genetic variation suggests rapid population expansion may be responsible for the high levels of variation.


Evolutionary constraints on chaperone-mediated folding provide an antiviral approach refractory to development of drug resistance.
The genome diversity of RNA viruses allows for rapid adaptation to a wide variety of adverse conditions. Accordingly, viruses can escape inhibition by most antiviral compounds targeting either viral or host factors. Here we exploited the capacity of RNA viruses for rapid adaptation to explore the evolutionary constraints of chaperone-mediated protein folding. We hypothesized that inhibiting a host molecular chaperone required for folding of a viral protein would force the virus to evolve an alternate folding strategy. We identified the chaperone Hsp90 as an essential factor for folding and maturation of picornavirus capsid proteins. Pharmacological inhibition of Hsp90 impaired the replication of poliovirus, rhinovirus, and coxsackievirus in cell culture. Strikingly, anti-Hsp90 treatment did not yield drug-resistant viruses, suggesting that the complexity of capsid folding precludes the emergence of alternate folding pathways. These results reveal tight evolutionary constraints on chaperone-mediated protein folding, which may be exploited for viral inhibition in vivo. Indeed, Hsp90 inhibitors drastically reduced poliovirus replication in infected animals without the emergence of drug-resistant escape mutants. We propose that targeting folding of viral proteins may provide a general antiviral strategy that is refractory to development of drug resistance.


Laser Self-Mixing Fiber Bragg Grating Sensor for Acoustic Emission Measurement.
Fiber Bragg grating (FBG) is considered a good candidate for acoustic emission (AE) measurement. The sensing and measurement in traditional FBG-based AE systems are based on the variation in laser intensity induced by the Bragg wavelength shift. This paper presents a sensing system by combining self-mixing interference (SMI) in a laser diode and FBG for AE measurement, aiming to form a new compact and cost-effective sensing system. The measurement model of the overall system was derived. The performance of the presented system was investigated from both aspects of theory and experiment. The results show that the proposed system is able to measure AE events with high resolution and over a wide dynamic frequency range.


Chalcogen-height dependent magnetic interactions and magnetic order switching in FeSexTe1-x.
Magnetic properties of iron chalcogenide superconducting materials are investigated using density-functional calculations. We find that the stability of magnetic phases is very sensitive to the height of chalcogen species from the Fe plane: while FeTe with optimized Te height has the double-stripe (pi, 0) magnetic ordering, the single-stripe (pi, pi) ordering becomes the ground state when Te is lowered below a critical height by, e.g., Se doping. This behavior is understood by opposite Te-height dependences of the superexchange interaction and a longer range magnetic interaction mediated by itinerant electrons. We also demonstrate a linear temperature dependence of the macroscopic magnetic susceptibility in the single-stripe phase in contrast with the constant behavior in the double-stripe phase. Our findings provide a comprehensive and unified view on the magnetism in FeSexTe1-x and iron pnictide superconductors.


Intake of red wine increases the number and functional capacity of circulating endothelial progenitor cells by enhancing nitric oxide bioavailability.
Red wine (RW) consumption has been associated with a reduction of cardiovascular events, but limited data are available on potential mediating mechanisms. This study tested the hypothesis that intake of RW may promote the circulating endothelial progenitor cell (EPC) level and function through enhancement of nitric oxide bioavailability. Eighty healthy, young subjects were randomized and assigned to consume water (100 mL), RW (100 mL), beer (250 mL), or vodka (30 mL) daily for 3 weeks. Flow cytometry was used to quantify circulating EPC numbers, and in vitro assays were used to evaluate EPC functions. After RW ingestion, endothelial function determined by flow-mediated vasodilation was significantly enhanced; however, it remained unchanged after water, beer, or vodka intake. There were significantly increased numbers of circulating EPC (defined as KDR(+)CD133(+), CD34(+)CD133(+), CD34(+)KDR(+)) and EPC colony-forming units only in the RW group (all P<0.05). Only RW ingestion significantly enhanced plasma levels of nitric oxide and decreased asymmetrical dimethylarginine (both P<0.01). Incubation of EPC with RW (but not beer or ethanol) and resveratrol in vitro attenuated tumor necrosis factor-alpha-induced EPC senescence and improved tumor necrosis factor-alpha-suppressed EPC functions and tube formation. Incubation with nitric oxide donor sodium nitroprusside significantly ameliorated the inhibition of tumor necrosis factor-alpha on EPC proliferation, but incubation with endothelial nitric oxide synthase inhibitor l-NAME and PI3K inhibitor markedly attenuated the effect of RW on EPC proliferation. The intake of RW significantly enhanced circulating EPC levels and improved EPC functions by modifying nitric oxide bioavailability. These findings may help explain the beneficial effects of RW on the cardiovascular system. This study demonstrated that a moderate intake of RW can enhance circulating levels of EPC in healthy subjects by increasing nitric oxide availability. Direct incubation of EPC with RW and resveratrol can modify the functions of EPC, including attenuation of senescence and promotion of EPC adhesion, migration, and tube formation. These data suggest that RW ingestion may alter the biology of EPC, and these alterations may contribute to its unique cardiovascular-protective effect.


Energy metabolism and contractile function after 15 beats of moderate myocardial ischemia.
Difficulties in studying myocardial metabolism with adequate time resolution have led to contradictory conclusions regarding the mechanisms causing contractile abnormalities during the early stages of ischemia. In acutely instrumented swine, we investigated whether abnormalities in subendocardial ATP, phosphocreatine, or lactate content develop rapidly enough during the first few heart beats after onset of partial myocardial ischemia to contribute to contractile failure. Within the first 15 beats of a 40-50% reduction in left anterior descending coronary artery blood flow, regional myocardial function was significantly reduced but continuing to deteriorate. Rapidly frozen transmural left ventricular biopsies obtained on the 15th heart beat (+/- 1.5 beats) after the onset of ischemia revealed significant decrements in subendocardial phosphocreatine and ATP levels to 77% (p less than 0.05) and 84% (p less than 0.005) of control values, respectively, but minimal change in lactate content. Metabolic effects as assessed by transmural averages took longer to become detectable; thus, there was a tendency to underestimate the importance of subendocardial metabolic effects on myocardial function. When left ventricular preload was assessed during this early time period, left ventricular end-diastolic wall thickness only decreased by 3%, and left ventricular end-diastolic pressure did not change significantly despite a large fall in coronary perfusion pressure. Thus, in an in vivo pig model with techniques optimized to detect subendocardial metabolic changes within the period of very early moderate myocardial ischemia, abnormalities in high energy phosphate compounds occurred rapidly enough to contribute to developing myocardial dysfunction, whereas preload-mediated mechanisms related to vascular distending pressure could not explain the functional deterioration under these conditions.


"Research" and "science" in the first half of the chiropractic century.
In the first 50 years of the chiropractic profession, a variety of unorthodox meanings for the terms "research," "science" and related words were in evidence. In harmony with popular conceptions of the day, science was constructed as a relatively static body of knowledge and was thought to reflect the will of God. Research was an ill-defined activity, and acquisition of new knowledge did not involve the experimental methodology that increasingly took hold in biology and medicine in the twentieth century. Chiropractors often viewed science and research as marketing strategies. Clinical data collection, when it occurred at all, was not described in sufficient detail to permit replication. Results were enthusiastically interpreted as indisputable proof of investigators' a priori assumptions about the effectiveness of chiropractic methods. A few in the profession recognized the general lack of understanding of the scientific method and sought reform from within. However, the colleges were unwilling to introduce coursework in research methods. At the end of World War II, the broad-scope national association of chiropractors in the United States established a nonprofit foundation for the purpose of raising funds for chiropractic research and education. Research plans were poorly conceived and grandiose: the first major initiative of the Chiropractic Research Foundation involved a nationwide publicity and fund-raising campaign modeled after the March of Dimes. When these efforts failed and the possibility of establishing free-standing research centers collapsed, the Foundation sought to shift responsibility for research to the schools. The poverty-stricken chiropractic colleges lacked the research sophistication for this task. Several more decades would pass before a sustained research effort and interest in clinical experimentation would become evident in chiropractic.


Toxoplasma gondii infection in meat animals from Africa: Systematic review and meta-analysis of sero-epidemiological studies.
Toxoplasma gondii is an ubiquitous apicomplexan parasite which causes toxoplasmosis in humans and animals. Felids especially cats are definitive hosts and almost all warm-blooded mammals, including livestock and human can serve as intermediate hosts. Food animals can be reservoirs for T. gondii and act as one of the sources for parasite transmission to humans. The objective of this study is to collect serological data on the prevalence of anti-T. gondii antibody, and risk factors for certain food animals from Africa to provide a quantitative estimate of T. gondii infection among these species from different African countries. Four databases were used to search seroepidemiological data on the prevalence of anti-T. gondii antibody in food animals between 1969 and 2016 from African countries. The search focused on data obtained by serologic test in food animals and meta-analyses were performed per species. A total of 30,742 individual samples from 24 countries, described in 68 articles were studied. The overall estimated prevalence for toxoplasmosis in chicken, camel, cattle, sheep, goat, pig were 37.4% (29.2-46.0%), 36% (18-56%), 12% (8-17%), 26.1% (17.0-37.0%), 22.9% (12.3-36.0%), and 26.0% (20-32.0%), respectively. Moreover, major risk factor of infection was age, farming system, and farm location. A significant variation in the seroepidemiological data was observed within each species and country. The results can aid in an updated epidemiological analysis but also can be used as an important input in quantitative microbial risk assessment models. Further studies are required for a better and continual evaluation of the occurrence of this zoonotic infection.


[Decentralization of workers' health surveillance in the city of Campinas, São Paulo State, Brazil: a process evaluation].
This study evaluates the decentralization of workers' health surveillance to district teams in Campinas, São Paulo State, Brazil, from 1990 to 2006. The history and context of this process were retrieved from documents at the Municipal Health Department in order to establish evaluation criteria and define facilitating factors and obstacles in guideline implementation, as well as to measure the gap between guidelines and reality. The evaluation used qualitative techniques with stakeholders' focus groups. The results showed that decentralization was defined according to the prevailing political options and according to the principles of the Unified National Health System (SUS). Facilitating factors included the increase in technical capability and integration among the teams, and obstacles included excessively centralized planning with limited participation, insufficient resources, and lack of clarity between the services concerning roles and responsibilities. The discussion of these results with health system administrators, social actors, and participating health professionals has contributed to the analysis and organization of transformative measures in regional public practices in workers' health surveillance.


A graphic elicitation technique to represent patient rights.
A patient charter is an explicit declaration of the rights of patients within a particular health care setting. In early 2020 the Save the Children Emergency Health Unit deployed to Cox's Bazar Bangladesh to support the establishment of a severe acute respiratory infection isolation and treatment centre as part of the COVID-19 response. We developed a charter of patient rights and had it translated into Bangla and Burmese; however, the charter remained inaccessible to Rohingya and members of the host community with low literacy. To both visualise and contextualise the patient charter we undertook a graphic elicitation method involving both the Rohingya and host communities. We carried out two focus group discussions during which we discussed the charter and agreed how best to illustrate the individual rights contained therein. Logistical constraints and infection prevention and control procedures limited our ability to follow up with the original focus group participants and to engage in back-translation as we had planned; however, we were able to elicit rich descriptions of each right. Reflecting on our method we were able to identify several key learnings relating to: 1) our technique for eliciting feedback on the charter verbatim versus a broader discussion of concepts referenced within each right, 2) our decision to include both men and women in the same focus group, 3) our decision to ask focus group participants to describe specific features of each illustration and how this benefited the inclusivity of our illustrations, and 4) the potential of the focus groups to act as a means to introduce the charter to communities. Though executing our method was operationally challenging we were able to create culturally appropriate illustrations to accompany our patient charter. In contexts of limited literacy it is possible to enable access to critical clinical governance and accountability tools.


Adaptation of redband trout in desert and montane environments.
Natural populations that evolve under extreme climates are likely to diverge because of selection in local environments. To explore whether local adaptation has occurred in redband trout (Oncorhynchus mykiss gairdneri) occupying differing climate regimes, we used a limited genome scan approach to test for candidate markers under selection in populations occurring in desert and montane streams. An environmental approach to identifying outlier loci, spatial analysis method and linear regression of minor allele frequency with environmental variables revealed six candidate markers (P < 0.01). Putatively neutral markers identified high genetic differentiation among desert populations relative to montane sites, likely due to intermittent flows in desert streams. Additionally, populations exhibited a highly significant pattern of isolation by temperature (P< 0.0001) and those adapted to the same environment had similar allele frequencies across candidate markers, indicating selection for differing climates. These results imply that many genes are involved in the adaptation of redband trout to differing environments, and selection acts to reinforce localization. The potential to predict genetic adaptability of individuals and populations to changing environmental conditions may have profound implications for species that face extensive anthropogenic disturbances.


National Authority for Health: France.
The French National Authority for Health (Haute Autorité de Santé, or HAS) was established to assist France's public institutions in optimizing the basket of reimbursable goods and services and to help health care professionals continuously improve their clinical practice by defining best-care standards and identifying relevant tools and methods. HAS carries out single technology assessment (STA) and multiple technology assessment (MTA), assessing both the intrinsic benefit of the new technology and its effectiveness compared with that of existing technologies. A new treatment may not be covered unless it provides either improved benefit or lower cost, and STA is mandatory before a new drug, device, or medical procedure can be added to the benefit list for sickness funds. While HAS recommendations are advisory, the decision-making bodies (the Ministry of Health or the union of sickness funds) accept its findings in most cases.


Predictors of Delirium Development in Older Medical Inpatients: Readily Identifiable Factors at Admission.
Identifying patients at high risk of delirium is crucial to facilitate prevention. Although dementia is the most consistent risk factor across populations, it remains under-diagnosed. Hence understanding other markers of delirium vulnerability on admission is important. We aimed to identify predictors of incident delirium development in older medical inpatients that were readily identifiable at presentation to the emergency department. Medical inpatients of ≥70 years were assessed on admission for delirium using the Revised Delirium Rating Scale (DRS-R98) and those with prevalent delirium were excluded. Consenting non-delirious patients were then assessed daily using the DRS-R98. Data pertaining to multiple baseline delirium risk factors were collected, including pre-morbid dementia. Multivariable logistic regression was used to examine which factors predicted the development of incident delirium. Of 555 patients approached, 184 (33.1%) had prevalent delirium. Following other exclusions, 191 were included in the study and 61 developed incident delirium. Predictors of incident delirium on multivariable analysis, controlling for confounders, were dementia (OR 2.54, 95% CI 1.01-6.43, p = 0.048); Barthel Index score (OR 1.15 for each unit decrease in score, 95% CI 1.06-1.25, p = 0.001), and Modified Cumulative Illness Rating Scale score (OR 1.13 for each unit increase in score, 95% CI 1.05-1.22, p = 0.001). Dementia is a well-known risk factor for delirium; however, it too is under-recognized and on admission can be missed. Conversely, the Barthel Index is a simple and widely used measure of functional ability that may prove useful in stratifying those at risk of in-hospital delirium on admission.


Biochemical and cytological analysis of the complex periplasmic flagella from Spirochaeta aurantia.
The periplasmic flagella of Spirochaeta aurantia were isolated and were found to be ultrastructurally and biochemically complex. Generally, flagellar filaments were 18 to 20 nm in diameter and appeared to consist of an 11 to 13-nm-wide inner region and an outer layer. The hook-basal body region consisted of two closely apposed disks connected to a hook by a rod. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of purified flagella together with a Western blot analysis of a motility mutant that produces hooks and basal bodies but not flagellar filaments revealed that the filaments were composed of three major polypeptides of 37,500, 34,000, and 31,500 apparent molecular weight (37.5K, 34K, and 31.5K polypeptides) and three minor polypeptides of 36,000, 33,000, and 32,000 apparent molecular weight (36K, 33K, and 32K polypeptides). Purified hook-basal body preparations were greatly enriched in three polypeptides in the range of 62,000 to 66,000 apparent molecular weight. Immunogold labeling experiments with a monoclonal antibody specific for the 37.5K flagellin and one that reacts with an epitope common to the 36K, 34K, 33K, 32K, and 31.5K flagellins revealed that the 37.5K major polypeptide was a component of the outer layer, whereas one or more of the other polypeptides constituted the core.


Imaging metal-like monoclinic phase stabilized by surface coordination effect in vanadium dioxide nanobeam.
In correlated systems, intermediate states usually appear transiently across phase transitions even at the femtosecond scale. It therefore remains an open question how to determine these intermediate states-a critical issue for understanding the origin of their correlated behaviour. Here we report a surface coordination route to successfully stabilize and directly image an intermediate state in the metal-insulator transition of vanadium dioxide. As a prototype metal-insulator transition material, we capture an unusual metal-like monoclinic phase at room temperature that has long been predicted. Coordinate bonding of L-ascorbic acid molecules with vanadium dioxide nanobeams induces charge-carrier density reorganization and stabilizes metallic monoclinic vanadium dioxide, unravelling orbital-selective Mott correlation for gap opening of the vanadium dioxide metal-insulator transition. Our study contributes to completing phase-evolution pathways in the metal-insulator transition process, and we anticipate that coordination chemistry may be a powerful tool for engineering properties of low-dimensional correlated solids.


Enteroendocrine Cells: Sensing Gut Microbiota and Regulating Inflammatory Bowel Diseases.
Host sensing in the gut microbiota has been crucial in the regulation of intestinal homeostasis. Although inflammatory bowel diseases (IBDs), multifactorial chronic inflammatory conditions of the gastrointestinal tract, have been associated with intestinal dysbiosis, the detailed interactions between host and gut microbiota are still not completely understood. Enteroendocrine cells (EECs) represent 1% of the intestinal epithelium. Accumulating evidence indicates that EECs are key sensors of gut microbiota and/or microbial metabolites. They can secrete cytokines and peptide hormones in response to microbiota, either in traditional endocrine regulation or by paracrine impact on proximal tissues and/or cells or via afferent nerve fibers. Enteroendocrine cells also play crucial roles in mucosal immunity, gut barrier function, visceral hyperalgesia, and gastrointestinal (GI) motility, thereby regulating several GI diseases, including IBD. In this review, we will focus on EECs in sensing microbiota, correlating enteroendocrine perturbations with IBD, and the underlying mechanisms.


Discrimination, Abuse, Harassment, and Burnout in Surgical Residency Training.
Physicians, particularly trainees and those in surgical subspecialties, are at risk for burnout. Mistreatment (i.e., discrimination, verbal or physical abuse, and sexual harassment) may contribute to burnout and suicidal thoughts. A cross-sectional national survey of general surgery residents administered with the 2018 American Board of Surgery In-Training Examination assessed mistreatment, burnout (evaluated with the use of the modified Maslach Burnout Inventory), and suicidal thoughts during the past year. We used multivariable logistic-regression models to assess the association of mistreatment with burnout and suicidal thoughts. The survey asked residents to report their gender. Among 7409 residents (99.3% of the eligible residents) from all 262 surgical residency programs, 31.9% reported discrimination based on their self-identified gender, 16.6% reported racial discrimination, 30.3% reported verbal or physical abuse (or both), and 10.3% reported sexual harassment. Rates of all mistreatment measures were higher among women; 65.1% of the women reported gender discrimination and 19.9% reported sexual harassment. Patients and patients' families were the most frequent sources of gender discrimination (as reported by 43.6% of residents) and racial discrimination (47.4%), whereas attending surgeons were the most frequent sources of sexual harassment (27.2%) and abuse (51.9%). Proportion of residents reporting mistreatment varied considerably among residency programs (e.g., ranging from 0 to 66.7% for verbal abuse). Weekly burnout symptoms were reported by 38.5% of residents, and 4.5% reported having had suicidal thoughts during the past year. Residents who reported exposure to discrimination, abuse, or harassment at least a few times per month were more likely than residents with no reported mistreatment exposures to have symptoms of burnout (odds ratio, 2.94; 95% confidence interval [CI], 2.58 to 3.36) and suicidal thoughts (odds ratio, 3.07; 95% CI, 2.25 to 4.19). Although models that were not adjusted for mistreatment showed that women were more likely than men to report burnout symptoms (42.4% vs. 35.9%; odds ratio, 1.33; 95% CI, 1.20 to 1.48), the difference was no longer evident after the models were adjusted for mistreatment (odds ratio, 0.90; 95% CI, 0.80 to 1.00). Mistreatment occurs frequently among general surgery residents, especially women, and is associated with burnout and suicidal thoughts.


Online exclusive: behavioral adjustment of children and adolescents with cancer: teacher, parent, and self-report.
To describe behavioral adjustment in children and adolescents with acute lymphoblastic leukemia (ALL) and to determine whether behavioral adjustment is correlated with cognitive and academic abilities. Descriptive, cross-sectional design. Two pediatric oncology treatment centers. 47 children and adolescents who had been receiving ALL therapy for at least one year or who were off therapy for no more than three years and their parents and teachers. Wechsler Intelligence Scale for Children-Revised (WISC-R) and Wide Range Achievement Test-Revised (WRAT-R) data were available on a subset of 17 subjects. Parent, teacher, and self-report Behavioral Assessment System for Children (BASC) ratings were used to measure behavioral adjustment. WISC-R measured cognitive abilities, and WRAT-R measured academic abilities. Demographic, family, and treatment-related data also were collected. Behavioral adjustment and cognitive and academic abilities. At least 20% of teacher ratings for somatization, learning problems, leadership, and study skills; parent ratings for somatization, adaptability, attention problems, withdrawal, anxiety, social skills, and depression; and self-report ratings for anxiety and attitude to school were in the at-risk range (i.e., presence of significant problems that require treatment). The majority of teacher BASC ratings were correlated significantly with WISC-R and WRAT-R scores. Self-report depression and social stress ratings were correlated significantly with some WISC-R and WRAT-R scores. Treatment-related experiences such as body image alterations and mental and emotional problems were associated with problematic behaviors, including depression, somatization, withdrawal, and social stress. Youth with ALL are at risk for some behavioral adjustment problems, particularly anxiety, somatization, adaptability, attention, and withdrawal. Cognitive and academic abilities are associated with some dimensions of behavioral adjustment. Findings suggest the need for ongoing assessment of behavioral adjustment and cognitive and academic abilities of children with ALL. Behavioral interventions that target at-risk mannerisms, such as somatization, depression, anxiety, and social stress, are needed. Central nervous system treatment may contribute to behavioral adjustment problems, as well as to cognitive and academic problems. Strategies to improve academic abilities also may have a positive effect on behavioral adjustment.


Medication coaching program for patients with minor stroke or TIA: a pilot study.
Patients who are hospitalized with a first or recurrent stroke often are discharged with new medications or adjustment to the doses of pre-admission medications, which can be confusing and pose safety issues if misunderstood. The purpose of this pilot study was to assess the feasibility of medication coaching via telephone after discharge in patients with stroke. Two-arm pilot study of a medication coaching program with 30 patients (20 intervention, 10 control). Consecutive patients admitted with stroke or TIA with at least 2 medications changed between admission and discharge were included. The medication coach contacted intervention arm patients post-discharge via phone call to discuss risk factors, review medications and triage patients' questions to a stroke nurse and/or pharmacist. Intervention and control participants were contacted at 3 months for outcomes. The main outcomes were feasibility (appropriateness of script, ability to reach participants, and provide requested information) and participant evaluation of medication coaching. The median lengths of the coaching and follow-up calls with requested answers to these questions were 27 minutes and 12 minutes, respectively, and participant evaluations of the coaching were positive. The intervention participants were more likely to have seen their primary care provider than were control participants by 3 months post discharge. This medication coaching study executed early after discharge demonstrated feasibility of coaching and educating stroke patients with a trained coach. Results from our small pilot showed a possible trend towards improved appointment-keeping with primary care providers in those who received coaching.


Potential Alzheimer's Disease Therapeutics Among Weak Cysteine Protease Inhibitors Exhibit Mechanistic Differences Regarding Extent of Cathepsin B Up-Regulation and Ability to Block Calpain.
Cysteine protease inhibitors have long been part of drug discovery programs for Alzheimer's disease (AD), traumatic brain injury (TBI), and other disorders. Select inhibitors reduce accumulating proteins and AD pathology in mouse models. One such compound, Z-Phe-Aladiazomethylketone (PADK), exhibits a very weak IC50 (9-11 μM) towards cathepsin B (CatB), but curiously PADK causes marked up-regulation of the Aβ-degrading CatB and improves spatial memory. Potential therapeutic and weak inhibitor E64d (14 μM IC50) also up-regulates CatB. PADK and E64d were compared regarding the blockage of calcium-induced cytoskeletal deterioration in brain samples, monitoring the 150-kDa spectrin breakdown product (SBDP) known to be produced by calpain. PADK had little to no effect on SBDP production at 10-100 μM. In contrast, E64d caused a dose-dependent decline in SBDP levels with an IC50 of 3-6 μM, closely matching its reported potency for inhibiting μ-calpain. Calpain also cleaves the cytoskeletal organizing protein gephyrin, producing 49-kDa (GnBDP49) and 18-kDa (GnBDP18) breakdown products. PADK had no apparent effect on calcium-induced gephyrin fragments whereas E64d blocked their production. E64d also protected the parent gephyrin in correspondence with reduced BDP levels. The findings of this study indicate that PADK's positive and selective effects on CatB are consistent with human studies showing exercise elevates CatB and such elevation correlates with improved memory. On the other hand, E64d exhibits both marginal CatB enhancement and potent calpain inhibition. This dual effect may be beneficial for treating AD. Alternatively, the potent action on calpain-related pathology may explain E64d's protection in AD and TBI models.


Crossing the taxonomic divide: conflict and its resolution in societies of reproductively totipotent individuals.
Reproduction in groups may be unequal, with one or a few individuals monopolizing direct reproduction assisted by nonbreeding helpers. In social insects this has frequently led to a pronounced queen-worker dichotomy and a loss of reproductive totipotency among workers. However, in some invertebrate and all vertebrate societies, all or most individuals remain reproductively totipotent. In these groups, conflicts of interest over reproduction are potentially greatest. Here, we synthesize previous analyses of reproductive conflict, aggression and breeder replacement in haplodiploid societies of totipotent individuals and extend them to cover diploid (vertebrate) examples. We test predictions arising from this approach using the best-studied invertebrate (Dinoponera queenless ants) and vertebrate (naked mole-rat, Heterocephalus glaber) examples, although in principle our analysis applies to all similar groups. We find that premature replacement of a parent breeder by nonbreeders (overthrow) is rare. Dominant coercive control of nonbreeders by the breeder is often unnecessary and honest signalling of breeder vitality can maintain group stability and resolve conflicts over reproduction. We hope that by providing an explicit transfer of social theory between ants and naked mole-rats we will stimulate further cross-taxonomic studies that will greatly broaden our understanding of sociality.


Topological frustration in beta alpha-repeat proteins: sequence diversity modulates the conserved folding mechanisms of alpha/beta/alpha sandwich proteins.
The thermodynamic hypothesis of Anfinsen postulates that structures and stabilities of globular proteins are determined by their amino acid sequences. Chain topology, however, is known to influence the folding reaction, in that motifs with a preponderance of local interactions typically fold more rapidly than those with a larger fraction of nonlocal interactions. Together, the topology and sequence can modulate the energy landscape and influence the rate at which the protein folds to the native conformation. To explore the relationship of sequence and topology in the folding of beta alpha-repeat proteins, which are dominated by local interactions, we performed a combined experimental and simulation analysis on two members of the flavodoxin-like, alpha/beta/alpha sandwich fold. Spo0F and the N-terminal receiver domain of NtrC (NT-NtrC) have similar topologies but low sequence identity, enabling a test of the effects of sequence on folding. Experimental results demonstrated that both response-regulator proteins fold via parallel channels through highly structured submillisecond intermediates before accessing their cis prolyl peptide bond-containing native conformations. Global analysis of the experimental results preferentially places these intermediates off the productive folding pathway. Sequence-sensitive Gō-model simulations conclude that frustration in the folding in Spo0F, corresponding to the appearance of the off-pathway intermediate, reflects competition for intra-subdomain van der Waals contacts between its N- and C-terminal subdomains. The extent of transient, premature structure appears to correlate with the number of isoleucine, leucine, and valine (ILV) side chains that form a large sequence-local cluster involving the central beta-sheet and helices alpha2, alpha 3, and alpha 4. The failure to detect the off-pathway species in the simulations of NT-NtrC may reflect the reduced number of ILV side chains in its corresponding hydrophobic cluster. The location of the hydrophobic clusters in the structure may also be related to the differing functional properties of these response regulators. Comparison with the results of previous experimental and simulation analyses on the homologous CheY argues that prematurely folded unproductive intermediates are a common property of the beta alpha-repeat motif.


Assault characteristics and posttraumatic stress disorder in rape victims.
A diagnosis of posttraumatic stress disorder (PTSD) encompasses several of the symptoms associated with the response to rape. The object of this study was to determine the incidence of PTSD among 51 rape victims and to compare the groups with and without PTSD with respect to the characteristics of the assaults. The results showed that 70% of the victims had PTSD and further suggested that PTSD was likely to be a long-term problem for these women. The results supported the view that psychological treatment approaches to rape victims should take into account the posttraumatic nature of the response. The incidence of rapes by strangers, of physical force being used, of weapons being displayed and of injuries being sustained by the victim were all higher in the group of women who had PTSD. These are all features that can be readily identified in the initial assessment of the rape victim and may help to identify the victims at risk of developing long-term psychological sequelae after rape. Appropriate therapeutic action taken early may influence the prognosis for a significant proportion of rape victims.


Evolutionarily conserved positive and negative cis-acting elements control the blastoderm-specific expression of the Drosophila serendipity alpha cellularisation gene.
The serendipity alpha (sry alpha) cellularisation gene is only transcribed at the blastoderm stage, when this morphogenetic event takes place. We show that a 95 bp sry alpha upstream region is sufficient for blastoderm-specific expression of a lacZ reporter gene. This region encompasses four nucleotide motifs (I-IV, 5' to 3') conserved at similar relative positions in several Drosophila species. Removal of motif I leads to ectopic expression of lacZ in precursor cells of the PNS. Deletion of motif IV decreases the level of lacZ transcripts and modifies their banded pattern of accumulation late in cycle 14, whereas deletion of motifs II and III abolishes the sry alpha promoter activity. Motif III includes a consensus recognition site for b-HLH proteins. A point mutation in this E-box both severely reduces lacZ expression at blastoderm and prevents its ectopic expression in the PNS upon deleting motif I. These two effects depend upon da+ activity, suggesting that daughterless may positively control sry alpha transcription.


Accelerated cerebral ischemic injury by activated macrophages/microglia after lipopolysaccharide microinjection into rat corpus callosum.
In cerebral ischemic insults, activated inflammatory cells such as microglia and macrophages may be implicated in the pattern and degree of ischemic injury by producing various bioactive mediators. In the present study, we provide the evidence that activated microglia/macrophages accelerate cerebral ischemic injury by overexpression of inducible nitric oxide synthase (iNOS). To activate microglia/macrophages, a potent inflammation inducer lipopolysaccharide (LPS, 5 microg/5 microl) was microinjected into rat corpus callosum. Isolectin B4-positive microglia/macrophages were abundantly observed in ipsilateral hemisphere at 1 day after LPS injection. RT-PCR showed that LPS injection induced iNOS mRNA expression mostly in microglia/macrophages, peaking in intensity at 15 h after LPS injection. While ischemic injury was little evoked in control rats by 2-h middle cerebral artery occlusion (MCAO) followed by 3-h reperfusion, it was markedly increased in rats pre-injected with LPS 1 day before MCAO. However, no significant difference between control and LPS-pretreated groups was observed after 24-h reperfusion. The increased ischemic injury in LPS-treated rats was well correlated with iNOS level expressed over 3 orders of magnitude than in LPS-untreated rats. Immunohistochemical studies showed that iNOS- and nitrotyrosine (a peroxynitrite marker)-positive cells were prominent throughout the infarct area. NOS inhibitors aminoguanidine or N(G)-nitro-L-arginine, simultaneously injected with LPS, reduced the iNOS immunoreactivity and infarct volume, especially in penumbra regions. Total glutathione levels in ischemic regions were decreased more in LPS pre-injected rats than in control ones. Further defining the role of NO in cerebral ischemic insults would provide the rationale for new therapeutic strategies based on modulation of microglial and macrophageal NO production in the brain.


Exploring knowledge and attitudes about emergency contraceptive pills among university students in Jamaica. A qualitative approach.
Emergency contraceptive pills (ECPs) are an important option for university students who may be at high risk for unplanned pregnancies. In the Caribbean, little research has been carried out on university student's knowledge and opinions of this method. This study uses qualitative methodology to explore knowledge and opinions on ECPs among university students attending The University of the West Indies (UWI) Mona campus. We conducted eight focus groups (n = 71) with female and male university students at The University of the West Indies, Kingston, Jamaica, in March 2000. The group discussions lasted approximately two hours. We tape-recorded discussions and then transcribed and analyzed them by coding responses according to themes. General knowledge of ECPs was high, but students lacked specific information about the method such as its time frame and its mechanism of action. Most students supported the method, especially after learning correct information. However, several students were concerned about its side effects and the potential for abuse or irresponsible use by young adults. Although the university students in this study lacked detailed information about ECPs, their opinion toward the method was favourable. We suggest further research to investigate the prevalence of misinformation about the method among other groups of Jamaicans.


What adaptation to research is needed following crises: a comparative, qualitative study of the health workforce in Sierra Leone and Nepal.
Health workers are critical to the performance of health systems; yet, evidence about their coping strategies and support needs during and post crisis is lacking. There is very limited discussion about how research teams should respond when unexpected crises occur during on-going research. This paper critically presents the approaches and findings of two health systems research projects that explored and evaluated health worker performance and were adapted during crises, and provides lessons learnt on re-orientating research when the unexpected occurs. Health systems research was adapted post crisis to assess health workers' experiences and coping strategies. Qualitative in-depth interviews were conducted with 14 health workers in a heavily affected earthquake district in Nepal and 25 frontline health workers in four districts in Ebola-affected Sierra Leone. All data were transcribed and analysed using the framework approach, which included developing coding frameworks for each study, applying the frameworks, developing charts and describing the themes. A second layer of analysis included analysis across the two contexts, whereas a third layer involved the research teams reflecting on the approaches used to adapt the research during these crises and what was learned as individuals and research teams. In Sierra Leone, health workers were heavily stigmatised by the epidemic, leading to a breakdown of trust. Coping strategies included finding renewed purpose in continuing to serve their community, peer and family support (in some cases), and religion. In Nepal, individual determination, a sense of responsibility to the community and professional duty compelled staff to stay or return to their workplace. The research teams had trusting relationships with policy-makers and practitioners, which brought credibility and legitimacy to the change of research direction as well as the relationships to maximise the opportunity for findings to inform practice. In both contexts, health workers demonstrated considerable resilience in continuing to provide services despite limited support. Embedded researchers and institutions are arguably best placed to navigate emerging ethical and social justice challenges and are strategically positioned to support the co-production of knowledge and ensure research findings have impact.


Early deprivation leads to long-term reductions in motivation for reward and 5-HT1A binding and both effects are reversed by fluoxetine.
Early life stress is a risk factor in aetiology of depression. In rats, early life stress can lead to pro-depressive biomarkers in adulthood. The present study in male Wistar rats investigated the effects of early life deprivation and fluoxetine on motivation for reward, activity in the forced swim test, and brain monoamine receptors, in adulthood. P1-14 pups were isolated for 4 h/day (early deprivation, ED) or were handled for 1 min (CON). They were weaned at PND21 and left undisturbed until 4-6 months old. The ED and CON groups were halved to receive either vehicle or fluoxetine (FLX, 10 mg/kg, 31 days). Thus, four treatment groups were studied: CON-VEH, CON-FLX, ED-VEH and ED-FLX, n = 8 each. On a progressive ratio schedule, ED-VEH animals showed significantly reduced motivation to obtain sucrose versus CON-VEH, and this reward-motivation deficit was reversed by FLX. Activity in the forced swim test was unaffected by ED and increased by FLX. Quantitative autoradiography was used to determine 5-HT1A and 5-HT2C receptor binding with [O-methyl-(3)H]WAY 100635 and [(3)H]mesulergine (added spiperone and 8-OH-DPAT), respectively. In ED-VEH versus CON-VEH, 5-HT1A receptor binding was significantly reduced in anterior cingulate, motor cortex, ventral hippocampal CA1 and dorsal raphé; this was reversed by chronic FLX. Concomitant ED-dependent reductions observed in 5-HT2C (motor and frontal cortices, ventral CA1 and dorsal raphé) and D2 (dorsolateral striatum and accumbens) binding were not reversed by FLX. Because chronic FLX treatment reversed the ED-induced behavioural and 5-HT1A binding deficits, the 5-HT1A receptor is implicated as a selective therapeutic target.


A variant of the mammalian somatotopic map in a bat.
Two ordered representations of the body surface, S-I and S-II, have been described on the cortical surface of the brains of a variety of mammals; additional separate topographical maps have been found in the somatosensory cortex of the cat and monkey. Except for minor variations in the placement of the body parts, the basic somatotopy of the maps is remarkably consistent across species. As the reasons for this consistency and the minor variations are unclear, we examined the somatotopy of the bat, whose body plan has been modified extensively so that the forelimb can be used for flight. We report here that in both S-I and S-II of the grey-headed flying fox, not only is the representation of the distal forelimb displaced from its usual position on the map, but the digits are directed caudally instead of rostrally as they are in all other mammals studied. The variant somatotopy appears to reflect the postural differences between flying and walking mammals, supporting the notion that topographical maps may have functional significance apart from their point-to-point connections with the sensory periphery.


Prediction of prostate cancer recurrence using quantitative phase imaging: Validation on a general population.
Prediction of biochemical recurrence risk of prostate cancer following radical prostatectomy is critical for determining whether the patient would benefit from adjuvant treatments. Various nomograms exist today for identifying individuals at higher risk for recurrence; however, an optimistic under-estimation of recurrence risk is a common problem associated with these methods. We previously showed that anisotropy of light scattering measured using quantitative phase imaging, in the stromal layer adjacent to cancerous glands, is predictive of recurrence. That nested-case controlled study consisted of specimens specifically chosen such that the current prognostic methods fail. Here we report on validating the utility of optical anisotropy for prediction of prostate cancer recurrence in a general population of 192 patients, with 17% probability of recurrence. Our results show that our method can identify recurrent cases with 73% sensitivity and 72% specificity, which is comparable to that of CAPRA-S, a current state of the art method, in the same population. However, our results show that optical anisotropy outperforms CAPRA-S for patients with Gleason grades 7-10. In essence, we demonstrate that anisotropy is a better biomarker for identifying high-risk cases, while Gleason grade is better suited for selecting non-recurrence. Therefore, we propose that anisotropy and current techniques be used together to maximize prediction accuracy.


TAp73alpha induces tau phosphorylation in HEK293a cells via a transcription-dependent mechanism.
p73 and tau both play roles in neurodevelopment and neurodegeneration. In this pilot study we show by Western blotting that TAp73alpha induces phosphorylation of human 2N4R tau at threonine-205 and at the PHF-1 epitope (serine366/serine404) in HEK293a cells. Neither the dominant negative isoform, DeltaNp73, nor a transcriptionally inactive mutant TAp73alpha(R292H) altered tau phosphorylation indicating that tau phosphorylation is dependent on the transcriptional activity of TAp73alpha. Consistent with this, confocal microscopy revealed that tau and TAp73alpha were spatially separated within the cell; tau being located in the cytoskeletal compartment whilst TAp73alpha was found in the nucleus. These findings have ramifications for microtubule dynamics associated with axonal growth during development and for neuronal death associated with Alzheimer's disease and other tauopathies.


A one-dimensional random laser based on artificial high-index contrast scatterers.
The realization of a one-dimensional (1D) random laser (RL) by using artificially fabricated scatterers is reported in this letter, and the lasing characteristic has also been investigated comprehensively. The manipulation of the lasing mode in the 1D microwire (MW) RL can be achieved through micro-pits prepared by the laser-ablation technique. Well-defined sharp lasing peaks were realized based on the coherence feedback process in the 1D optical waveguide. The near- and far-field images exhibit excellent spatial intensity distribution, and the stability of lasing modes has also been investigated. Our results represent a novel method towards the development of a manipulated-RL, which will highlight the application of disordered systems in optoelectronic devices.


Views of junior doctors about whether their medical school prepared them well for work: questionnaire surveys.
The transition from medical student to junior doctor in postgraduate training is a critical stage in career progression. We report junior doctors' views about the extent to which their medical school prepared them for their work in clinical practice. Postal questionnaires were used to survey the medical graduates of 1999, 2000, 2002 and 2005, from all UK medical schools, one year after graduation, and graduates of 2000, 2002 and 2005 three years after graduation. Summary statistics, chi-squared tests, and binary logistic regression were used to analyse the results. The main outcome measure was the level of agreement that medical school had prepared the responder well for work. Response rate was 63.7% (11610/18216) in year one and 60.2% (8427/13997) in year three. One year after graduation, 36.3% (95% CI: 34.6, 38.0) of 1999/2000 graduates, 50.3% (48.5, 52.2) of 2002 graduates, and 58.2% (56.5, 59.9) of 2005 graduates agreed their medical school had prepared them well. Conversely, in year three agreement fell from 48.9% (47.1, 50.7) to 38.0% (36.0, 40.0) to 28.0% (26.2, 29.7). Combining cohorts at year one, percentages who agreed that they had been well prepared ranged from 82% (95% CI: 79-87) at the medical school with the highest level of agreement to 30% (25-35) at the lowest. At year three the range was 70% to 27%. Ethnicity and sex were partial predictors of doctors' level of agreement; following adjustment for them, substantial differences between schools remained. In years one and three, 30% and 34% of doctors specified that feeling unprepared had been a serious or medium-sized problem for them (only 3% in each year regarded it as serious). The vast knowledge base of clinical practice makes full preparation impossible. Our statement about feeling prepared is simple yet discriminating and identified some substantial differences between medical schools. Medical schools need feedback from graduates about elements of training that could be improved.


Surgical options for intrahepatic cholangiocarcinoma.
Intrahepatic cholangiocarcinoma (ICC) is the second most common primary liver cancer, accounting for 10-15% of primary hepatic malignancy. The incidence and cancer-related mortality of ICC continue to increase worldwide. At present, hepatectomy is still the most effective treatment for ICC patients to achieve long-term survival, although its overall efficacy may not be as good as that for patients with hepatocellular carcinoma (HCC) due to the unique pathogenesis and clinical-pathological profiles of ICC. Viral infection, lithiasis and metabolic factors may all be associated with the pathogenesis of ICC. Poor blood supply, cirrhosis (in rare cases), surrounding organ invasion, and lymph node/distal metastasis have significant impacts on the selection of surgical strategies, surgical resection rate, postoperative complications, recurrence and metastasis. Surgical treatment for ICC includes R0 resection, lymphadenectomy, total gross resection of the involved biliary tracts, blood vessels and surrounding tissues in adjacent organs, and reconstruction. Postoperative adjuvant therapy and local-regional therapy after recurrence may improve survival. Liver transplantation (LT) is reported to have a moderate treatment effect on early ICC although its efficacy remains controversial. In this article, we reviewed the epidemiology and staging of ICC and highlighted the selection of surgical modalities and postoperative outcomes of ICC patients via literature review.


Social, economic, and health impact of the respiratory syncytial virus: a systematic search.
Bronchiolitis caused by the respiratory syncytial virus (RSV) and its related complications are common in infants born prematurely, with severe congenital heart disease, or bronchopulmonary dysplasia, as well as in immunosuppressed infants. There is a rich literature on the different aspects of RSV infection with a focus, for the most part, on specific risk populations. However, there is a need for a systematic global analysis of the impact of RSV infection in terms of use of resources and health impact on both children and adults. With this aim, we performed a systematic search of scientific evidence on the social, economic, and health impact of RSV infection. A systematic search of the following databases was performed: MEDLINE, EMBASE, Spanish Medical Index, MEDES-MEDicina in Spanish, Cochrane Plus Library, and Google without time limits. We selected 421 abstracts based on the 6,598 articles identified. From these abstracts, 4 RSV experts selected the most relevant articles. They selected 65 articles. After reading the full articles, 23 of their references were also selected. Finally, one more article found through a literature information alert system was included. The information collected was summarized and organized into the following topics: 1. Impact on health (infections and respiratory complications, mid- to long-term lung function decline, recurrent wheezing, asthma, other complications such as otitis and rhino-conjunctivitis, and mortality; 2. Impact on resources (visits to primary care and specialists offices, emergency room visits, hospital admissions, ICU admissions, diagnostic tests, and treatments); 3. Impact on costs (direct and indirect costs); 4. Impact on quality of life; and 5. Strategies to reduce the impact (interventions on social and hygienic factors and prophylactic treatments). We concluded that 1. The health impact of RSV infection is relevant and goes beyond the acute episode phase; 2. The health impact of RSV infection on children is much better documented than the impact on adults; 3. Further research is needed on mid- and long-term impact of RSV infection on the adult population, especially those at high-risk; 4. There is a need for interventions aimed at reducing the impact of RSV infection by targeting health education, information, and prophylaxis in high-risk populations.


Inflammation and oxidative stress are lower in physically fit and active adults.
The objective of this study was to determine if the inverse relationship between perceived physical fitness (pFIT) and exercise frequency (ExFreq) levels and chronic inflammation and oxidative stress exists after making statistical adjustments for confounders including body mass index (BMI), age, gender, and cigarette smoking. Study participants (60% female and 40% male; n = 998) varied widely in age (18-85 years) and BMI (16.7-52.7 kg/m(2)) completed an extensive medical/health and lifestyle questionnaire, and data were used to establish pFIT and ExFreq tertiles. Biomarkers included serum C-reactive protein (CRP), total blood leukocytes, five plasma cytokines [interleukin (IL)-6, IL-10, tumor necrosis factor-α (TNF-α), monocyte chemoattractant protein-1 (MCP1), and granulocyte colony-stimulating factor (GCSF)], F2 -isoprostanes, ferric reducing ability of plasma (FRAP), and oxygen radical absorbance capacity (ORAC). A general linear model was used to examine relationships between pFIT and ExFreq with inflammation and oxidative stress while controlling for age, gender, BMI, and smoking. Benjamini-Hochberg method for false discovery rate correction was used for multiple testing corrections. Significant tests (P < 0.05) for trend were found for the effect of pFIT and ExFreq on CRP, white blood cell, IL-6, TNF-α, GCSF, and F2 -isoprostanes, but not MCP1, IL-10, FRAP, and ORAC, after adjustment for confounders. These data indicate that an inverse relationship exists among chronic inflammation, oxidative stress, and pFIT and ExFreq at the community level even after adjustment for important confounders.


Early infant multivitamin supplementation is associated with increased risk for food allergy and asthma.
Dietary vitamins have potent immunomodulating effects in vitro. Individual vitamins have been shown to skew T cells toward either T-helper 1 or T-helper 2 phenotypic classes, suggesting that they may participate in inflammatory or allergic disease. With the exception of antioxidant protection, there has been little study on the effect of early vitamin supplementation on the subsequent risk for asthma and allergic disease. The objective of this study was to determine whether early vitamin supplementation during infancy affects the risk for asthma and allergic disease during early childhood. Cohort data were analyzed from the National Center for Health Statistics 1988 National Maternal-Infant Health Survey, which followed pregnant women and their newborns, and the 1991 Longitudinal Follow-up of the same patients, which measured health and disease outcomes. Patients were stratified by race and breastfeeding status. Factors that are known to be associated with alteration of risk for asthma or food allergies were identified using univariate logistic regression. Those factors were then analyzed in multivariate logistic regression models. Early vitamin supplementation was defined as vitamin use within the first 6 months. There were >8000 total patients in the study. The overall incidence of asthma was 10.5% and of food allergy was 4.9%. In univariate analysis, male gender, smoker in the household, child care, prematurity (<37 weeks), being black, no history of breastfeeding, lower income, and lower education were associated with higher risk for asthma. Child care, higher levels of education, income, and history of breastfeeding were associated with a higher risk for food allergies. In multivariate logistic analyses, a history of vitamin use within the first 6 months of life was associated with a higher risk for asthma in black infants (odds ratio [OR]: 1.27; 95% confidence interval [CI]: 1.04-1.56). Early vitamin use was also associated with a higher risk for food allergies in the exclusively formula-fed population (OR: 1.63; 95% CI: 1.21-2.20). Vitamin use at 3 years of age was associated with increased risk for food allergies but not asthma in both breastfed (OR: 1.62; 95% CI: 1.19-2.21) and exclusively formula-fed infants (OR: 1.39; 95% CI: 1.03-1.88). Early vitamin supplementation is associated with increased risk for asthma in black children and food allergies in exclusively formula-fed children. Additional study is warranted to examine which components most strongly contribute to this risk.


Energy and fibre intake in a group of captive giraffe (Giraffa camelopardalis) offered increasing amounts of browse.
We investigated the effect of diet on intake of energy and fibre in a group of three captive adult giraffe by weighing offered diet items and leftovers for 7 days after an adaptation period of 7 days. Digestion coefficients were calculated using, as internal marker, the acid detergent lignin content of a faecal sample pooled from subsamples taken during the last 5 days of intake measurement. Two lucerne hay-only diets of differing quality (L1, L2) were fed, as well as the regular diet of lucerne hay and concentrates (L2C), and the regular diet supplemented with 3 or 6 kg of edible, fresh browse material (L2CB3, L2CB6). The proportion of roughage in the ingested diets L2CB6 (45 +/- 5% dry matter), L2CB3 (35 +/- 3%) and L2C (37 +/- 10%) did not differ significantly. Digestible energy intake was low on the hay-only diets [L1: 0.28 +/- 0.06 MJ/kg body weight (BW)0.75; L2: 0.33 +/- 0.10 MJ/kg BW0.75] and increased from L2C (0.60 +/- 0.13 MJ/kg BW0.75) to a significant increase with L2CB3 (0.72 +/- 0.17 MJ/kg BW0.75); no further increase was obvious for L2CB6 (0.63 +/- 0.15MJ/kg BW0.75). The results confirm that giraffes are unlikely to meet energy requirements on lucerne hay-only diets. In a feeding scenario where both lucerne hay and the concentrate component of the diet are fed ad libitum, the animals tended to exchange hay for browse when browse was added. Only the higher level of browse supplementation led to a potentially beneficial increase in fibre intake. Whether additional browse supplementation will lead to increased intakes in a feeding scenario with restricted concentrate provision can be suspected but remains to be demonstrated.


[Immune checkpoint inhibitor-induced anti-striational antibodies in myasthenia gravis and myositis: a case report].
A 78-year-old man was treated with ipilimumab and nivolumab for advanced renal cell carcinoma with liver and lymph node metastasis. He developed diplopia, ptosis, dysphagia, and weakness of the limbs and neck, 1 month after treatment. Serum creatine kinase (CK) levels were elevated, and neck MRI revealed inflammation of the deep trunk muscles. Although anti-acetylcholine receptor antibody was negative, the edrophonium test was positive. Anti-striational antibodies such as the anti-titin and the anti-muscular voltage-gated potassium channel (Kv 1.4) antibodies (which serve as biomarkers of immune checkpoint inhibitors associated with myasthenia gravis and myositis) were positive (anti-titin antibody titer 11.51, normal <1 index; anti-Kv 1.4 antibody titer 15.13, normal <1 index). Intravenous methylprednisolone pulse therapy (1,000 mg/day for 3 days), plasmapheresis, and oral prednisolone (PSL) (20 mg/day) administration improved the patient's neurological function and normalized the serum CK levels. The PSL dosage was tapered without any worsening of clinical signs. The antibody titers decreased but remained positive (anti-titin antibody 5.00, anti-Kv 1.4 antibody 3.83) one year after the initial evaluation. Therefore, low-dose PSL (5 mg/day) administration was continued, and the patient was in remission.


The Therapeutic Benefits of Nanoencapsulation in Drug Delivery to the Anterior Segment of the Eye: A Systematic Review.
Background: Although numerous nanoparticle formulations have been developed for ocular administration, concerns are being raised about a possible mismatch between potential promises made by the field of nanoparticle research and demonstration of actual therapeutic benefit. Therefore, the primary focus of this present review was to critically assess to what extent nanoencapsulation of ocular drugs improved the therapeutic outcome when treating conditions in the anterior segment of the eye. Methods: A systematic search was conducted using Medline, PubMed, and Embase databases as well as Google Scholar for published peer-reviewed articles in English focusing on conventional nanoparticles used as drug delivery systems to the anterior segment of the eye in in vivo studies. The major therapeutic outcomes were intraocular pressure, tear secretion, number of polymorphonuclear leucocytes and pupil size. The outcome after encapsulation was compared to the non-encapsulated drug. Results: From the search, 250 results were retrieved. Thirty-eight studies met the inclusion criteria. Rabbits were used as study subjects in all but one study, and the number of animals ranged from 3 to 10. Coated and uncoated liposomes, lipid-based and polymeric nanoparticles, as well as micelles, were studied, varying in both particle size and surface charge, and encapsulating a total of 24 different drugs, including 6 salts. The majority of the in vivo studies demonstrated some improvement after nanoencapsulation, but the duration of the benefit varied from less than 1 h to more than 20 h. The most common in vitro methods performed in the studies were drug release, transcorneal permeation, and mucin interaction. Discussion: Nanoparticles that are small and mucoadhesive, often due to positive surface charge, appeared beneficial. Although in vitro assays can unravel more of the hidden and sophisticated interplay between the encapsulated drug and the nanoparticle structure, they suffered from a lack of in vitro-in vivo correlation. Therefore, more research should be focused towards developing predictive in vitro models, allowing rational design and systematic optimization of ocular nanoparticles with minimal animal experimentation.


Single penile incision for combined hypospadias and inguinal surgery: A comparative study.
We sought to compare the surgical outcomes of hypospadias repair with correction of inguinal pathology using a single penile incision vs. conventional approach using two incisions. This is a retrospective study that reviewed all patients who underwent concurrent surgical repair for both hypospadias and inguinal pathologies between January 2003 and November 2015. Patients were classified into Group A, conventional (inguinal or scrotal and penile incision) approach; or Group B, single penile incision approach. Baseline characteristics, including age, degree of hypospadias, type and laterality of inguinal pathology, operative time, and surgical outcomes, were collected. Between groups, variable comparisons were analyzed using Mann-Whitney U-Test and Fisher-exact test. Statistical significant set at <0.05. Seventy-six patients (Group A: 40; Group B: 36) were eligible for study. Baseline characteristics of both groups were comparable, with no significant statistical difference. Overall mean operative time for Group A was 139.3 ± 56.2 minutes, while Group B was 107.8 ± 46.7 minutes (Z=2.6; U=470.5; p=0.009). Two patients in Group A and two patients in Group B had testicular ascension, all of which also had hypospadias-related complications (p=1.0). Hypospadias-related complications in Group A included seven urethrocutaneous fistulae and two repair dehiscence. Eight urethrocutaneous fistulae, one urethral stricture, and two repair dehiscence occurred in Group B (p=0.448). Surgical outcome appearance in both groups were comparable, with no statistically significant difference (p=0.466). Single penile incision for both hypospadias repair and correction of inguinal pathology is a feasible technique and comparable to the conventional approach, with similar surgical outcomes and shorter overall operative time.


The kinetics of L-selectin tethers and the mechanics of selectin-mediated rolling.
Two mechanisms have been proposed for regulating rolling velocities on selectins. These are (a) the intrinsic kinetics of bond dissociation, and (b) the reactive compliance, i.e., the susceptibility of the bond dissociation reaction to applied force. To determine which of these mechanisms explains the 7.5-11.5-fold faster rolling of leukocytes on L-selectin than on E- and P-selectins, we have compared the three selectins by examining the dissociation of transient tethers. We find that the intrinsic kinetics for tether bond dissociation are 7-10-fold more rapid for L-selectin than for E- and P-selectins, and are proportional to the rolling velocities through these selectins. The durations of pauses during rolling correspond to the duration of transient tethers on low density substrates. Moreover, applied force increases dissociation kinetics less for L-selectin than for E- and P-selectins, demonstrating that reactive compliance is not responsible for the faster rolling through L-selectin. Further measurements provide a biochemical and biophysical framework for understanding the molecular basis of rolling. Displacements of tethered cells during flow reversal, and measurements of the distance between successive pauses during rolling provide estimates of the length of a tether and the length of the adhesive contact zone, and suggest that rolling occurs with as few as two tethers per contact zone. Tether bond lifetime is an exponential function of the force on the bond, and the upper limit for the tether bond spring constant is of the same order of magnitude as the estimated elastic spring constant of the lectin-EGF unit. Shear uniquely enhances the rate of L-selectin transient tether formation, and conversion of tethers to rolling adhesions, providing further understanding of the shear threshold requirement for rolling through L-selectin.


DNA rearrangements in the alpha 5(IV) collagen gene (COL4A5) of individuals with Alport syndrome: further refinement using pulsed-field gel electrophoresis.
Alport syndrome (AS), an X-linked kidney disorder, has been shown to be caused by mutations in the gene for the alpha 5-chain of type IV collagen (COL4A5), which maps to Xq22. On the basis of the results of conventional Southern blot analysis of AS patient DNAs, we employed pulsed-field gel electrophoresis to characterize further three gene rearrangements at the 3'-end of alpha 5(IV). We were able to construct long-range restriction maps for all three of these patients and deduce the extent and nature of each rearrangement. One of these mutations is a 450-kb simple deletion that includes 12 kb of the alpha 5(IV) gene. A second mutation has been shown to be a direct duplication of 35 kb of alpha 5(IV) genomic DNA, and a third mutation involves a complex insertion/deletion event resulting in an overall loss of 25 kb.