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Introduction Super-utilizers (patients with 4 or more emergency department [ED] visits a year) account for 10% to 26% of all ED visits and are responsible for a growing proportion of healthcare expenditures. Patients recognize the ED as a reliable provider of acute care, as well as a timely resource for diagnosis and treatment. The value of ED care is indisputable in critical and emergent conditions, but in the case of non-urgent conditions, ED utilization may represent an inefficiency in the healthcare system. We sought to identify patient and clinical characteristics associated with ED super-utilization in a universally insured population. Material and Methods We performed a retrospective cohort study using TRICARE claims data from the Military Health System Data Repository (2011-2015). We reviewed the claims data of all adult patients (aged 18-64 years) who had at least one encounter at the ED for any cause. Multivariable logistic regression was used to determine independent factors associated with ED super-utilization. Results Factors associated with increased odds of ED super-utilization included Charlson Score ≥2 (adjusted odds ratio [aOR] 1.98, 95% confidence interval [CI]: 1.90-2.06), being eligible for Medicare (aOR 1.95, 95% CI: 1.90-2.01), and female sex (aOR 1.35, 95% CI: 1.33-1.37). Active duty service members (aOR 0.69, 95% CI 0.68-0.72) and beneficiaries with higher sponsor-rank (Officers: aOR 0.50, 95% CI: 0.55-0.57; Senior enlisted: aOR 0.82, 95% CI: 0.81-0.83) had lower odds of ED super-utilization. The most common primary diagnoses for ED visits among super-utilizers were abdominal pain, headache and migraine, chest pain, urinary tract infection, nausea and vomiting, and low back pain. Conclusions Risk of ED super-utilization appears to increase with age and diminished health status. Patient demographic and clinical characteristics of ED super-utilization identified in this study can be used to formulate healthcare policies addressing gaps in primary care in diagnoses associated with ED super-utilization and develop interventions to address modifiable risk factors of ED utilization.

INTRODUCTION: During the Global War on Terrorism, many US Military service members sustained injuries with potentially long-lasting functional limitations and chronic pain. We sought to understand the patterns of prescription opioid use among service members injured in combat. METHODS: We queried the Military Health System Data Repository to identify service members injured in combat between 2007 and 2011. Sociodemographics, injury characteristics, treatment information, and costs of care were abstracted for all eligible patients. We surveyed for prescription opioid utilization subsequent to hospital discharge and through 2018. Negative binomial regression was used to identify factors associated with cumulative prescription opioid use. RESULTS: We identified 3,981 service members with combat-related injuries presenting during the study period. The median age was 24 years (interquartile range [IQR], 22-29 years), 98.5% were male, and the median follow-up was 3.3 years. During the study period, 98% (n = 3,910) of patients were prescribed opioids at least once and were prescribed opioids for a median of 29 days (IQR, 9-85 days) per patient-year of follow-up. While nearly all patients (96%; n = 3,157) discontinued use within 6 months, 91% (n = 2,882) were prescribed opioids again after initially discontinuing opioids. Following regression analysis, patients with preinjury opioid exposure, more severe injuries, blast injuries, and enlisted rank had higher cumulative opioid use. Patients who discontinued opioids within 6 months had an unadjusted median total health care cost of US $97,800 (IQR, US $42,364-237,135) compared with US $230,524 (IQR, US $134,387-370,102) among those who did not discontinue opioids within 6 months (p < 0.001). CONCLUSION: Nearly all service members injured in combat were prescribed opioids during treatment, and the vast majority experienced multiple episodes of prescription opioid use. Only 4% of the population met the criteria for sustained prescription opioid use at 6 months following discharge. Early discontinuation may not translate to long-term opioid cessation in this population. LEVEL OF EVIDENCE: Epidemiology study, level III.

Background In prior reports from population-based databases, black patients with extremity soft tissue sarcoma (ESTS) have lower reported rates of limb-sparing surgery and adjuvant treatment. The objective of this study was to compare the multimodality treatment of ESTS between black and white patients within a universally insured and equal-access health care system. Methods Claims data from TRICARE, the US Department of Defense insurance plan that provides health care coverage for 9 million active-duty personnel, retirees, and dependents, were queried for patients younger than 65 y with ESTS who underwent limb-sparing surgery or amputation between 2006 and 2014 and identified as black or white race. Multivariable logistic regression analysis was used to evaluate the impact of race on the utilization of surgery, chemotherapy, and radiation. Results Of the 719 patients included for analysis, 605 patients (84%) were white and 114 (16%) were black. Compared with whites, blacks had the same likelihood of receiving limb-sparing surgery (odds ratio [OR], 0.861; 95% confidence interval [95% CI], 0.284-2.611; P = 0.79), neoadjuvant radiation (OR, 1.177; 95% CI, 0.204-1.319; P = 0.34), and neoadjuvant (OR, 0.852; 95% CI, 0.554-1.311; P = 0.47) and adjuvant (OR, 1.211; 95% CI, 0.911-1.611; P = 0.19) chemotherapy; blacks more likely to receive adjuvant radiation (OR, 1.917; 95% CI, 1.162-3.162; P = 0.011). Conclusions In a universally insured population, racial differences in the rates of limb-sparing surgery for ESTS are significantly mitigated compared with prior reports. Biologic or disease factors that could not be accounted for in this study may contribute to the increased use of adjuvant radiation among black patients.

Only 25% of dog owners report receiving behavioral advice from their veterinarian. Preadoption counseling and general behavioral information sharing in routine puppy visits have been shown to significantly reduce the number of dogs that are euthanized or surrendered. Clinicians often categorize small dogs as being more difficult to house-train; however, there have been no published studies to date to confirm the veracity of this assumption. Some authors have proposed that smaller dogs have both a higher metabolism and a smaller bladder associated with more frequent urination which may lead to incomplete house-training. It is crucial to elucidate the underlying cause(s) of incomplete house-training because lack of house-training has been identified as a common reason for relinquishment. The purpose of this study was to evaluate the relationship between body size and completeness of house-training. An internet survey was made readily available on www.surveymonkey.com and posted on several general practice and referral center webpages and social media pages encouraging dog owners to participate. Questions regarding age at the time of adoption or purchase, level of obedience training, the role of the pet in the family, and medical disorders were included to rule in or out other contributing factors. Seven hundred thirty-five surveys submitted by private dog owners were fully completed and included in the study. A chi-square test of independence was performed which revealed a statistically significant difference between large dogs and small dogs regarding the completeness of house-training. Specifically, large dogs were more likely to be fully house-trained than small dogs. There was also a correlation between the level of training and completeness of house-training. Further research is warranted to understand why these differences are present. Clinically, this information provides knowledge which animal care professionals can use to advise clients that dogs of different sizes may require varied types and lengths of training to accomplish the goal of complete house-training.

Background In the move toward value-based care, bundled payments are believed to reduce waste and improve coordination. Some commercial insurers have addressed this through the use of bundled payment, the provision of one fee for all care associated with a given index procedure. This system was pioneered by Medicare, using a population generally over 65 years of age, and despite its adoption by mainstream insurers, little is known of bundled payments’ ability to reduce variation or cost in a working-age population. This study uses a universally-insured, nationally-representative population of adults aged 18–65 to examine the effect of bundled payments for five high-cost surgical procedures which are known to vary widely in Medicare reimbursement: hip replacement, knee replacement, coronary artery bypass grafting (CABG), lumbar spinal fusion, and colectomy. Methods Five procedures conducted on adults aged 18–65 were identified from the TRICARE database from 2011 to 2014. A 90-day period from index procedure was used to determine episodes of associated post-acute care. Data was sorted by Zip code into hospital referral regions (HRR). Payments were determined from TRICARE reimbursement records, they were subsequently price standardized and adjusted for patient and surgical characteristics. Variation was assessed by stratifying the HRR into quintiles by spending for each index procedure. Results After adjusting for case mix, significant inter-quintile variation was observed for all procedures, with knee replacement showing the greatest variation in both index surgery (107%) and total cost of care (75%). Readmission was a driver of variation for colectomy and CABG, with absolute cost variation of $17,257 and $13,289 respectively. Other post-acute care spending was low overall (≤$1606, for CABG). Conclusions This study demonstrates significant regional variation in total spending for these procedures, but much lower spending for post-acute care than previously demonstrated by similar procedures in Medicare. Targeting post-acute care spending, a common approach taken by providers in bundled payment arrangements with Medicare, may be less fruitful in working aged populations.

Utility-implemented load-shedding aims to balance the supply and demand of a constrained power grid for a scheduled period of time by switching off supply to selected regions. The unintended consequence of this strategy occurs at the return of supply, where an instantaneous peak demand is experienced as an impulse at localised points of the power system. This results in frequent trips on medium voltage breakers that can damage infrastructure and result in delayed return of supply to end-users. The aim of this paper is to examine demand-side management interventions using intelligent control of electric water heaters (EWHs) to reduce the instantaneous peak demand at return of supply. Domestic water heating load was chosen as it is a deferrable, dispatchable, passive load with a high power requirement. The approach taken simulates the loading effect of 3 600 EWHs using three measured tank states and the combination of the Monte Carlo algorithm for probabilistic population behaviour. A typical 150-litre, 3 kW EWH in horizontal orientation is used, with a maximum draw volume of 50-litre per tank per day. It is shown that system changes to load-shedding schedules from a period of 4.5-hours to 2.5-hours beneficially reduces the instantaneous peak demand with the shorter period. For a 3 600 EWH-population, simulated results comparing uniformly distributed stochastic delays for activation time windows within 15, 30, 45, and 60 minutes show that a 15-minute activation time window could reduce the expected peak demand by a factor of two. This is important because intelligent controller retrofits onto EWHs is becoming more ubiquitous and if this simple sub-routine is built-in to produce a short delay after a period of no supply, it could result in significant passive impact on the power grid.

Bacterial vaginosis (BV) has been reported in one-third of women worldwide at different life stages, due to the complex balance in the ecology of the vaginal microbiota. It is a common cause of abnormal vaginal discharge and is associated with other health issues. Since the first description of anaerobic microbes associated with BV like Gardnerella vaginalis in the 1950s, researchers have stepped up the game by incorporating advanced molecular tools to monitor and evaluate the extent of dysbiosis within the vaginal microbiome, particularly on how specific microbial population changes compared to a healthy state. Moreover, treatment failure and BV recurrence rate remain high despite the standard antibiotic treatment. Consequently, researchers have been probing into alternative or adjunct treatments, including probiotics or even vaginal microbiota transplants, to ensure successful treatment outcomes and reduce the colonization by pathogenic microbes of the female reproductive tract. The current review summarizes the latest findings in probiotics use for BV and explores the potential of vaginal microbiota transplants in restoring vaginal health.

Objective: Mindfulness-based intervention (MBI) has been proposed to alleviate loneliness and improve social connectedness. Several randomized controlled trials (RCTs) have been conducted to evaluate the effectiveness of MBI. This study aimed to critically evaluate and determine the effectiveness and safety of MBI in alleviating the feeling of loneliness.Methods: We searched Medline, Embase, PsycInfo, Cochrane CENTRAL, and AMED for publications from inception to May 2020. We included RCTs with human subjects who were enrolled in MBI with loneliness as an outcome. The quality of evidence was assessed using Cochrane's Risk of Bias (ROB) tool and Grading of Recommendations Assessment, Development, and Evaluation (GRADE). A random-effects model was used for meta-analysis.Results: Out of 92 articles identified, eight studies involving 815 participants were included in this study. Most (7/8) trials conducted a minimum of 8 weeks of MBI. Most of the trials (5/8) used UCLA-Loneliness Scale. A pooled analysis combining three trials and compared with wait-list showed significant improvement in loneliness score reduction using the UCLA-R scale with MD of −6.33 [95% confidence interval (CI): −9.39, −3.26]. Subgroup analysis with only two Cognitively-Based Compassion Training (CBCT) trials also showed similar MD of −6.05 (95% CI: −9.53, 2.58). The overall quality of evidence (GRADE) was low.Conclusions: Mindfulness intervention with an average length of 8-week duration significantly improved the population's loneliness level with no mental health issue. However, this evidence had a low GRADE level.

Introduction Emergency department (ED) utilization represents an expensive and growing means of accessing care for a variety of conditions. Prior studies have characterized ED utilization in the general population. We aim to identify the clinical conditions that drive ED utilization in a universally insured population and the impacts of care setting on ED use and admissions in the U.S. Military Health System. Methods We queried TRICARE claims data from October 1, 2012, to September 30, 2015, to identify all ED visits for adult patients (age 18-64). The primary presenting diagnoses of all ED visits and those leading to admission are presented with descriptive statistics. Logistic regression was used to identify clinical and sociodemographic factors associated with admission from the ED. Results A total of 4,687,205 ED visits were identified, of which 46% took place in the DoD healthcare facilities (direct care). The most common diagnoses across all ED visits were abdominal pain, chest pain, headache, nausea and vomiting, and urinary tract infection. A total of 270,127 (5.8%) ED visits led to inpatient admission. The most common diagnoses leading to admission were chest pain, abdominal pain, depression, conditions relating to acute psychological stress, and pneumonia. For patients presenting with 1 of the 10 most common ED diagnoses, those who were seen at a civilian ED were significantly less likely to be admitted (3.4%) compared to direct care facilities (4.1%) in an adjusted logistic regression model (Adjusted Odds Ratio 0.40 [95% CI: 0.40-0.41], P Conclusions Ultimately, we show that abdominal pain and chest pain are the most common reasons for presentation to the ED in the Military Health System and the most common presenting diagnoses for admission from the ED. Among patients presenting with the most common ED conditions, direct care EDs were significantly more likely to admit patients than civilian facilities.

Across group-living animals, linear dominance hierarchies lead to disparities in access to resources, health outcomes, and reproductive performance. Studies of how dominance rank affects these outcomes typically employ one of several dominance rank metrics without examining the assumptions each metric makes about its underlying competitive processes. Here we compare the ability of two dominance rank metrics—ordinal rank and proportional or ‘standardized’ rank—to predict 20 distinct traits in a well-studied wild baboon population in Amboseli, Kenya. We propose that ordinal rank best predicts outcomes when competition is density-dependent, while proportional rank best predicts outcomes when competition is density-independent. We found that for 75% (15/20) of the traits, one of the two rank metrics performed better than the other. Strikingly, all male traits were better predicted by ordinal than by proportional rank, while female traits were evenly split between being better predicted by proportional or ordinal rank. Hence, male and female traits are shaped by different competitive regimes: males’ competitive environments are largely driven by density-dependent resource access (e.g., access to estrus females), while females’ competitive environments are shaped by both density-independent resource access (e.g. distributed food resources) and density-dependent resource access. However, traits related to competition for social and mating partners are an exception to this sex-biased pattern: these traits were better predicted by ordinal rank than by proportional rank for both sexes. We argue that this method of comparing how different rank metrics predict traits of interest can be used as a way to distinguish between different competitive processes operating in animal societies.

The risk for female students in academia of sexual harassment by male faculty and staff remains a national crisis. This study examined effects of severity of coercive sexual harassment (CSH) by a male professor of a female student on peer bystander intervention responses. A total of 180 undergraduate and graduate college students who volunteered for an online survey were randomly assigned to one of three vignette conditions that varied severity of CSH. Following Bowes-Sperry’s ethical model of bystander behaviors, it was predicted that severity of CSH would affect cognitive appraisals and emotional reactions, which would further predict intentions for intervention behaviors. Predictions generally were supported. Logistic regression analyses indicated that severity of CSH, fears of consequences, and negative emotions toward the professor were significant predictors of direct confrontation of the professor by the student observer, whereas only severity and negative emotions toward the professor were predictors of the indirect intervention (finding an excuse to remove the victim) or delegation (going to get help) while the incident was occurring. Fears of personal consequences suppressed direct intervention in the moderate condition. Findings suggest ongoing need within academia for clarification of behaviors that constitute CSH, as well as safety for bystanders who intervene.

Ecoimmunological patterns and processes remain understudied in wild primates, in part because of the lack of non-invasive methods to measure immunity. Secretory immunoglobulin A (sIgA) is the most abundant antibody present at mammalian mucosal surfaces and provides an important first line of defense against pathogens. Recent studies show that sIgA can be measured non-invasively in feces and is a good marker of mucosal immunity. Here we validated a commercial ELISA kit to measure fecal IgA in baboons, tested the robustness of its results to variation in collection and storage conditions, and developed a cost-effective in-house ELISA for baboon fecal IgA. Using data from the custom ELISA, we assessed the relationship between fecal IgA concentrations and gastrointestinal parasite burden, and tested how sex, age, and reproductive effort predict fecal IgA in wild baboons. We find that IgA concentrations can be measured in baboon feces using an in-house ELISA and are highly correlated to the values obtained with a commercial kit. Fecal IgA concentrations are stable when extracts are stored for up to 22 months at −20°C. Fecal IgA concentrations were negatively correlated with parasite egg counts (Trichuris trichiura), but not parasite richness. Fecal IgA did not vary between the sexes, but for males, concentrations were higher in adults versus adolescents. Lactating females had significantly lower fecal IgA than pregnant females, but neither pregnant nor lactating female concentrations differed significantly from cycling females. Males who engaged in more mate-guarding exhibited similar IgA concentrations to those who engaged in little mate-guarding. These patterns may reflect the low energetic costs of mucosal immunity, or the complex dependence of IgA excretion on individual condition. Adding a non-invasive measure of mucosal immunity will promote a better understanding of how ecology modulates possible tradeoffs between the immune system and other energetically costly processes in the wild.

Across group-living animals, linear dominance hierarchies lead to disparities in access to resources, health outcomes and reproductive performance. Studies of how dominance rank predicts these traits typically employ one of several dominance rank metrics without examining the assumptions each metric makes about its underlying competitive processes. Here, we compare the ability of two dominance rank metrics—simple ordinal rank and proportional or ‘standardized’ rank—to predict 20 traits in a wild baboon population in Amboseli, Kenya. We propose that simple ordinal rank best predicts traits when competition is density-dependent, whereas proportional rank best predicts traits when competition is density-independent. We found that for 75% of traits (15/20), one rank metric performed better than the other. Strikingly, all male traits were best predicted by simple ordinal rank, whereas female traits were evenly split between proportional and simple ordinal rank. Hence, male and female traits are shaped by different competitive processes: males are largely driven by density-dependent resource access (e.g. access to oestrous females), whereas females are shaped by both density-independent (e.g. distributed food resources) and density-dependent resource access. This method of comparing how different rank metrics predict traits can be used to distinguish between different competitive processes operating in animal societies.

Background: This study explored the knowledge of attitudes to and perception of consumption of antibiotics, and understanding of antibiotic resistance among students and general population across South and East Asia region. Method: A systematic review was performed through PubMed, Scopus, EBSCOhost (Medline) and EMBASE scientific databases. Cross-sectional studies aimed to investigate the knowledge of, attitude to and perception of antibiotic use as well as studies concerning the demand of antibiotics, antibiotic resistance were evaluated. Other inclusion criteria were: studies published in English prior to July 2016; survey that examined one of the 7 topics proposed by Gualano et al. (2015) in their systematic review on knowledge and attitudes about antibiotics. Results: A total of 9 studies were selected for systematic review. A lack of knowledge about antibiotics was identified. Approximately 50% of the sample mistakenly believed that antibiotics could treat viral infections which are commonly responsible for the common cold, sore throat and cough. The majority (above 60%) of the sample were compliant with completing a full course of antibiotics but unfortunately 40% of the sample still selfadministered antibiotics without prescription. Conclusion: Knowledge of, attitudes to and perceptions of antibiotic use are found to be inappropriate among the student and general public in the South and East Asia region. Although antibiotic resistance is a global issue, every country in the South and East Asia region is encouraged to take their own initiative to curb antibiotic resistance. Key words: Antimicrobial resistance; Prescribing practice; Anti-infective agents; Antibacterials; Pharmacoepidemiology.

Objective: To compare disparities in postoperative outcomes for African Americans after surgical intervention in the universally insured military system, versus the civilian setting in California. Background: Health reform proponents cite the reduction of disparities for African Americans and minorities as an expected benefit. The impact of universal health insurance on reducing surgical disparities for African Americans has not previously been examined. Methods: We used Department of Defense health insurance (Tricare) data (2006–2010) to measure outcomes for African Americans as compared with Whites after 12 major surgical procedures across multiple specialties. The experience of African Americans in the Tricare system was compared with a similar cohort undergoing surgery in the state of California using the State Inpatient Database (2007–2011). Results: No significant difference in postoperative complications [odds ratio (OR) 0.91; 95% confidence interval (CI) 0.81, 1.03] or mortality (OR 0.98; 95% CI 0.43, 2.25) were encountered between African Americans and Whites receiving surgery at hospitals administered by the Department of Defense. African Americans in California who were uninsured or on Medicaid had significantly increased odds of mortality (OR 4.76; 95% CI 2.82, 8.05), complications (OR 1.67; 95% CI 1.34, 2.08), failure to rescue (OR 2.72; 95% CI 1.25, 5.94), and readmission (OR 1.78; 95% CI 1.45, 2.19). Conclusions: In the equal access military healthcare system, African Americans have outcomes similar to Whites. Disparities were evident in California, especially among those without private insurance. These facts point toward the potential benefits of a federally administered system in which all patients are treated uniformly.

Background Recently, many reports indicated the role of the gut microbiome in the development of autism in young children. Microbes are capable of synthesizing small molecules like fatty acids and sugars which act as signalling molecules to activate/deactivate nervous system or even trigger an inflammatory response. Thus, the current review aims to explore the role of microbes in the development of autism, summarizing data from animal models and human studies. Methods Referring to the preferred reporting items for systematic reviews and meta-analyses (PRISMA) guidelines, searches were performed in three databases (PubMed, ScienceDirect, Web of knowledge and; database inception to 31/12/2018) using ‘microbiome’ OR ‘microbiota’ combined with ‘autism’ as MeSH terms. All the titles and Abstracts retrieved were screened based on the inclusion and exclusion criteria. Studies reporting gut microbiome data in relation to the development of autism were included. Studies without gut microbiome data and/or reports on the evaluation on autism were excluded, along with reviews, conference Abstracts, case studies, and comments. Results Out of the 2237 articles were accessed, seven studies were eligible for the qualitative analysis according to the inclusion criteria. Two studies described the murine model of autism, while the remaining five studies focused on children. One study revealed that there are no significant differences in gut microbiome among the three groups--severe, mild and healthy children, while three studies indicated that there is a higher abundance of Proteobacteria and Bacteroides in autistic children, with a reduced population of Firmicutes and Actinobacteria (figure 1). One study highlighted that higher Clostridiaceae species present in autistic children which explain for the development of autism in children as these species could produce toxic metabolic products (e.g., phenols, p-cresol, indole derivatives). Similar results were also observed in autistic animal models - there is increase abundance of Bacteroides, Parabacteroides, Sutterella, Dehalobacterium and Oscillospira genera. Conclusions Altogether, these results revealed a positive correlation between dysbiosis and autism. Microbiome alterations may contribute to the development of autism, particularly via the production of toxic bacterial metabolites and alteration in immune function. By keeping the ‘healthy’ gut bacteria in check, these efforts could improve intestinal and mental health, easing and reducing autistic behaviour in children.

In the class Actinobacteria, the renowned genus Streptomyces comprised of a group of uniquely complex bacteria that capable of synthesizing a great variety of bioactive metabolites. Streptomycetes are noted to possess several special qualities such as multicellular life cycle and large linearized chromosomes. The significant contribution of Streptomyces in microbial drug discovery as witnessed through the discovery of many important antibiotic drugs has undeniably encourage the exploration of these bacteria from different environments, especially the mangrove environments. This review emphasizes on the genus Streptomyces and reports on the diversity of actinobacterial population from mangroves at different regions of the world as well as discovery of mangrove-derived novel Streptomyces species. Overall, the research on diversity of Actinobacteria in the mangrove environments remains limited. A total of 19 novel Streptomyces spp. isolated from mangroves between the year 2009 - early 2019, notably from China, India, Malaysia, and Thailand. Hence, it will be worthwhile to encourage the study of these bacteria from mangroves of different locations.

Contains fulltext : 170424.pdf (Publisher’s version ) (Closed access) Cerebral small vessel disease (cSVD) and amyloid-beta (Abeta) deposition often co-exist in (prodromal) dementia, and both types of pathology have been associated with neurodegeneration. We examined whether cSVD and Abeta have independent or interactive effects on hippocampal volume (HV) in a memory clinic population. We included 87 individuals with clinical diagnoses of Alzheimer's disease (AD) (n = 24), mild cognitive impairment (MCI) (n = 26), and subjective cognitive complaints (SCC) (n = 37). cSVD magnetic resonance imaging markers included white matter hyperintensity (WMH) volume, lacunar infarct presence, and microbleed presence. Abeta pathology was assessed as cerebrospinal fluid-derived Abeta1 - 42 levels and dichotomized into normal or abnormal, and HV was determined by manual volumetric measurements. A linear hierarchical regression approach was applied for the detection of additive or interaction effects between cSVD and Abeta on HV in the total participant group (n = 87) and in the non-demented group (including SCC and MCI individuals only, n = 63). The results revealed that abnormal Abeta and lacunar infarct presence were independently associated with lower HV in the non-demented individuals. Interestingly, Abeta and WMH pathology interacted in the non-demented individuals, such that WMH had a negative effect on HV in individuals with abnormal CSF Abeta42 levels, but not in individuals with normal CSF Abeta42 levels. These associations were not present when individuals with AD were included in the analyses. Our observations suggest that relatively early on in the disease process older individuals with abnormal Abeta levels are at an increased risk of accelerated disease progression when concomitant cSVD is present.

Background Recently researchers speculated that the gut microbiota might contribute to the development of chronic diseases like Alzheimer’s disease (AD), given that some bacteria are capable of synthesising amyloids and inducing inflammation via production of endotoxins. Thus, this study aims to understand the role of gut microbiota in AD development, obtaining clues on the regulation of gut microbes to prevent and tackle AD. Methods Following the preferred reporting items for systematic reviews and meta-analyses (PRISMA) guidelines, searches were performed in 4 databases (PubMed, Medline, ScienceDirect, EBSCO; database inception to February 2018) using ‘gut’, ‘microbiome’ combined with ‘Alzheimer’s’ or ‘amyloid’ as MeSH terms. All titles and abstracts retrieved were screened based on the inclusion and exclusion criteria. Studies reporting gut microbiome data in relation to the development of AD were included. Studies without gut microbiome data and/or AD biomarker(s) were excluded, along with reviews, conference abstracts, systematic reviews, meta-analyses, comments, and letters to the editor. Results After removing duplicate records, a total of 155 articles were accessed, which resulted in 70 articles excluded based on their titles and abstracts. Seven out of 85 studies were eligible for the qualitative analysis according to the inclusion criteria. Among these, six studies involve murine model, while one study was conducted in humans. Using transgenic murine model of AD, all studies reported a remarkable shift in the microbiota composition as compared to controls, with significant changes in phyla like Firmicutes and Bacteroidetes. The high abundance of pro-inflammation bacteria like Proteobacteria may induce immunological reactions and neuroinflammation, which are known aetiology of AD. Complementing with in vivo studies, Cattaneo et al. (2017) recorded higher abundance of pro-inflammatory bacteria (e.g. Escherichia/Shigella, Pseudomonas aeruginosa) and lower distribution of anti-inflammatory bacteria (e.g. Bacillus fragilis, Eubacterium rectale, Eubacterium hallii, Faecalibacterium prausnitzii and Bacteroides fragilis) in amyloid-positive patients as compared to healthy subjects. Conclusions As depicted in figure 1: The dysbiosis of the gut microbiota plays a significant role in the development of AD. Evidence showed the high abundance of pro-inflammatory bacteria with a reduced population of anti-inflammatory bacteria could promote neuroinflammation, which can exacerbate amyloid beta plaques formation.

Background Racial disparities in colorectal cancer (CRC) screening are frequently attributed to variations in insurance status. The objective of this study was to ascertain whether universal insurance would lead to more equitable utilization of CRC screening for black patients in comparison with white patients. Methods Claims data from TRICARE (insurance coverage for active, reserve, and retired members of the US Armed Services and their dependents) for 2007-2010 were queried for adults aged 50 years in 2007, and they were followed forward in time for 4 years (ages, 50-53 years) to identify their first lower endoscopy and/or fecal occult blood test (FOBT). Variations in CRC screening were compared with descriptive statistics and multivariate logistic regression. Results Among the 24,944 patients studied, 69.2% were white, 20.3% were black, 4.9% were Asian, and 5.6% were other. Overall, 54.0% received any screening: 83.7% received endoscopy, and 16.3% received FOBT alone. Compared with whites, black patients had higher screening rates (56.5%) and had 20% higher risk-adjusted odds of being screened (95% confidence interval [CI], 1.11-1.29). Asian patients had a likelihood of screening similar to that of white patients (odds ratio [OR], 1.06; 95% CI, 0.92-1.23). Females (OR, 1.20; 95% CI, 1.10-1.33), active-duty personnel (OR, 1.15; 95% CI, 1.06-1.25), and officers (OR, 1.28; 95% CI, 1.18-1.37) were also more likely to be screened. Conclusion Within an equal-access, universal health care system, black patients had higher rates of CRC screening in comparison with prior reports and even in comparison with white patients within the population. These findings highlight the need to understand and develop meaningful approaches for promoting more equitable access to preventative care. Moreover, equal-access, universal health insurance for both the military and civilian populations can be presumed to improve access for underserved minorities.

Significance Chimpanzees, bonobos, and gorillas harbor close relatives of human Plasmodium vivax, but current knowledge of these parasites is limited to a small number of gene fragments derived almost exclusively from mitochondrial DNA. We compared nearly full-length genomes of ape parasites with a global sample of human P. vivax and tested the function of human and ape P. vivax proteins believed to be important for erythrocyte binding. The results showed that ape parasites are 10-fold more diverse than human P. vivax and exhibit no evidence of species specificity, whereas human P. vivax represents a bottlenecked lineage that emerged from within this parasite group. Thus, African apes represent a large P. vivax reservoir whose impact on human malaria eradication requires careful monitoring., Wild-living African apes are endemically infected with parasites that are closely related to human Plasmodium vivax, a leading cause of malaria outside Africa. This finding suggests that the origin of P. vivax was in Africa, even though the parasite is now rare in humans there. To elucidate the emergence of human P. vivax and its relationship to the ape parasites, we analyzed genome sequence data of P. vivax strains infecting six chimpanzees and one gorilla from Cameroon, Gabon, and Côte d’Ivoire. We found that ape and human parasites share nearly identical core genomes, differing by only 2% of coding sequences. However, compared with the ape parasites, human strains of P. vivax exhibit about 10-fold less diversity and have a relative excess of nonsynonymous nucleotide polymorphisms, with site-frequency spectra suggesting they are subject to greatly relaxed purifying selection. These data suggest that human P. vivax has undergone an extreme bottleneck, followed by rapid population expansion. Investigating potential host-specificity determinants, we found that ape P. vivax parasites encode intact orthologs of three reticulocyte-binding protein genes (rbp2d, rbp2e, and rbp3), which are pseudogenes in all human P. vivax strains. However, binding studies of recombinant RBP2e and RBP3 proteins to human, chimpanzee, and gorilla erythrocytes revealed no evidence of host-specific barriers to red blood cell invasion. These data suggest that, from an ancient stock of P. vivax parasites capable of infecting both humans and apes, a severely bottlenecked lineage emerged out of Africa and underwent rapid population growth as it spread globally.

INTRODUCTION The factors that contribute to variation in utilization of laparoscopic inguinal hernia repair are unknown. We sought to determine the current usage patterns of laparoscopic and open surgery in the elective repair of uncomplicated unilateral inguinal hernia in a large population with universal health care coverage comprised of Department of Defense (DoD) beneficiaries. MATERIALS AND METHODS The DoD Military Health System Data Repository (MDR) tracks health care delivered to a universally insured population of active/reserve/retired members of the U.S. Armed Services and their dependents. The MDR was queried for elective unilateral inguinal hernia repair among adult patients between 2008 and 2014. The primary outcome was laparoscopic (vs. open) approach to hernia repair. We conducted univariable and multivariable analyses of patient- and systems-level factors associated with approach to inguinal hernia repair. This research was approved by our institutional review board prior to commencement of the study and need for informed consent was waived given the design of this study. RESULTS Among 37,742 elective uncomplicated unilateral inguinal hernia repairs, 35% (n = 13,114) were performed laparoscopically. In 2014, 40% of inguinal hernia repairs were performed laparoscopically, compared with 27% of repairs in 2008 (P < 0.01). In multivariable analysis, laparoscopic hernia repair was more likely for male patients (OR = 1.38, 95% CI = 1.23-1.54, P < 0.01), military (vs. civilian) institutions (OR = 1.34, 95% CI = 1.28-1.41, P < 0.01), active-duty officers (vs. active-duty enlisted; OR = 1.21, 95% CI = 1.12-1.30, P < 0.01), and more recent year of surgery (P < 0.01). Laparoscopic repair was significantly less likely among patients with greater than one comorbidity (vs. none; OR = 0.68, 95% CI = 0.61-0.76, P < 0.01). CONCLUSION In a large, universally insured population of military service members and their dependents, laparoscopic inguinal repair is increasingly used and was preferred over open repair for younger, healthier, active-duty patients and those treated within the military (vs. non-military) care system.

Rattus argentiventer (Rice Field Rat) is responsible for destruction of paddy. The mammal is known for its rapid reproductive potential which can be a target for biological control of this species. Phospholipase C-zeta (PLC ζ) is a specific enzyme found in sperm of mammals responsible for triggering calcium oscillations leading to egg activation during fertilization. It facilitates the first step of egg activation causing egg division and subsequent development into an embryo. The method used in this study was to identify PLC gene fragments from the testis of Rattus argentiventer using two-step Reverse Transcriptase Polymerase Chain Reaction (PCR) and quantified with Real Time Reverse Transcriptase Polymerase Chain Reaction (qRT-PCR). Following that, sequencing of PLC was performed. The identified sequence was then keyed in into the BLAST portal in NCBI (National Centre of Bioinformatics) for comparison with a standard PLC sequence from Rattus norvegicus for sequence alignment. The result showed that PLCζ was present in Rattus argentiventer and qRTPCR could quantify the amount of PLCζ available.As such this would give additional information of detection and quantification techniques for rapid identification and detection of PLCζ to be carried out in the approach of controlling the rapid growth of Rattus argentiventer population.

HIV-1 can evolve HLA-specific escape variants in response to HLA-mediated cellular immunity. HLA alleles that are common in the host population may increase the frequency of such escape variants at the population level. When loss of viral fitness is caused by immune escape variation, these variants may revert upon infection of a new host who does not have the corresponding HLA allele. Furthermore, additional escape variants may appear in response to the nonconcordant HLA alleles. Because individuals with rare HLA alleles are less likely to be infected by a partner with concordant HLA alleles, viral populations infecting hosts with rare HLA alleles may undergo a greater amount of evolution than those infecting hosts with common alleles due to the loss of preexisting escape variants followed by new immune escape. This hypothesis was evaluated using maximum likelihood phylogenetic trees of each gene from 272 full-length HIV-1 sequences. Recent viral evolution, as measured by the external branch length, was found to be inversely associated with HLA frequency in nef (p < 0.02), env (p < 0.03), and pol (p < or = 0.05), suggesting that rare HLA alleles provide a disproportionate force driving viral evolution compared to common alleles, likely due to the loss of preexisting escape variants during early stages postinfection.

HIV transmission via breastfeeding accounts for a considerable proportion of infant HIV acquisition. However, the origin and evolution of the virus population in breast milk, the likely reservoir of transmitted virus variants, are not well characterized. In this study, HIV envelope ( env ) genes were sequenced from virus variants amplified by single-genome amplification from plasmas and milk of 12 chronically HIV-infected, lactating Malawian women. Maximum likelihood trees and statistical tests of compartmentalization revealed interspersion of plasma and milk HIV env sequences in the majority of subjects, indicating limited or no compartmentalization of milk virus variants. However, phylogenetic tree analysis further revealed monotypic virus variants that were significantly more frequent in milk (median proportion of identical viruses, 29.5%; range, 0 to 61%) than in plasma (median proportion of identical viruses, 0%; range, 0 to 26%) ( P = 0.002), suggesting local virus replication in the breast milk compartment. Moreover, clonally amplified virus env genes in milk produced functional virus Envs that were all CCR5 tropic. Milk and plasma virus Envs had similar predicted phenotypes and neutralization sensitivities to broadly neutralizing antibodies in both transmitting and nontransmitting mothers. Finally, phylogenetic comparison of longitudinal milk and plasma virus env sequences revealed synchronous virus evolution and new clonal amplification of evolved virus env genes in milk. The limited compartmentalization and the clonal amplification of evolving, functional viruses in milk indicate continual seeding of the mammary gland by blood virus variants, followed by transient local replication of these variants in the breast milk compartment.

Diabetic retinopathy (DR) is among the leading causes of new onset blindness in adults. Effective treatment may delay the onset and progression of this disease provided it is diagnosed early. At present retinopathy can only be diagnosed via formal examination of the eye by a trained specialist, which limits the population that can be effectively screened. An easily accessible, reliable screening biomarker of diabetic retinopathy would be of tremendous benefit in detecting the population in need of further assessment and treatment. This review highlights specific biomarkers that show promise as screening markers to detect early diabetic retinopathy or even to detect patients at increased risk of DR at the time of diagnosis of diabetes. The pathobiology of DR is complex and multifactorial giving rise to a wide array of potential biomarkers. This review provides an overview of these pathways and looks at older markers such as advanced glycation end products (AGEs), inflammatory markers, vascular endothelial growth factor (VEGF) as well as other newer proteins with a role in the pathogenesis of DR including neuroprotective factors such as brain derived neurotrophic factor (BDNF) and Pigment Epithelium Derived Factor (PEDF); SA100A12, pentraxin 3, brain natriuretic peptide, apelin 3, and chemerin as well as various metabolites such as lipoprotein A, folate, and homocysteine. We also consider the possible role of proteins identified through proteomics work whose levels are altered in the sera of patients with DR as screening markers though their role in pathophysiology remains to be characterized. The role of microRNA as a promising new screening marker is also discussed.

In response to the increased seafood demand from the ever-going human population, aquaculture has become the fastest growing animal food-producing sector. However, the indiscriminate use of antibiotics as a biological control agents for fish pathogens has led to the emergence of antibiotic resistance bacteria. Probiotics are defined as living microbial supplement that exert beneficial effects on hosts as well as improvement of environmental parameters. Probiotics have been proven to be effective in improving the growth, survival and health status of the aquatic livestock. This review aims to highlight the genus Streptomyces can be a good candidate for probiotics in aquaculture. Studies showed that the feed supplemented with Streptomyces could protect fish and shrimp from pathogens as well as increase the growth of the aquatic organisms. Furthermore, the limitations of Streptomyces as probiotics in aquaculture is also highlighted and solutions are discussed to these limitations.

This study investigates the effectiveness of olfactory aversion (OA) and directed masturbation (DM) as forms of conditioning techniques in an adult male convicted of downloading child pornography. The use of OA was employed as an aversive form of conditioning to reduce the occurrence of deviant fantasy. DM techniques were employed as a method of reward to increase the occurrence of appropriate sexual behaviour. The effectiveness of the modifying techniques was measured against a reduction in the frequency and length of time spent masturbating to deviant and non-deviant sexual fantasies, following the olfactory stimuli when paired with DM. After 26-weeks of intervention, results indicate a significant decrease in the frequency of deviant fantasy and time spent masturbating to the deviant fantasy. The reduction in the use of deviant fantasy corresponded to an increase in the frequency and time spent masturbating to a non-deviant fantasy, although the increase was not statistically significant. The use of OA and DM as a method of fantasy control is discussed along with the relative strengths and weaknesses.

Hepatitis C virus (HCV) infection is characterized by persistent replication of a complex mixture of viruses termed a “quasispecies.” Transmission is generally associated with a stringent population bottleneck characterized by infection by limited numbers of “transmitted/founder” (T/F) viruses. Characterization of T/F genomes of human immunodeficiency virus type 1 (HIV-1) has been integral to studies of transmission, immunopathogenesis, and vaccine development. Here, we describe the identification of complete T/F genomes of HCV by single-genome sequencing of plasma viral RNA from acutely infected subjects. A total of 2,739 single-genome-derived amplicons comprising 10,966,507 bp from 18 acute-phase and 11 chronically infected subjects were analyzed. Acute-phase sequences diversified essentially randomly, except for the poly(U/UC) tract, which was subject to polymerase slippage. Fourteen acute-phase subjects were productively infected by more than one genetically distinct virus, permitting assessment of recombination between replicating genomes. No evidence of recombination was found among 1,589 sequences analyzed. Envelope sequences of T/F genomes lacked transmission signatures that could distinguish them from chronic infection viruses. Among chronically infected subjects, higher nucleotide substitution rates were observed in the poly(U/UC) tract than in envelope hypervariable region 1. Fourteen full-length molecular clones with variable poly(U/UC) sequences corresponding to seven genotype 1a, 1b, 3a, and 4a T/F viruses were generated. Like most unadapted HCV clones, T/F genomes did not replicate efficiently in Huh 7.5 cells, indicating that additional cellular factors or viral adaptations are necessary for in vitro replication. Full-length T/F HCV genomes and their progeny provide unique insights into virus transmission, virus evolution, and virus-host interactions associated with immunopathogenesis., IMPORTANCE Hepatitis C virus (HCV) infects 2% to 3% of the world’s population and exhibits extraordinary genetic diversity. This diversity is mirrored by HIV-1, where characterization of transmitted/founder (T/F) genomes has been instrumental in studies of virus transmission, immunopathogenesis, and vaccine development. Here, we show that despite major differences in genome organization, replication strategy, and natural history, HCV (like HIV-1) diversifies essentially randomly early in infection, and as a consequence, sequences of actual T/F viruses can be identified. This allowed us to capture by molecular cloning the full-length HCV genomes that are responsible for infecting the first hepatocytes and eliciting the initial immune responses, weeks before these events could be directly analyzed in human subjects. These findings represent an enabling experimental strategy, not only for HCV and HIV-1 research, but also for other RNA viruses of medical importance, including West Nile, chikungunya, dengue, Venezuelan encephalitis, and Ebola viruses.

Selection of T-cell vaccine antigens for chronic persistent viral infections has been largely empirical. To define the relationship, at the population level, between the specificity of the cellular immune response and viral control for a relevant human pathogen, we performed a comprehensive analysis of the 160 dominant CD8(+) T-cell responses in 578 untreated HIV-infected individuals from KwaZulu-Natal, South Africa. Of the HIV proteins targeted, only Gag-specific responses were associated with lowering viremia. Env-specific and Accessory/Regulatory protein-specific responses were associated with higher viremia. Increasing breadth of Gag-specific responses was associated with decreasing viremia and increasing Env breadth with increasing viremia. Association of the specific CD8(+) T-cell response with low viremia was independent of HLA type and unrelated to epitope sequence conservation. These population-based data, suggesting the existence of both effective immune responses and responses lacking demonstrable biological impact in chronic HIV infection, are of relevance to HIV vaccine design and evaluation.

Selecting human immunodeficiency virus (HIV) sequences for inclusion within vaccines has been a difficult problem, as circulating HIV strains evolve relentlessly and become increasingly divergent over time. We report an assessment of this divergence from three perspectives: (i) across different hosts as a function of time of infection, (ii) between donors and recipients in known transmission pairs, and (iii) within individual hosts over time in relation to the initially replicating virus and to the deduced ancestral sequence of the intrahost viral population. Surprisingly, we consistently found less divergence between viruses from different individuals sampled in primary infection than in individuals sampled at more advanced stages of illness. Furthermore, longitudinal analysis of intrahost divergence revealed a 2- to 3-year period of evolution toward a common ancestral sequence at the start of infection, indicating that HIV recovers certain ancestral features when infecting a new host. These results have important implications for the study of HIV population genetics and rational vaccine design, including favoring the inclusion of viral gene sequences taken early in infection.

Background: Teaching research at the undergraduate level can be a challenge. This is particularly true for distance education courses, in which students often feel isolated from each other and from faculty. Faculty teaching on-campus and distance education research courses designed a student research project and a method for bringing distance and on-campus students together to present their research findings. Method: By actively engaging students in the research process and providing an on-campus research day for the presentation of student posters (Year 1) and research papers (Year 2), course faculty were able to achieve their two goals of directly involving students in the nursing research process and creating a greater student sense of belonging within the college learning community. Results: Statistical analysis of an evaluation survey among on-site and off-site registered nurse to bachelor of science in nursing students indicated both groups ranked the Year 1 poster research day as good to very good. Students attending the Year 2 research day, where they presented papers, ranked the day as good. Statistical analysis indicated a significant difference between the two research days, with a clear student preference for posters. Conclusion: Students valued their active participation in the research process and the opportunity to get together and present their work. However, students clearly preferred a poster presentation format to presenting their findings as a research paper.

Although HIV-1 infection reached Asia late relative to other regions and countries the degree of HIV infection climbed to epidemic proportions over the past 5 years. In 1995 2.5 million of the 4.7 million new HIV infections worldwide occurred in southeast Asia. The Philippines National HIV Sentinel Surveillance Program operational since 1993 has estimated that the current HIV prevalence among a representative population of male and female prostitutes men having sex with men IV drug users and male sexually transmitted disease patients is about 1%. The transmission pattern among 71% of reported cases follows that of a typical heterosexually transmitted disease. Findings are reported from a study conducted to determine the genetic variability of HIV-1 among infected Filipinos and to analyze phylogenetic relationships temporal introductions and transmission dynamics of identified variants. Findings are based upon polymerase chain reaction amplification and the direct sequencing of a 204 base-pair fragment of the env C2-V3 region from uncultured peripheral blood mononuclear cells obtained from 51 HIV-1-positive Filipinos infected between 1987 and mid-1996. The evolutionary distance and phylogenetic relationships among the DNA sequences were estimated. The strains were classified as follows: 37 subtype B 8 subtype E 3 subtype A 2 subtype C and 1 subtype D. The overall env nucleotide divergence ranged from 11.7% to 32.2% while the nucleotide variation appeared to be random and no temporal ordering was observed. Variation of the sequences at the tip of the V3 loop was very broad. Subtypes B and C isolates showed no close genetic relationship to other Asian variants and only 3 of the subtype E strains had close affinity to known Asian sequences. About 66% of cases are thought to have been infected outside of the Philippines.

The Malagarasi River has long been thought to be a barrier to chimpanzee movements in western Tanzania. This potential geographic boundary could affect chimpanzee ranging behavior, population connectivity and pathogen transmission, and thus has implications for conservation strategies and government policy. Indeed, based on mitochondrial DNA sequence comparisons it was recently argued that chimpanzees from communities to the north and to the south of the Malagarasi are surprisingly distantly related, suggesting that the river prevents gene flow. To investigate this, we conducted a survey along the Malagarasi River. We found a ford comprised of rocks that researchers could cross on foot. On a trail leading to this ford, we collected 13 fresh fecal samples containing chimpanzee DNA, two of which tested positive for SIVcpz. We also found chimpanzee feces within the riverbed. Taken together, this evidence suggests that the Malagarasi River is not an absolute barrier to chimpanzee movements and communities from the areas to the north and south should be considered a single population. These results have important consequences for our understanding of gene flow, disease dynamics and conservation management.

Hepatitis C virus (HCV) is present in the host with multiple variants generated by its error prone RNA-dependent RNA polymerase. Little is known about the initial viral diversification and the viral life cycle processes that influence diversity. We studied the diversification of HCV during acute infection in 17 plasma donors, with frequent sampling early in infection. To analyze these data, we developed a new stochastic model of the HCV life cycle. We found that the accumulation of mutations is surprisingly slow: at 30 days, the viral population on average is still 46% identical to its transmitted viral genome. Fitting the model to the sequence data, we estimate the median in vivo viral mutation rate is 2.5×10−5 mutations per nucleotide per genome replication (range 1.6–6.2×10−5), about 5-fold lower than previous estimates. To confirm these results we analyzed the frequency of stop codons (N = 10) among all possible non-sense mutation targets (M = 898,335), and found a mutation rate of 2.8–3.2×10−5, consistent with the estimate from the dynamical model. The slow accumulation of mutations is consistent with slow turnover of infected cells and replication complexes within infected cells. This slow turnover is also inferred from the viral load kinetics. Our estimated mutation rate, which is similar to that of other RNA viruses (e.g., HIV and influenza), is also compatible with the accumulation of substitutions seen in HCV at the population level. Our model identifies the relevant processes (long-lived cells and slow turnover of replication complexes) and parameters involved in determining the rate of HCV diversification., Author Summary Hepatitis C virus (HCV) is a RNA virus that infects over 170 million people across the world. It leads to a chronic infection in the majority of people who are infected (>70%). Most people only discover that they are infected long after initial infection. Thus, it is difficult to study the very early events in infection. Here we study 17 individuals during the earliest possible stages of infection, from before the virus is detectable in the plasma to around 35 days post-infection. We focus on understanding the viral kinetics and the diversification of HCV during this acute phase of infection. During chronic infection HCV is present in the host as a swarm of multiple variants generated by its error prone copying. We studied the early diversification of HCV during acute infection using a new mathematical model of HCV replication. We found that after a phase of fast increase in viral load, accompanied by viral diversification, there is a stabilization of viral load and diversity levels. Using our model, we were able to estimate for the first time the HCV mutation rate during acute infection. We estimated the median in vivo viral mutation rate is 2.5×10−5 mutations per nucleotide per genome replication (range 1.6–6.2×10−5), about 5-fold lower than previous estimates. We also used a different approach, based on results of classical genetics, to calculate HCV's mutation rate and obtained consistent results (2.8–3.2×10−5).

HIV-1 groups M and N emerged within the last century following two independent cross-species transmissions of SIVcpz from chimpanzees to humans. In contrast to pandemic group M strains, HIV-1 group N viruses are exceedingly rare, with only about a dozen infections identified, all but one in individuals from Cameroon. Poor adaptation to the human host may be responsible for this limited spread of HIV-1 group N in the human population. Here, we analyzed the function of Vpu proteins from seven group N strains from Cameroon, the place where this zoonosis originally emerged. We found that these N-Vpus acquired four amino acid substitutions (E15A, V19A and IV25/26LL) in their transmembrane domain (TMD) that allow efficient interaction with human tetherin. However, despite these adaptive changes, most N-Vpus still antagonize human tetherin only poorly and fail to down-modulate CD4, the natural killer (NK) cell ligand NTB-A as well as the lipid-antigen presenting protein CD1d. These functional deficiencies were mapped to amino acid changes in the cytoplasmic domain that disrupt putative adaptor protein binding sites and an otherwise highly conserved ßTrCP-binding DSGxxS motif. As a consequence, N-Vpus exhibited aberrant intracellular localization and/or failed to recruit the ubiquitin-ligase complex to induce tetherin degradation. The only exception was the Vpu of a group N strain recently discovered in France, but originally acquired in Togo, which contained intact cytoplasmic motifs and counteracted tetherin as effectively as the Vpus of pandemic HIV-1 M strains. These results indicate that HIV-1 group N Vpu is under strong host-specific selection pressure and that the acquisition of effective tetherin antagonism may lead to the emergence of viral variants with increased transmission fitness., Author Summary Differences in their degree of adaptation to humans may explain why only one of four ape-derived SIV zoonoses spawned the AIDS pandemic. Specifically, only HIV-1 strains of the pandemic M group evolved a fully functional Vpu that efficiently antagonizes human tetherin and degrades CD4. In comparison, the rare group N viruses gained some anti-tetherin activity but lost the CD4 degradation function. Here, we show that the N-Vpu transmembrane domain has adapted to interact with human tetherin and identified the mutations that enable this interaction. However, we also show that most N-Vpus remain poor tetherin antagonists and fail to reduce the surface expression of CD4, the natural killer cell ligand NTB-A and the lipid-antigen presenting protein CD1d. This is due to mutations in their cytoplasmic region that are associated with aberrant protein localization and impaired interaction with the ubiquitin/proteasome pathway. A remarkable exception is the Vpu of the first HIV-1 N strain known to be transmitted outside of Cameroon, which contains a functional cytoplasmic domain and is a highly effective tetherin antagonist. These data indicate that group N viruses are still adapting to humans and that the acquisition of potent anti-tetherin activity may eventually lead to the emergence of viral variants that exhibit increased transmission fitness.

The adult congenital heart disease (CHD) population is becoming increasingly prevalent. Approximately 6 per 1,000 live births in the United States are affected by complex CHD, and as many as 75 per 1,000 live births have simple lesions such as ventricular septal defects [1]. Improved medical and surgical care has decreased early and late mortality resulting in the increased number of patients surviving well into adulthood with both simple and complex forms of CHD. As a result of these improvements, the number of adult survivors with CHD surpassed the number of pediatric patients with CHD in the year 2000. Currently, it is estimated there are approximately 800,000 adults with CHD in the United States [2].

Full-length HIV-1 genome sequencing provides important data needed to address several vaccine design, molecular epidemiologic and pathogenesis questions. A protocol is presented for obtaining near full-length genomes (NFLGs) from subjects infected with HIV-1 subtype C. This protocol was used to amplify NFLGs from 244 of 366 (67%) samples collected at two clinics in Durban, South Africa (SK and PS). Viral load was directly associated with frequency of successful NFLG amplification for both cohorts (PS; p = 0.005 and SK; p < 0.001). Seventeen of 38 initially NFLG-negative SK samples had variation within the PCR primer binding sites, however only 3 of these were successfully re-amplified using re-designed primers homologous to the target viruses. NFLGs were obtained from 7 of 24 PBMC samples processed from subjects whose plasma did not yield a NFLG. Stable plasmid clones were obtained from all 244 NFLG-positive PCR products, and both strands of each genome were sequenced, using a primary set of 46 primers. These methods thus allow the large-scale collection of HIV-1 NFLGs from populations infected primarily with subtype C. The methods are readily adaptable to other HIV-1 subtypes, and provide materials for viral functional analyses and population-based molecular epidemiology studies that include analysis of viral genome chimerization.

Cytotoxic T lymphocytes (CTLs) are critical for the control of human immunodeficiency virus, but containment of virus replication can be undermined by mutations in CTL epitopes that lead to virus escape. We analyzed the evolution in vivo of an immunodominant, HLA-A2–restricted CTL epitope and found two principal, diametrically opposed evolutionary pathways that exclusively affect T cell–receptor contact residues. One pathway was characterized by acquisition of CTL escape mutations and the other by selection for wild-type amino acids. The pattern of CTL responses to epitope variants shaped which variant(s) prevailed in the virus population. The pathways notably influenced the amount of plasma virus, as patients with efficient CTL selection had lower plasma viral loads than did patients without efficient selection. Thus, viral escape from CTL responses does not necessarily correlate with disease progression.

Understanding the properties of human immunodeficiency virus type 1 (HIV-1) variants capable of establishing infection is critical to the development of a vaccine against AIDS. Previous studies of men have shown that the HIV-1 env gene is homogeneous early in infection, leading to the suggestion that infection is established by a single transmitted variant. However, we report here that all of eight homosexual men evaluated beginning 3.7 to 9 weeks following onset of symptoms of acute infection harbored diverse virus populations in their blood, with median genetic distances averaging 1.08% in the env C2V5 region and 0.81% in the gag p17 gene. Within another 4.7 to 11 weeks, the variant lineage in env became more homogeneous, while gag sequences continued to diversify. Thus, the homogenization that has been reported to characterize acute infection is actually preceded by the replication of multiple virus variants. This early selective process focuses on viral properties within Env but not Gag p17. Hence, the viral homogeneity observed early in HIV-1 infection results from a selective process that occurs during the establishment of infection.

The development of viral diversity during the course of human immunodeficiency virus type 1 (HIV-1) infection may significantly influence viral pathogenesis. The paradigm for HIV-1 evolution is based primarily on studies of male cohorts in which individuals were presumably infected with a single virus variant of subtype B HIV-1. In this study, we evaluated virus evolution based on sequence information of the V1, V2, and V3 portions of HIV-1 clade A envelope genes obtained from peripheral blood and cervical secretions of three women with genetically heterogeneous viral populations near seroconversion. At the first sample following seroconversion, the number of nonsynonymous substitutions per potential nonsynonymous site (dn) significantly exceeded substitutions at potential synonymous sites (ds) in plasma viral sequences from all individuals. Generally, values of dn remained higher than values of ds as sequences from blood or mucosa evolved. Mutations affected each of the three variable regions of the envelope gene differently; insertions and deletions dominated changes in V1, substitutions involving charged amino acids occurred in V2, and sequential replacement of amino acids over time at a small subset of positions distinguished V3. The relationship among envelope nucleotide sequences obtained from peripheral blood mononuclear cells, plasma, and cervical secretions was evaluated for each individual by both phylogenetic and phenetic analyses. In all subjects, sequences from within each tissue compartment were more closely related to each other than to sequences from other tissues (phylogenetic tissue compartmentalization). At time points after seroconversion in two individuals, there was also greater genetic identity among sequences from the same tissue compartment than among sequences from different tissue compartments (phenetic tissue compartmentalization). Over time, temporal phylogenetic and phenetic structure was detectable in mucosal and plasma viral samples from all three women, suggesting a continual process of migration of one or a few infected cells into each compartment followed by localized expansion and evolution of that population.

Genital secretions are the source of a majority of human immunodeficiency virus type 1 (HIV-1) infections. Culturable HIV-1 (23, 59) and proviral DNA (21, 37) have been detected in the nonspermatozoal mononuclear cell fraction of semen, which includes CD4+ lymphoid cells, monocytes, and macrophages (2, 54). Cell-free virions, measured as viral RNA, are also found in seminal fluid (20, 21, 37). Heteroduplex tracking analyses of PCR-amplified HIV-1 env sequences from donor-recipient sexual transmission pairs revealed distinctions between viral variant populations in plasma, peripheral blood mononuclear cells (PBMC), seminal cells, and seminal fluid (62). In three of five cases, virus in semen mononuclear cells was the most closely related to that transmitted to the recipient, and hence these cells were implicated as a likely vehicle for transmission (62). Drug resistance mutations in HIV populations are also unequally distributed in the blood and semen (29). Genetic differences were also found between proviral variants in peripheral blood and those in genital secretions in one study of women infected with HIV-1 envelope sequence subtypes A and D (41). Distinctions between viral populations in other anatomical sites have also been noted (14, 24, 27). Early following infection with envelope subtype B virus via homosexual or perinatal contact, intravenous drug use, and blood transfusion, individuals harbor viruses in their blood that are homogeneous over the V3 region of the envelope protein (8, 10, 30, 36, 39, 47, 57, 60, 61). V3 loop sequences are also relatively conserved between individuals early following infection and correspond to a macrophage-tropic or non-syncytium-inducing (NSI) signature sequence (6, 35, 47, 53, 60), despite the fact that the donor may harbor a highly differentiated virus population in the blood (56, 62). A simultaneously greater amount of viral genetic diversity has been reported in the gag gene at these early times (60, 61). This observation led to the hypothesis that selection may occur for the dissemination of a subset of variants with a particular envelope-mediated phenotype, possibly macrophage tropism. With the recognition of the existence of HIV coreceptors, this could also correspond to a tropism for coreceptors of the CC-chemokine receptor class or exclusion of the viruses that utilize CXCR-4 (1, 12, 13, 18). It is also possible that macrophage-tropic HIV variants are more likely to be found in, and thereby transmitted from, the genital compartment, since, in contrast to the balance in the blood, a greater number of monocytes and macrophages are found in semen (2, 54). In contrast to other modes of transmission, complex HIV-1 envelope sequence populations have been reported in one study of recently seroconverting women heterosexually infected with subtype A or D (41). Sexual transmission from men requires infection of cells in or migrating to seminal fluid or cell-free transfer of virus to this compartment. A comparison of blood and semen viral populations may therefore indicate whether virus in the male genital tract commingles with that of blood or corresponds to a compartmentalized subset or distinct population of variants. In this report, we describe a cross-sectional study of proviral sequences in peripheral blood and nonspermatozoal semen cells from five HIV-1 subtype B-infected individuals. Strong evidence for nonrandom distribution of virus variants between the two compartments and of a more restricted founder population accounting for the semen provirus population was found in all three of the patients who also harbored virus with mutations characteristic of a syncytium-inducing (SI) viral genotype. However, no evidence was found for restriction of the SI genotype from the semen.

To determine the distribution of HIV-1 subtypes in Sao Paulo, Brazil.Samples were obtained from 80 consecutive HIV-1-infected individuals attending the Immunodeficiency Clinic at the University of Sao Paulo in 1993. Peripheral blood mononuclear cells (PBMC) were separated by Ficoll-Hypaque gradient and a portion was used for routine CD4 counts; the remainder were frozen. PBMC were proteinase-K-digested and DNA-purified by organic extraction. Samples were amplified for the env region of HIV, and envelope sequence subtypes determined by heteroduplex mobility analysis using prototypic subtypes as references. A subset of these were also sequenced through the C2-V3 region of env.A total 69 of 80 samples yielded env polymerase chain reaction product enabling subtype determination; samples that did not amplify were those with low DNA yields. Among 12 injecting drug users (IDU) or sexual partners of IDU, four were typed as clade F and eight as clade B. Forty-three homosexual men or female sexual partners of bisexual men were typed as clade B. The 14 additional cases without known risk factors were typed as clade B.These data suggest that subtype F is related to injecting drug use in Brazil.Serum samples from 80 consecutive HIV-1-infected individuals presenting to the Immunodeficiency Clinic at the University of Sao Paulo in 1993 were analyzed to determine the distribution of HIV-1 subtypes in the city. Peripheral blood mononuclear cells (PBMC) were separated using Ficoll-Hypaque gradient, a portion was used for routine CD4 counts, and the rest were frozen. PBMC were proteinase-K-digested and DNA-purified by organic extraction. The samples were amplified for the env region of HIV, and envelope sequence subtypes determined by heteroduplex mobility analysis using prototypic subtypes as references. A subset was also sequenced through the C2-V3 region of env. 69 samples yielded env polymerase chain reaction product enabling subtype determination. The samples which did not amplify had low DNA yields. Among 12 IV-drug users or their sex partners, four were typed as clade F and eight as clade B. 43 homosexual men or female sex partners of bisexual men were typed as clade B. The 14 additional cases with no known risk factor were typed as clade B.

Like human immunodeficiency virus type 1 (HIV-1), simian immunodeficiency virus of chimpanzees (SIVcpz) can cause CD4+ T cell loss and premature death. Here, we used molecular surveillance tools and mathematical modeling to estimate the impact of SIVcpz infection on chimpanzee population dynamics. Habituated (Mitumba and Kasekela) and non-habituated (Kalande) chimpanzees were studied in Gombe National Park, Tanzania. Ape population sizes were determined from demographic records (Mitumba and Kasekela) or individual sightings and genotyping (Kalande), while SIVcpz prevalence rates were monitored using non-invasive methods. Between 2002–2009, the Mitumba and Kasekela communities experienced mean annual growth rates of 1.9% and 2.4%, respectively, while Kalande chimpanzees suffered a significant decline, with a mean growth rate of −6.5% to −7.4%, depending on population estimates. A rapid decline in Kalande was first noted in the 1990s and originally attributed to poaching and reduced food sources. However, between 2002–2009, we found a mean SIVcpz prevalence in Kalande of 46.1%, which was almost four times higher than the prevalence in Mitumba (12.7%) and Kasekela (12.1%). To explore whether SIVcpz contributed to the Kalande decline, we used empirically determined SIVcpz transmission probabilities as well as chimpanzee mortality, mating and migration data to model the effect of viral pathogenicity on chimpanzee population growth. Deterministic calculations indicated that a prevalence of greater than 3.4% would result in negative growth and eventual population extinction, even using conservative mortality estimates. However, stochastic models revealed that in representative populations, SIVcpz, and not its host species, frequently went extinct. High SIVcpz transmission probability and excess mortality reduced population persistence, while intercommunity migration often rescued infected communities, even when immigrating females had a chance of being SIVcpz infected. Together, these results suggest that the decline of the Kalande community was caused, at least in part, by high levels of SIVcpz infection. However, population extinction is not an inevitable consequence of SIVcpz infection, but depends on additional variables, such as migration, that promote survival. These findings are consistent with the uneven distribution of SIVcpz throughout central Africa and explain how chimpanzees in Gombe and elsewhere can be at equipoise with this pathogen., Author Summary It is now well established that human immunodeficiency virus type 1 (HIV-1), which causes acquired immune deficiency syndrome (AIDS), emerged following cross-species transmission of a simian immunodeficiency virus that infects chimpanzees (SIVcpz). Although SIVcpz was originally believed not to be pathogenic, a recent study conducted in Gombe National Park, Tanzania, found that infected chimpanzees can develop AIDS-like symptoms and have a high mortality. To examine the impact of SIVcpz on chimpanzee survival, we examined and then modeled the effect of virus infection on chimpanzee population growth. We studied three communities, including the little-studied Kalande community, located in the south of the park. We found that 46% of Kalande apes were infected with SIVcpz, compared to less than 13% of the other communities. Interestingly, the communities with lower infection rates grew, while Kalande suffered a catastrophic decline. Mathematical modeling indicated that chimpanzee populations infected with SIVcpz are likely to decline, although intercommunity migration counteracted this negative effect, allowing many simulated populations to persist. These results suggest that SIVcpz infection, together with other variables, contributed to the Kalande population decline. However, population extinction is not a necessary consequence of SIVcpz infection, but can be counteracted by other factors, such as migration, that promote population survival. These findings are consistent with the uneven distribution of SIVcpz across Africa and may explain how chimpanzees as a species have survived this pathogen.

Genetic analysis of human immunodeficiency virus type 1 (HIV-1) from cases of mother-to-infant transmission were analyzed in an effort to provide insights into the viral selection that may occur during transmission, as well as the timing and source of transmitted viruses. HIV-1 env genes obtained from seven mothers and their perinatally infected infants in Sweden were studied. Five envelope sequence clades (A to E) were found to be represented. We used a heteroduplex tracking assay (HTA) to assess the genetic relatedness between early viral isolates from the infants and serial maternal virus populations taken during pregnancy and at delivery. HTA findings were used to select for DNA sequence analysis maternal virus populations that were either closely or more distantly related to the infant virus. In each case, nucleotide sequence analysis confirmed the genetic relationships inferred by the HTA. Only maternal peripheral blood was sampled, and large sets of maternal specimens throughout pregnancy were generally not available. However, no consistent correlation was found to support the hypothesis that infant viruses should match blood-derived maternal virus genotypes found early in pregnancy if infants were found to be infected at birth or, conversely, that infant viruses should match blood-derived maternal virus genotypes found at delivery if infants were found to be infected only some time later.

Interpersonal connection is a fundamental human motivation, and the extent to which it is fulfilled is a strong predictor of symptoms of internalizing disorders such as social anxiety and depression, perhaps especially during the “social reorienting” period of adolescence. However, little is known about the contribution to this effect of the individual’s social motivations, which are intensified during adolescence. Furthermore, social goal orientation – an individual’s priorities and intentions in social interactions – is an important predictor of vulnerability to internalizing symptoms. Adolescents spend most of their waking lives in classrooms, bounded social networks with a limited pool of candidates for befriending. This study investigated whether friendships within one’s class protects against internalizing symptoms in part by reducing the desire for more classmate friendships, which may tend to promote maladaptive social goals. Participants were 423 young adolescents (M age = 13.2, sd = 0.52 years; 49.4% girls). As predicted, adolescents’ number of reciprocated classroom friendships had a protective effect on internalizing symptoms which was serially mediated by desire for more such friendships, and social goal orientation. However, only demonstration-avoidance goals significantly predicted internalizing symptoms. Unreciprocated friendship nominations were unexpectedly associated with stronger desire and more social anxiety symptoms. The results suggest that the effect of number of friends is mediated by the individual’s thoughts and feelings about their number of friendships, such that a strong desire for more friendships promotes maladaptive goals, oriented toward social status and consequently less oriented toward the cultivation of interpersonal intimacy with the friends they already have.

Growth mindset, the belief that personal attributes such as intelligence are malleable, has previously been related to more effort investment. Here, we investigated how undergraduates' mindset (N = 114) relates to the choice to invest effort during an arithmetic task, indexed by whether they make low vs. high effort-related choices. Social cognitive theory suggests that past performance experiences (mastery vs. failure) and physiological state are important sources for competence self-evaluations. Therefore, in addition to mindset, we also investigated how effort-related choices are influenced more dynamically, by failures and physiological responses during the task. Growth mindset and physiological effort mobilization did not predict effort-related choices but making mistakes did predict lower effort choices in the subsequent round. This study further supports the importance of mastery experiences for effort investment and provides a novel approach for integrating different levels of influence on effort-related choices during an educationally-relevant task.