Injection Drug Use Alters Plasma Regulation of the B Cell Response.

The opioid epidemic continues to be a major public health issue that includes millions of people who inject drugs (PWID). PWID have increased incidence of serious infections, including HIV as well as metabolic and inflammatory sequelae. We sought to discern the extent of systemic alterations in humoral immunity associated with injection drug use, including alterations in the plasma proteome and its regulation of B cell responsiveness. Comprehensive plasma proteomics analysis of HIV negative/hepatitis C negative individuals with a history of recent injection heroin use was performed using mass spectrometry and ELISA. The effects of plasma from PWID and healthy controls on the in vitro proliferation and transcriptional profile of B cell responses to stimulation were determined by flow cytometry and RNA-Seq. The plasma proteome of PWID was distinct from healthy control individuals, with numerous immune-related analytes significantly altered in PWID, including complement (C3, C5, C9), immunoglobulin (IgD, IgM, kappa light chain), and other inflammatory mediators (CXCL4, LPS binding protein, C-reactive protein). The plasma of PWID suppressed the in vitro proliferation of B cells. Transcriptome analysis indicated that PWID plasma treatment increased B cell receptor and CD40 signaling and shifted B cell differentiation from plasma cell-like toward germinal center B cell-like transcriptional profiles. These results indicate that the systemic inflammatory milieu is substantially altered in PWID and may impact their B cell responses.

Proteobacteria impair anti-tumor immunity in the omentum by consuming arginine.

Gut microbiota influence anti-tumor immunity, often by producing immune-modulating metabolites. However, microbes consume a variety of metabolites that may also impact host immune responses. We show that tumors grow unchecked in the omenta of microbe-replete mice due to immunosuppressive Tregs. By contrast, omental tumors in germ-free, neomycin-treated mice or mice colonized with altered Schaedler's flora (ASF) are spontaneously eliminated by CD8+ T cells. These mice lack Proteobacteria capable of arginine catabolism, causing increases in serum arginine that activate the mammalian target of the rapamycin (mTOR) pathway in Tregs to reduce their suppressive capacity. Transfer of the Proteobacteria, Escherichia coli (E. coli), but not a mutant unable to catabolize arginine, to ASF mice reduces arginine levels, restores Treg suppression, and prevents tumor clearance. Supplementary arginine similarly decreases Treg suppressive capacity, increases CD8+ T cell effectiveness, and reduces tumor burden. Thus, microbial consumption of arginine alters anti-tumor immunity, offering potential therapeutic strategies for tumors in visceral adipose tissue.

A SEROLOGIC SURVEY OF FRANCISELLA TULARENSIS EXPOSURE IN WILDLIFE ON THE ARCTIC COASTAL PLAIN OF ALASKA, USA.

Tularemia is an infectious zoonotic disease caused by one of several subspecies of Francisella tularensis bacteria. Infections by F. tularensis are common throughout the northern hemisphere and have been detected in more than 250 wildlife species. In Alaska, US, where the pathogen was first identified in 1938, studies have identified F. tularensis antibodies in a diverse suite of taxa, including insects, birds, and mammals. However, few such investigations have been conducted recently and knowledge about the current distribution and disease ecology of F. tularensis is limited, particularly in Arctic Alaska, an area undergoing rapid environmental changes from climate warming. To help address these information gaps and provide insights about patterns of exposure among wildlife, we assessed the seroprevalence of F. tularensis antibodies in mammals and tundra-nesting geese from the Arctic Coastal Plain of Alaska, 2014-17. With a commercially available slide agglutination test, we detected antibodies in 14.7% of all individuals sampled (n=722), with titers ranging from 1:20 to 1:320. We detected significant differences in seroprevalence between family groups, with Canidae (foxes, Vulpes spp.) and Sciuridae (Arctic ground squirrel, Spermophilus parryii) having the highest seroprevalence at 21.5% and 33.3%, respectively. Mean seroprevalence for Ursidae (polar bears, Ursus maritimus) was 13.3%, whereas Cervidae (caribou, Rangifer tarandus) had comparatively low seroprevalence at 6.5%. Antibodies were detected in all Anatidae species sampled, with Black Brant (Branta bernicla nigricans) having the highest seroprevalence at 13.6%. The detection of F. tularensis antibodies across multiple taxa from the Arctic Coastal Plain and its nearshore marine region provides evidence of exposure to this pathogen throughout the region and highlights the need for renewed surveillance in Alaska.

Effects of radiant exposure and distance on resin-based composite polymerization.

PURPOSE: To evaluate the hardness profile of three resin-based restorative composites (RBC) (Filtek Z250XT, Filtek One Bulk Fill, Filtek Bulk Fill Flow) polymerized by a multi-wave curing light. METHODS: Specimens (n= 12) were prepared by inserting 2 mm RBC increments into a split-mold and polymerized from the top using either 20- or 40-second exposure times. Specimen curing was performed directly at a 1 mm distance (control-group) or through an ivorine-tooth slot preparation at a 5 mm distance (experimental-group). Specimens were stored (37 ± 1°C/24 hours), then subjected to Knoop indenter (25g/5 seconds). Specimens' KHN values were obtained from the upper and lower surfaces. Relative hardness (RH) (lower-to-upper ratio) was calculated for each specimen. Data were analyzed with three-way ANOVA and Tukey's HSD (α= 0.05). RESULTS: There was no significant RH difference among RBCs in the control group, regardless of the exposure time (P> 0.05). Average RH ratios for all RBCs tested in this group were greater than 0.80. However, the average RH values of the experimental RBC group were significantly lower. The RH for Z250 was 0.39 in the 20-second group, while RH was 0.63 in the 40-second group. BF had an RH ratio of 0.70 in the 20-second and 0.72 in the 40-second group, while One Bulk had a ratio of 0.65 in the 20-second and 0.71 in the 40-second groups. Doubled exposure time substantially increased RH of all tested materials at a 1 mm tip-to-material distance. Clinically relevant 5 mm light-tip to material-surface distance significantly reduced polymerization efficacy of RBC specimens, regardless of the exposure time. CLINICAL SIGNIFICANCE: Adequate light-polymerization of resin-based direct restoratives is necessary for long-term clinical success. Polymerizing Class 2 restorations is challenging due to a hard-to-reach location and an increased distance between the light source and the restorative material. Insufficient polymerization is often seen at the bottom of the proximal box of the Class 2 cavity, with a detrimental effect on restoration longevity.

CROPSR: an automated platform for complex genome-wide CRISPR gRNA design and validation.

BACKGROUND: CRISPR/Cas9 technology has become an important tool to generate targeted, highly specific genome mutations. The technology has great potential for crop improvement, as crop genomes are tailored to optimize specific traits over generations of breeding. Many crops have highly complex and polyploid genomes, particularly those used for bioenergy or bioproducts. The majority of tools currently available for designing and evaluating gRNAs for CRISPR experiments were developed based on mammalian genomes that do not share the characteristics or design criteria for crop genomes. RESULTS: We have developed an open source tool for genome-wide design and evaluation of gRNA sequences for CRISPR experiments, CROPSR. The genome-wide approach provides a significant decrease in the time required to design a CRISPR experiment, including validation through PCR, at the expense of an overhead compute time required once per genome, at the first run. To better cater to the needs of crop geneticists, restrictions imposed by other packages on design and evaluation of gRNA sequences were lifted. A new machine learning model was developed to provide scores while avoiding situations in which the currently available tools sometimes failed to provide guides for repetitive, A/T-rich genomic regions. We show that our gRNA scoring model provides a significant increase in prediction accuracy over existing tools, even in non-crop genomes. CONCLUSIONS: CROPSR provides the scientific community with new methods and a new workflow for performing CRISPR/Cas9 knockout experiments. CROPSR reduces the challenges of working in crops, and helps speed gRNA sequence design, evaluation and validation. We hope that the new software will accelerate discovery and reduce the number of failed experiments.

Association between health system specialty pharmacy use and health care costs among national sample of Medicare Advantage beneficiaries.

BACKGROUND: Health care expenditures are growing rapidly. There is a growing body of literature showing that health system specialty pharmacy is associated with improvement in clinical outcomes; however, there is a lack of data on its effect on health care costs and utilization. OBJECTIVE: To perform exploratory research assessing the association between health system specialty pharmacy use and health care costs and utilization. METHODS: A retrospective cohort study was conducted examining medical and pharmacy claims from 2018 and 2019 of Medicare Advantage beneficiaries. Optum Advisory Service's proprietary deidentified Normative Health Information database was used, which includes claims, membership, and provider data for 12.6 million Medicare Advantage members. Members who filled a prescription at a health system specialty pharmacy and had a specialty provider participating in the health system specialty pharmacy care model in clinic were assigned to the intervention group. Members who did not use a health system specialty pharmacy but had the same provider (provider benchmark group) or different provider (network benchmark group) were considered as comparisons. The network benchmark group was further refined to match variation in health care cost due to geography. The primary outcome measure was total health care costs (across the medical and pharmacy benefit) on a per-patient per-month basis. Secondary outcomes were selected utilization drivers and cost subcomponents. Cost and utilization metrics were calculated on a risk-adjusted basis using Centers for Medicare & Medicaid Services Hierarchical Condition Categories (CMS-HCC) risk score methodology. Differences were assessed for categorical variables with chi-square tests, and 2-tailed t-tests were used for continuous variables. RESULTS: Of the analytic sample of 9,780 members used in this study, 208 (2.1%) used health system specialty pharmacy services. During the 2018 baseline period, total health care costs and utilization were similar after CMS-HCC risk score adjustment ($9,520 among health system specialty pharmacy users; $8,691 among the provider benchmark group; $9,510 among the network benchmark group) but lower in 2019 ($7,060, $7,683, and $8,152, respectively). The differences in 2019 were primarily driven by savings in pharmacy and free-standing physician-related costs. CONCLUSIONS: Use of a health system specialty pharmacy is associated with a lower health care cost. Further study is required to analyze how drug and disease-specific interactions influence total health care costs and utilization for health system specialty pharmacy populations. DISCLOSURES: This study was funded by Shields Health Solutions and completed with Optum Advisory Services, which provided all analysis and was the sole source of data. University of Massachusetts Medical School investigators were independent context experts, who volunteered their time for this study. Hellems is employed by Optum Advisory Services; Fasching and Smith are employed by Shields Health Solutions; and Soni and McManus are employed by the University of Massachusetts Medical School. Soni received support from the National Institute of General Medical Science (T32GM107000), National Center for Advancing Translational Sciences (TL1-TR001454), and the Eunice Kennedy Shriver National Institute of Child Health and Human Development (1F30HD091975-03). McManus's time was supported by R01HL126911, R01HL137734, R01HL137794, R01HL135219, R01HL136660, U54HL143541 from the National Heart, Lung and Blood Institute. McManus has received research support from Bristol Myers Squibb, Care Evolution, Samsung, Apple Computer, Pfizer, Biotronik, Boehringer Ingelheim, Philips Research Institute, Flexcon, Fitbit; has consulted for Bristol Myers Squibb, Pfizer, Philips, Samsung Electronics, Rose Consulting, Boston Biomedical Associates, and FlexCon; and is also a member of the Operations Committee and Steering Committee for the GUARD-AF Study (NCT04126486), sponsored by Bristol Meyers Squibb and Pfizer. The other authors have nothing additional to disclose. The content is solely the responsibility of the authors and does not necessarily represent the official views of the NIH. The funders played no role in the design and conduct of the study; collection, management, analysis, and interpretation of the data; preparation, review, or approval of the manuscript; and decision to submit the manuscript for publication.

Mating Competitiveness of Transgenic Aedes aegypti (Diptera: Culicidae) Males Against Wild-Type Males Reared Under Simulated Field Conditions.

Efforts directed at genetic modification of mosquitoes for population control or replacement are highly dependent on the initial mating success of transgenic male mosquitoes following their release into natural populations. Adult mosquito phenotypes are influenced by the environmental conditions experienced as larvae. Semifield studies conducted to date have not taken that under consideration when testing male mating fitness, and have compared mating success of males reared under identical environmental conditions. We performed pairwise mating challenges between males from a genetically modified laboratory strain (BF2) versus males from a recent Trinidad field isolate of Aedes aegypti (L.), a major vector of multiple arboviruses. We utilized larval density and nutrition to simulate environmental stress experienced by the Trinidad males and females. Our results indicated that environmental stress during larval development negatively influenced the competitiveness and reproductive success of males from the Trinidad population when paired with optimum reared BF2 males. Small (0.027 m3) and large (0.216 m3) trials were conducted wherein stressed or optimum Trinidad males competed with optimum BF2 males for mating with stressed Trinidad females. When competing with stress reared Trinidad males, optimum reared BF2 males were predominant in matings with stress reared Trinidad females, and large proportions of these females mated with males of both strains. When competing with optimum reared Trinidad males, no difference in mating success was observed between them and BF2 males, and frequencies of multiple matings were low. Our results indicate that future mating competition studies should incorporate appropriate environmental conditions when designing mating fitness trials of genetically modified males.

Co-Producing Interdisciplinary Knowledge and Action for Sustainable Water Governance: Lessons from the Development of a Water Resources Decision Support System in Pernambuco, Brazil.

One of the most pressing global challenges for sustainable development is freshwater management. Sustainable water governance requires interdisciplinary knowledge about environmental and social processes as well as participatory strategies that bring scientists, managers, policymakers, and other stakeholders together to cooperatively produce knowledge and solutions, promote social learning, and build enduring institutional capacity. Cooperative production of knowledge and action is designed to enhance the likelihood that the findings, models, simulations, and decision support tools developed are scientifically credible, solutions-oriented, and relevant to management needs and stakeholders' perspectives. To explore how interdisciplinary science and sustainable water management can be co-developed in practice, the experiences of an international collaboration are drawn on to improve local capacity to manage existing and future water resources efficiently, sustainably, and equitably in the State of Pernambuco in northeastern Brazil. Systems are developed to model and simulate rainfall, reservoir management, and flood forecasting that allow users to create, save, and compare future scenarios. A web-enabled decision support system is also designed to integrate models to inform water management and climate adaptation. The challenges and lessons learned from this project, the transferability of this approach, and strategies for evaluating the impacts on management decisions and sustainability outcomes are discussed.

Identification of Host Blood Meals of Mosquitoes (Diptera: Culicidae) Collected at the Aripo Savannas Scientific Reserve in Trinidad, West Indies.

Surveillance for blood-fed female mosquitoes was performed between August 2015 and February 2016 at sites along the periphery of the Aripo Savannas Environmentally Reserve (ASSR) located in northeastern Trinidad, West Indies. We collected engorged female mosquitoes representing 13 species. DNA extractions from dissected abdomens were subjected to PCR amplification with three primer pairs targeting the mitochondrial cytochrome oxidase I and cytochrome b gene sequences. High-quality sequence information and host identification were obtained for 42 specimens representing eight mosquito species with at least one primer combination. A broad range of vertebrates including humans were identified, but the majority were nonhuman mammals, both domestic and wild. Domestic dogs were the most common host and may represent potential sentinel species for monitoring local enzootic arbovirus activity in Trinidad. Culex declarator Dyer and Knab and Culex nigripalpus Theobald were the most common blood-fed mosquito species comprising 79.1% of the total number identified. These species obtained blood meals from birds, nonhuman mammals, and human hosts, and therefore pose significant risks as potential bridge vectors for epizootic arbovirus transmission in the ASSR area as well as other sylvan areas in Trinidad. These data represent the first such results for Trinidad.

Stunting, food security, markets and food policy in Rwanda.

BACKGROUND: Over the past two decades, Rwanda has experienced impressive economic growth, resulting in considerable improvements in living standards and poverty reduction. Despite these gains, progress on reducing the level of stunting in smallholder rural children, particularly boys, continues to be a serious concern. METHODS: Policies, dietary diversity and socio-economic factors that may influence stunting in rural Rwandan children were evaluated using a logit model with clustered variance-covariance estimators based on village membership of the household. RESULTS: Stunting of rural children was found to be multidimensionally related to the child's gender, weight and age; the dietary diversity, marriage status and education level of the head of household; mother's height; presence of a family garden or if they owned livestock; environmental factors such as altitude and soil fertility and location relative to a main road en route to a market; and a policy that promoted food production. CONCLUSIONS: Findings suggest that agricultural policies may be subsidizing poor dietary behavior in that the aggregation of production encourages households to sell high quality nutritious food such as fruit and vegetables, for more voluminous amounts of nutritionally substandard goods, hence low dietary diversity. However, it is less clear if rural food markets are capable of supplying diverse and nutritious foods at affordable prices on a consistent basis, resulting in a lack of diversity and hence, low nutrient quality diets. Rwanda's next round of food security policies should focus on nutrition insecurity with special emphasis on the lack of protein, micronutrients and calories. Multipronged policies and programs focused on income growth, food security, enhanced access to markets and gender-related nutrition risks from inception through 2 years of age in the rural areas are required to improve rural household health outcomes, stunting in particular.

Cancer-related fatigue and sleep deficiency in cancer care continuum: concepts, assessment, clusters, and management.

Throughout the cancer continuum, patients are faced with the cancer- and treatment-related side effects that can have a negative impact on their overall quality of life. Cancer-related fatigue (CRF) and sleep deficiency are among the symptoms that patients and their caregivers most often experience. An increasing body of literature suggests that a strong correlation between CRF and sleep deficiency exists, indicating that they may be reciprocally related and that they may have similar underlying etiology. This paper aims at bringing together the opinions of leading cancer control (i.e., CRF and sleep) and oncology experts in order to increase the understanding of CRF and sleep deficiency's assessment, associated symptom clustering, symptom burden shared by caregivers, and CRF and sleep deficiency management in the cancer care context.

QTL Determining Diel Flight Activity in Male Culex pipiens Mosquitoes.

Members of the Culex pipiens complex differ in physiological traits that facilitate their survival in diverse environments. Assortative mating within the complex occurs in some regions where autogenous (the ability to lay a batch of eggs without a blood meal) and anautogenous populations are sympatric, and differences in mating behaviors may be involved. For example, anautogenous populations mate in flight/swarms, while autogenous populations often mate at rest. Here, we characterized flight activity of males and found that anautogenous strain males were crepuscular, while autogenous strain males were crepuscular and nocturnal, with earlier activity onset times. We conducted quantitative trait locus (QTL) mapping to explore the genetic basis of circadian chronotype (crepuscular vs. crepuscular and nocturnal) and time of activity onset. One major-effect QTL was identified for chronotype, while 3 QTLs were identified for activity onset. The highest logarithm of the odds (LOD) score for the chronotype QTL coincides with a chromosome 3 marker that contains a 15-nucleotide indel within the coding region of the canonical clock gene, cryptochrome 2. Sequencing of this locus in 7 different strains showed that the C-terminus of CRY2 in the autogenous forms contain deletions not found in the anautogenous forms. Consequently, we monitored activity in constant darkness and found males from the anautogenous strain exhibited free running periods of ~24 h while those from the autogenous strain were ~22 h. This study provides novel insights into the genetic basis of flight behaviors that likely reflect adaptation to their distinct ecological niches.

The Supplemental Nutrition Assistance Program: Current and Proposed Restricted Food Expenditures.

INTRODUCTION: Between 2000 and 2017, a total of 23 states proposed legislation to further restrict Supplemental Nutrition Assistance Program (SNAP) purchases. In the absence of a pilot program, the potential effect of such restrictions is unclear. The objective of this study is to provide insight on the proposed restrictions' effectiveness by characterizing SNAP households' expenditures on current and proposed restricted foods, and comparing them with their cash expenditures. Restrictions on sugar-sweetened beverages, snack foods, and foods ineligible under the Special Supplemental Program for Women, Infants and Children (WIC) are considered. METHODS: The National Household Food Acquisition and Purchase Survey (collected 2012-2013) provided weekly food expenditures for 1,234 SNAP households. Descriptive statistics and t-tests (completed in 2017) were used to characterize and compare households' cash and restricted food expenditures. RESULTS: On average, SNAP households' allocated 7%, 6%, 17%, and 66% of their food expenditures to currently restricted foods, sugar-sweetened beverages, snack foods, and WIC-ineligible foods. Given a sugar-sweetened beverage or snack food restriction, the average SNAP household can cover their restricted expenditures with cash. However, the average household's expenditures on current restricted and WIC-ineligible foods exceed their cash expenditures by a mean of $40.84 (SE=$2.44). Note that results characterize the impact of proposed SNAP restrictions on the average SNAP household. Individually, it is likely that some SNAP households would be affected by a sugar-sweetened beverage or snack food restriction. CONCLUSIONS: Legislation restricting specific foods will likely be less effective at altering SNAP households' food expenditures than legislation seeking to restrict all WIC-ineligible foods.

A transcriptomic survey of the impact of environmental stress on response to dengue virus in the mosquito, Aedes aegypti.

Populations of Aedes aegypti naturally exhibit variable susceptibility to dengue viruses. This natural variation can be impacted by nutritional stress resulting from larval-stage crowding, indicating the influence of environment components on the adult mosquito immune response. In particular, larval crowding was previously shown to reduce the susceptibility of adult females of a Trinidad field isolate of A. aegypti to the dengue serotype 2 (JAM1409) virus. Here, we present the first whole transcriptome study to address the impact of environmental stress on A. aegypti response to dengue virus. We examined expression profiles of adult females resulting from crowded and optimum reared larvae from the same Trinidad isolate at two critical early time points-3 and 18 hours post dengue virus infected blood meal. We exposed specimens to either a dengue or naïve blood meal, and then characterized the response in ten gene co-expression modules based on their transcriptional associations with environmental stress and time. We further analyzed the top 30 hub or master regulatory genes in each of the modules, and validated our results via qRT-PCR. These hub genes reveal which functions are critical to the mechanisms that confer dengue virus refractoriness or susceptibility to stress conditioned A. aegypti, as well as the time points at which they are most important.

Larval stress alters dengue virus susceptibility in Aedes aegypti (L.) adult females.

In addition to genetic history, environmental conditions during larval stages are critical to the development, success and phenotypic fate of the Aedes aegypti mosquito. In particular, previous studies have shown a strong genotype-by-environment component to adult mosquito body size in response to optimal vs stressed larval conditions. Here, we expand upon those results by investigating the effects of larval-stage crowding and nutritional limitation on the susceptibility of a recent field isolate of Aedes aegypti to dengue virus serotype-2. Interestingly, female mosquitoes from larvae subjected to a stressed regime exhibited significantly reduced susceptibility to disseminated dengue infection 14days post infection compared to those subjected to optimal regimes. Short term survivorship post-infected blood feeding was not significantly different. As with body size, dengue virus susceptibility of a mosquito population is determined by a combination of genetic and environmental factors and is likely maintained by balancing selection. Here, we provide evidence that under different environmental conditions, the innate immune response of field-reared mosquitoes exhibits a large range of phenotypic variability with regard to dengue virus susceptibility. Further, as with body size, our results suggest that mosquitoes reared under optimal laboratory conditions, as employed in all mosquito-pathogen studies to date, may not always be realistic proxies for natural populations.

Description of Brachymeria philornisae sp. n. (Hymenoptera: Chalcididae), a parasitoid of the bird parasite Philornis trinitensis (Diptera: Muscidae) in Tobago, with a review of the sibling species.

In this paper, we describe and illustrate a new species of parasitoid wasp as Brachymeria philornisae Delvare, sp. nov. The new species was reared from Philornis trinitensis Dodge & Aitken (Diptera: Muscidae) puparia that were found in the nests of the bird species Mimus gilvus (Vieillot) (Mimidae) and Tiaris bicolor (L.) (Thraupidae) in Tobago. The new species is of particular interest as it may be considered a potential biological control agent in locations where Philornis species are invasive, such as the Galapagos Islands. Closely related Brachymeria species had taxonomically ambiguous relationship in the past and are compared and reviewed. The species have been classified in the subgenus Pseudobrachymeria, but are here treated within the newly defined subconica species-group of Brachymeria as part of a sibling species complex designated as the subrugosa complex. Species assigned to the subconica species-group are listed and five, one unnamed, are assigned to the subrugosa complex and their females keyed. Species are separated by qualitative characters and morphometry using distance measurements. Trigonura annulipes Costa Lima is renamed as Brachymeria costalimai Delvare nom. nov. because of secondary homonymy.

Patient specific methods for room-mounted x-ray imagers for monoscopic/stereoscopic prostate motion monitoring.

PURPOSE: To investigate the improvement of combined monoscopic/stereoscopic prostate motion monitoring with room-mounted dual x-ray systems by adopting patient specific methods. METHODS: The linac couch was used as a motion stage to simulate 40 highly dynamic real patient prostate trajectories. For each trajectory, 40 s pretreatment and 120 s treatment periods were extracted to represent a typical treatment fraction. Motion was monitored via continuous stereoscopic x-ray imaging of a single gold fiducial and images were retrospectively divided into periods of stereoscopic and monoscopic imaging to simulate periodic blocking of the room-mounted system by the gantry during arc-based therapy. The accuracy of the combined motion monitoring was assessed by comparison with the linac couch log files. To estimate 3-D marker position during monoscopic imaging, the use of population statistics was compared to both maximum likelihood estimation and stereoscopic localization based estimation of individualized prostate probability density functions (PDFs) from the pretreatment period. The inclusion of intrafraction updating was compared to pretreatment initialization alone. RESULTS: Combined mono/stereoscopic localization was successfully implemented. During the transitions from stereoscopic to monoscopic imaging, fiducial localization exhibits sharp discontinuities when population PDFs were employed. Patient specific PDFs successfully reduced the localization error when estimated from stereoscopic localizations, whereas maximum likelihood estimation (MLE) was too unstable in the room-mounted geometry. Intrafraction stereoscopic updating provided further increases in accuracy. Residual error tended to decrease throughout the treatment fraction, as the patient-specific PDFs became more refined. CONCLUSIONS: This is the first demonstration of toggled monoscopic/stereoscopic localization using room-mounted dual x-ray imagers, enabling continuous intrafraction motion monitoring for these systems. We showed that both pretreatment individualization and intrafraction updating should be used to provide the most accurate motion monitoring.

Aedes aegypti (L.) mosquitoes in Trinidad, West Indies: longevity case studies.

Few laboratory and field studies have reported long survival periods for Ae. aegypti females and even fewer have designed experiments to characterize this important life history trait. This study was conducted under laboratory conditions to determine the number of blood meals taken by individual females, the number of eggs laid per individual female, the length of the gonotrophic cycle, and the duration of female survival. The results showed individual females oviposited between 670 and 1,500 eggs throughout their lifetimes, females undergoing large numbers of gonotrophic cycles and surviving up to 224 days. These results are discussed in the context of vector competence, unique alternating high and low oviposition patterns observed after week 14, and resource partitioning/allocation by older Ae. aegypti females after blood feeding.

First reported enterovirus D68 infection in pediatric patients from the Caribbean region: evidence of spread from the U.S. outbreak.

The 2014 enterovirus D68 (EV-D68) outbreak in the United States raised concerns about the introduction of the virus in the Caribbean region. The objective of this study was to provide rapid evidence of the introduction of EV-D68 strains in the Caribbean region during the 2014 outbreak in the United States, using a relatively simple phylogenetic approach. From October 2014 to May 2015, four EV-D68 cases from two countries (Bermuda and Dominica) were detected at the regional referral laboratory at the Caribbean Public Health Agency (Port of Spain, Trinidad and Tobago) based on molecular testing of respiratory specimens. All cases were children presenting to hospitals with moderate respiratory distress. No cases of acute flaccid paralysis were detected. Phylogenetic analysis of the Caribbean strains showed more than 99% similarity with the 2014 U.S.-outbreak strain, providing evidence of the introduction and circulation of the virus in the region.