Pupal Exuviae of Culex Pipiens L. (Diptera: Culicidae) Can be Utilised as a Non-Invasive Method of Biotype Differentiation.

BACKGROUND: Culex pipiens L. is a principal vector of zoonotic arboviruses in Europe, acting in both an amplification role in enzootic transmission between avian hosts and as a bridge vector between avian hosts and mammals. The species consists of two forms which are indistinguishable using morphological methods but possess varying ecological and physiological traits that influence their vector capacity. In this study we validate methods that can be used to extract trace DNA from single pupal exuviae of Cx. pipiens for use in molecular speciation of samples. These DNA extraction methods are compared using measurement of the total yield and successful identification using a real-time polymerase chain reaction (PCR) assay. RESULTS: Genomic DNA was initially extracted from colony-derived individuals using an ethanol precipitation method, two commercially available DNA extraction kits: DNeasy® Blood & Tissue Kit (Qiagen, UK) and Wizard® SV Genomic DNA Purification System (Promega, UK) and a direct real-time PCR method. Time elapsed between eclosion and processing of pupae significantly influenced Cx. pipiens form identification as nucleic acid concentration and PCR amplification success decreased with increased time elapsed. Real-time PCR amplification success, however, was not shown to vary significantly between the three extraction methods, with all methods successfully identifying all samples, but the direct real-time PCR method achieved a lesser amplification success rate of 70% (n = 20 for each treatment). More variable results were produced when field-derived exuviae were used, with no significant difference in real-time PCR amplification success found across the four methods and a lower overall rate of successful identification of 55-80%. CONCLUSIONS: This study shows that both colony and field derived Cx. pipiens pupal exuviae can be a useful non-invasive source of trace DNA permitting accurate biotype differentiation for at least twenty-four hours post-eclosion. The significance and utility of this technique in ecological and behavioural studies of Cx. pipiens is discussed and recommendations made for use according to experimental scenario.

Smooth vs Textured Expanders: Patient Factors and Anatomic Plane Are Greater Factors in Determining First-Stage Breast Reconstruction Outcomes.

BACKGROUND: Textured implants and expanders are associated with an increased risk of breast implant-associated anaplastic large cell lymphoma (BIA-ALCL). As a result, plastic surgeons are utilizing smooth expanders, but many perceive these produce undesirable outcomes including infection, seroma, and lateral displacement. OBJECTIVES: The aim of this study was to compare clinical outcomes of smooth and textured expanders. METHODS: Breast reconstruction patients from January 2018 to May 2021 were retrospectively reviewed. Included patients underwent placement of tissue expanders at the time of mastectomy. Primary outcomes included postoperative seroma, infection, malposition, days to final reconstruction, explantation, and the need for capsulorrhaphy. RESULTS: In total, 233 patients were reviewed, of whom 167 met both inclusion and exclusion criteria. There was no statistically significant difference in poor outcomes comparing smooth and textured expanders. Days to final reconstruction was lower with smooth expanders per breast (P = .0424). The subpectoral group was associated with an increased likelihood of undergoing capsulorrhaphy (P = .004). Prepectoral placement was associated with more seromas (P = .0176) and infections (P = .0245). Demographic factors included older age as a protective factor for undergoing capsulorrhaphy (odds ratio [OR] = 0.962, P = .038), obesity increased the risk of infection (OR = 5.683, P = .0279) and malposition (OR = 6.208, P = .0222), and radiation was associated with malposition (OR = 3.408, P = .0246). CONCLUSIONS: There was no significant difference in poor outcomes between smooth and textured expanders. Patient demographics and anatomical plane placement had greater effects on infection, seroma, and the need for capsulorrhaphy compared with tissue expander texturing.

Visualisation of Bluetongue Virus in the Salivary Apparatus of Culicoides Biting Midges Highlights the Accessory Glands as a Primary Arboviral Infection Site.

BACKGROUND: Arthropods transmit a wide range of pathogens of importance for the global health of humans, animals, and plants. One group of these arthropod vectors, Culicoides biting midges (Diptera: Ceratopogonidae), is the biological vector of several human and animal pathogens, including economically important livestock viruses like bluetongue virus (BTV). Like other arthropod-borne viruses (arboviruses), Culicoides-borne viruses must reach and replicate in the salivary apparatus, from where they can be transmitted to susceptible hosts through the saliva during subsequent blood feeding. Despite the importance of the salivary gland apparatus for pathogen transmission to susceptible animals from the bite of infected Culicoides, these structures have received relatively little attention, perhaps due to the small size and fragility of these vectors. RESULTS: In this study, we developed techniques to visualize the infection of the salivary glands and other soft tissues with BTV, in some of the smallest known arbovirus vectors, Culicoides biting midges, using three-dimensional immunofluorescence confocal microscopy. We showed BTV infection of specific structures of the salivary gland apparatus of female Culicoides vectors following oral virus uptake, related visualisation of viral infection in the salivary apparatus to high viral RNA copies in the body, and demonstrated for the first time, that the accessory glands are a primary site for BTV replication within the salivary apparatus. CONCLUSIONS: Our work has revealed a novel site of virus-vector interactions, and a novel role of the accessory glands of Culicoides in arbovirus amplification and transmission. Our approach would also be applicable to a wide range of arbovirus vector groups including sand flies (Diptera: Psychodidae), as well as provide a powerful tool to investigate arbovirus infection and dissemination, particularly where there are practical challenges in the visualization of small size and delicate tissues of arthropods.

Field-Reassortment of Bluetongue Virus Illustrates Plasticity of Virus Associated Phenotypic Traits in the Arthropod Vector and Mammalian Host In Vivo.

Segmented RNA viruses are a taxonomically diverse group that can infect plant, wildlife, livestock and human hosts. A shared feature of these viruses is the ability to exchange genome segments during coinfection of a host by a process termed "reassortment." Reassortment enables rapid evolutionary change, but where transmission involves a biological arthropod vector, this change is constrained by the selection pressures imposed by the requirement for replication in two evolutionarily distant hosts. In this study, we use an in vivo, host-arbovirus-vector model to investigate the impact of reassortment on two phenotypic traits, virus infection rate in the vector and virulence in the host. Bluetongue virus (BTV) (Reoviridae) is the causative agent of bluetongue (BT), an economically important disease of domestic and wild ruminants and deer. The genome of BTV comprises 10 linear segments of dsRNA, and the virus is transmitted between ruminants by Culicoides biting midges (Diptera: Ceratopogonidae). Five strains of BTV representing three serotypes (BTV-1, BTV-4, and BTV-8) were isolated from naturally infected ruminants in Europe and ancestral/reassortant lineage status assigned through full genome sequencing. Each strain was then assessed in parallel for the ability to replicate in vector Culicoides and to cause BT in sheep. Our results demonstrate that two reassortment strains, which themselves became established in the field, had obtained high replication ability in C. sonorensis from one of the ancestral virus strains, which allowed inferences of the genome segments conferring this phenotypic trait. IMPORTANCE Reassortment between virus strains can lead to major shifts in the transmission parameters and virulence of segmented RNA viruses, with consequences for spread, persistence, and impact. The ability of these pathogens to adapt rapidly to their environment through this mechanism presents a major challenge in defining the conditions under which emergence can occur. Utilizing a representative mammalian host-insect vector infection and transmission model, we provide direct evidence of this phenomenon in closely related ancestral and reassortant strains of BTV. Our results demonstrate that efficient infection of Culicoides observed for one of three ancestral BTV strains was also evident in two reassortant strains that had subsequently emerged in the same ecosystem.

The Acquisition and Retention of Lumpy Skin Disease Virus by Blood-Feeding Insects Is Influenced by the Source of Virus, the Insect Body Part, and the Time since Feeding.

Lumpy skin disease virus (LSDV) is a poxvirus that causes severe systemic disease in cattle and is spread by mechanical arthropod-borne transmission. This study quantified the acquisition and retention of LSDV by four species of Diptera (Stomoxys calcitrans, Aedes aegypti, Culex quinquefasciatus, and Culicoides nubeculosus) from cutaneous lesions, normal skin, and blood from a clinically affected animal. The acquisition and retention of LSDV by Ae. aegypti from an artificial membrane feeding system was also examined. Mathematical models of the data were generated to identify the parameters which influence insect acquisition and retention of LSDV. For all four insect species, the probability of acquiring LSDV was substantially greater when feeding on a lesion compared with feeding on normal skin or blood from a clinically affected animal. After feeding on a skin lesion LSDV was retained on the proboscis for a similar length of time (around 9 days) for all four species and for a shorter time in the rest of the body, ranging from 2.2 to 6.4 days. Acquisition and retention of LSDV by Ae. aegypti after feeding on an artificial membrane feeding system that contained a high titer of LSDV was comparable to feeding on a skin lesion on a clinically affected animal, supporting the use of this laboratory model as a replacement for some animal studies. This work reveals that the cutaneous lesions of LSD provide the high-titer source required for acquisition of the virus by insects, thereby enabling the mechanical vector-borne transmission. IMPORTANCE Lumpy skin disease virus (LSDV) is a high consequence pathogen of cattle that is rapidly expanding its geographical boundaries into new regions such as Europe and Asia. This expansion is promoted by the mechanical transmission of the virus via hematogenous arthropods. This study quantifies the acquisition and retention of LSDV by four species of blood-feeding insects and reveals that the cutaneous lesions of LSD provide the high titer virus source necessary for virus acquisition by the insects. An artificial membrane feeding system containing a high titer of LSDV was shown to be comparable to a skin lesion on a clinically affected animal when used as a virus source. This promotes the use of these laboratory-based systems as replacements for some animal studies. Overall, this work advances our understanding of the mechanical vector-borne transmission of LSDV and provides evidence to support the design of more effective disease control programmes.

Quantifying and Modeling the Acquisition and Retention of Lumpy Skin Disease Virus by Hematophagus Insects Reveals Clinically but Not Subclinically Affected Cattle Are Promoters of Viral Transmission and Key Targets for Control of Disease Outbreaks.

Lumpy skin disease virus (LSDV) is a vector-transmitted poxvirus that causes disease in cattle. Vector species involved in LSDV transmission and their ability to acquire and transmit the virus are poorly characterized. Using a highly representative bovine experimental model of lumpy skin disease, we fed four model vector species (Aedes aegypti, Culex quinquefasciatus, Stomoxys calcitrans, and Culicoides nubeculosus) on LSDV-inoculated cattle in order to examine their acquisition and retention of LSDV. Subclinical disease was a more common outcome than clinical disease in the inoculated cattle. Importantly, the probability of vectors acquiring LSDV from a subclinical animal (0.006) was very low compared with that from a clinical animal (0.23), meaning an insect feeding on a subclinical animal was 97% less likely to acquire LSDV than one feeding on a clinical animal. All four potential vector species studied acquired LSDV from the host at a similar rate, but Aedes aegypti and Stomoxys calcitrans retained the virus for a longer time, up to 8 days. There was no evidence of virus replication in the vector, consistent with mechanical rather than biological transmission. The parameters obtained in this study were combined with data from studies of LSDV transmission and vector life history parameters to determine the basic reproduction number of LSDV in cattle mediated by each of the model species. This reproduction number was highest for Stomoxys calcitrans (19.1), followed by C. nubeculosus (7.1) and Ae. aegypti (2.4), indicating that these three species are potentially efficient transmitters of LSDV; this information can be used to inform LSD control programs.IMPORTANCE Lumpy skin disease virus (LSDV) causes a severe systemic disease characterized by cutaneous nodules in cattle. LSDV is a rapidly emerging pathogen, having spread since 2012 into Europe and Russia and across Asia. The vector-borne nature of LSDV transmission is believed to have promoted this rapid geographic spread of the virus; however, a lack of quantitative evidence about LSDV transmission has hampered effective control of the disease during the current epidemic. Our research shows subclinical cattle play little part in virus transmission relative to clinical cattle and reveals a low probability of virus acquisition by insects at the preclinical stage. We have also calculated the reproductive number of different insect species, therefore identifying efficient transmitters of LSDV. This information is of utmost importance, as it will help to define epidemiological control measures during LSDV epidemics and of particular consequence in resource-poor regions where LSD vaccination may be less than adequate.

Protein modeling to assess the pathogenicity of rare variants of SERPINA1 in patients suspected of having Alpha 1 Antitrypsin Deficiency.

BACKGROUND: Alpha 1 Antitrypsin (AAT) is a key serum proteinase inhibitor encoded by SERPINA1. Sequence variants of the gene can cause Alpha 1 Antitrypsin Deficiency (AATD), a condition associated with lung and liver disease. The majority of AATD cases are caused by the 'Z' and 'S' variants - single-nucleotide variations (SNVs) that result in amino acid substitutions of E342K and E264V. However, SERPINA1 is highly polymorphic, with numerous potentially clinically relevant variants reported. Novel variants continue to be discovered, and without reports of pathogenicity, it can be difficult for clinicians to determine the best course of treatment. METHODS: We assessed the utility of next-generation sequencing (NGS) and predictive computational analysis to guide the diagnosis of patients suspected of having AATD. Blood samples on serum separator cards were submitted to the DNA1 Advanced Screening Program (Biocerna LLC, Fulton, Maryland, USA) by physicians whose patients were suspected of having AATD. Laboratory analyses included quantification of serum AAT levels, qualitative analysis by isoelectric focusing, and targeted genotyping and NGS of the SERPINA1 gene. Molecular modeling software UCSF Chimera (University College of San Francisco, CA) was used to visualize the positions of amino acid changes as a result of rare/novel SNVs. Predictive software was used to assess the potential pathogenicity of these variants; methods included a support vector machine (SVM) program, PolyPhen-2 (Harvard University, Cambridge, MA), and FoldX (Centre for Genomic Regulation, Barcelona, Spain). RESULTS: Samples from 23 patients were analyzed; 21 rare/novel sequence variants were identified by NGS, including splice variants (n = 2), base pair deletions (n = 1), stop codon insertions (n = 2), and SNVs (n = 16). Computational modeling of protein structures caused by the novel SNVs showed that 8 were probably deleterious, and two were possibly deleterious. For the majority of probably/possibly deleterious SNVs (I50N, P289S, M385T, M221T, D341V, V210E, P369H, V333M and A142D), the mechanism is probably via disruption of the packed hydrophobic core of AAT. Several deleterious variants occurred in combination with more common deficiency alleles, resulting in very low AAT levels. CONCLUSIONS: NGS and computational modeling are useful tools that can facilitate earlier, more precise diagnosis, and consideration for AAT therapy in AATD.

Understanding Long-Term Trajectories in Web-Based Happiness Interventions: Secondary Analysis From Two Web-Based Randomized Trials.

BACKGROUND: A critical issue in understanding the benefits of Web-based interventions is the lack of information on the sustainability of those benefits. Sustainability in studies is often determined using group-level analyses that might obscure our understanding of who actually sustains change. Person-centric methods might provide a deeper knowledge of whether benefits are sustained and who tends to sustain those benefits. OBJECTIVE: The aim of this study was to conduct a person-centric analysis of longitudinal outcomes, examining well-being in participants over the first 3 months following a Web-based happiness intervention. We predicted we would find distinct trajectories in people's pattern of response over time. We also sought to identify what aspects of the intervention and the individual predicted an individual's well-being trajectory. METHODS: Data were gathered from 2 large studies of Web-based happiness interventions: one in which participants were randomly assigned to 1 of 14 possible 1-week activities (N=912) and another wherein participants were randomly assigned to complete 0, 2, 4, or 6 weeks of activities (N=1318). We performed a variation of K-means cluster analysis on trajectories of life satisfaction (LS) and affect balance (AB). After clusters were identified, we used exploratory analyses of variance and logistic regression models to analyze groups and compare predictors of group membership. RESULTS: Cluster analysis produced similar cluster solutions for each sample. In both cases, participant trajectories in LS and AB fell into 1 of 4 distinct groups. These groups were as follows: those with high and static levels of happiness (n=118, or 42.8%, in Sample 1; n=306, or 52.8%, in Sample 2), those who experienced a lasting improvement (n=74, or 26.8% in Sample 1; n=104, or 18.0%, in Sample 2), those who experienced a temporary improvement but returned to baseline (n=37, or 13.4%, in Sample 1; n=82, or 14.2%, in Sample 2), and those with other trajectories (n=47, or 17.0%, in Sample 1; n=87, or 15.0% in Sample 2). The prevalence of depression symptoms predicted membership in 1 of the latter 3 groups. Higher usage and greater adherence predicted sustained rather than temporary benefits. CONCLUSIONS: We revealed a few common patterns of change among those completing Web-based happiness interventions. A noteworthy finding was that many individuals began quite happy and maintained those levels. We failed to identify evidence that the benefit of any particular activity or group of activities was more sustainable than any others. We did find, however, that the distressed portion of participants was more likely to achieve a lasting benefit if they continued to practice, and adhere to, their assigned Web-based happiness intervention.

The Effects of Inflammation on Alpha 1 Antitrypsin Levels in a National Screening Cohort.

Alpha 1 Antitrypsin (AAT) is a highly polymorphic serum protein. Several genetic variants are associated with varying degrees of decreased serum levels; however, these levels can rise in response to infection, inflammation, injury and estrogen levels. Although the effect of inflammation is well established, it has never been studied quantitatively with respect to specific genotypes in a large representative sample. Using data from a national AAT deficiency-targeted screening cohort, we evaluated AAT levels of patients with normal and deficiency genotypes in response to inflammation, indicated by elevated serum C-reactive protein (CRP). Additionally, we utilized a regression analysis to adjust for the effect of inflammation for each genotype. Across all stratified genotype groups, increased AAT levels were observed in patients with CRP ≥5 mg/L. Different AAT phenotypes reacted differently in the acute phase; M showed a strong response and Z a reduced reaction. Nevertheless, we discovered that inflammation significantly masked clinically relevant base AAT levels in some PI*MZ individuals; approximately a quarter of PI*MZ samples showed signs of inflammation. Median AAT levels (mg/dL) in the presence of inflammation are given for several genotypes; numbers in parentheses are levels from the cohort without inflammation/adjusted levels from the cohort with inflammation using the newly devised algorithm: PI*MM: 162 (142/140); PI*MS: 136 (117/115); PI*MZ: 104 (85/89); PI*MF: 161 (132/141); PI*SS: 115 (96/91); PI*SZ: 66 (54/50). We conclude that simultaneous determinations of CRP and AAT levels, and genotyping are clinically valuable in defining AAT variants and that the effect of inflammation can be adjusted for.

State-of-the-art testing for alpha-1 antitrypsin deficiency.

BACKGROUND: Alpha-1 antitrypsin deficiency (AATD) is a genetic condition characterized by low serum levels of the protein alpha-1 antitrypsin. Because there are no unique clinical symptoms that point to a definitive diagnosis of AATD, laboratory testing is crucial to differentiate this disease from others. OBJECTIVE: To summarize advances in laboratory techniques used to test for AATD. METHODS: Data were sourced from a nonsystematic literature review of MEDLINE and the author's personal literature collection, and by checking reference lists of sourced articles. RESULTS: Since the original description of AATD by Laurell and Eriksson in 1963, testing methods have undergone major changes. Currently, alpha-1 antitrypsin protein is quantified by immunologic measurement in serum, and the phenotype is characterized by isoelectric focusing and/or targeted genotyping of predefined mutations. In addition, whole-gene sequencing of the gene SERPINA1 can be undertaken. However, this is costly and generally used only if targeted genotyping cannot conclusively identify the variant. The introduction of next-generation sequencing (NGS), which enables rapid and accurate sequencing of large quantities of DNA fragments in a single reaction, may help reduce costs. With its increasing availability, NGS may begin to appear in testing protocols. Clinical guidelines recommend that patients are tested for AATD if they have chronic irreversible airflow obstruction, especially those with early onset disease or a positive family history of AATD. Despite this, AATD is still underrecognized, and significant delays exist between symptom onset and diagnosis. CONCLUSION: Traditional testing practices have limitations. Screening programs that incorporate NGS are the most comprehensive methods available for accurate diagnosis of AATD.

Bluetongue, Schmallenberg - what is next? Culicoides-borne viral diseases in the 21st Century.

In the past decade, two pathogens transmitted by Culicoides biting midges (Diptera: Ceratopogonidae), bluetongue virus and Schmallenberg virus, have caused serious economic losses to the European livestock industry, most notably affecting sheep and cattle. These outbreaks of arboviral disease have highlighted large knowledge gaps on the biology and ecology of indigenous Culicoides species. With these research gaps in mind, and as a means of assessing what potential disease outbreaks to expect in the future, an international workshop was held in May 2013 at Wageningen University, The Netherlands. It brought together research groups from Belgium, France, Germany, Spain, Switzerland, United Kingdom and The Netherlands, with diverse backgrounds in vector ecology, epidemiology, entomology, virology, animal health, modelling, and genetics. Here, we report on the key findings of this workshop.

Target Happiness Attenuates Perceivers' Moral Condemnation of Prejudiced People.

Five experiments (combined N = 4,915) tested the prediction that the moral boost of happiness would persist for social targets with moral failings. In Studies 1 and 2, White and Black participants, respectively, judged happy (versus unhappy) racist targets more morally good. In Study 3, happy (versus unhappy) racist targets were judged more morally good and less (more) likely to engage in racist (good) behavior. Behavioral expectations explained the link between happiness and moral evaluations. Study 4 replicated Studies 1 to 3 in the context of sexism. In Study 5, happy (versus unhappy) targets who engaged in racially biased behavior were evaluated as more morally good, and this effect was explained by behavioral forecasts. Happiness boosts attributions of moral goodness for prejudiced people and does so via expectations for future behavior. Future directions are discussed.

Arthropod-Borne Viruses of Human and Animal Importance: Overwintering in Temperate Regions of Europe during an Era of Climate Change.

The past three decades have seen an increasing number of emerging arthropod-borne viruses in temperate regions This process is ongoing, driven by human activities such as inter-continental travel, combined with the parallel emergence of invasive arthropods and an underlying change in climate that can increase the risk of virus transmission and persistence. In addition, natural events such as bird migration can introduce viruses to new regions. Despite the apparent regularity of virus emergence, arthropod-borne viruses circulating in temperate regions face the challenge of the late autumn and winter months where the arthropod vector is inactive. Viruses therefore need mechanisms to overwinter or they will fail to establish in temperate zones. Prolonged survival of arthropod-borne viruses within the environment, outside of both vertebrate host and arthropod vector, is not thought to occur and therefore is unlikely to contribute to overwintering in temperate zones. One potential mechanism is continued infection of a vertebrate host. However, infection is generally acute, with the host either dying or producing an effective immune response that rapidly clears the virus. There are few exceptions to this, although prolonged infection associated with orbiviruses such as bluetongue virus occurs in certain mammals, and viraemic vertebrate hosts therefore can, in certain circumstances, provide a route for long-term viral persistence in the absence of active vectors. Alternatively, a virus can persist in the arthropod vector as a mechanism for overwintering. However, this is entirely dependent on the ecology of the vector itself and can be influenced by changes in the climate during the winter months. This review considers the mechanisms for virus overwintering in several key arthropod vectors in temperate areas. We also consider how this will be influenced in a warming climate.

Interventions to promote medical student well-being: an overview of systematic reviews.

OBJECTIVE: To conduct an overview of systematic reviews that explore the effectiveness of interventions to enhance medical student well-being. DESIGN: Overview of systematic reviews. DATA SOURCES: The Cochrane Library of Systematic Reviews, MEDLINE, APA PsychInfo, CINAHL and Scopus were searched from database inception until 31 May 2023 to identify systematic reviews of interventions to enhance medical student well-being. Ancestry searching and citation chasing were also conducted. DATA EXTRACTION AND SYNTHESIS: The Assessing the Methodological Quality of Systematic Reviews V.2 tool was used to appraise the quality of the included reviews. A narrative synthesis was conducted, and the evidence of effectiveness for each intervention was rated. RESULTS: 13 reviews (with 94 independent studies and 17 616 students) were included. The reviews covered individual-level and curriculum-level interventions. Individual interventions included mindfulness (n=12), hypnosis (n=6), mental health programmes (n=7), yoga (n=4), cognitive and behavioural interventions (n=1), mind-sound technology (n=1), music-based interventions (n=1), omega-3 supplementation (n=1), electroacupuncture (n=1) and osteopathic manipulative treatment (n=1). The curriculum-level interventions included pass/fail grading (n=4), problem-based curriculum (n=2) and multicomponent curriculum reform (n=2). Most interventions were not supported by sufficient evidence to establish effectiveness. Eleven reviews were rated as having 'critically low' quality, and two reviews were rated as having 'low' quality. CONCLUSIONS: Individual-level interventions (mindfulness and mental health programmes) and curriculum-level interventions (pass/fail grading) can improve medical student well-being. These conclusions should be tempered by the low quality of the evidence. Further high-quality research is required to explore additional effective interventions to enhance medical student well-being and the most efficient ways to implement and combine these for maximum benefit.

Specific T-cell subsets have a role in anti-viral immunity and pathogenesis but not viral dynamics or onwards vector transmission of an important livestock arbovirus.

Introduction: Bluetongue virus (BTV) is an arthropod-borne Orbivirus that is almost solely transmitted by Culicoides biting midges and causes a globally important haemorrhagic disease, bluetongue (BT), in susceptible ruminants. Infection with BTV is characterised by immunosuppression and substantial lymphopenia at peak viraemia in the host. Methods: In this study, the role of cell-mediated immunity and specific T-cell subsets in BTV pathogenesis, clinical outcome, viral dynamics, immune protection, and onwards transmission to a susceptible Culicoides vector is defined in unprecedented detail for the first time, using an in vivo arboviral infection model system that closely mirrors natural infection and transmission of BTV. Individual circulating CD4+, CD8+, or WC1+ γδ T-cell subsets in sheep were depleted through the administration of specific monoclonal antibodies. Results: The absence of cytotoxic CD8+ T cells was consistently associated with less severe clinical signs of BT, whilst the absence of CD4+ and WC1+ γδ T cells both resulted in an increased clinical severity. The absence of CD4+ T cells also impaired both a timely protective neutralising antibody response and the production of IgG antibodies targeting BTV non-structural protein, NS2, highlighting that the CD4+ T-cell subset is important for a timely protective immune response. T cells did not influence viral replication characteristics, including onset/dynamics of viraemia, shedding, or onwards transmission of BTV to Culicoides. We also highlight differences in T-cell dependency for the generation of immunoglobulin subclasses targeting BTV NS2 and the structural protein, VP7. Discussion: This study identifies a diverse repertoire of T-cell functions during BTV infection in sheep, particularly in inducing specific anti-viral immune responses and disease manifestation, and will support more effective vaccination strategies.

Large-Bore Mechanical Thrombectomy of Acute Pulmonary Embolism at a Community-Based Hospital: A Case Series.

Acute pulmonary embolism (PE) is a common, and sometimes fatal, diagnosis that results in over 11,000 deaths in hospitalized patients in the USA annually. In patients with high-risk or high-intermediate-risk PE and especially in whom thrombolytic therapy is contraindicated, mechanical thrombectomy can be an effective treatment option. This case series outlines three patients with PE who were successfully treated with large bore aspiration thrombectomy by Interventional Radiology at a community-based military treatment facility (MTF). Two patients had presented to the emergency department with acute PE and a third of patients with acute PE were transferred from an outside hospital specifically for mechanical thrombectomy due to a complication from systemic anticoagulation. The patients were categorized as good candidates for immediate large-bore aspiration thrombectomy, a recently added capability at the MTF. The patients showed immediate improvement post-procedure and required only one night admission for observation to the intensive care unit. Implementation of this new capability for patients with acute high-risk or high-intermediate-risk PE or with contraindications to thrombolysis provides an alternative treatment with immediate, life-saving capability.

Investigation of diel activity of Culicoides biting midges (Diptera: Ceratopogonidae) in the United Kingdom by using a vehicle-mounted trap.

Truck trap collections of Culicoides biting midges (Diptera: Ceratopogonidae) were made during 2 yr of sampling from 2008 to 2009 at a farm site in southern England. Samples were collected from 810 sample runs carried out over 52 d and contained 7,095 Culicoides of which more than half (50.3%) were identified as Culicoides obsoletus Meigen by using a multiplex polymerase chain reaction assay. Other commonly encountered species included Culicoides scoticus Downes & Kettle (14.7% of total Culicoides caught), Culicoides dewulfi Goetghebuer (3.7%), and Culicoides chiopterus Meigen (4.2%). The activity rates of these species were examined with regard to both meteorological factors (light intensity, humidity, temperature, and wind speed and direction) and other potentially contributing variables (lunar phase and brightness, sunset time, and year) by using generalized linear models. All the species examined were collected in greater abundance at sunset, although the relationship between underlying light intensity and numbers was less pronounced in C. dewulfi and C. chiopterus. Collections of Culicoides were reduced at temperatures above 21 degrees C and were inversely related to wind speed. Variation between species was recorded, however, in response to wind direction: C. dewulfi and C. chiopterus were associated with prevailing winds passing through fields containing livestock, whereas C. obsoletus and C. scoticus demonstrated no such relationship. A male:female ratio of 1:3.56 was observed in catches, and male populations were protandrous. These results are discussed with reference both to the ecology of these species and methods currently used to predict adult Culicoides movement and abundance in Europe.

Emergence dynamics of adult Culicoides biting midges at two farms in south-east England.

BACKGROUND: Culicoides biting midges (Diptera: Ceratopogonidae) are biological vectors of livestock arboviruses that cause diseases with significant economic, social and welfare impacts. Within temperate regions, livestock movement during arbovirus outbreaks can be facilitated by declaring a 'seasonal vector-free period' (SVFP) during winter when adult Culicoides are not active. In this study we carry out long-term monitoring of Culicoides adult emergence from larval development habitats at two farms in the UK to validate current definitions of the SVFP and to provide novel bionomic data for known vector species. METHODS: Standard emergence traps were used to collect emerging adult Culicoides from dung heaps at two cattle farms in the south-east of England from June to November 2016 and March 2017 to May 2018. Culicoides were morphologically identified to species or complex level and count data were analysed using a simple population dynamic model for pre-adult Culicoides that included meteorological components. RESULTS: More than 96,000 Culicoides were identified from 267 emergence trapping events across 2 years, revealing clear evidence of bivoltinism from peaks of male populations of Culicoides obsoletus emerging from dung heaps. This pattern was also reflected in the emergence of adult female Obsoletus complex populations, which dominated the collections (64.4% of total catch) and emerged throughout the adult active period. Adult male C. obsoletus were observed emerging earlier than females (protandry) and emergence of both sexes occurred throughout the year. Culicoides chiopterus and Culicoides scoticus were also identified in spring emergence collections, providing the first evidence for the overwintering of larvae in dung heaps for these species. CONCLUSIONS: This study demonstrates continual and highly variable rates of emergence of Culicoides throughout the year. A lack of evidence for mass emergence in spring along with the ability to observe male generations highlights the need for complementary surveillance techniques in addition to light-trap data when investigating seasonality and phenology. Evidence was found of other vector species, C. chiopterus and C. scoticus, utilising cattle dung heaps as an overwintering habitat, further highlighting the importance of these habitats on farms.

Hernia Following Rectus Sheath Hematoma.

Rectus sheath hematomas (RSH) are increasing in prevalence, presumably correlating with increased use of anticoagulation medications and an aging population. Comorbidities such as blood dyscrasias, atherosclerosis, and hypertension are associated with an increased risk of developing an RSH. Iatrogenic origin of RSH, secondary to treatment of various abdominal pathologies, is not uncommon. Due to its exceptionally non-specific array of clinical signs and symptoms, RSH can be challenging to diagnose in the clinical setting without the aid of radiological images. Abdominal computed tomography (CT) is generally the modality of choice through which the RSH can be successfully identified and characterized. CT imaging can play an important role in the planning of RSH management, as effective management varies depending on the size and position of the RSH. Recurrent bleeding, hypovolemic shock, abdominal compartment syndrome, myonecrosis, and infection have been traditionally considered as the more prominent complications of RSH. However, with more cases occurring, more complications are being described in the literature. The following case presents a previously unreported complication of RSH, that of bowel herniation into a potential space created by a previously treated RSH.

Before/after Bayes: A comparison of frequentist and Bayesian mixed-effects models in applied psychological research.

Bayesian methods are becoming increasingly used in applied psychological research. Previous researchers have thoroughly written about much of the details already, including the philosophy underlying Bayesian methods, computational issues associated with Bayesian model estimation, Bayesian model development and summary, and the role of Bayesian methods in the so-called replication crisis. In this paper, we seek to provide case studies comparing the use of frequentist methods to the use of Bayesian methods in applied psychological research. These case studies are intended to 'illustrate by example' the ways that Bayesian modelling differs from frequentist modelling and the differing conclusions that one may arrive at using the two methods. The intended audience is applied psychological researchers who have been trained in the traditional frequentist framework, who are familiar with mixed-effects models and who are curious about how statistical results might look in a Bayesian context. Along with our case studies, we provide general opinions and guidance on the use of Bayesian methods in applied psychological research.