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Abstract Coronavirus disease 2019 (COVID-19) is associated with extreme inflammatory response, disordered hemostasis and high thrombotic risk. A high incidence of thromboembolic events has been reported despite thromboprophylaxis, raising the question of a more effective anticoagulation. First-line hemostasis tests such as activated partial thromboplastin time, prothrombin time, fibrinogen and D-dimers are proposed for assessing thrombotic risk and monitoring hemostasis, but are vulnerable to many drawbacks affecting their reliability and clinical relevance. Specialized hemostasis-related tests (soluble fibrin complexes, tests assessing fibrinolytic capacity, viscoelastic tests, thrombin generation) may have an interest to assess the thrombotic risk associated with COVID-19. Another challenge for the hemostasis laboratory is the monitoring of heparin treatment, especially unfractionated heparin in the setting of an extreme inflammatory response. This review aimed at evaluating the role of hemostasis tests in the management of COVID-19 and discussing their main limitations.

Background: Obesity interventions rely predominantly on managing dietary intake and/or increasing physical activity but sustained adherence to behavioural regimens is often poor. Avatar technology is well established within the computer gaming industry and evidence suggests that virtual representations of self may impact real-world behaviour, acting as a catalyst for sustained weight loss behaviour modification. However, the effectiveness of avatar technology in promoting weight loss is unclear. Aims: We aimed to assess the quantity and quality of empirical support for the use of avatar technologies in adult weight loss interventions. Method: A systematic review of empirical studies was undertaken. The key objectives were to determine if: (i) the inclusion of avatar technology leads to greater weight loss achievement compared to routine intervention; and (ii) whether weight loss achievement is improved by avatar personalisation (avatar visually reflects self). Results: We identified 6 papers that reported weight loss data. Avatar-based interventions for weight loss management were found to be effective in the short (4–6 weeks) and medium (3–6 months) term and improved weight loss maintenance in the long term (12 months). Only 2 papers included avatar personalisation, but results suggested there may be some added motivational benefit. Conclusions: The current evidence supports that avatars may positively impact weight loss achievement and improve motivation. However, with only 6 papers identified the evidence base is limited and therefore findings need to be interpreted with caution. Keywords: Obesity, Weight loss, Avatars, Virtual reality, Second life, Systematic review

Small-cell lung cancer (SCLC) is often associated with paraneoplastic syndromes. To assess the role of anti-neuronal autoantibodies (NAAs) as biomarkers of treatment outcome, we assessed NAAs in serial samples from SCLC patients treated with chemoimmunotherapy compared to chemotherapy alone. We evaluated 2 cohorts: in cohort 1 (C1), 47 patients received standard platinum/etoposide, and in cohort 2 (C2), 38 patients received ipilimumab, carboplatin and etoposide. Serum samples at baseline and subsequent time points were analyzed for the presence of NAAs. NAAs were detected at baseline in 25 patients (53.2%) in C1 and in 20 patients (52.6%) in C2 (most frequently anti-Sox1). NAA at baseline was associated with limited disease (75% vs 50%; p: 0.096) and better overall survival (15.1 m vs 11.7 m; p: 0.032) in C1. Thirteen patients (28.9%) showed 2 or more reactivities before treatment; this was associated with worse PFS (5.5 m vs 7.3 m; p: 0.005) in patients treated with chemoimmunotherapy. NAA titers decreased after therapy in 68.9% patients, with no differential patterns of change between cohorts. Patients whose NAA titer decreased after treatment, showed longer OS [18.5 m (95% CI: 15.8 – 21.2)] compared with those whose NAA increased [12.3 m (95% CI: 8.1 – 16.5; p 0.049)], suggesting that antibody levels correlate to tumor load. Our findings reinforce the role of NAAs as prognostic markers and tumor activity/burden in SCLC, warrant further investigation in their predictive role for immunotherapy and raise concern over the use of immunotherapy in patients with more than one anti-NAA reactivity.

Different plants are known to have different soil microbial communities associated with them. Agricultural management practices such as fertiliser and pesticide addition, crop rotation, and grazing animals can lead to different microbial communities in the associated agricultural soils. Soil dilution plates, most-probable-number (MPN), community level physiological profiling (CLPP), and buried slide technique as well as some measured soil physicochemical parameters were used to determine changes during the growing season in the ecosystem profile in wheat fields subjected to wheat monoculture or wheat in annual rotation with medic/clover pasture. Statistical analyses showed that soil moisture had an over-riding effect on seasonal fluctuations in soil physicochemical and microbial populations. While within season soil microbial activity could be differentiated between wheat fields under rotational and monoculture management, these differences were not significant.

Biting midges (Culicoides spp.) are important vectors of several insect borne arboviruses but are underrepresented in terms of availability of high-resolution genomic resources. We assembled and annotated complete mitochondrial genomes for two Culicoides species, namely C. brevitarsis and C. imicola which are proven vectors for Bluetongue Virus (BTV). We used both long read and short read sequencing technologies to assemble the circular genomes. The genome sizes are 17,100 bp and 17,031 bp, respectively, all comprising 37 genes, including 13 protein, 22 tRNA, two rRNA coding genes, and one non-coding AT rich control region. The gene organizations and orientations are comparable to other available Culicoides mitogenomes, except for a translocation of ND2 and six tRNA genes in both C. brevitarsis and C. imicola. Eleven protein-coding genes encode a full TAA stop codon, while two (ND5, COX3) are completed by mRNA polyadenylation. Phylogenetic analysis of the mitogenomes showed C. brevitarsis and C. imicola form a monophyletic group. The sequences of these mitogenomes contribute to a baseline of molecular tools for diagnostics and surveillance for use by World Organisation for Animal Health (WOAH) reference laboratories for monitoring vectors of emerging diseases.

Abstract Background In Alabama only 4% of those eligible have been screened for lung cancer. The ALCASE project focused on navigating eligible individuals to lung cancer screening. Methods Trained local staff enrolled screen eligible individuals from seven rural counties and one urban county. Demographics and knowledge of and barriers to lung cancer screening were collected using questionnaires. Education was provided and individuals were navigated to undergo screening. Descriptive statistics for enrolled and screened participants were computed using SAS 9.4. Debriefing interviews were conducted with the ALCASE staff regarding facilitators/barriers to implementing this project and lessons learned. Using NVivo, themes were identified through a combined deductive and inductive process. Results In total, 447 people were contacted of which 257 were enrolled. Participants were predominantly African American (86.8%), female (56.8%), and 86.4% had health insurance. Study participants acknowledged the need for more education of lung cancer/screening procedures; help navigating clinics for screening services and having healthcare facilities close to home. The top five barriers to getting screened were transportation, financial issues, emotional concerns, healthcare insurance, and COVID‐19. Of the 257, 106 participants (41%) completed a primary care referral and were screened for lung cancer. Debriefing interviews revealed: (i) Overall impressions of implementing ALCASE were positive. (ii) Barriers in implementing ALCASE were identified predominantly at the physician and institutional level. (iii) Facilitators in implementing ALCASE were identified at multiple levels. (iv) Suggestions on improving lung cancer screening leaned toward mitigating barriers at the institutional and structural level. Conclusion Ability to get screened is severely challenged by both personal and structural barriers.

Abstract Thrombopoietin (Tpo) is the primary regulator of megakaryocyte and platelet numbers and is required for haematopoetic stem cell maintenance. Tpo functions by binding its receptor (TpoR, a homodimeric Class I cytokine receptor) and initiating cell proliferation or differentiation. Here we characterise the murine Tpo:TpoR signalling complex biochemically and structurally, using cryo-electron microscopy. Tpo uses opposing surfaces to recruit two copies of receptor, forming a 1:2 complex. Although it binds to the same, membrane-distal site on both receptor chains, it does so with significantly different affinities and its highly glycosylated C-terminal domain is not required. In one receptor chain, a large insertion, unique to TpoR, forms a partially structured loop that contacts cytokine. Tpo binding induces the juxtaposition of the two receptor chains adjacent to the cell membrane. The therapeutic agent romiplostim also targets the cytokine-binding site and the characterisation presented here supports the future development of improved TpoR agonists.

Abstract Mosquitoes have profoundly affected human history and continue to threaten human health through the transmission of a diverse array of pathogens. The phylogeny of mosquitoes has remained poorly characterized due to difficulty in taxonomic sampling and limited availability of genomic data beyond the most important vector species. Here, we used phylogenomic analysis of 709 single copy ortholog groups from 256 mosquito species to produce a strongly supported phylogeny that resolves the position of the major disease vector species and the major mosquito lineages. Our analyses support an origin of mosquitoes in the early Triassic (217 MYA [highest posterior density region: 188–250 MYA]), considerably older than previous estimates. Moreover, we utilize an extensive database of host associations for mosquitoes to show that mosquitoes have shifted to feeding upon the blood of mammals numerous times, and that mosquito diversification and host-use patterns within major lineages appear to coincide in earth history both with major continental drift events and with the diversification of vertebrate classes.

Background and Significance: Achievable signal to noise ratios (SNR) in magnetic resonance microscopy images are limited by acquisition times and the decreasing number of spins in smaller voxels. A common method of enhancing SNR is to cool the RF receiver coil. Significant SNR gains are realized only when the Johnson noise generated within the RF hardware is large compared to the electromagnetic noise produced by the sample. Cryogenic cooling of imaging probes is in common use in high field systems, but it is difficult to insulate a sample from the extreme temperatures involved and in practice imaging cryoprobes have been limited to surface or partial volume designs only. In order to use a solenoid in which the windings were not directly cooled and in close proximity to the sample, we designed a chamber to cool only the tune and match circuitry and show experimentally it is possible to achieve much of the theoretically available SNR gain. Methods: A microcoil circuit consisting of two tuning capacitors, one fixed capacitor, and SMB coaxial cable was designed to resonate at 650 MHz for imaging on a Bruker 15.2 T scanner. Sample noise increases with the sample diameter, so surface loops and solenoids of varying diameters were tested on the bench to determine the largest diameter coil that demonstrated significant SNR gains from cooling. A liquid N2 cryochamber was designed to cool the tune and match circuit, coaxial cable, and connectors, while leaving the RF coil in ambient air. As the cryochamber was filled with liquid N2, quality factors were measured on the bench while monitoring the coil's surface temperature. Improvements of SNR on images of ionic solutions were demonstrated via cooling the tune and match circuit in the magnet bore. Results: At 650 MHz, loops and solenoids < 3 mm in diameter showed significant improvements in quality factor on the bench. The resistance of the variable capacitors and the coaxial cable were measured to be 45% and 32% of room temperature values near the Larmor frequency. Images obtained with a 2 turn, 3 mm diameter loop with the matching circuit at room temperature and then cooled with liquid nitrogen demonstrated SNR improvements of a factor of 2. Conclusions: By cooling the tune and match circuit and leaving the surface loop in ambient air, SNR was improved by a factor of 2. The results are significant because it allows for more space to insulate the sample from the extreme temperatures used in imaging cryoprobes.

Abstract PEAK pseudokinases regulate cell migration, invasion and proliferation by recruiting key signaling proteins to the cytoskeleton. Despite lacking catalytic activity, alteration in their expression level is associated with several aggressive cancers. Here, we elucidate the molecular details of key PEAK signaling interactions with the adapter proteins CrkII and Grb2 and the scaffold protein 14-3-3. Our findings rationalize why the dimerization of PEAK proteins has a crucial function in signal transduction and provide biophysical and structural data to unravel binding specificity within the PEAK interactome. We identify a conserved high affinity 14-3-3 motif on PEAK3 and demonstrate its role as a molecular switch to regulate CrkII binding and signaling via Grb2. Together, our studies provide a detailed structural snapshot of PEAK interaction networks and further elucidate how PEAK proteins, especially PEAK3, act as dynamic scaffolds that exploit adapter proteins to control signal transduction in cell growth/motility and cancer.

Circulating cell-free nucleic acids (ccfNAs) of plasma are a remarkable source of genetic, epigenetic and transcriptomic materials originating from different cells, tissues and organs of an individual. They have been increasingly studied over the past decade as they can carry several important pieces of information about the health status of an individual, which makes them biomarkers of choice for non-invasive diagnosis of numerous diseases and health conditions. However, few studies have investigated variations of plasma ccfNAs in healthy subjects, particularly in relation to aging, healthy aging and longevity, despite the great variability of these biological processes among individuals. Here, we reviewed several studies that focused on the analysis of circulating cell-free DNA (ccfDNA) and microRNAs (ccfmiRNAs) during aging and in the elderly, including some on exceptionally long-lived individuals, i.e., centenarians. After a brief overview of the types, origins and functions of plasma ccfNAs, we described the variations of both ccfDNA and ccfmiRNAs during aging as well as the identification of several potential ccfDNA-based and ccfmiRNA-based biomarkers of aging, healthy aging and/or longevity. We finally highlighted some prospects offered by ccfNAs for the understanding and improvement of healthy aging and longevity.

Abstract Knowledge gaps regarding the potential role of pesticides in the loss of agricultural biodiversity worldwide and mixture-related issues hamper proper risk assessment of unintentional impacts of pesticides, rendering essential the monitoring of wildlife exposure to these compounds. Free-ranging mammal exposure to legacy (Banned and Restricted: BRPs) and currently used (CUPs) pesticides was investigated, testing the hypotheses of: (1) a background bioaccumulation for BRPs whereas a “hot-spot” pattern for CUPs, (2) different contamination profiles between carnivores and granivores/omnivores, and (3) the role of non-treated areas as refuges towards exposure to CUPs. Apodemus mice (omnivore) and Crocidura shrews (insectivore) were sampled over two French agricultural landscapes (n = 93). The concentrations of 140 parent chemicals and metabolites were screened in hair samples. A total of 112 compounds were detected, showing small mammal exposure to fungicides, herbicides and insecticides with 32 to 65 residues detected per individual (13–26 BRPs and 18–41 CUPs). Detection frequencies exceeded 75% of individuals for 13 BRPs and 25 CUPs. Concentrations above 10 ng/g were quantified for 7 BRPs and 29 CUPs (in 46% and 72% of individuals, respectively), and above 100 ng/g for 10 CUPs (in 22% of individuals). Contamination (number of compounds or concentrations) was overall higher in shrews than rodents and higher in animals captured in hedgerows and cereal crops than in grasslands, but did not differ significantly between conventional and organic farming. A general, ubiquitous contamination by legacy and current pesticides was shown, raising issues about exposure pathways and impacts on ecosystems. We propose a concept referred to as “biowidening”, depicting an increase of compound diversity at higher trophic levels. This work suggests that wildlife exposure to pesticide mixtures is a rule rather than an exception, highlighting the need for consideration of the exposome concept and questioning appropriateness of current risk assessment and mitigation processes.

Post-transplant lymphoproliferative disorder (PTLD) is a potentially fatal complication following kidney transplantation, and there is a critical and unmet need for PTLD treatments associated with more pronounced and durable responses. To date, reports on the use of CD19-targeted chimeric antigen receptor (CAR) T (CAR-T) cells in patients after solid organ transplant (SOT) have been anecdotal, clinical presentations and outcomes have been heterogenous, and a longitudinal analysis of CAR-T cell expansion and persistence in PTLD patients has not been reported. Our report describes a patient with a history of renal transplant who received CD19-directed CAR-T cell therapy for the treatment of refractory PTLD, diffuse large B cell lymphoma (DLBCL)-type. We show that even with the background of prolonged immunosuppression for SOT, it is possible to generate autologous CAR-T products capable of expansion and persistence in vivo, without evidence of excess T-cell exhaustion. Our data indicate that CAR-T cells generated from a SOT recipient with PTLD can yield deep remissions without increased toxicity or renal allograft dysfunction. Future clinical studies should build on these findings to investigate CAR-T therapy, including longitudinal monitoring of CAR-T phenotype and function, for PTLD in SOT recipients.

Disparities in physical activity (PA) exist in rural regions and prior research suggests environmental features and community resources likely contribute. It is important to identify the opportunities and barriers that influence activity to appropriately inform PA interventions in such areas. Thus, we assessed the built environment, programs and policies related to PA opportunity in six rural Alabama counties that were purposively selected to inform a PA randomized controlled trial. Assessments were conducted August 2020-May 2021 using the Rural Active Living Assessment. Town characteristics and recreational amenities were captured using the Town Wide Assessment (TWA). PA programs and policies were examined with the Program and Policy Assessment. Walkability was evaluated using the Street Segment Assessment (SSA). Using the scoring system (0–100), the overall TWA score was 49.67 (range: 22–73), indicating few schools within walking distance (≤5 miles of the town’s center) and town-wide amenities (e.g., trails, water/recreational activities) for PA. The Program and Policy Assessment showed a paucity of programming and guidelines to support activity (overall average score of 24.67, [range: 22–73]). Only one county had a policy requiring walkways/bikeways in new public infrastructure projects. During assessment of 96 street segments, few pedestrian-friendly safety features [sidewalks (32%), crosswalks (19%), crossing signals (2%), and public lighting (21%)] were observed. Limited opportunities for PA (parks and playgrounds) were identified. Barriers such as few policies and safety features (crossing signals, speed bumps) were indicated as factors that should be addressed when developing PA interventions and informing future policy efforts.

Abstract Background Speciation genes contribute disproportionately to species divergence, but few examples exist, especially in vertebrates. Here we test whether Zan, which encodes the sperm acrosomal protein zonadhesin that mediates species-specific adhesion to the egg’s zona pellucida, is a speciation gene in placental mammals. Results Genomic ontogeny reveals that Zan arose by repurposing of a stem vertebrate gene that was lost in multiple lineages but retained in Eutheria on acquiring a function in egg recognition. A 112-species Zan sequence phylogeny, representing 17 of 19 placental Orders, resolves all species into monophyletic groups corresponding to recognized Orders and Suborders, with

Abstract Lymphoblastoid cell lines (LCLs) derive from blood infected in vitro by Epstein–Barr virus and were used in several genetic, transcriptomic and epigenomic studies. Although few changes were shown between LCL and blood genotypes (SNPs) validating their use in genetics, more were highlighted for other genomic features and/or in their transcriptome and epigenome. This could render them less appropriate for these studies, notably when blood DNA could still be available. Here we developed a simple, high-throughput and cost-effective real-time PCR approach allowing to distinguish blood from LCL DNA samples based on the presence of EBV relative load and rearranged T-cell receptors γ and β. Our approach was able to achieve 98.5% sensitivity and 100% specificity on DNA of known origin (458 blood and 316 LCL DNA). It was further applied to 1957 DNA samples from the CEPH Aging cohort comprising DNA of uncertain origin, identifying 784 blood and 1016 LCL DNA. A subset of these DNA was further analyzed with an epigenetic clock indicating that DNA extracted from blood should be preferred to LCL for DNA methylation-based age prediction analysis. Our approach could thereby be a powerful tool to ascertain the origin of DNA in old collections prior to (epi)genomic studies.

Introduction/Objectives: Despite the introduction of lung cancer screening using low dose computed tomography (LDCT), overall screening rates in the U.S. remain low, with certain populations including Black and rural communities experiencing additional disparities. The primary objective of this study was to understand the facilitators of lung cancer screening initiation and retention in Alabama reported by people at risk from mostly rural, mostly Black populations in Jefferson County—including the urban center of Birmingham—and 6 rural counties: Choctaw, Dallas, Greene, Hale, Marengo, and Sumter. Methods: We conducted semi-structured telephone interviews with 58 people who underwent lung cancer screening between December 2019 and January 2022. Participant responses were recorded by the interviewer for analysis. Open-ended responses were coded to identify emergent themes. Results: The most reported influences to initiate screening were information or suggestion from a Community Health Advisor (CHAs) or the supervising county coordinator, suggestion from a friend, or consideration of a personal history of smoking. Most participants reported multiple influences. Physicians were not very influential in decisions to initiate screening, but they were extremely influential in participants’ intent to continue screening, both positively and negatively. Knowing the recommended timeline for their annual scans was also a predictor of intention to continue screening. Participants screened during the COVID-19 state of emergency expressed less certainty about dates of next scans and more ambivalence about intention to continue screening. Conclusions: This study shows the benefit of using multiple methods to support increased awareness of and interest in lung cancer screening, particularly when educational messaging through CHAs is used. Clear guideline-based messages from healthcare providers about recommended screening is important for increasing retention. COVID-19 related implementation challenges impacted screening recruitment and retention. Future research is warranted to further explore use of CHAs in lung cancer screening.

The culture of embryos in the non-essential amino acid L-proline (Pro) or its analogues pipecolic acid (PA) and L-4-thiazolidine carboxylic acid (L4T) improves embryo development, increasing the percentage that develop to the blastocyst stage and hatch. Staining of 2-cell and 4-cell embryos with tetramethylrhodamine methyl ester and 2′,7′-dichlorofluorescein diacetate showed that the culture of embryos in the presence of Pro, or either of these analogues, reduced mitochondrial activity and reactive oxygen species (ROS), respectively, indicating potential mechanisms by which embryo development is improved. Inhibition of the Pro metabolism enzyme, proline oxidase, by tetrahydro-2-furoic-acid prevented these reductions and concomitantly prevented the improved development. The ways in which Pro, PA and L4T reduce mitochondrial activity and ROS appear to differ, despite their structural similarity. Specifically, the results are consistent with Pro reducing ROS by reducing mitochondrial activity while PA and L4T may be acting as ROS scavengers. All three may work to reduce ROS by contributing to the GSH pool. Overall, our results indicate that reduction in mitochondrial activity and oxidative stress are potential mechanisms by which Pro and its analogues act to improve pre-implantation embryo development.

The extraordinary morphological diversity among extant mammals poses a challenge for studies of speciation, adaptation, molecular evolution, and reproductive isolation. Despite the recent wealth of molecular studies on mammalian phylogenetics, uncertainties remain surrounding both ancestral and more recent divergence events that have proven difficult to resolve. Multi-gene datasets, especially including genes that are highly divergent, often provide increased support for higher-level affinities within Mammalia; however, such analyses require vast amounts of genomic sequence data and at times, intensive, high-performance computational effort. Furthermore, despite the large-scale efforts dedicated to comprehensive, multi-gene phylogenetic analyses using a combination of mitochondrial, nuclear, and other sequences (e.g., tRNA, ultra-conserved elements, and transposable elements), many relationships across Mammalia remain highly controversial. To offer another approach and provide a phylogenetic solution to this longstanding issue, here we present a phylogenetic tool based on a single reproductive molecular marker, zonadhesin (gene: Zan), one of two known mammalian speciation genes, which encodes the rapidly evolving sperm protein zonadhesin that mediates species-specific adhesion to the egg and thereby promotes reproductive isolation among placental mammals (Eutheria). Topological comparison of Zan Maximum Likelihood phylogenies to a nearly complete mammalian supertree confirmed Zan’s striking phylogenetic utility and resolution at both deeper and more terminal nodes in the placental mammalian phylogeny. This single gene marker yielded an equivalent and/or superiorly supported topology in comparison to a supertree generated using DNA sequences from a supermatrix of 31 genes from 5911 species (extinct and extant). Resolution achieved with this new phylogenetic approach provides unique insights into the divergence of both early and recent mammalian radiations. Finally, and perhaps most importantly, the utility of zonadhesin as a singular molecular marker was especially useful in clades where sufficient taxon sampling is impossible to achieve, and where only a subset of members of the mammalian species tree is available. The eutherian relationships presented here provide a foundation for future studies in the reconstruction of mammalian classifications, including reproductive isolation, hybridization, and biodiversification of species.

Patel et al. report crystal structures of the kinase domain of Doublecortin-like kinase 1 (DCLK1) in complex with a potent inhibitor and its precursors. Their structural study reveals insights into the mode of action of the inhibitor DCLK1-IN-1 and sets the basis for the design of isoform-specific inhibitors of DCLK1, which plays a role in tumour growth and development.

Abstract Background Culex (Culex) tritaeniorhynchus is an important vector of Japanese encephalitis virus (JEV) affecting feral pigs, native mammals and humans. The mosquito species is widely distributed throughout Southeast Asia, Africa and Europe, and thought to be absent in Australia. Methods In February and May, 2020 the Medical Entomology unit of the Northern Territory (NT) Top End Health Service collected Cx. tritaeniorhynchus female specimens (n = 19) from the Darwin and Katherine regions. Specimens were preliminarily identified morphologically as the Vishnui subgroup in subgenus Culex. Molecular identification was performed using cytochrome c oxidase subunit 1 (COI) barcoding, including sequence percentage identity using BLAST and tree-based identification using maximum likelihood analysis in the IQ-TREE software package. Once identified using COI, specimens were reanalysed for diagnostic morphological characters to inform a new taxonomic key to related species from the NT. Results Sequence percentage analysis of COI revealed that specimens from the NT shared 99.7% nucleotide identity to a haplotype of Cx. tritaeniorhynchus from Dili, Timor-Leste. The phylogenetic analysis showed that the NT specimens formed a monophyletic clade with other Cx. tritaeniorhynchus from Southeast Asia and the Middle East. We provide COI barcodes for most NT species from the Vishnui subgroup to aid future identifications, including the first genetic sequences for Culex (Culex) crinicauda and the undescribed species Culex (Culex) sp. No. 32 of Marks. Useful diagnostic morphological characters were identified and are presented in a taxonomic key to adult females to separate Cx. tritaeniorhynchus from other members of the Vishnui subgroup from the NT. Conclusions We report the detection of Cx. tritaeniorhynchus in Australia from the Darwin and Katherine regions of the NT. The vector is likely to be already established in northern Australia, given the wide geographical spread throughout the Top End of the NT. The establishment of Cx. tritaeniorhynchus in Australia is a concern to health officials as the species is an important vector of JEV and is now the sixth species from the subgenus Culex capable of vectoring JEV in Australia. We suggest that the species must now be continuously monitored during routine mosquito surveillance programmes to determine its current geographical spread and prevent the potential transmission of exotic JEV throughout Australia. Graphical abstract

Understanding virus assembly could identify potential drug targets. Here the authors use a safe and efficient method to solve pathogenic flavivirus structures, revealing two lipid-like ligands within highly conserved pockets of the stem region of envelope protein that are important for virus maturation.

Here the authors describe a simple reverse genetics method that relies on overlapping cDNA fragments for generation of positive-strand viruses including SARS-CoV-2 and characterize them in vitro and in vivo.

Anti-SARS-CoV-2 antibodies are crucial for protection from future COVID-19 infections, limiting disease severity, and control of viral transmission. While patients with the most common type of hematologic malignancy, B cell lymphoma, often develop insufficient antibody responses to messenger RNA (mRNA) vaccines, vaccine-induced T cells would have the potential to ‘rescue’ protective immunity in patients with B cell lymphoma. Here we report the case of a patient with B cell lymphoma with profound B cell depletion after initial chemoimmunotherapy who received a total of six doses of a COVID-19 mRNA vaccine. The patient developed vaccine-induced anti-SARS-CoV-2 antibodies only after the fifth and sixth doses of the vaccine once his B cells had started to recover. Remarkably, even in the context of severe treatment-induced suppression of the humoral immune system, the patient was able to mount virus-specific CD4+ and CD8+ responses that were much stronger than what would be expected in healthy subjects after two to three doses of a COVID-19 mRNA vaccine and which were even able to target the Omicron ‘immune escape’ variant of the SARS-CoV-2 virus. These findings not only have important implications for anti-COVID-19 vaccination strategies but also for future antitumor vaccines in patients with cancer with profound treatment-induced immunosuppression.

Abstract Intragenomic rRNA variation is a critical concern for eukaryotic metabarcoding studies, due to its potentially confounding effects on species delimitation and biodiversity estimates derived from ‐Omics data. In the present study, we assessed patterns associated with 18S rRNA metabarcoding loci in marine nematodes, including characterization of intragenomic rRNA gene variants (number of variants and abundance profiles) and aspects of datasets that can obscure biological signals (e.g., amplification of nontarget DNA, ambiguous taxonomy assignments). We estimated amplicon sequence variants (ASVs) using DADA2 from an 18S rRNA metabarcoding dataset (Illumina MiSeq) generated from individual marine nematodes. Illumina data were analyzed in conjunction with nematode morphological identifications and nearly full‐length 18S reference sequences (~1,600 bp Sanger barcodes) generated for a subset of the same specimens. Our results indicated that levels of intragenomic rRNA variation appeared to vary widely across nematode taxa (irrespective of phylogenetic clades or ecological feeding groups) and that coamplification of nontarget DNA was common (relic DNA, gut contents, etc.). The DADA2 pipeline appeared to produce a biologically accurate profile of intragenomic rRNA variants in nematodes that was consistent with “Head‐Tail” patterns (of dominant vs. minor rRNA gene variants) identified in previous studies. Although intragenomic rRNA variation appears to be ubiquitous in marine nematodes, nematode identifications were highly congruent across our three methods for species delimitation (traditional morphological taxonomy, Sanger DNA barcoding, and high‐throughput metabarcoding). In spite of pervasive intragenomic variation and high copy number of rRNA genes, the most abundant ASVs in metabarcoding datasets are likely to represent true species barcodes and thus confer an accurate view of extant biodiversity. However, our findings also emphasize the importance of applying bioinformatic filtering techniques and developing well‐curated reference databases in order to better link rRNA molecules with specimen‐level data and alleviate the confounding effects of intragenomic gene variants in studies of microbial eukaryotes.

Flagellotropic phages spin down flagella to reach the bacterial surface and must withstand remarkable drag forces. Here authors show how two nested sets of chainmail stabilise the viral head and a beta-hairpin regulates the formation of the robust yet pliable tail, characteristic of siphoviruses.

Continuous exposure of tissue antigen (Ag) to the autoantigen-specific regulatory T cells (Treg) is required to maintain Treg-dependent systemic tolerance. Thus, testis autoantigens, previously considered as sequestered, may not be protected by systemic tolerance. We now document that the complete testis antigen sequestration is not valid. The haploid sperm Ag lactate dehydrogenase 3 (LDH3) is continuously exposed and not sequestered. It enters the residual body (RB) to egress from the seminiferous tubules and interact with circulating antibody (Ab). Some LDH3 also remains inside the sperm cytoplasmic droplets (CD). Treg-depletion in the DEREG mice that express diphtheria toxin receptor on the Foxp3 promoter results in spontaneous experimental autoimmune orchitis (EAO) and Ab to LDH3. Unlike the wild-type male mice, mice deficient in LDH3 (wild-type female or LDH3 NULL males) respond vigorously to LDH3 immunization. However, partial Treg depletion elevated the wild-type male LDH3 responses to the level of normal females. In contrast to LDH3, zonadhesin (ZAN) in the sperm acrosome displays properties of a sequestered Ag. However, when ZAN and other sperm Ag are exposed by vasectomy, they rapidly induce testis Ag-specific tolerance, which is terminated by partial Treg-depletion, leading to bilateral EAO and ZAN Ab response. We conclude that some testis/sperm Ag are normally exposed because of the unique testicular anatomy and physiology. The exposed Ag: 1) maintain normal Treg-dependent systemic tolerance, and 2) are pathogenic and serve as target Ag to initiate EAO. Unexpectedly, the sequestered Ags, normally non-tolerogenic, can orchestrate de novo Treg-dependent, systemic tolerance when exposed in vasectomy.

Doublecortin-like kinase 1 (DCLK1) is a functional serine/threonine (S/T)-kinase and a member of the doublecortin family of proteins which are characterized by their ability to bind to microtubules (MTs). DCLK1 is a proposed cancer driver gene, and its upregulation is associated with poor overall survival in several solid cancer types. However, how DCLK1 associates with MTs and how its kinase function contributes to pro-tumorigenic processes is poorly understood. This review builds on structural models to propose not only the specific functions of the domains but also attempts to predict the impact of individual somatic missense mutations on DCLK1 functions. Somatic missense mutations in DCLK1 are most frequently located within the N-terminal MT binding region and likely impact on the ability of DCLK1 to bind to αβ-tubulin and to polymerize and stabilize MTs. Moreover, the MT binding affinity of DCLK1 is negatively regulated by its auto-phosphorylation, and therefore mutations that affect kinase activity are predicted to indirectly alter MT dynamics. The emerging picture portrays DCLK1 as an MT-associated protein whose interactions with tubulin heterodimers and MTs are tightly controlled processes which, when disrupted, may confer pro-tumorigenic properties.

Abstract Objectives Solid organ transplant recipients (SOTR) receiving post‐transplant immunosuppression show increased COVID‐19‐related mortality. It is unclear whether an additional dose of COVID‐19 vaccines can overcome the reduced immune responsiveness against severe acute respiratory syndrome coronavirus 2 (SARS‐CoV‐2) variants. Methods We analysed humoral immune responses against SARS‐CoV‐2 and its variants in 53 SOTR receiving SARS‐CoV‐2 vaccination. Results Following the initial vaccination series, 60.3% of SOTR showed no measurable neutralisation and only 18.9% demonstrated neutralising activity of > 90%. More intensive immunosuppression, antimetabolites in particular, negatively impacted antiviral immunity. While absolute IgG levels were lower in SOTR than controls, antibody titres against microbial recall antigens were higher. By contrast, SOTR showed reduced vaccine‐induced IgG/IgA antibody titres against SARS‐CoV‐2 and its delta variants and fewer linear B‐cell epitopes, indicating reduced B‐cell diversity. Importantly, a third vaccine dose led to an increase in anti‐SARS‐CoV‐2 antibody titres and neutralising activity across alpha, beta and delta variants and to the induction of anti‐SARS‐CoV‐2 CD4+ T cells in a subgroup of patients analysed. By contrast, we observed significantly lower antibody titres after the third dose with the omicron variant compared to the ancestral SARS‐CoV‐2 and the improvement in neutralising activity was much less pronounced than for all the other variants. Conclusion Only a small subgroup of solid organ transplant recipients is able to generate functional antibodies after an initial vaccine series; however, an additional vaccine dose resulted in dramatically improved antibody responses against all SARS‐CoV‐2 variants except omicron where antibody responses and neutralising activity remained suboptimal.

Individual identification is required for long-term investigations that examine population-level changes in survival or abundance, and mechanisms associated with these changes in wild populations. Such identification generally requires the application of a unique mark, or the documentation of characteristics distinctive to each individual animal. To minimize impacts to often declining populations, scientific and ethical concerns encourage marking strategies that minimize handling time (i.e., stress) for captured individuals. We examined the relative efficacy of passive integrated transponder (PIT)-tagging and photo-identification to identify individual Boreal toads (Anaxyrus boreas boreas) in field and indoor settings. We evaluated whether initial handling time was influenced by identification method (PIT-tag or photo-identification) or environment (field or indoor) and assessed the applicability of each method in long-term monitoring programs. Initial handling time was higher for PIT-tagging than photo-identification and higher in the field than in an indoor environment; however, handling time for previously PIT-tagged individuals was greatly reduced such that photo-identification led to > 5.5 times more handling time than PIT-tagging over the course of a toad's lifetime. Investigators must determine the trade-off between initial and subsequent handling times to minimize the expected (mean) cumulative handling time for an individual over the course of a study. Cumulative handling time is a function of the study design and the species’ survival and detection probabilities. We developed a Shiny Application to allow investigators to determine the identification method that minimizes handling time for their own study system.

Rhabdomyomatous mesenchymal hamartoma (RMH) is an interesting, often unexpected, benign cutaneous lesion that can present in children. Surgical excision is the intervention of choice. This case report describes a case of RMH and is unique in that it is the first instance of a recurrence. A review of the literature is included.

Environmental exposure of humans to pollutants has been associated with adverse health outcomes, but few studies have evaluated the multiple exposure of general populations. In the present study, we used hair analysis to assess the exposure of a general adult population (n = 497) in Luxembourg to 34 persistent and 33 non-persistent organic pollutants from 11 chemical families, such as polychlorinated biphenyls (PCBs), polybrominated diphenyl ethers (PBDEs), organochlorine pesticides (OCPs), organophosphate pesticides (OPPs), and pyrethroid pesticides (PYRs). We detected 24 persistent and 29 non-persistent organic pollutants, with 17 pollutants being detected in more than 50% of hair samples. The median concentrations for pollutants detected in 100% of the samples were 0.37 pg/mg for lindane (γ-HCH), 0.15 pg/mg for hexachlorobenzene (HCB), 14.1 pg/mg for p-nitrophenyl (PNP), and 0.10 pg/mg for trifluralin. Each participant in this study had detectable levels of at least 10 of the pollutants analyzed, and 50% of participants had 19 or more, suggesting the simultaneous exposure to numerous different pollutants among our study population. Significant correlations were often found between pollutants from the same family, with the strongest being found between two PYR metabolites, trans/cis-3-(2,2-dichlorovinyl)-2,2-dimethylcyclopropane-carboxylic acid (Cl2CA) and 3-phenoxybenzoic acid (3-PBA). Results from multiple linear regression analyses showed that sex, age and/or body mass index were significantly associated with 15 out of the 17 frequently detected pollutants. The current study is the first nationwide biomonitoring investigating organic contaminants in the Luxembourg population using hair analysis.

Background/Aim: The quantitative assessment of human exposure to contaminants such as pesticides is a crucial step in the characterization of exposure-associated risk. For this purpose, biomonitoring is often privileged as it presents the advantage of integrating all the possible sources and routes of exposure and of being representative of the internal dose resulting from exposure. Although biological fluids such as urine and blood have been used to date for this purpose, increasing interest has also been observed over the past decade for hair analysis. The present work aimed at comparing the information obtained from the analysis of urine versus hair regarding exposure to pesticides in a pilot cohort of pregnant women. Methods: In ninety-three pregnant women included in the pilot of the French cohort ELFE, one urine and one hair sample were collected simultaneously from each subject at the maternity. Samples were analyzed using GC–MS/MS analytical methods allowing for the detection of both parent pesticides and metabolites, and designed to be as similar as possible between urine and hair for reliable inter-matrix comparison. Fifty-two biomarkers of exposure were targeted, including parents and metabolites of organochlorines, organophosphates, pyrethroids, carbamates, phenylpyrazoles and other pesticides. Results: The number of different compounds detected ranged from 16 to 27 (median = 22) in hair, and from 3 to 22 (median = 12) in urine. In hair, 24 compounds were found in > 40% of the individuals, whereas only 12 compounds presented the same frequency of detection in urine. Among the chemicals detected in > 80% of both hair and urine samples, only one (pentachlorophenol) showed a signification correlation between hair and urine concentrations. Conclusions: The present results highlight the multiple exposure of the pregnant women included in this cohort and suggest that hair provides more comprehensive information on pesticide exposure than urine analysis. This study thus supports the relevance of hair analysis in future epidemiological studies investigating association between exposure and adverse health effects.

The shallow Pacific Arctic shelf has historically acted as an effective carbon sink, characterized by tight benthic pelagic coupling. However, the strength of the biological carbon pump in the Arctic has been predicted to weaken with climate change due to increased duration of the open-water period for primary production, enhanced nutrient limitation, and increased pelagic heterotrophy. In order to gain insights into how the biological carbon pump is functioning under the recent conditions of extreme warming and sea ice loss on the Pacific Arctic shelf, we measured sinking particulate organic carbon (POC) fluxes with drifting and moored sediment traps, as well as rates of primary production and particle-associated microbial respiration during June 2018. In Bering Shelf/Anadyr Water masses, sinking POC fluxes ranged from 0.8 to 2.3 g C m–2 day–1, making them among the highest fluxes ever documented in the global oceans. Furthermore, high export ratios averaging 82% and low rates of particle-associated microbial respiration also indicated negligible recycling of sinking POC in the water column. These results highlight the extraordinary strength of the biological carbon pump on the Pacific Arctic shelf during an unusually warm and low-sea ice year. While additional measurements and time are needed to confirm the ultimate trajectory of these fluxes in response to ongoing climate change, these results do not support the prevailing hypothesis that the strength of the biological carbon pump in the Pacific Arctic will weaken under these conditions.

The influence of partisan identification infiltrates all aspects of a democracy. This study employs an innovative design to explore the presidential debate-viewing experience among young citizens. Data were collected from across the United States for all three 2016 presidential debates between Hillary Clinton and Donald Trump using pretest/posttest surveys and debate viewers’ Twitter posts. Examining Twitter expression as a form of political elaboration, the study employs a social identity theoretical perspective to better understand the process through which viewers form political attitudes. Applying the theory of identity-motivated elaboration (TIME) to presidential debates, the current research illuminates how partisan social identification changes the way viewers think about political issues and, resultantly, evaluate candidates and form political opinions. A strong partisan social identification results in greater identity-consistent elaboration and Twitter expression throughout one’s presidential debate viewing, which results in more biased candidate image evaluations and, subsequently, stronger preference for the in-party candidate.

Background:. As the national opioid epidemic escalates, rates of the Hepatitis C (HCV) infection have similarly risen. Surgeons exposed intraoperatively secondary to sharp instrument or needle-sticks are affected both socioeconomically and physically. Current treatment strategies involve antiretroviral agents that have not been universally available. This study evaluates the current risk of surgeon exposure to HCV. Methods:. CDC data regarding state-by-state HCV diagnosis reporting were combined with the plastic surgery workforce data from the ASPS. Proxy variables for exposure risk to HCV were generated for each state and compared. Results:. West Virginia plastic surgeons were found to have a significantly elevated risk of exposure (60.0 versus 18.7, P < 0.0001). Their exposure risk is a notable outlier compared with the rest of the country (Risk >3 × IQR + 75th percentile). Similarly, states within the Ohio Valley were found to be at increased risk (34.8 versus 16.0, P = 0.05). States most heavily burdened by the opioid crisis were found to be at an increased risk for HCV exposure (40.8 versus 13.6, P = 0.0003). Conclusions:. Plastic surgeons employed in states within the Ohio Valley were found to be at an increased risk of exposure to HCV. Plastic surgeons operating in states severely impacted by the opioid crisis were found to be at an increased risk of exposure. These findings underscore the importance of reducing the risk in the operating room and the need for better data collection to better understand this association and mitigate the risk to the operating surgeon.

The high use of pesticides worldwide and the constant exposure of humans to these toxic-by-design chemicals have drawn the attention on the possible consequences on human health. However, information on the exposure of the general population to pesticides remain very limited in most countries, especially in urban areas. In the present work, hair analysis was conducted to investigate the exposure of 204 urban women living in two Chinese cities (Baoding and Dalian) to 110 pesticides and 30 metabolites of the following families: organochlorines, organophosphates, carbamates, pyrethroids, neonicotinoids, phenylpyrazoles, acid herbicides, urea herbicides and azoles. Results showed that 71 pesticides and 23 metabolites were found in the hair samples, with concentrations ranging up to 1070 pg/mg in hair. In each hair sample, the number of detected chemicals ranged from 25 to 50, demonstrating the cumulative exposure to pesticides among Chinese women in the studied regions. The concentrations of 38 chemicals (e.g., p-nitrophenol, diethyldithiophosphate, λ-cyhalothrin, permethrin, carbendazim and tebuconazole) were significantly different between women in Baoding and Dalian, indicating the regional differences in exposure to pesticide. Using a multiple regression analysis, we found that concentrations of a few dominant pesticides were associated with age, body mass index (BMI), cooking frequency and regions. These results can provide baseline information on exposure of female adult Chinese population to multiple pesticides and support future studies focused on the health effects associated with pesticide exposure. Keywords: Multiclass pesticides, Metabolites, Human biomonitoring, Hair analysis, Urban residents, Environmental pollution

Monitoring aquatic biodiversity through DNA extracted from environmental samples (eDNA) combined with high-throughput sequencing, commonly referred to as eDNA metabarcoding, is increasing in popularity within the scientific community. However, sampling strategies, laboratory protocols and analytical pipelines can influence the results of eDNA metabarcoding surveys. While the impact of laboratory protocols and analytical pipelines have been extensively studied, the importance of sampling strategies on eDNA metabarcoding surveys has not received the same attention. To avoid underestimating local biodiversity, adequate sampling strategies (i.e. sampling intensity and spatial sampling replication) need to be implemented. This study evaluated the impact of sampling strategies along an altitudinal and biodiversity gradient in the upper section of the Murrumbidgee River (Murray-Darling Basin, Australia). An eDNA metabarcoding survey was used to determine the local fish biodiversity and evaluate the influence of sampling intensity and spatial sampling replication on the biodiversity estimates. The results show that optimal eDNA sampling strategies varied between sites and indicate that river morphology, species richness and species abundance affect the optimal sampling intensity and spatial sampling replication needed to accurately assess the fish biodiversity. While the generality of the patterns will need to be confirmed through future studies, these findings provide a basis to guide future eDNA metabarcoding surveys in river systems.

Polycyclic aromatic hydrocarbons (PAH) are produced from incomplete combustion of organic matter and released as environmental contaminants from activities such as transports, wood combustion, coal-fired power plants. In numerous urban areas worldwide, the levels of PAH exposure are considered critical regarding public health issues. The possibility to detect PAH and PAH metabolites biologically incorporated in human hair was demonstrated and proposed as biomarkers of exposure. Nevertheless, the possibility to distinguish different levels of exposure between different populations is still needed to validate the relevance of hair analysis in epidemiological studies. In this work, hair samples were collected from 204 women from two cities in China based on one year Air Quality Index history from governmental data (Baoding as polluted city and Dalian less polluted city). 8 out of the 15 parent PAH and 7 out of the 56 metabolites analyzed in this study were detected in all the samples. The highest concentrations in hair were observed for phenanthrene (4.2 to 889 pg/mg) > fluoranthene (1.05 to 204 pg/mg) > pyrene (3.2 to 124 pg/mg) for parent PAH, and for 9-OH-fluorene (0.04 to 1.78 pg/mg) > 2-OH-naphthalene (0.68 to 811 pg/mg) > 1-OH-anthracene (0.24 to 10.9 pg/mg) for metabolites. 14 parent PAH and 15 metabolites presented a significantly higher concentration in the hair samples collected from Baoding, as compared to Dalian. The median concentration of parent PAH was from 1.5 to 2.8 times higher in the hair of the subjects from Baoding than in subjects from Dalian and that of PAH metabolites was from 1 to 2.3 times higher. The study of inter-chemical associations revealed similarities and differences between the two areas, suggesting common and different sources of exposure depending on PAH respectively. The results confirmed the relevance of hair analysis to identify qualitative and quantitative differences in PAH exposure between populations from different areas. This study is the first one to investigate both parent PAH and their metabolites in a biological matrix. Keywords: Polycyclic aromatic hydrocarbons, Hair analysis, Cumulative exposure, Environmental exposure, Biomarkers

Healthy aging does not affect all features of language processing equally. In this study, we investigated the effects of aging on different processes involved in fluent sentence production, a complex task that requires the successful execution and coordination of multiple processes. In Experiment 1, we investigated age-related effects on the speed of syntax selection using a syntactic priming paradigm. Both young and older adults produced target sentences quicker following syntactically related primes compared to unrelated primes, indicating that syntactic facilitation effects are preserved with age. In Experiment 2, we investigated age-related effects in syntactic planning and lexical retrieval using a planning scope paradigm: participants described moving picture displays designed to elicit sentences with either initial coordinate or simple noun phrases and, on half of the trials, the second picture was previewed. Without preview, both age groups were slower to initiate sentences with larger coordinate phrases, suggesting a similar phrasal planning scope. However, age-related differences did emerge relating to the preview manipulation: while young adults displayed speed benefits of preview in both phrase conditions, older adults only displayed speed preview benefits within the initial phrase (coordinate condition). Moreover, preview outside the initial phrase (simple condition) caused older adults to become significantly more error-prone. Thus, while syntactic planning scope appears unaffected by aging, older adults do appear to encounter problems with managing the activation and integration of lexical items into syntactic structures. Taken together, our findings indicate that healthy aging disrupts the lexical, but not the syntactic, processes involved in sentence production.

Children represent one of the most vulnerable parts of the population regarding the effects of pollutants exposure on health. In this study, hair samples were collected between October 2013 and August 2015 from 142 French children originating from different geographical areas (urban and rural) and analysed with a GC/MS-MS method, allowing for the detection of 55 biomarkers for pesticides and metabolites both persistent and non-persistent from different families, including: organochlorines, organophosphates, pyrethroids, azoles, dinitroanilines, oxadiazines, phenylpyrazoles and carboxamidas; 4 polychlorobiphenyls (PCBs) and 5 polybromodiphenylethers (PBDEs). The number of compounds detected in each sample ranged from 9 up to 37 (21 on average), which clearly highlighted the cumulative exposure of the children. The results also showed a wide range of concentration of the pollutants in hair (often more than 100 times higher in the most exposed child compared to the less exposed), suggesting significant disparities in the exposure level, even in children living in the same area. In addition to the detection of currently used chemicals, the presence of persistent organic pollutants (POPs) in children also suggests that the French population is still exposed to POPs nowadays. PCP, DEP, PNP, 3Me4NP, trans-Cl2CA, 3PBA, fipronil and fipronil sulfone, presented statistically significant higher concentration in the hair of boys compared to girls. PCP, PNP and 3Me4NP presented statistically significant higher concentration in younger children. Finally, this study also suggests that local environmental contamination would not be the main source of exposure, and that individual specificities (habits, diet…) would be the main contributors to the exposure to the pollutants analysed here. The present study strongly supports the relevance of hair for the biomonitoring of exposure and provides the first values of organic pollutant concentration in the hair of French children. Keywords: Organic pollutants, Children, Hair analysis, Exposure

Patients after autologous (autoSCT) and allogeneic stem cell transplantation (alloSCT) are at an increased risk of COVID-19-related morbidity and mortality, compounded by an immune system weakened by the underlying malignancy and prior treatments. Allogeneic transplantation, including stem cell and solid organ transplants, requires intensive immunosuppressive prophylaxis, which may further undermine the development of a protective vaccine-induced anti-viral immunity. Herein, we report on short- and long-term antiviral immune responses in two peri-stem cell transplant recipients and a third patient who received a COVID-19 vaccination after kidney transplantation. Our data indicate that: (1) patients post-alloSCT may be able to mount an anti-COVID-19 immune response; however, a sufficient time interval between transplant and exposure may be of critical importance; (2) alloSCT recipients with preexisting anti-SARS-CoV-2 immunity are at risk for losing protective humoral immunity following transplantation, particularly if the stem-cell donor lacks antiviral immunity, e.g., vaccine-derived immunity; and (3) some post-transplant patients are completely unable to build an immune response to a COVID-19 vaccine, perhaps based on the prophylactic suppression of T cell immunity.

Carotenoids are lipophilic pigments which have been associated with a number of health benefits, partly related to antioxidant effects. However, due to their poor solubility during digestion, carotenoid bioavailability is low and variable. In this study, we investigated the effect of frequently consumed proteins on carotenoid bioaccessibility and cellular uptake. Whey protein isolate (WPI), soy protein isolate (SPI), sodium caseinate (SC), gelatin (GEL), turkey and cod, equivalent to 0/10/25/50% of the recommended dietary allowance (RDA, approx. 60g/d), were co-digested gastro-intestinally with carotenoid-rich food matrices (tomato and carrot juice, spinach), and digesta further studied in Caco-2 cell models. Lipid digestion, surface tension and microscopic visualization were also carried out. Co-digested proteins positively influenced the micellization of carotenes (up to 3-fold, depending on type and concentration), especially in the presence of SPI (p < 0.001). An increased cellular uptake was observed for xanthophylls/carotenes (up to 12/33%, p < 0.001), which was stronger for matrices with an initially poor carotenoid micellization (i.e., tomato juice, p < 0.001), similar to what was encountered for bioaccessibility. Turkey and cod had a weaker impact. Significant interactions between carotenoids, lipids and proteins were observed during digestion. Co-digested proteins generally improved lipid digestion in all matrices (p < 0.001), especially for carrot juice, though slight decreases were observed for GEL. Protein impact on the surface tension was limited. In conclusion, proteins generally improved both carotenoid bioaccessibility and cellular uptake, depending on the matrices and carotenoid-type (i.e., carotene vs. xanthophylls), which may be relevant under specific circumstances, such as intake of carotenoid-rich food items low in lipids.

Circoviruses are the simplest viruses known to autonomously replicate in vertebrates. Here the authors present three structures for distinct macromolecular assemblies of the capsid protein from the beak and feather disease virus that provides insights into the regulation of viral capsid assembly.

Background:. Orofacial clefting (OFC) is the most common developmental craniofacial malformation, and causal etiologies largely remain unknown. The opioid crisis has led to a large proportion of infants recovering from neonatal abstinence syndrome (NAS) due to in-utero narcotics exposure. We sought to characterize the prevalence of OFC in infants with NAS. Methods:. This cohort study analyzed live births at our institution from 2013 to 2017 to identify any association between OFC and NAS. Results:. Prevalence of OFC was 6.79 and 1.63 (per 1,000 live births) in the NAS and general population, respectively. Odds ratios for NAS patients having developed OFC, isolated cleft palate, isolated cleft lip, and combined cleft lip and palate compared with the general population were found to be 4.18 (P = 0.001), 5.92 (P = 0.001), 3.79 (P = 0.05), and 2.94 (P = 0.35), respectively. Analyses performed comparing the NAS and general populations to control for potential confounding variables influencing the NAS population yielded no significant differences with exception of in-utero exposure to physician prescribed opioids. Conclusions:. Prevalence of OFC in infants with NAS was higher than the general live birth population. Isolated cleft palate and isolated cleft lip, specifically, were significantly more prevalent in NAS patients compared with the general population and were associated with in-utero opioid exposure.

Background: Considerable resources have been invested in both low- and high-fidelity simulators in surgical training. The purpose of this study was to investigate if the Fundamentals of Laparoscopic Surgery (FLS, low-fidelity box trainer) and LapVR (high-fidelity virtual reality) training systems correlate with operative performance on the Global Operative Assessment of Laparoscopic Skills (GOALS) global rating scale using a porcine cholecystectomy model in a novice surgical group with minimal laparoscopic experience. Methods: Fourteen postgraduate year 1 surgical residents with minimal laparoscopic experience performed tasks from the FLS program and the LapVR simulator as well as a live porcine laparoscopic cholecystectomy. Performance was evaluated using standardized FLS metrics, automatic computer evaluations, and a validated global rating scale. Results: Overall, FLS score did not show an association with GOALS global rating scale score on the porcine cholecystectomy. None of the five LapVR task scores were significantly associated with GOALS score on the porcine cholecystectomy. Conclusions: Neither the low-fidelity box trainer or the high-fidelity virtual simulator demonstrated significant correlation with GOALS operative scores. These findings offer caution against the use of these modalities for brief assessments of novice surgical trainees, especially for predictive or selection purposes.

Sperm competence in animal fertilization requires the collective activities of numerous sperm-specific proteins that are typically alloimmunogenic in females. Consequently, sperm membrane alloantigens are potential targets for contraceptives that act by blocking the proteins' functions in gamete interactions. Here we used a targeted proteomics approach to identify the major alloantigens in swine sperm membranes and lipid rafts, and thereby systematically defined the repertoire of these sperm-specific proteins in a single species. Gilts with high alloantibody reactivity to proteins in sperm membranes or lipid rafts produced fewer offspring (73% decrease) than adjuvant-only or nonimmune control animals. Alloantisera recognized more than 20 potentially unique sperm membrane proteins and five sperm lipid raft proteins resolved on two-dimensional immunoblots with or without prior enrichment by anion exchange chromatography. Dominant sperm membrane alloantigens identified by mass spectrometry included the ADAMs fertilin α, fertilin ß, and cyritestin. Less abundant alloantigens included ATP synthase F1 β subunit, myo-inositol monophosphatase-1, and zymogen granule membrane glycoprotein-2. Immunodominant sperm lipid raft alloantigens included SAMP14, lymphocyte antigen 6K, and the epididymal sperm protein E12. Of the fifteen unique membrane alloantigens identified, eleven were known sperm-specific proteins with uncertain functions in fertilization, and four were not previously suspected to exist as sperm-specific isoforms. De novo sequences of tryptic peptides from sperm membrane alloantigen "M6" displayed no evident homology to known proteins, so is a newly discovered sperm-specific gene product in swine. We conclude that alloimmunizing gilts with sperm membranes or lipid rafts evokes formation of antibodies to a relatively small number of dominant alloantigens that include known and novel sperm-specific proteins with possible functions in fertilization and potential utility as targets for immunocontraception.

The nutrition environment, including food store type, may influence dietary choices, which in turn can affect risk of obesity and related chronic diseases such as CHD, diabetes and cancer. The objective of the present study was to elucidate the extent to which healthy foods are available and affordable in various rural food outlets. A subset of the nutrition environment was assessed using the Nutrition Environment Measures Survey in Stores (NEMS-S). The NEMS-S instrument assessed the availability and price of healthy foods (e.g. low-fat/non-fat milk, lean meats and reduced-fat dinner entrées) compared with less healthy counterparts (e.g. whole milk, non-lean meats and regular dinner entrées). The NEMS-S also assessed the quality of fresh fruits and vegetables. Availability, prices and quality of healthy foods were compared between grocery stores (n 24) and convenience stores (n 67) in nine rural counties in Alabama. Mean availability subscale score (possible range 0 to 30; higher score indicates a greater number of healthier foods were available) for grocery stores was 22·6 (sd 8·1), compared with 6·6 (sd 5·2) in convenience stores (P < 0·0001); and mean price subscale score (possible range −9 to 18; higher score indicates that healthier options were less expensive than the less healthy options) for grocery stores was 2·4 (sd 2·7), compared with 0·7 (sd 1·2) in convenience stores (P = 0·0080). Mean total NEMS-S score (possible range −9 to 54) in grocery stores was 29·8 (sd 10·9) compared with 7·3 (sd 7·1) in convenience stores (P < 0·0001). Both grocery and convenience stores could be strategic points of intervention to improve the nutrition environment in the counties that were surveyed.