Search

A vast literature analyzes how Bengali identities developed in colonial India. This article steps away from both celebratory approaches and a focus on the colonial period. Instead, it explores how non-Bengalis increasingly challenged Bengali superiority in more recent times. As the colonial incarnation of a genteel Bengaliness lost its bearings and split into competing territorial manifestations in East Pakistan (later Bangladesh) and India, it encountered rising hostility and developed both assertive and timid configurations. This article offers an exploratory overview of how various groups of non-Bengalis have been rebuffing Bengali dominance by means of cultural distancing, graphic resistance, the ideology of indigeneity, insurgency, and the legal and military force of postcolonial states.

As the aging population increases, it is critical to find ways to sustain older adults' health and well-being. Mindfulness-Based Stress Reduction (MBSR) may be one approach, but its effects are difficult to discern because few studies have conducted randomized controlled trials with an active control group and blinded examiners. We begin to address these gaps with a pilot study examining the feasibility of conducting an MBSR intervention with an active control condition in healthy older adults.Participants were randomly assigned to one of two classes, MBSR or Brain Health education. Classes were matched for time, format, and instructor. The study examined acceptability, practicality, implementation, and preliminary efficacy using a range of participant questionnaires, instructor ratings, cognitive measures assessed by blinded examiners, and attendance.Both MBSR and the Brain Health class evidenced high rates of recruitment, participant satisfaction, and retention. Implementation procedures were successful, and preliminary results revealed similar levels of efficacy across both classes.This study demonstrates the feasibility of an MBSR intervention in healthy older adults.MBSR, with its focus on improving stress and self-awareness, has the potential to be an approach that can improve aging adults' health and coping skills.

Background: The efficacy and safety of prophylactic full-dose anticoagulation and antiplatelet therapy in critically ill COVID-19 patients remain uncertain. Methods: COVID-PACT (Prevention of Arteriovenous Thrombotic Events in Critically-ill COVID-19 Patients Trial) was a multicenter, 2×2 factorial, open-label, randomized-controlled trial with blinded end point adjudication in intensive care unit–level patients with COVID-19. Patients were randomly assigned to a strategy of full-dose anticoagulation or standard-dose prophylactic anticoagulation. Absent an indication for antiplatelet therapy, patients were additionally randomly assigned to either clopidogrel or no antiplatelet therapy. The primary efficacy outcome was the hierarchical composite of death attributable to venous or arterial thrombosis, pulmonary embolism, clinically evident deep venous thrombosis, type 1 myocardial infarction, ischemic stroke, systemic embolic event or acute limb ischemia, or clinically silent deep venous thrombosis, through hospital discharge or 28 days. The primary efficacy analyses included an unmatched win ratio and time-to-first event analysis while patients were on treatment. The primary safety outcome was fatal or life-threatening bleeding. The secondary safety outcome was moderate to severe bleeding. Recruitment was stopped early in March 2022 (≈50% planned recruitment) because of waning intensive care unit–level COVID-19 rates. Results: At 34 centers in the United States, 390 patients were randomly assigned between anticoagulation strategies and 292 between antiplatelet strategies (382 and 290 in the on-treatment analyses). At randomization, 99% of patients required advanced respiratory therapy, including 15% requiring invasive mechanical ventilation; 40% required invasive ventilation during hospitalization. Comparing anticoagulation strategies, a greater proportion of wins occurred with full-dose anticoagulation (12.3%) versus standard-dose prophylactic anticoagulation (6.4%; win ratio, 1.95 [95% CI, 1.08–3.55]; P =0.028). Results were consistent in time-to-event analysis for the primary efficacy end point (full-dose versus standard-dose incidence 19/191 [9.9%] versus 29/191 [15.2%]; hazard ratio, 0.56 [95% CI, 0.32–0.99]; P =0.046). The primary safety end point occurred in 4 (2.1%) on full dose and in 1 (0.5%) on standard dose ( P =0.19); the secondary safety end point occurred in 15 (7.9%) versus 1 (0.5%; P =0.002). There was no difference in all-cause mortality (hazard ratio, 0.91 [95% CI, 0.56–1.48]; P =0.70). There were no differences in the primary efficacy or safety end points with clopidogrel versus no antiplatelet therapy. Conclusions: In critically ill patients with COVID-19, full-dose anticoagulation, but not clopidogrel, reduced thrombotic complications with an increase in bleeding, driven primarily by transfusions in hemodynamically stable patients, and no apparent excess in mortality. Registration: URL: https://www.clinicaltrials.gov ; Unique identifier: NCT04409834.

Museums throughout the United States house undocumented collections with variable access. With the technological developments over the past twenty years, digital documentation, particularly photogrammetry, has become more accessible and affordable which can bring these collections to a wider audience. This tool can help enable collaboration among researchers, institutions, and museum professionals, contribute to the stewardship of these collections, and overcome the physical and financial barriers that hinder open-access, allowing museum professionals to display their artifacts in new, engaging ways and make them more accessible to the public and other scholars. With the goal of using digital documentation to make archaeology and anthropology collections more accessible, five small artifacts from disparate collections in the Evansville Museum of Arts, History & Science were recorded, drawn, and modeled: a Jerash type oil lamp, a Samaritan type oil lamp, a Roman oil lamp from Meidum, Egypt, an Ottoman ceramic smoking pipe, and a ceramic sherd. This case study illustrates how small to medium size institutions can construct a photogrammetry toolkit with minimal financial and staff resources.

Groin ultrasonography (US) has been used as an adjunct to inguinal hernia diagnosis, but there is limited evidence as to whether its use affects surgical decision-making. The primary aim of this study was to examine whether groin US affects surgical management of inguinal hernia; the secondary goal was to estimate the frequency of groin US ordered before surgical consultation.We performed a retrospective chart review of 400 consecutive patients aged older than 18 years referred to 1 of 4 general surgeons in Calgary, Alberta, for inguinal hernia between January 2014 and January 2015. Bilateral groin examinations were entered as separate entries into the database. Outcomes assessed included the frequency of groin US examinations performed within 1 year before the general surgery consultation, presence of inguinal hernia on clinical examination (CE), presence of inguinal hernia on groin US, and whether the hernia proceeded to herniorrhaphy.A total of 476 groins in the 400 patients (354 [88.5%] male; mean age 53.5 yr [standard deviation 15.2 yr]) were evaluated for a hernia during the study period. Groin US was performed before general surgery consultation in 336 cases (70.6%). Overall, 364 (76.5%) of the hernias were clinically palpable; of the 364, 220 (60.4%) had preconsultation US, even in the presence of a positive CE finding. Of the 112 groins that did not have a clinically palpable hernia, 103 (92.0%) underwent preconsultation US. Of the 476 groins, 315 (66.2%) underwent inguinal hernia repair: 310 (85.2%) of the 364 with clinically palpable hernias and 5 (4.8%) of the 103 with clinically negative findings but positive groin US findings. Surgical decision-making based on CE findings occurred in 390 cases (81.9%) overall, whereas surgery based on groin US findings alone occurred in 5 of 336 cases (1.5%).Routine groin US was frequently performed before general surgery consultation, whether a hernia was detectable on clinical examination or not. Positive groin US results alone infrequently affected whether the patient proceeded to surgery. Clinical examination findings played a larger role in surgical decision-making than groin US results. Eliminating the practice of routine groin US may provide considerable health care cost savings.

Rheumatoid arthritis (RA) synovitis is dominated by monocytes/macrophages with inflammatory patterns resembling microbial stimulation. In search of triggers, we reduced the intestinal microbiome in 20 RA patients (open label study DRKS00014097) by bowel cleansing and 7-day fasting (≤250 kcal/day) and performed immune monitoring and microbiome sequencing. Patients with metabolic syndrome (n = 10) served as a non-inflammatory control group. Scores of disease activity (DAS28/SDAI) declined within a few days and were improved in 19 of 20 RA patients after breaking the fast (median ∆DAS28 = −1.23; ∆SDAI = −43%) or even achieved remission (DAS28 < 2.6/n = 6; SDAI < 3.3/n = 3). Cytometric profiling with 46 different surface markers revealed the most pronounced phenomenon in RA to be an initially increased monocyte turnover, which improved within a few days after microbiota reduction and fasting. Serum levels of IL-6 and zonulin, an indicator of mucosal barrier disruption, decreased significantly. Endogenous cortisol levels increased during fasting but were insufficient to explain the marked improvement. Sequencing of the intestinal microbiota indicated that fasting reduced potentially arthritogenic bacteria and changed the microbial composition to species with broader metabolic capabilities. More eukaryotic, predominantly fungal colonizers were observed in RA, suggesting possible involvement. This study demonstrates a direct link between the intestinal microbiota and RA-specific inflammation that could be etiologically relevant and would support targeted nutritional interventions against gut dysbiosis as a causal therapeutic approach.

The integrity and proper expression of genomes are safeguarded by DNA and RNA surveillance pathways. While many RNA surveillance factors have additional functions in the nucleus, little is known about the incidence and physiological impact of converging RNA and DNA signals. Here, using genetic screens and genome-wide analyses, we identified unforeseen SMG-1-dependent crosstalk between RNA surveillance and DNA repair in living animals. Defects in RNA processing, due to viable THO complex or PNN-1 mutations, induce a shift in DNA repair in dividing and non-dividing tissues. Loss of SMG-1, an ATM/ATR-like kinase central to RNA surveillance by nonsense-mediated decay (NMD), restores DNA repair and radio-resistance in THO-deficient animals. Mechanistically, we find SMG-1 and its downstream target SMG-2/UPF1, but not NMD per se, to suppress DNA repair by non-homologous end-joining in favour of single strand annealing. We postulate that moonlighting proteins create short-circuits in vivo, allowing aberrant RNA to redirect DNA repair.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infections are initiated by attachment of the receptor-binding domain (RBD) on the viral Spike protein to angiotensin-converting enzyme-2 (ACE2) on human host cells. This critical first step occurs in dynamic environments, where external forces act on the binding partners and avidity effects play an important role, creating an urgent need for assays that can quantitate SARS-CoV-2 interactions with ACE2 under mechanical load. Here, we introduce a tethered ligand assay that comprises the RBD and the ACE2 ectodomain joined by a flexible peptide linker. Using magnetic tweezers and atomic force spectroscopy as highly complementary single-molecule force spectroscopy techniques, we investigate the RBD:ACE2 interaction over the whole physiologically relevant force range. We combine the experimental results with steered molecular dynamics simulations and observe and assign fully consistent unbinding and unfolding events across the three techniques, enabling us to establish ACE2 unfolding as a molecular fingerprint. Measuring at forces of 2 to 5 pN, we quantify the force dependence and kinetics of the RBD:ACE2 bond in equilibrium. We show that the SARS-CoV-2 RBD:ACE2 interaction has higher mechanical stability, larger binding free energy, and a lower dissociation rate compared to SARS-CoV-1, which helps to rationalize the different infection patterns of the two viruses. By studying how free ACE2 outcompetes tethered ACE2, we show that our assay is sensitive to prevention of bond formation by external binders. We expect our results to provide a way to investigate the roles of viral mutations and blocking agents for targeted pharmaceutical intervention., This study was supported by German Research Foundation Projects 386143268 and 111166240, a Human Frontier Science ProgramCross Disciplinary Fellowship (LT000395/2020C) and European Molecular Biology Organization Non-Stipendiary long-term fellowship (ALTF 1047-2019) to L.F.M., and the Physics Department of LMU Munich. R.C.B. and P.S.F.C.G. are supported by start-up funds provided by Auburn University, and D.L. acknowledges support from the Spanish Ministry of Science, Innovation and Universities for the Spanish State Research Agency Retos Grant RTI2018- 099318-B-I00, cofunded by the European Regional Development Fund.

The doctrine of the beatifying vision of God may be derived not only from Sacred Scripture’s promise to the faithful that they shall ‘see God’ (Matt 5:8; 1 Cor 13:12; 1 John 3:2), but also (and perhaps more fundamentally) from Scripture’s testimony that God ‘is not the God of disorder but of peace’ (1 Cor 14:33). An analysis of the concepts of nature, on the part of humanity, and agency, on the part of divinity, within a broadly Aristotelian framework elucidates why early modern Reformed scholastics exposited the beatific vision as the ultimate end of humankind as a fitting implicate of the biblical testimony that God is a God of order.

Purpose: Adoptive therapy with genetically engineered T cells carrying redirected antigen specificity is a new option for the treatment of cancer. This approach is not yet available for metastatic renal cell carcinoma (RCC), due to the scarcity of therapeutically useful reagents. We analyzed tumor-infiltrating lymphocytes (TIL) from RCC to identify T-cell specificities with shared tumor-specific recognition to develop T-cell receptor (TCR)-engineered T lymphocytes for adoptive therapy of RCC.Experimental Design: We established a T-cell clone from TIL that recognized a human leukocyte antigen (HLA)-A2–restricted tumor antigen. The TCR α- and β-chain genes were isolated, modified by codon optimization and murinization, and retrovirally transduced into peripheral blood lymphocytes (PBL). A TCR-expressing indicator line (B3Z-TCR53) was established to screen for antigen prevalence in RCC, other malignancies, and normal cell counterparts.Results: TCR53-engineered PBL recapitulated the specificity of the TIL and showed tumor-specific HLA-A2–restricted effector activities (IFN-γ, tumor necrosis factor-α, interleukin-2, macrophage inflammatory protein-1β, cytotoxicity). PBL-TCR53 of healthy donors and RCC patients exhibited similar transduction efficiency, expansion, and polyfunctional profile. Using B3Z-TCR53 cells, 130 tumor and normal cells were screened and shared TCR53 peptide: MHC expression was found in >60% of RCC and 25% of tumor lines of other histology, whereas normal tissue cells were not recognized.Conclusions: To date, TCR53 is the only TCR with shared HLA-A2–restricted recognition of RCC. It fulfills the criteria for utilization in TCR gene therapy and advances T cell–based immunotherapy to patients with RCC and other malignancies expressing the TCR ligand. Clin Cancer Res; 16(8); 2333–43. ©2010 AACR.

Supplementary Data from T-Cell Receptor Gene–Modified T Cells with Shared Renal Cell Carcinoma Specificity for Adoptive T-Cell Therapy

constructs, murine HCC model, spheroid and imaging, RCC tissue pathology

Inherent intermediate- to low-affinity T-cell receptors (TCR) that develop during the natural course of immune responses may not allow sufficient activation for tumor elimination, making the majority of T cells suboptimal for adoptive T-cell therapy (ATT). TCR affinity enhancement has been implemented to provide stronger T-cell activity but carries the risk of creating undesired cross-reactivity leading to potential serious adverse effects in clinical application. We demonstrate here that engineering of low-avidity T cells recognizing a naturally processed and presented tumor-associated antigen with a chimeric PD-1:28 receptor increases effector function to levels seen with high-avidity T cells of identical specificity. Upgrading the function of low-avidity T cells without changing the TCR affinity will allow a large arsenal of low-avidity T cells previously thought to be therapeutically inefficient to be considered for ATT. PD-1:28 engineering reinstated Th1 function in tumor-infiltrating lymphocytes that had been functionally disabled in the human renal cell carcinoma environment without unleashing undesired Th2 cytokines or IL10. Involved mechanisms may be correlated to restoration of ERK and AKT signaling pathways. In mouse tumor models of ATT, PD-1:28 engineering enabled low-avidity T cells to proliferate stronger and prevented PD-L1 upregulation and Th2 polarization in the tumor milieu. Engineered T cells combined with checkpoint blockade secreted significantly more IFNγ compared with T cells without PD-1:28, suggesting a beneficial combination with checkpoint blockade therapy or other therapeutic strategies. Altogether, the supportive effects of PD-1:28 engineering on T-cell function make it an attractive tool for ATT. Cancer Res; 77(13); 3577–90. ©2017 AACR.

Agrobacterium tumefaciens, a pathogenic bacterium capable of transforming plants through horizontal gene transfer, is nowadays the preferred vector for plant genetic engineering. The vehicle for transfer is the T-strand, a single-stranded DNA molecule bound by the bacterial protein VirD2, which guides the T-DNA into the plant's nucleus where it integrates. How VirD2 is removed from T-DNA, and which mechanism acts to attach the liberated end to the plant genome is currently unknown. Here, using newly developed technology that yields hundreds of T-DNA integrations in somatic tissue of Arabidopsis thaliana, we uncover two redundant mechanisms for the genomic capture of the T-DNA 5' end. Different from capture of the 3' end of the T-DNA, which is the exclusive action of polymerase theta-mediated end joining (TMEJ), 5' attachment is accomplished either by TMEJ or by canonical non-homologous end joining (cNHEJ). We further find that TMEJ needs MRE11, whereas cNHEJ requires TDP2 to remove the 5' end-blocking protein VirD2. As a consequence, T-DNA integration is severely impaired in plants deficient for both MRE11 and TDP2 (or other cNHEJ factors). In support of MRE11 and cNHEJ specifically acting on the 5' end, we demonstrate rescue of the integration defect of double-deficient plants by using T-DNAs that are capable of forming telomeres upon 3' capture. Our study provides a mechanistic model for how Agrobacterium exploits the plant's own DNA repair machineries to transform it.

Most published autism research, and the funding that supports it, remains focused on basic and clinical science. However, the public health impact of autism drives a compelling argument for utilizing a public health approach to autism research. Fundamental to the public health perspective is a focus on health determinants to improve quality of life and to reduce the potential for adverse outcomes across the general population, including in vulnerable subgroups. While the public health research process can be conceptualized as a linear, 3-stage path consisting of discovery - testing - translation/dissemination/implementation, in this paper we propose an integrated, cyclical research framework to advance autism public health objectives in a more comprehensive manner. This involves discovery of primary, secondary and tertiary determinants of health in autism; and use of this evidence base to develop and test detection, intervention, and dissemination strategies and the means to implement them in 'real world' settings. The proposed framework serves to facilitate identification of knowledge gaps, translational barriers, and shortfalls in implementation; guides an iterative research cycle; facilitates purposeful integration of stakeholders and interdisciplinary researchers; and may yield more efficient achievement of improved health and well-being among persons on the autism spectrum at the population-level. LAY SUMMARY: Scientists need better ways to identify and address gaps in autism research, conduct research with stakeholders, and use findings to improve the lives of autistic people. We recommend an approach, based in public health science, to guide research in ways that might impact lives more quickly.

Funding: Open access funded by Max Planck Society. The authors acknowledge the Deutsche Forschungsgemeinschaft (DFG, German Research Foundation) under Germany’s Excellence Strategy-EXC-2123 Quantum Frontiers - 390837967; Core program PC2-PN23080202 and the PN-III-P2-2.1-PED-2021-0378 (contract no. 575PED/2022) granted projects, financed by the Romanian Ministry of Research, Innovation and Digitalization/UEFISCDI; and the generous allocation of computer time at the computing center of Donostia International Physics Center and at the Red Española de Supercomputación (project QHS-2021-2-0019). A.A. acknowledges support from Project No. PID2019-103910GB-I00, funded by MCIN/AEI/10.13039/501100011033/ and FEDER Una manera de hacer Europa, and Project No. IT-1527-22 funded by the Basque Government. The changes of properties and preferential interactions based on subtle energetic differences are important characteristics of organic molecules, particularly for their functionalities in biological systems. Only slightly energetically favored interactions are important for the molecular adsorption and bonding to surfaces, which define their properties for further technological applications. Here, prochiral tetracenothiophene molecules are adsorbed on the Cu(111) surface. The chiral adsorption configurations are determined by Scanning Tunneling Microscopy studies and confirmed by first-principles calculations. Remarkably, the selection of the adsorption sites by chemically different moieties of the molecules is dictated by the arrangement of the atoms in the first and second surface layers. Furthermore, we have investigated the thermal effects on the direct desulfurization reaction that occurs under the catalytic activity of the Cu substrate. This reaction leads to a product that is covalently bound to the surface in chiral configurations. Publisher PDF

Monoclonal antibodies can provide important pre- or post-exposure protection against infectious disease for those not yet vaccinated or in individuals that fail to mount a protective immune response after vaccination. Inmazeb (REGN-EB3), a three-antibody cocktail against Ebola virus, lessened disease and improved survival in a controlled trial. Here, we present the cryo-EM structure at 3.1 Å of the Ebola virus glycoprotein, determined without symmetry averaging, in a simultaneous complex with the antibodies in the Inmazeb cocktail. This structure allows the modeling of previously disordered portions of the glycoprotein glycan cap, maps the non-overlapping epitopes of Inmazeb, and illuminates the basis for complementary activities and residues critical for resistance to escape by these and other clinically relevant antibodies. We further provide direct evidence that Inmazeb protects against the rapid emergence of escape mutants, whereas monotherapies even against conserved epitopes do not, supporting the benefit of a cocktail versus a monotherapy approach.

The SARS-CoV-2 Omicron variant of concern (VoC) and its sublineages contain 31-36 mutations in spike and escape neutralization by most therapeutic antibodies. In a pseudovirus neutralization assay, 66 of the nearly 400 candidate therapeutics in the Coronavirus Immunotherapeutic Consortium (CoVIC) panel neutralize Omicron and multiple Omicron sublineages. Among natural immunoglobulin Gs (IgGs), especially those in the receptor-binding domain (RBD)-2 epitope community, nearly all Omicron neutralizers recognize spike bivalently, with both antigen-binding fragments (Fabs) simultaneously engaging adjacent RBDs on the same spike. Most IgGs that do not neutralize Omicron bind either entirely monovalently or have some (22%-50%) monovalent occupancy. Cleavage of bivalent-binding IgGs to Fabs abolishes neutralization and binding affinity, with disproportionate loss of activity against Omicron pseudovirus and spike. These results suggest that VoC-resistant antibodies overcome mutagenic substitution via avidity. Hence, vaccine strategies targeting future SARS-CoV-2 variants should consider epitope display with spacing and organization identical to trimeric spike. ispartof: CELL REPORTS vol:42 issue:1 ispartof: location:United States status: published

Accurately determining the macronutrient profile of mare milk is a precursor to studying how milk composition affects foals’ growth and development. This study optimized and validated an extraction and quantification method for mare milk oligosaccharides, which make up a portion of the carbohydrate fraction of mare milk. Mare milk was extracted with chloroform and methanol, and oligosaccharides were selectively isolated from the carbohydrate fraction using porous-graphitized carbon solid-phase-extraction (SPE). Good recovery rates for milk oligosaccharides (between 70 and 100%) were achieved with the optimized method. This study also compared the use of Fourier-Transform infrared (FTIR) spectroscopy versus wet chemistry quantification methods for protein, fat, and lactose. The FTIR method produced statistically equivalent protein contents to the wet chemistry method, along with substantial savings in both analyst time and consumable consumption. FTIR analysis slightly underestimated the fat content of mare milk relative to the official wet chemistry method, with the difference between the methods increasing at higher fat contents. FTIR also overestimated the lactose content of mare milk and appeared to generate “lactose” values that included the milk oligosaccharides and thus represented the total carbohydrate (lactose and milk oligosaccharides) content of mare milk.

Om mogelijke risico’s van chemische stoffen in consumentenproducten te kunnen beoordelen, is een goede schatting nodig van de mate waarin mensen eraan blootstaan als zij het product gebruiken. Voor deze schatting heeft het RIVM het computerprogramma ConsExpo ontwikkeld, waarvoor recentelijk een webapplicatie is gemaakt (ConsExpo Web). Hiermee kan bijvoorbeeld de blootstelling van een bepaalde chemische stof binnenshuis tijdens het gebruik van bijvoorbeeld verf, schoonmaakmiddelen of cosmetica worden berekend. Voor de gebruikers van ConsExpo Web zijn Factsheets geschreven waarin standaardmodellen en standaardwaarden (defaults) staan voorgeschreven. Door deze modellen en waarden te gebruiken, wordt de blootstellingsschatting op een transparante en gestandaardiseerde manier uitgevoerd. Er zijn meerdere Factsheets, waarvan nu de Factsheet over doe-het-zelfproducten is herzien. In de Factsheet over doe-het-zelfproducten staan standaardwaarden die bruikbaar zijn om de blootstelling aan een stof in een doe-het-zelfproduct te schatten. Voorbeelden van die waarden zijn de frequentie van het gebruik en hoeveelheden van het gebruikte product. In de herziene versie staan de nieuwste beschikbare databronnen beschreven, is de nieuwe informatie beoordeeld en waar nodig zijn de standaardwaarden aangepast. Parallel aan de publicatie van de herziene doe-het-zelfproducten Factsheet zullen ook de gepubliceerde standaardwaarden in de ConsExpo-database van onder meer lijmen, kitten, vullers, en coatings worden vernieuwd.

Background Eusociality is widely considered to evolve through kin selection, where the reproductive success of an individual’s close relative is favored at the expense of its own. High genetic relatedness is thus considered a prerequisite for eusociality. While ants are textbook examples of eusocial animals, not all ants form colonies of closely related individuals. One such example is the ectatommine ant Rhytidoponera metallica, which predominantly forms queen-less colonies that have such a low intra-colony relatedness that they have been proposed to represent a transient, unstable form of eusociality. However, R. metallica is among the most abundant and widespread ants on the Australian continent. This apparent contradiction provides an example of how inclusive fitness may not by itself explain the maintenance of eusociality and raises the question of what other selective advantages maintain the eusocial lifestyle of this species. Results We provide a comprehensive portrait of the venom of R. metallica and show that the colony-wide venom consists of an exceptionally high diversity of functionally distinct toxins for an ant. These toxins have evolved under strong positive selection, which is normally expected to reduce genetic variance. Yet, R. metallica exhibits remarkable intra-colony variation, with workers sharing only a relatively small proportion of toxins in their venoms. This variation is not due to the presence of chemical castes, but has a genetic foundation that is at least in part explained by toxin allelic diversity. Conclusions Taken together, our results suggest that the toxin diversity contained in R. metallica colonies may be maintained by a form of group selection that selects for colonies that can exploit more resources and defend against a wider range of predators. We propose that increased intra-colony genetic variance resulting from low kinship may itself provide a selective advantage in the form of an expanded pharmacological venom repertoire. These findings provide an example of how group selection on adaptive phenotypes may contribute to maintaining eusociality where a prerequisite for kin selection is diminished.

Additional file 4: Figure S1. Activity of Rm1a and Rm4a. Figure S2. Cytotoxicity of R. metallica aculeatoxin peptides. Figure S3. Gene structures of some ant aculeatoxins. Figure S4. Presence of aculeatoxin alleles among workers from the same R. metallica colony. Figure S5. Phylogeny of R. metallica aculeatoxins estimated from full-length or only signal- and propeptide domains. Table S4. Recombination analysis of R. metallica clades 1–3.Table S5. Primers used to amplify aculeatoxins from R. metallica genomic DNA.

Additional file 2: Table S1. Peptides and proteins identified in the pooled venom of R. metallica. Peptide and protein names and corresponding encoding contig id are provided for each, along with the best BLAST hit against UniProtKB, evidence supporting their presence in the venom, mature molecular weight, expression level (TPM), and the sequence of the mature domain and full prepropeptide. Complete (1) evidence includes identification of the complete mature peptide sequence from either the bottom-up or top-down proteomic experiments, while “partial” includes only partial identification from the bottom-up experiment.

Voltage-gated sodium (NaV) channels are critical regulators of neuronal excitability and are targeted by many toxins that directly interact with the pore-forming α subunit, typically via extracellular loops of the voltage-sensing domains, or residues forming part of the pore domain. Excelsatoxin A (ExTxA), a pain-causing knottin peptide from the Australian stinging tree Dendrocnide excelsa, is the first reported plant-derived NaV channel modulating peptide toxin. Here we show that TMEM233, a member of the dispanin family of transmembrane proteins expressed in sensory neurons, is essential for pharmacological activity of ExTxA at NaV channels, and that co-expression of TMEM233 modulates the gating properties of NaV1.7. These findings identify TMEM233 as a previously unknown NaV1.7-interacting protein, position TMEM233 and the dispanins as accessory proteins that are indispensable for toxin-mediated effects on NaV channel gating, and provide important insights into the function of NaV channels in sensory neurons. This work was supported by the University of Queensland Protein Production Facility and the Australian Cancer Research Foundation (ACRF) Cancer Biology Imaging Facility. Financial support was provided by the Australian Research Council (ARC; DP200102377, CE200100012) and the Australian National Health and Medical Research Council (NHMRC) fellowship to I.V. (APP1162503), Project grant (APP1162597) to M.M. and L.D.R., and Medical Research Council grant to J.J.C. and A.L.O. (MR/R011737/1). B.C.A. received funding from an NHMRC CJ Martin Fellowship (APP1162427). J.R.D. received funding through an ARC DECRA fellowship (DE210100422). S.D.H. and S.G.W. were supported by Merit Review Award B9253-C from the U.S. Dept. of Veterans Affairs Rehabilitation Service; The Center for Neuroscience & Regeneration Research is a Collaboration of the Paralyzed Veterans of America with Yale University. M.R.I. and D.A.A. received grants from the UK Medical Research Council (MR/S003428/1, MR/W002426/1). J.J.C., S.L., A.M.H., J.Zhao. and J.N.W. were supported by the Wellcome Trust (200183). A.M.H. receives funding from Qatar University (QUSD-CMED-2018/9-3; QUCG-CMED-19/20-4). We thank Dr. Patrick Walsh and Vincent Truong from Anatomic Inc. for helpful discussions on the gene expression profile of Anatomic Inc. iPSC-derived sensory neurons; Virginia Nink (Queensland Brain Institute, University of Queensland) for expert assistance with flow cytometry; and Prof Jenny Stow and Dr Lin Luo (Institute for Molecular Bioscience, University of Queensland) for provision of FANTOM Riken cDNA plasmids. Scopus

Gene editing through repair of CRISPR-Cas9-induced chromosomal breaks offers a means to correct a wide range of genetic defects. Directing repair to produce desirable outcomes by modulating DNA repair pathways holds considerable promise to increase the efficiency of genome engineering. Here, we show that inhibition of non-homologous end joining (NHEJ) or polymerase theta-mediated end joining (TMEJ) can be exploited to alter the mutational outcomes of CRISPR-Cas9. We show robust inhibition of TMEJ activity at CRISPR-Cas9-induced double-strand breaks (DSBs) using ART558, a potent polymerase theta (Polϴ) inhibitor. Using targeted sequencing, we show that ART558 suppresses the formation of microhomology-driven deletions in favor of NHEJ-specific outcomes. Conversely, NHEJ deficiency triggers the formation of large kb-sized deletions, which we show are the products of mutagenic TMEJ. Finally, we show that combined chemical inhibition of TMEJ and NHEJ increases the efficiency of homology-driven repair (HDR)-mediated precise gene editing. Our work reports a robust strategy to improve the fidelity and safety of genome engineering.

This entry looks at the commons (land and other resources that are subject to common use) and wildlife conservation (protecting plant and animal species and their habitats). Historically, the societies of Northeast India had developed sophisticated ideas about shared resources. These were challenged by colonial and postcolonial concepts, an encounter that has many dimensions. Here, this entry focuses on just one: the emergence of zones that the state set aside for the protection of wildlife. ‘Vertical’ conservation spread throughout Northeast India, and this led to tensions between humans over who actually own the local habitat and has the right to fashion it. It also created new confrontations between humans and protected ‘flagship species’. Social scientists suggest that we can mitigate these complications by imagining ‘interspecies communities’ that embrace humans, animals, and plants.

DNA double-strand breaks are a major threat to cellular survival and genetic integrity. In addition to high fidelity repair, three intrinsically mutagenic DNA break repair routes have been described, i.e. single-strand annealing (SSA), polymerase theta-mediated end-joining (TMEJ) and residual ill-defined microhomology-mediated end-joining (MMEJ) activity. Here, we identify C. elegans Helicase Q (HELQ-1) as being essential for MMEJ as well as for SSA. We also find HELQ-1 to be crucial for the synthesis-dependent strand annealing (SDSA) mode of homologous recombination (HR). Loss of HELQ-1 leads to increased genome instability: patchwork insertions arise at deletion junctions due to abortive rounds of polymerase theta activity, and tandem duplications spontaneously accumulate in genomes of helq-1 mutant animals as a result of TMEJ of abrogated HR intermediates. Our work thus implicates HELQ activity for all DSB repair modes guided by complementary base pairs and provides mechanistic insight into mutational signatures common in HR-defective cancers., Microhomology-mediated end-joining (MMEJ) is a poorly defined mutagenic DNA break repair pathway. Here the authors show that the helicase HELQ is essential for polymerase theta-independent MMEJ, single-strand annealing and homologous recombination through synthesis dependent strand annealing in C. elegans.

The COVID-19 pandemic has limited in-lab cognitive testing. While at-home alternatives exist (testing over the phone), differences in test design and delivery complicate direct comparison of most in-lab and at-home tests. Here we describe the design, infrastructure, and implementation of the California Cognitive Assessment Battery (CCAB), a cognitive test battery and administration system. Using automated remote administration over cellular networks, identical computerized cognitive tests can be administered at-home or in the lab.The CCAB comprises 30+ computerized cognitive tests. Test delivery is automated through text, text-to-speech, and graphical instructions; test scoring is also automated, using automatic speech recognition for verbal responses. The test delivery infrastructure consists of two components: the test kit (MS Surface Pro tablet computer with mouse, headset, and cellular dongle) , and a web-browser application "CCAB Examiner," used by the test administrator to control test delivery, view real-time progress and scores, and monitor/chat with the participant via AV feeds. The system is coordinated by a cloud-based server which also delivers the CCAB Examiner application to any browser. Once connected, the "CCAB Test Station" application and CCAB Examiner communicate over a commercial Communication as a Service (CAAS) provider, which transmits AV streams and a text messaging protocol. CCAB Test Station is designed to isolate control, communications, and testing functions for maximal fault tolerance. For example, test administration continues automatically even if Examiner and Test Station lose connection. Test data is uploaded to the server during sessions and re-synced when the test kit returns.442 participants underwent three 90-minute CCAB test sessions on successive days (72 total tests per participant). More than 98% of participants finished all three sessions, and99% of tests were completed. Cellular connection failures occurred occasionally, but98% resulted only in the loss of A/V feeds for the examiner, while tests continued automatically and examiners were able to monitor real-time performance data.The CCAB permits identical tests to be administered at home or in the laboratory, while cellular-network based remote administration allows for nationwide access for all socioeconomic groups, even for people in remote areas without Wi-Fi, or those with limited technical expertise.

BackgroundMany studies have reported an increased risk of autism spectrum disorder (ASD) associated with some maternal diagnoses in pregnancy. However, such associations have not been studied systematically, accounting for comorbidity between maternal disorders. Therefore our aim was to comprehensively test the associations between maternal diagnoses around pregnancy and ASD risk in offspring.MethodsThis exploratory case–cohort study included children born in Israel from 1997 to 2008, and followed up until 2015. We used information on all ICD-9 codes received by their mothers during pregnancy and the preceding year. ASD risk associated with each of those conditions was calculated using Cox proportional hazards regression, adjusted for the confounders (birth year, maternal age, socioeconomic status and number of ICD-9 diagnoses during the exposure period).ResultsThe analytic sample consisted of 80 187 individuals (1132 cases, 79 055 controls), with 822 unique ICD-9 codes recorded in their mothers. After extensive quality control, 22 maternal diagnoses were nominally significantly associated with offspring ASD, with 16 of those surviving subsequent filtering steps (permutation testing, multiple testing correction, multiple regression). Among those, we recorded an increased risk of ASD associated with metabolic [e.g. hypertension; HR = 2.74 (1.92–3.90), p = 2.43 × 10−8], genitourinary [e.g. non-inflammatory disorders of cervix; HR = 1.88 (1.38–2.57), p = 7.06 × 10−5] and psychiatric [depressive disorder; HR = 2.11 (1.32–3.35), p = 1.70 × 10−3] diagnoses. Meanwhile, mothers of children with ASD were less likely to attend prenatal care appointment [HR = 0.62 (0.54–0.71), p = 1.80 × 10−11].ConclusionsSixteen maternal diagnoses were associated with ASD in the offspring, after rigorous filtering of potential false-positive associations. Replication in other cohorts and further research to understand the mechanisms underlying the observed associations with ASD are warranted.

Agrobacterium tumefaciens-mediated transformation has been for decades the preferred tool to generate transgenic plants. During this process, a T-DNA carrying transgenes is transferred from the bacterium to plant cells, where it randomly integrates into the genome via polymerase theta (Pol theta)-mediated end joining (TMEJ). Targeting of the T-DNA to a specific genomic locus via homologous recombination (HR) is also possible, but such gene targeting (GT) events occur at low frequency and are almost invariably accompanied by random integration events. An additional complexity is that the product of recombination between T-DNA and target locus may not only map to the target locus (true GT), but also to random positions in the genome (ectopic GT). In this study, we have investigated how TMEJ functionality affects the biology of GT in plants, by using Arabidopsis thaliana mutated for the TEBICHI gene, which encodes for Pol theta. Whereas in TMEJ-proficient plants we predominantly found GT events accompanied by random T-DNA integrations, GT events obtained in the teb mutant background lacked additional T-DNA copies, corroborating the essential role of Pol theta in T-DNA integration. Pol theta deficiency also prevented ectopic GT events, suggesting that the sequence of events leading up to this outcome requires TMEJ. Our findings provide insights that can be used for the development of strategies to obtain high-quality GT events in crop plants.

Objectives Mindfulness-Based Stress Reduction (MBSR) involves training in mindful meditation and has been shown to improve functioning across a range of different disorders. However, little research has focused on the use of MBSR in stroke patients, and previous MBSR studies typically have not included an active control condition to account for non-specific factors that could contribute to the observed benefits. Methods We conducted a pilot study of MBSR in chronic stroke patients, comparing MBSR to an active control condition. Half of participants were randomly assigned to a standard 8-week MBSR class, and the other half of participants were assigned to an 8-week Brain Health class matched for schedule, instructor, and format. Participants were assessed pre- and post-intervention by blinded examiners on a neuropsychological battery that included primary outcome measures of psychological and cognitive functioning. Participants were also given an anonymous questionnaire following the post-intervention testing session to measure class satisfaction. Results Both the MBSR and Brain Health classes were rated favorably by participants. Recruitment and retention rates were high, and methods for participant randomization and examiner blinding were successful. Class implementation in terms of execution was also successful, as rated by outside experts. Conclusions This study established the feasibility of conducting MBSR and Brain Health classes in a chronic stroke population. Trial Registration https://ClinicalTrials.gov, NCT #: 02600637

Mitteilungen zur Kirchlichen Zeitgeschichte, Bd. 4 (2010)

Developing potent therapeutics and effective vaccines are the ultimate goals in controlling infectious diseases. Lassa virus (LASV), the causative pathogen of Lassa fever (LF), infects hundreds of thousands annually, but effective antivirals or vaccines against LASV infection are still lacking. Furthermore, neutralizing antibodies against LASV are rare. Here, we describe biochemical analyses and high-resolution cryo-electron microscopy structures of a therapeutic cocktail of three broadly protective antibodies that target the LASV glycoprotein complex (GPC), previously identified from survivors of multiple LASV infections. Structural and mechanistic analyses reveal compatible neutralizing epitopes and complementary neutralization mechanisms that offer high potency, broad range, and resistance to escape. These antibodies either circumvent or exploit specific glycans comprising the extensive glycan shield of GPC. Further, they require mammalian glycosylation, native GPC cleavage, and proper GPC trimerization. These findings guided engineering of a next-generation GPC antigen suitable for future neutralizing antibody and vaccine discovery. Together, these results explain protective mechanisms of rare, broad, and potent antibodies and identify a strategy for the rational design of therapeutic modalities against LF and related infectious diseases.

Monoclonal antibodies can provide important pre- or post-exposure protection against disease for those not yet vaccinated or in individuals that fail to mount a protective immune response after vaccination. A key concern in use of monotherapy monoclonal antibody products lies in the high risk of mutagenic escape. Inmazeb (REGN-EB3), a three-antibody cocktail against Ebola virus, demonstrated efficacy in lessening disease course and improving survival in a randomized, controlled trial. Here we present the cryoEM structure at 3.1 Å of the Ebola virus glycoprotein, determined without symmetry averaging, in a simultaneous complex with eight Fab fragments of antibodies in the Inmazeb cocktail. This structure allows modeling of previously disordered portions of the glycan cap, maps the non-overlapping epitopes of Inmazeb, and illuminates the basis for complementary activities, as well as residues that are critical for resistance to escape by each component of this cocktail and other clinically relevant antibodies. We also provide direct evidence that, unlike monotherapy treatments, including those targeting conserved epitopes, the Inmazeb protects against the rapid emergence of EBOV escape mutants and supports the benefit of the combination approach.