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Cornelia de Lange Syndrome (CdLS) is a rare, dominantly inherited multisystem developmental disorder characterized by highly variable manifestations of growth and developmental delays, upper limb involvement, hypertrichosis, cardiac, gastrointestinal, craniofacial, and other systemic features. Pathogenic variants in genes encoding cohesin complex structural subunits and regulatory proteins (NIPBL, SMC1A, SMC3, HDAC8, and RAD21) are the major pathogenic contributors to CdLS. Heterozygous or hemizygous variants in the genes encoding these five proteins have been found to be contributory to CdLS, with variants in NIPBL accounting for the majority (>60%) of cases, and the only gene identified to date that results in the severe or classic form of CdLS when mutated. Pathogenic variants in cohesin genes other than NIPBL tend to result in a less severe phenotype. Causative variants in additional genes, such as ANKRD11, EP300, AFF4, TAF1, and BRD4, can cause a CdLS-like phenotype. The common role that these genes, and others, play as critical regulators of developmental transcriptional control has led to the conditions they cause being referred to as disorders of transcriptional regulation (or DTRs). Here, we report the results of a comprehensive molecular analysis in a cohort of 716 probands with typical and atypical CdLS in order to delineate the genetic contribution of causative variants in cohesin complex genes as well as novel candidate genes, genotype-phenotype correlations, and the utility of genome sequencing in understanding the mutational landscape in this population.

Abstract The advanced stage at diagnosis of colorectal cancer (CRC) may be related to individual factors, socioeconomic conditions, and healthcare service availability. The objective of the study was to analyze the prevalence of advanced stage CRC at the time of diagnosis and its association with individual, contextual, socioeconomic, and healthcare service indicators. An observational, cross-sectional study was conducted, analyzing cases of malignant neoplasms of the colon and rectum in individuals of both sexes, aged between 18 and 99 years, diagnosed between 2010 and 2019 in Brazil (n = 69,047). Data were collected from the Hospital Cancer Registry (HCR), Atlas of Human Development in Brazil, and from the National Registry of Health Institutions (NRHI). A Multilevel Poisson Regression model with random intercept was used. The prevalence of advanced stage CRC at diagnosis was 65.6%. Advanced stage was associated with older age groups prevalence ratio (PR) 4.40 and younger age groups (PR 1.84), low Human Development Index (HDI) (PR 1.22), and low density of family health strategy teams (PR 1.10). The study highlights the unequal distribution of social determinants of health in the diagnosis CRC in Brazil, revealing the need to evaluate and redirect public policies aimed at improving early detection and prevention of CRC in the country.

Sawsan HA Abu El Hassan,1 Engy A Wahsh,2 Abdelmaksod Mohammed Mousa,3 Ahmed RN Ibrahim,4 Emad Lotfy Mohammed5 1Anesthesiology, Intensive Care and Pain Therapy, Faculty of Medicine, Ain Shams University, Cairo, Egypt; 2Department of Clinical Pharmacy, Faculty of Pharmacy, October 6 University, Giza, 12585, Egypt; 3Neurological and Spine Surgery Department, Faculty of Medicine, October 6 University, Giza, 12585, Egypt; 4Department of Clinical Pharmacy, College of Pharmacy, King Khalid University, Abha, 61421, Saudi Arabia; 5Anesthesiology, Intensive Care and Pain Therapy, Faculty of Medicine, Misr University for Science and Technology, Giza, EgyptCorrespondence: Engy A Wahsh; Ahmed RN Ibrahim, Email engywahsh@o6u.edu.eg; aribrahim@kku.edu.saBackground: As posterior lumbosacral spine fixation surgeries are common spine procedures done nowadays due to different causes and mostly accompanied with moderate-to-severe postoperative pain, so should find effective postoperative analgesia for these patients. This study aimed to observe analgesic effect of dexmedetomidine combined with bupivacaine versus bupivacaine alone for erector spinae plane block ESPB for postoperative pain control of posterior lumbosacral spine fixation surgeries.Methods: Double-blind randomized controlled study including 90 patients who were randomly allocated into 3 groups (30 patients for each): Dexmedetomidine combined with bupivacaine (DB group), bupivacaine (B group), and saline (control) (S group). US-guided ESPB was performed preoperatively bilaterally in all patients of the 3 groups. All patients received intravenous patient-controlled postoperative analgesia with morphine and 1 gm intravenous paracetamol every 8 hours. Primary clinical outcomes were active (while mobilization) and passive (at rest) visual analog scale (VAS) pain score at first 24 hours measured every 2 hours, opioid consumption (number of PCA presses), and need for rescue analgesia. Other clinical outcomes included active and passive VAS pain score at second 24 hours, measured every 4 hours, opioid consumption, need for rescue analgesia, postoperative opioid side effects, and intraoperative dexmedetomidine side effects as bradycardia and hypotension.Results: Active and passive VAS pain scores, postoperative opioid consumption, need for rescue analgesia, and postoperative opioid side effects were significantly lower in DB group when compared to other groups (B and S groups). There were no additional intraoperative dexmedetomidine side effects as bradycardia and hypotension. The estimated effect-size r was − 0.58 and Cohen’s d was − 1.46.Conclusion: Addition of dexmedetomidine to bupivacaine 0.25% in ESPB for postoperative pain control in patients of posterior lumbosacral spine fixation surgeries resulted in lower active and passive VAS pain scores, decreased postoperative opioid consumption, need for rescue analgesia and postoperative opioid side effects without additional intraoperative dexmedetomidine side effects.Clinicaltrials.gov Identifier: NCT05590234.Keywords: lumbosacral spine fixation surgeries, erector spinae plane block, ESPB, dexmedetomidine, postoperative analgesia, opioid consumption

Resumo: A detecção precoce é uma das estratégias para o controle do câncer de mama e, para tanto, é fundamental garantir o acesso à investigação dos casos suspeitos para continuidade do cuidado e tratamento oportuno. Este estudo tem por objetivo estimar a necessidade de procedimentos para detecção precoce dessa neoplasia e avaliar a sua adequação no atendimento às mulheres rastreadas e sintomáticas no Sistema Único de Saúde (SUS), no ano de 2019. Foi realizado um estudo descritivo transversal para analisar a oferta de exames de detecção precoce do câncer de mama, comparando a necessidade estimada com os procedimentos realizados no SUS. Foram utilizados os parâmetros disponibilizados pelo Instituto Nacional de Câncer para estimar a população e a necessidade de exames para a detecção precoce. No Sistema de Informações Ambulatoriais do SUS, obteve-se o número de procedimentos realizados em 2019. Observou-se um déficit de mamografias de rastreamento no país (-45,1%), variando entre -31,4% na Região Sul a -70,5% na Região Norte. Se a oferta desse exame fosse direcionada para a população-alvo do rastreamento, o déficit no país reduziria para -14,8% e haveria sobreoferta no Sul (6,2%). Os procedimentos de investigação diagnóstica apresentaram variações entre as regiões, com maiores déficits de punção por agulha grossa (-90,8%) e biópsia/exérese de nódulo da mama (-80,6%) observados no Centro-oeste, e o maior déficit de exames anatomopatológicos no Norte (-88,5%). A comparação entre a produção e a necessidade de procedimentos para detecção precoce do câncer de mama no Brasil identificou déficits e inadequações que devem ser melhor conhecidos e equacionados em nível estadual e municipal.

Abstract Background Chronic migraine, a highly disabling migraine subtype, affects nearly 2% of the general population. Understanding migraine chronification is vital for developing better treatment and prevention strategies. An important factor in the chronification of migraine is the overuse of acute headache medication. However, the mechanisms behind the transformation of episodic migraine to chronic migraine and vice versa have not yet been elucidated. We performed a longitudinal epigenome-wide association study to identify DNA methylation (DNAm) changes associated with treatment response in patients with chronic migraine and medication overuse as part of the Chronification and Reversibility of Migraine clinical trial. Blood was taken from patients with chronic migraine (n = 98) at baseline and after a 12-week medication withdrawal period. Treatment responders, patients with ≥ 50% reduction in monthly headache days (MHD), were compared with non-responders to identify DNAm changes associated with treatment response. Similarly, patients with ≥ 50% versus

Abstract Background and context The scale of land degradation worldwide has led to nearly one billion hectares committed to restoration globally. However, achieving such restoration targets will necessitate complex trade-offs against limited time, competing knowledge, costs, resources and varying stakeholder and societal preferences. Participatory scenarios allow a way to identify collaborative solutions for restoration planning and implementation best suited for the local cultures and societies they are tied to. They can be used to navigate uncertainties surrounding future trajectories of restored areas by evaluating trade-offs in outcomes. This research aims to systematically map the evidence on the use of participatory scenarios in restoration planning. We use the following research question: What evidence exists on the use of participatory scenarios in ecological restoration? This is answered by examining the characteristics of the evidence base, types of study design, types of outcomes, trade-offs in outcomes, and the role of participants. Methods A comprehensive and reproducible search strategy was followed using bibliographic databases, web-based searches, and targeted searching. Search results underwent a two-step screening process according to eligibility criteria. Metadata on key areas of interest were extracted from included texts and were narratively synthesised alongside data visualisations to answer the research questions. Review findings 18,612 records were initially identified, and 106 articles were included in the final map. Most studies were conducted in Europe and North America, focusing on restoring agricultural land or forests. Most texts used mixed methods and explored multiple outcome types, but environmental outcomes were the most assessed. Within environmental outcomes, indicators for ecological function were assessed more frequently than structural or compositional indicators. The most common reason for choosing outcomes and indicators was stakeholder interest. Trade-offs in social, ecological, and economic outcomes were mainly examined across space using mapping techniques, while far fewer studies looked at trade-offs across stakeholders and time. Participants were mostly included in the scenario creation step and were usually chosen purposefully by the research team. Conclusions It is difficult to understand how useful scenarios are for restoration planning because few texts reported how scenarios fed into the process. Despite this, the range of outcomes used and different method types adopted suggests participatory scenarios allow for integrating different knowledge and approaches, alongside facilitating the use of qualitative or semi-quantitative data when this is more appropriate or quantitative data is not widely available. To better use participatory scenarios as a tool for ecological restoration planning, decision-makers can push for greater levels and definitions of participation from the offset of restoration projects with specified, regular, and structured communication and participation channels. We also recommend more systematic methods of participant selection, such as stakeholder analysis. Further research is needed to understand the effectiveness of participatory scenarios in restoration planning and whether the participation of stakeholders was successful in meeting objectives. To improve the evidence base, future studies should clearly evaluate their effectiveness in the restoration planning process and their success in meeting their participatory objectives.

BackgroundMigraine has an increased prevalence in several immune disorders, but genetic cause-effect relationships remain unclear. Mendelian randomization (MR) was used in this study to explore whether immune diseases are causally associated with migraine and its subtypes.MethodsWe conducted a two-sample bidirectional multivariate Mendelian randomization study. Single-nucleotide polymorphisms (SNP) for six immune diseases, including rheumatoid arthritis (RA), systemic lupus erythematosus (SLE), type 1 diabetes mellitus (T1D), allergic rhinitis (AR), asthma and psoriasis, were used as genetic instrumental variables. Summary statistics for migraine were obtained from 3 databases: the International Headache Genetics Consortium (IHGC), UK Biobank, and FinnGen study. MR analyses were performed per outcome database for each exposure and subsequently meta-analyzed. Reverse MR analysis was performed to determine whether migraine were risk factors for immune diseases. In addition, we conducted a genetic correlation to identify shared genetic variants for these two associations.ResultsNo significant causal relationship was found between immune diseases and migraine and its subtypes. These results were robust with a series of sensitivity analyses. Using the linkage disequilibrium score regression method (LDSC), we detected no genetic correlation between migraine and immune diseases.ConclusionThe evidence from our study does not support a causal relationship between immune diseases and migraine. The mechanisms underlying the frequent comorbidity of migraine and several immune diseases need to be further elucidated.

Mechanisms underlying the migraine aura are incompletely understood, which to large extent is related to a lack of models in which cortical spreading depolarization (CSD), the correlate of the aura, occurs spontaneously. Here, we investigated electrophysiological and behavioural CSD features in freely behaving mice expressing mutant CaV2.1 Ca2+ channels, either with the milder R192Q or the severer S218L missense mutation in the α1 subunit, known to cause familial hemiplegic migraine type 1 (FHM1) in patients. Very rarely, spontaneous CSDs were observed in mutant but never in wildtype mice. In homozygous Cacna1aR192Q mice exclusively single-wave CSDs were observed whereas heterozygous Cacna1aS218L mice displayed multiple-wave events, seemingly in line with the more severe clinical phenotype associated with the S218L mutation. Spontaneous CSDs were associated with body stretching, one-directional slow head turning, and rotating movement of the body. Spontaneous CSD events were compared with those induced in a controlled manner using minimally invasive optogenetics. Also in the optogenetic experiments single-wave CSDs were observed in Cacna1aR192Q and Cacna1aS218L mice (whereas the latter also showed multiple-wave events) with movements similar to those observed with spontaneous events. Compared to wildtype mice, FHM1 mutant mice exhibited a reduced threshold and an increased propagation speed for optogenetically induced CSD with a more profound CSD-associated dysfunction, as indicated by a prolonged suppression of transcallosal evoked potentials and a reduction of unilateral forepaw grip performance. When induced during sleep, the optogenetic CSD threshold was particularly lowered, which may explain why spontaneous CSD events predominantly occurred during sleep. In conclusion, our data show that key neurophysiological and behavioural features of optogenetically induced CSDs mimic those of rare spontaneous events in FHM1 R192Q and S218L mutant mice with differences in severity in line with FHM1 clinical phenotypes seen with these mutations.

ABSTRACT Bacterial infections are a growing global healthcare concern, as an estimated annual 4.95 million deaths are associated with antimicrobial resistance (AMR). Methicillin-resistant Staphylococcus aureus is one of the deadliest pathogens and a high-priority pathogen according to the World Health Organization. Peptidoglycan hydrolases (PGHs) of phage origin have been postulated as a new class of antimicrobials for the treatment of bacterial infections, with a novel mechanism of action and no known resistances. The modular architecture of PGHs permits the creation of chimeric PGH libraries. In this study, the chimeric enzyme MEndoB was selected from a library of staphylococcal PGHs based on its rapid and sustained activity against staphylococci in human serum. The benefit of the presented screening approach was illustrated by the superiority of MEndoB in a head-to-head comparison with other PGHs intended for use against staphylococcal bacteremia. MEndoB displayed synergy with antibiotics and rapid killing in human whole blood with complete inhibition of re-growth over 24 h at low doses. Successful treatment of S. aureus-infected zebrafish larvae with MEndoB provided evidence for its in vivo effectiveness. This was further confirmed in a lethal systemic mouse infection model in which MEndoB significantly reduced S. aureus loads and tumor necrosis factor alpha levels in blood in a dose-dependent manner, which led to increased survival of the animals. Thus, the thorough lead candidate selection of MEndoB resulted in an outstanding second-generation PGH with in vitro, ex vivo, and in vivo results supporting further development.IMPORTANCEOne of the most pressing challenges of our era is the rising occurrence of bacteria that are resistant to antibiotics. Staphylococci are prominent pathogens in humans, which have developed multiple strategies to evade the effects of antibiotics. Infections caused by these bacteria have resulted in a high burden on the health care system and a significant loss of lives. In this study, we have successfully engineered lytic enzymes that exhibit an extraordinary ability to eradicate staphylococci. Our findings substantiate the importance of meticulous lead candidate selection to identify therapeutically promising peptidoglycan hydrolases with unprecedented activity. Hence, they offer a promising new avenue for treating staphylococcal infections.

Abstract Tissues are complex environments where different cell types are in constant interaction with each other and with non-cellular components. Preserving the spatial context during proteomics analyses of tissue samples has become an important objective for different applications, one of the most important being the investigation of the tumor microenvironment. Here, we describe a multiplexed protein biomarker detection method on the COMET instrument, coined sequential ImmunoFluorescence (seqIF). The fully automated method uses successive applications of antibody incubation and elution, and in-situ imaging enabled by an integrated microscope and a microfluidic chip that provides optimized optical access to the sample. We show seqIF data on different sample types such as tumor and healthy tissue, including 40-plex on a single tissue section that is obtained in less than 24 h, using off-the-shelf antibodies. We also present extensive characterization of the developed method, including elution efficiency, epitope stability, repeatability and reproducibility, signal uniformity, and dynamic range, in addition to marker and panel optimization strategies. The streamlined workflow using off-the-shelf antibodies, data quality enabling downstream analysis, and ease of reaching hyperplex levels make seqIF suitable for immune-oncology research and other disciplines requiring spatial analysis, paving the way for its adoption in clinical settings.

Abstract Background Cortical spreading depolarization (CSD), the neurophysiological correlate of the migraine aura, can activate trigeminal pain pathways, but the neurobiological mechanisms and behavioural consequences remain unclear. Here we investigated effects of optogenetically-induced CSDs on headache-related behaviour and neuroinflammatory responses in transgenic mice carrying a familial hemiplegic migraine type 1 (FHM1) mutation. Methods CSD events (3 in total) were evoked in a minimally invasive manner by optogenetic stimulation through the intact skull in freely behaving wildtype (WT) and FHM1 mutant mice. Related behaviours were analysed using mouse grimace scale (MGS) scoring, head grooming, and nest building behaviour. Neuroinflammatory changes were investigated by assessing HMGB1 release with immunohistochemistry and by pre-treating mice with a selective Pannexin-1 channel inhibitor. Results In both WT and FHM1 mutant mice, CSDs induced headache-related behaviour, as evidenced by increased MGS scores and the occurrence of oculotemporal strokes, at 30 min. Mice of both genotypes also showed decreased nest building behaviour after CSD. Whereas in WT mice MGS scores had normalized at 24 h after CSD, in FHM1 mutant mice scores were normalized only at 48 h. Of note, oculotemporal stroke behaviour already normalized 5 h after CSD, whereas nest building behaviour remained impaired at 72 h; no genotype differences were observed for either readout. Nuclear HMGB1 release in the cortex of FHM1 mutant mice, at 30 min after CSD, was increased bilaterally in both WT and FHM1 mutant mice, albeit that contralateral release was more pronounced in the mutant mice. Only in FHM1 mutant mice, contralateral release remained higher at 24 h after CSD, but at 48 h had returned to abnormal, elevated, baseline values, when compared to WT mice. Blocking Panx1 channels by TAT-Panx308 inhibited CSD-induced headache related behaviour and HMGB1 release. Conclusions CSDs, induced in a minimally invasive manner by optogenetics, investigated in freely behaving mice, cause various migraine relevant behavioural and neuroinflammatory phenotypes that are more pronounced and longer-lasting in FHM1 mutant compared to WT mice. Prevention of CSD-related neuroinflammatory changes may have therapeutic potential in the treatment of migraine.

Abstract Continuous monitoring of tissue microphysiology is a key enabling feature of the organ-on-chip (OoC) approach for in vitro drug screening and disease modeling. Integrated sensing units are particularly convenient for microenvironmental monitoring. However, sensitive in vitro and real-time measurements are challenging due to the inherently small size of OoC devices, the characteristics of commonly used materials, and external hardware setups required to support the sensing units. Here we propose a silicon-polymer hybrid OoC device that encompasses transparency and biocompatibility of polymers at the sensing area, and has the inherently superior electrical characteristics and ability to house active electronics of silicon. This multi-modal device includes two sensing units. The first unit consists of a floating-gate field-effect transistor (FG-FET), which is used to monitor changes in pH in the sensing area. The threshold voltage of the FG-FET is regulated by a capacitively-coupled gate and by the changes in charge concentration in close proximity to the extension of the floating gate, which functions as the sensing electrode. The second unit uses the extension of the FG as microelectrode, in order to monitor the action potential of electrically active cells. The layout of the chip and its packaging are compatible with multi-electrode array measurement setups, which are commonly used in electrophysiology labs. The multi-functional sensing is demonstrated by monitoring the growth of induced pluripotent stem cell-derived cortical neurons. Our multi-modal sensor is a milestone in combined monitoring of different, physiologically-relevant parameters on the same device for future OoC platforms.

Oil palm plantations growing on peat soil are associated with high soil CO2 emissions. Oil palm plantations are set up with regular spatial patterns consisting of different surface management microforms: bare soil harvest paths, frond piles, cover plants and drainage ditches. Currently, there is limited understanding about the extent that this spatial variation impacts soil carbon losses, in part due to the challenges of partitioning peat oxidation from total soil respiration. We explored this spatial variation by measuring total soil respiration (Rtot), root density and environmental variables at 210 locations. Measurements were taken along transects going from the base of oil palms into the different microforms. Rtot was partitioned into root respiration (Ra) and heterotrophic respiration (Rh) using two different methods: (i) a “distance from palm” method (which utilizes the fluxes taken from soil with minimal root density) and (ii) a “linear regression” method (which models root density and Rtot, using the regression intercept for Rh). Here, the distance from palm partitioning method gave higher Rh estimates than the linear regression method. Rh varied significantly between the different palms used in the assessment but did not show significant spatial variation aside from this. Rtot and Ra were highest next to the palm and decreased with increasing distance from the palm. Rtot and Ra also showed significant spatial variation between the different surface management microforms, with each giving significantly higher fluxes below the frond piles near the drainage ditches than from below the frond piles near the cover plants. Area-weighted upscaling gave plantation best estimates of Rtot, Rh, Ra of 0.158 ± 0.016, and 0.130 ± 0.036 and 0.029 ± 0.030 g CO2-C m−2 h−1, respectively. We conclude that spatial patterns impact root density, Ra and Rtot fluxes but not Rh fluxes.

Introdução: O rastreamento do câncer de mama no Brasil é recomendado para as mulheres de 50 a 69 anos, conforme diretrizes nacionais para a detecção precoce do câncer de mama do Instituto Nacional de Câncer/Ministério da Saúde. Embora ele seja tradicionalmente difundido como prática apenas benéfica, as evidências científicas apontam sua complexidade e a necessidade de comunicar o balanço entre os riscos e os benefícios, sobretudo em mulheres mais jovens. Objetivo: Descrever o processo participativo de elaboração de uma ferramenta de apoio à decisão para o rastreamento do câncer de mama quando buscado por mulheres com idade entre 40 e 49 anos no Brasil. Métodos: Estudo qualitativo-participativo que envolveu nove médicos de diferentes estados do Brasil e 104 mulheres na etapa de elaboração e 40 na de avaliação da ferramenta, seguindo as recomendações do International Patient Decision Aid Standards, em quatro etapas: rodas de conversa, síntese de evidências, revisão de ferramentas e avaliação da ferramenta por médicos e mulheres. Resultados: A ferramenta elaborada é inédita no Brasil e todos os médicos que a utilizaram a consideraram útil na conversa sobre os riscos e benefícios do rastreamento; 88,9% avaliaram que as informações facilitaram o entendimento — visão compartilhada por 80% das mulheres — e 77,8% consideraram que reduziu ou não interferiu no tempo de consulta. A ferramenta foi posteriormente aprimorada conforme as críticas e sugestões. Conclusões: O estudo mostrou o alcance do objetivo da ferramenta em oferecer suporte à decisão compartilhada e boa aceitação entre médicos e mulheres.

ImportanceThe reopening of colleges and universities in the US during the coronavirus disease 2019 (COVID-19) pandemic is a significant public health challenge. The development of accessible and practical approaches for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) detection in the college population is paramount for deploying recurrent surveillance testing as an essential strategy for virus detection, containment, and mitigation.ObjectiveTo determine the prevalence of SARS-CoV-2 in asymptomatic participants in a university community by using CREST (Cas13-based, rugged, equitable, scalable testing), a CRISPR-based test developed for accessible and large-scale viral screening.Design, setting, and participantsFor this cohort study, a total of 1808 asymptomatic participants were screened for SARS-CoV-2 using a CRISPR-based assay and a point-of-reference reverse transcriptase-quantitative polymerase chain reaction (RT-qPCR) test. Viral prevalence in self-collected oropharyngeal swab samples collected from May 28 to June 11, 2020, and from June 23 to July 2, 2020, was evaluated.ExposuresTesting for SARS-CoV-2.Main outcomes and measuresSARS-CoV-2 status, viral load, and demographic information of the study participants were collected.ResultsAmong the 1808 participants (mean [SD] age, 27.3 [11.0] years; 955 [52.8%] female), 732 underwent testing from May to early June (mean [SD] age, 28.4 [11.7] years; 392 [53.6%] female). All test results in this cohort were negative. In contrast, 1076 participants underwent testing from late June to early July (mean [SD] age, 26.6 [10.5] years; 563 [52.3%] female), with 9 positive results by RT-qPCR. Eight of these positive samples were detected by the CRISPR-based assay and confirmed by Clinical Laboratory Improvement Amendments-certified diagnostic testing. The mean (SD) age of the positive cases was 21.7 (3.3) years; all 8 individuals self-identified as students. These metrics showed that a CRISPR-based assay was effective at capturing positive SARS-CoV-2 cases in this student population. Notably, the viral loads detected in these asymptomatic cases resemble those seen in clinical samples, highlighting the potential of covert viral transmission. The shift in viral prevalence coincided with the relaxation of stay-at-home measures.Conclusions and relevanceThese findings reveal a shift in SARS-CoV-2 prevalence in a young and asymptomatic population and uncover the leading edge of a local outbreak that coincided with rising case counts in the surrounding county and the state of California. The concordance between CRISPR-based and RT-qPCR testing suggests that CRISPR-based assays are reliable and offer alternative options for surveillance testing and detection of SARS-CoV-2 outbreaks, as is required to resume operations in higher-education institutions in the US and abroad.

Abstract Background Engaging the endocannabinoid system through inhibition of monoacylglycerol lipase (MAGL) and fatty acid amide hydrolase (FAAH), degrading endocannabinoids (endoCBs) 2-arachidonoylglycerol (2-AG) and anandamide (AEA), was proposed as a promising approach to ameliorate migraine pain. However, the activity of MAGL and FAAH and action of endoCB on spiking activity of meningeal afferents, from which migraine pain originates, has not been explored thus far. Therefore, we here explored the analgesic effects of endoCB enhancement in rat and human meningeal tissues. Methods Both MAGL and FAAH activity and local 2-AG and AEA levels were measured by activity-based protein profiling (ABPP) and LC–MS/MS, respectively, in rat meninges obtained from hemiskulls of P38-P40 Wistar rats and human meninges from elderly patients undergoing non-migraine related neurosurgery. The action on endoCBs upon administration of novel dual MAGL/FAAH inhibitor AKU-005 on meningeal afferents excitability was tested by investigating paired KCl-induced spiking and validation with local (co-)application of either AEA or 2-AG. Finally, the specific TRPV1 agonist capsaicin and blocker capsazepine were tested. Results The basal level of 2-AG exceeded that of AEA in rat and human meninges. KCl-induced depolarization doubled the level of AEA. AKU-005 slightly increased spontaneous spiking activity whereas the dual MAGL/FAAH inhibitor significantly decreased excitation of nerve fibres induced by KCl. Similar inhibitory effects on meningeal afferents were observed with local applications of 2-AG or AEA. The action of AKU-005 was reversed by CB1 antagonist AM-251, implying CB1 receptor involvement in the anti-nociceptive effect. The inhibitory action of AEA was also reversed by AM-251, but not with the TRPV1 antagonist capsazepine. Data cluster analysis revealed that both AKU-005 and AEA largely increased long-term depression-like meningeal spiking activity upon paired KCl-induced spiking. Conclusions In the meninges, high anti-nociceptive 2-AG levels can tonically counteract meningeal signalling, whereas AEA can be engaged on demand by local depolarization. AEA-mediated anti-nociceptive effects through CB1 receptors have therapeutic potential. Together with previously detected MAGL activity in trigeminal ganglia, dual MAGL/FAAH inhibitor AKU-005 appears promising as migraine treatment. Graphical Abstract

Three-dimensional (3D) deformity assessment and leg realignment planning is emerging. The aim of this study was to (1) validate a novel 3D planning modality that incorporates the weight-bearing (WB) state (3D WB) by comparing it to existing modalities (3D non-weight-bearing (NWB), 2D WB) and (2) evaluate the influence of the modality (2D vs. 3D) and the WB condition on the measurements. Three different planning and deformity measurement protocols were analyzed in 19 legs that underwent medial open-wedge high tibial osteotomy (HTO): (1) a 3D WB protocol, after 2D/3D registration of 3D CT models onto the long-leg radiograph (LLR) (3D WB), (2) a 3D NWB protocol based on the 3D surface models obtained in the supine position (3D NWB), and (3) a 2D WB protocol based on the LLR (2D WB). The hip–knee–ankle angle (HKA), joint line convergence angle (JLCA), and the achieved surgical correction were measured for each modality and patient. All the measurement protocols demonstrated excellent intermodal agreement for the achieved surgical correction, with an ICC of 0.90 (95% CI: 0.76–0.96)) (p < 0.001). Surgical correction had a higher mean absolute difference compared to the 3D opening angle (OA) when measured with the WB protocols (3D WB: 2.7 ± 1.8°, 3D NWB: 1.9 ± 1.3°, 2D WB: 2.2 ± 1.3°), but it did not show statistical significance. The novel planning modality (3D WB) demonstrated excellent agreement when measuring the surgical correction after HTO compared to existing modalities.

ABSTRACT Objective: To characterize associated factors and overall survival of women with metastatic breast cancer treated with trastuzumab after its incorporation into the SUS, and additionally to present the direct costs of this technology. Methods: This is a retrospective cohort, based on data from computerized medical records from one of the units of the National Cancer Institute (INCA), in Rio de Janeiro-RJ, Brazil. Women with HER-2 positive metastatic breast cancer undergoing trastuzumab treatment from September 2017 to August 2018 were included. Overall survival was estimated using the Kaplan-Meier method and compared between groups using the log-rank test. Results: 136 women were selected, whose median age at diagnosis was 51 years (range: 21–81 years). The median OS was 43.63 months (95%CI 33.92–53.34). It is observed that the median OS for the population already diagnosed with metastatic disease (stage IV) was significantly lower than for patients diagnosed in stages I-III (37.43 months vs. 48.6 months, p

The data set contains images of leaf venation networks obtained from tree species in Malaysian Borneo. The data set contains 726 leaves from 295 species comprising 50 families, sampled from eight forest plots in Sabah. Image extents are approximately 1 × 1 cm, or 50 megapixels. All images contain a region of interest in which all veins have been hand traced. The complete data set includes over 30 billion pixels, of which more than 600 million have been validated by hand tracing. These images are suitable for morphological characterization of these species, as well as for training of machine-learning algorithms that segment biological networks from images. Data are made available under the Open Data Commons Attribution License. You are free to copy, distribute, and use the database; to produce works from the database; and to modify, transform, and build upon the database. You must attribute any public use of the database, or works produced from the database, in the manner specified in the license. For any use or redistribution of the database, or works produced from it, you must make clear to others the license of the database and keep intact any notices on the original database.

Sultan M Alshahrani,1 Khalid Orayj,1 Ali M Alqahtani,2 Adel Alfatease,3 Atheer Alshahrani,4 Ahmed RN Ibrahim1,5 1Clinical Pharmacy Department, College of Pharmacy, King Khalid University, Abha, Saudi Arabia; 2Pharmacology Department, College of Pharmacy, King Khalid University, Abha, Saudi Arabia; 3Pharmaceutics Department, College of Pharmacy, King Khalid University, Abha, Saudi Arabia; 4Alnahdi Pharmacies, Khamis Mushait, Saudi Arabia; 5Department of Biochemistry, Faculty of Pharmacy, Minia University, Minia, EgyptCorrespondence: Ahmed RN Ibrahim, Clinical Pharmacy Department, College of Pharmacy, King Khalid University, Abha, Saudi Arabia, Tel +966 554088979, Email aribrahim@kku.edu.saIntroduction: Community pharmacists play a key role as vaccinators for COVID-19. They can reduce the burden of the disease worldwide.Objective: This study used a cross-sectional questionnaire to determine whether the Saudi Arabian public was willing to obtain the COVID-19 vaccine via community pharmacists.Results: The questionnaire focused on the satisfaction, concerns, and opinions towards providing vaccination by community pharmacists. The study featured 415 individuals aged 18 and older (eligible for the COVID-19 vaccine). Of the participants in this study, 58.1% were aged 18– 25, with 55.4% female. Most participants (72.8%) have not been exposed to COVID-19 and are not aware of the approval of COVID-19 vaccination by community pharmacists. Of the 415 complete questionnaires, 45% believed that community pharmacists are not experienced in administering vaccines. However, 63% of participants are satisfied with getting the COVID-19 vaccination by a community pharmacist if no other option is available. More than 68% of the respondents agree that community pharmacies should expand their health care services to include vaccinations, prescriptions, checkups, and other forms of preventative medicine.Discussion: The availability of community pharmacist-administered vaccination in Saudi Arabia could be a significant factor in the success of the country’s vaccination program. This study may serve as a model to expand the role of pharmacists in other countries’ vaccination programs.Keywords: SARS-CoV-2, community pharmacy, immunization, perspective, adherence, patient satisfaction

Abstract Objetive: to analyze breast cancer screening monitoring indicators in the female population using the Brazilian National Health System, from 2018 to 2019. Methods: this was a descriptive study based on Cancer Information System (SISCAN) data; screening indicators were calculated following deterministic linkage of the mammography and histopathology databases. Results: in 2018, 807,430 women aged 50 to 69 years were screened for breast cancer, 91% of whom had a benign result, 1.8% probably benign, 6.7% inconclusive results and 0.5% results suggestive of cancer; the positive mammogram rate was 9.0%; biopsy was estimated to be indicated for 1.6% of the women, 33.9% of whom had a malignant result, and the cancer confirmation rate was 5.4 per 1,000 women. Conclusion: high benign lesion loss to follow-up was identified; the positive mammogram rate was lower than the international parameter, but the cancer detection rate was adequate and the percentage of inconclusive mammograms was acceptable.

An ideal wound dressing not only needs to absorb excess exudate but should also allow for a moist wound-healing environment as well as being mechanically strong. Such a dressing can be achieved by combining both a natural (alginate) and synthetic (poly(ethylene glycol) polymer. Interestingly, using an electron beam on (meth)acrylated polymers allows their covalent crosslinking without the use of toxic photo-initiators. The goal of this work was to crosslink alginate at different methacrylation degrees (26.1 and 53.5% of the repeating units) with diacrylated poly(ethylene glycol) (PEGDA) using electron-beam irradiation at different doses to create strong, transparent hydrogels. Infrared spectroscopy showed that both polymers were homogeneously distributed within the irradiated hydrogel. Rheology showed that the addition of PEGDA into alginate with a high degree of methacrylation and a polymer concentration of 6 wt/v% improved the storage modulus up to 15,867 ± 1102 Pa. Gel fractions > 90% and swelling ratios ranging from 10 to 250 times its own weight were obtained. It was observed that the higher the storage modulus, the more limited the swelling ratio due to a more crosslinked network. Finally, all species were highly transparent, with transmittance values > 80%. This may be beneficial for the visual inspection of healing progression. Furthermore, these polymers may eventually be used as carriers of photosensitizers, which is favorable in applications such as photodynamic therapy.