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Background: Antimicrobials cause perturbations in the composition and diversity of the host microbiome. We aimed to compare gut microbiome perturbations caused by oral tebipenem pivoxil hydrobromide (a novel carbapenem) and by amoxicillin-clavulanic acid (an orally administered β-lactam-β-lactam inhibitor combination widely used in clinical practice)., Methods: We did a phase 1, single-centre, randomised, parallel-group, active-control trial to evaluate the effect of tebipenem pivoxil hydrobromide on the human gut microbiota. Healthy participants aged 18 years or older with no documented illnesses during recruitment were enrolled at Karolinska University Hospital (Stockholm, Sweden). Study participants were stratified by sex and block-randomised in a 1:1 ratio to treatment with either tebipenem pivoxil hydrobromide (600 mg orally every 8 h) or amoxicillin-clavulanic acid (500 mg amoxicillin and 125 mg clavulanic acid orally every 8 h). The study included 10 days of treatment (days 1-10) and four follow-up visits (days 14, 21, 90, and 180). The trial was open-label for clinical investigators and patients, but masked for microbiology investigators. Faecal samples were collected at all visits. Sequencing of 16S rDNA was used to measure the diversity metrics, and quantitative culture to quantify selected taxa. The primary outcomes were changes in the α and β diversity and log count of colony-forming units for selected taxa between samples compared with baseline (day 1), and whether any changes reverted during the follow-up period. The analyses were done in the intention-to-treat population. This study was registered with ClinicalTrials.gov (NCT04376554)., Findings: The study was conducted between Jan 23, 2020, and April 6, 2021. 49 volunteers were screened for eligibility, among whom 30 evaluable participants (14 men and 16 women) were assigned: 15 (50%) to the tebipenem pivoxil hydrobromide group and 15 (50%) to the amoxicillin-clavulanic acid group. Baseline characteristics were similar between groups. Complete follow-up was available for all participants, and all participants except one completed treatment as assigned. The diversity metrics showed significant changes from baseline during the treatment period. Significant decreases in richness were observed on days 4-10 (p≤0·0011) in the amoxicillin-clavulanic acid group and on days 4-14 (p≤0·0019) in the tebipenem pivoxil hydrobromide group. Similarly, evenness was significantly decreased during treatment in the amoxicillin-clavulanic acid group (day 4, p=0·030) and the tebipenem pivoxil hydrobromide group (days 4-10, p<0·0001) compared with baseline. Quantitative cultures showed significant decreases in Enterobacterales (days 4-7, p≤0·0030), Enterococcus spp (days 4-14, p=0·025 to p<0·0001), Bifidobacterium spp (days 2-4, p≤0·026), and Bacteroides spp (days 4-10, p≤0·030) in the tebipenem pivoxil hydrobromide group. Similarly, in amoxicillin-clavulanic acid recipients, significant changes were observed in Enterobacterales (days 4-10, p≤0·048), Bifidobacterium spp (days 2-4, p≤0·013), and Lactobacillus spp (days 2-4, p≤0·020). Samples from the follow-up period were not significantly different from those at baseline in β diversity analysis (PERMANOVA, p>0·99). By the end of the study, no significant change was observed compared with baseline in either group. There were no deaths or severe adverse events., Interpretation: The impact of tebipenem pivoxil hydrobromide on the gut microbiome was similar to that of amoxicillin-clavulanic acid. The safety of antibiotic use with regard to the microbiome should be given attention, as dysbiosis is associated with health and disease., Funding: Spero Therapeutics., Competing Interests: Declaration of interests MN, DM, PBE, and IAC were employees of Spero Therapeutics and LBG was a consultant at Spero Therapeutics during the study. No personal honoraria were paid out to any of the academic investigators. All other authors declare no competing interests., (Copyright © 2023 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

Objective: Dual carbapenemase-producing organisms (DCPOs) are an emerging threat that expands the spectrum of antimicrobial resistance. There is limited literature on the clinical and genetic epidemiology of DCPOs., Methods: DCPO isolates were identified by Xpert ® Carba-R PCR testing of routine diagnostic cultures performed from 2018 to 2021 at a New York City health system. WGS was performed by Illumina and/or PacBio. Medical records of patients were reviewed for clinical and epidemiological data., Results: Twenty-six DCPO isolates were obtained from 13 patients. Klebsiella pneumoniae ( n  = 22) was most frequent, followed by Pseudomonas aeruginosa ( n  = 2), Escherichia coli ( n  = 1) and Enterobacter cloacae ( n  = 1). The most common DCPO combination was bla NDM / bla OXA-48-like ( n  = 16). Notably, 1.05% (24/2290) of carbapenem-resistant Enterobacterales isolates were identified as DCPOs. The susceptibility profiles matched the identified resistance genes, except for a K. pneumoniae ( bla KPC / bla OXA-48-like ) isolate that was phenotypically susceptible to meropenem. Eleven patients were hospitalized within the year prior to admission, and received antibiotic(s) 1 month prior. Seven patients were originally from outside the USA. Hypertension, kidney disease and diabetes were frequent comorbidities. Death in two cases was attributed to DCPO infection. WGS of eight isolates showed that carbapenemases were located on distinct plasmids, except for one K. pneumoniae isolate where NDM and KPC carbapenemases were located on a single IncC-type plasmid backbone., Conclusions: Here we characterized a series of DCPOs from New York City. Foreign travel, prior hospitalization, antibiotic usage and comorbidities were common among DCPO cases. All carbapenemases were encoded on plasmids, which may facilitate horizontal transfer., (© The Author(s) 2023. Published by Oxford University Press on behalf of British Society for Antimicrobial Chemotherapy.)

In equine dentistry, the physiological incisor occlusal surface is visually perceived as a plane with a distinct inclination to the head's coronal plane, extending rostro-ventrally to caudo-dorsally. To better understand the formation of this inclined plane and its connection to dental wear, we investigated the hypothesis that it arises from masticatory movements and the considerable distance between mandibular articular heads and the incisor occlusal surfaces, acting as the three points of support for the mandibles. Leveraging data from a large-scale clinical study involving static and dynamic orthodontic measurements in horses, we approximated the mandibular movement range where incisor occlusion and dental wear occur. By introducing and testing a segment coordinate system, we explored possible angular deviations from the occlusal plane caused by mandibular roll and pitch rotations during two lateral mandibular movement patterns, protrusion and retrusion. Theoretical biomechanical calculations and simulations confirmed the visual perception of the incisor occlusal surface as a plane. To further examine our assumptions, we employed a simple mechanical simulator to assess incisor normal occlusion and provoked malocclusions (diagonal, smile, and frown bite) by modifying temporomandibular joint (TMJ) movement patterns. The results from clinical investigations were corroborated by both the theoretical analysis and mechanical simulations, strengthening our understanding of the biomechanical basis behind the physiological incisor occlusal plane maintenance in horses. These findings have significant implications for equine dental health and contribute to a thorough understanding of TMJ dynamics., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Sterkenburgh, Hartl, Peham, Nowak, Kyllar and Kau.)

Background: We assessed the efficacy and safety of dalbavancin, a long-acting lipoglycopeptide with activity against Gram-positive pathogens, for treatment of acute bacterial skin and skin structure infections (ABSSSI) in patients with high body mass index (BMI) and/or diabetes., Methods: Data from two phase 3 trials of dalbavancin (1000 mg intravenous [IV], day 1; 500 mg IV, day 8) versus comparator and one phase 3b trial of single-dose (1500 mg IV, day 1) versus 2-dose (1000 mg IV, day 1; 500 mg IV, day 8) dalbavancin in adults with ABSSSI were pooled and summarized separately by baseline BMI and diabetes status. Clinical success at 48 to 72 hours (≥20% reduction in lesion size), end of treatment ([EOT] day 14), and day 28 was evaluated in the intent-to-treat (ITT) and microbiological ITT (microITT) populations. Safety data were reported in patients who received ≥1 dose of study drug., Results: In the dalbavancin ITT population (BMI, n = 2001; diabetes, n = 2010), at 48 to 72 hours (and EOT) clinical success was achieved in 89.3% (EOT, 90.9%) of patients with normal BMI and 78.9% to 87.6% (EOT, 91.0% to 95.2%) of patients with elevated BMI. Clinical success after dalbavancin treatment was achieved in 82.4% (EOT, 90.8%) of patients with diabetes and 86.0% (EOT, 91.6%) of patients without diabetes. Similar trends were observed for infections due to methicillin-resistant Staphylococcus aureus or methicillin-susceptible S aureus (microITT population)., Conclusions: Dalbavancin is effective, with sustained clinical success rates in patients with obesity or diabetes, with a similar safety profile across patient groups., Competing Interests: Potential conflicts of interest. The design, study conduct, and financial support for the study were provided by AbbVie. AbbVie participated in the interpretation of data, review, and approval of the publication. No honoraria or payments were made for authorship. TR, JL, RDL, and JP are employees of AbbVie and own AbbVie stock. BG, MN, and UR were employees of Allergan before its acquisition by AbbVie, at the time of study conduct and analysis. PLG was an employee of AbbVie and held stock in the company at the time of study conduct. Currently, BG is an employee of Shionogi Inc., MN is an employee of Spero Therapeutics, PLG is an employee of Becton Dickinson, and UR is an employee of BiomX Inc. Conflicts that the editors consider relevant to the content of the manuscript have been disclosed., (© The Author(s) 2023. Published by Oxford University Press on behalf of Infectious Diseases Society of America.)

Introduction: Omadacycline is approved for the treatment of community-acquired bacterial pneumonia (CABP) and acute bacterial skin and soft tissue infection (ABSSSI). The integration of newer agents into clinical use involves understanding the nuances of clinical decision-making. This review will provide an in-depth focus on omadacycline in clinical practice., Areas Covered: Literature review of omadacycline utilizing PubMed was performed to provide a comprehensive evaluation of omadacycline pharmacology, microbiology, registrational Phase 3 clinical trials, and post-marketing clinical studies. In addition, the immunomodulatory and other attributes of tetracycline class of antibiotics, of which omadacycline is a member, are reviewed, introducing the concept of antibiotic selection with attention to the bacterial pathogen and human host relationship., Expert Opinion: Omadacycline builds upon the favorable attributes of tetracycline antibiotics and provides very reliable empiric coverage for both Staphylococcus aureus and Streptococcus spp. Clinicians require a more robust understanding of antibiotics, including omadacycline, in order to optimize patient outcomes, streamline care, and reduce medical costs.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) variants are characterized by differences in transmissibility and response to therapeutics. Therefore, discriminating among them is vital for surveillance, infection prevention, and patient care. While whole-genome sequencing (WGS) is the "gold standard" for variant identification, molecular variant panels have become increasingly available. Most, however, are based on limited targets and have not undergone comprehensive evaluation. We assessed the diagnostic performance of the highly multiplexed Agena MassARRAY SARS-CoV-2 Variant Panel v3 to identify variants in a diverse set of 391 SARS-CoV-2 clinical RNA specimens collected across our health systems in New York City, USA and Bogotá, Colombia (September 2, 2020 to March 2, 2022). We demonstrated almost perfect levels of interrater agreement between this assay and WGS for 9 of 11 variant calls (κ ≥ 0.856) and 25 of 30 targets (κ ≥ 0.820) tested on the panel. The assay had a high diagnostic sensitivity (≥93.67%) for contemporary variants (e.g., Iota, Alpha, Delta, and Omicron [BA.1 sublineage]) and a high diagnostic specificity for all 11 variants (≥96.15%) and all 30 targets (≥94.34%) tested. Moreover, we highlighted distinct target patterns that could be utilized to identify variants not yet defined on the panel, including the Omicron BA.2 and other sublineages. These findings exemplified the power of highly multiplexed diagnostic panels to accurately call variants and the potential for target result signatures to elucidate new ones. IMPORTANCE The continued circulation of SARS-CoV-2 amid limited surveillance efforts and inconsistent vaccination of populations has resulted in the emergence of variants that uniquely impact public health systems. Thus, in conjunction with functional and clinical studies, continuous detection and identification are quintessential to informing diagnostic and public health measures. Furthermore, until WGS becomes more accessible in the clinical microbiology laboratory, the ideal assay for identifying variants must be robust, provide high resolution, and be adaptable to the evolving nature of viruses like SARS-CoV-2. Here, we highlighted the diagnostic capabilities of a highly multiplexed commercial assay to identify diverse SARS-CoV-2 lineages that circulated from September 2, 2020 to March 2, 2022 among patients seeking care in our health systems. This assay demonstrated variant-specific signatures of nucleotide/amino acid polymorphisms and underscored its utility for the detection of contemporary and emerging SARS-CoV-2 variants of concern.

Little is known about the evolution of cold tolerance in polar plant species and how they differ from temperate relatives. To gain insight into their biology and the evolution of cold tolerance, we compared the molecular basis of cold response in three Arctic Brassicaceae species. We conducted a comparative time series experiment to examine transcriptional responses to low temperature. RNA was sampled at 22°C, and after 3, 6, and 24 at 2°C. We then identified sets of genes that were differentially expressed in response to cold and compared them between species, as well as to published data from the temperate Arabidopsis thaliana. Most differentially expressed genes were species-specific, but a significant portion of the cold response was also shared among species. Among thousands of differentially expressed genes, ~200 were shared among the three Arctic species and A. thaliana, while ~100 were exclusively shared among the three Arctic species. Our results show that cold response differs markedly between Arctic Brassicaceae species, but probably builds on a conserved basis found across the family. They also confirm that highly polygenic traits such as cold tolerance may show little repeatability in their patterns of adaptation., (© 2022 The Authors. Molecular Ecology published by John Wiley & Sons Ltd.)

As severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) continues to circulate, multiple variants of concern have emerged. New variants pose challenges for diagnostic platforms because sequence diversity can alter primer/probe-binding sites (PBSs), causing false-negative results. The MassARRAY SARS-CoV-2 Panel (Agena Bioscience) uses RT-PCR and mass spectrometry to detect five multiplex targets across N and ORF1ab genes. Herein, we use a data set of 256 SARS-CoV-2-positive specimens collected between April 11, 2021, and August 28, 2021, to evaluate target performance with paired sequencing data. During this time frame, two targets in the N gene (N2 and N3) were subject to the greatest sequence diversity. In specimens with N3 dropout, 69% harbored the Alpha-specific A28095U polymorphism that introduces a 3'-mismatch to the N3 forward PBS and increases risk of target dropout relative to specimens with 28095A (relative risk, 20.02; 95% CI, 11.36 to 35.72; P < 0.0001). Furthermore, among specimens with N2 dropout, 90% harbored the Delta-specific G28916U polymorphism that creates a 3'-mismatch to the N2 probe PBS and increases target dropout risk (relative risk, 11.92; 95% CI, 8.17 to 14.06; P < 0.0001). These findings highlight the robust capability of MassARRAY SARS-CoV-2 Panel target results to reveal circulating virus diversity, and they underscore the power of multitarget design to capture variants of concern., (Copyright © 2022 Association for Molecular Pathology and American Society for Investigative Pathology. Published by Elsevier Inc. All rights reserved.)

The repeated, rapid and often pronounced patterns of evolutionary divergence observed in insular plants, or the 'plant island syndrome', include changes in leaf phenotypes, growth, as well as the acquisition of a perennial lifestyle. Here, we sequence and describe the genome of the critically endangered, Galápagos-endemic species Scalesia atractyloides Arnot., obtaining a chromosome-resolved, 3.2-Gbp assembly containing 43,093 candidate gene models. Using a combination of fossil transposable elements, k-mer spectra analyses and orthologue assignment, we identify the two ancestral genomes, and date their divergence and the polyploidization event, concluding that the ancestor of all extant Scalesia species was an allotetraploid. There are a comparable number of genes and transposable elements across the two subgenomes, and while their synteny has been mostly conserved, we find multiple inversions that may have facilitated adaptation. We identify clear signatures of selection across genes associated with vascular development, growth, adaptation to salinity and flowering time, thus finding compelling evidence for a genomic basis of the island syndrome in one of Darwin's giant daisies., (© 2022. The Author(s).)

We reported the frequency of resistance gene detection in Gram-negative blood culture isolates and correlated these findings with corresponding antibiograms. Data were obtained from 1045 isolates tested on the GenMark Dx ePlex Blood Culture Identification Gram-Negative Panels at the Mount Sinai Hospital Clinical Microbiology Laboratory in New York from March 2019 to February 2021. Susceptibilities were performed using Vitek 2 (bioMérieux Clinical Diagnostics) or Microscan (Beckman Coulter Inc.). bla CTX-M was detected in 26.4% Klebsiella pneumoniae, 23.5% Escherichia coli, and 16.4% Proteus mirabilis isolates. As would be expected, both bla CTX-M and bla CTX-M negative isolates were likely to be susceptible to newer agents while bla CTX-M positive isolates were more likely to be resistant to earlier generations of beta-lactam antibiotics. 3/204 bla CTX-M -positive isolates were found to be ceftriaxone-susceptible. Conversely, 2.8% ceftriaxone nonsusceptible strains were negative for all β-lactamase genes on the ePlex BCID-GN panel, including bla CTX-M . The prevalence of CTX-M-producing Enterobacterales remains high in the United States. A small number of bla CTX-M -positive isolates were susceptible to ceftriaxone, and a small number of ceftriaxone nonsusceptible isolates were negative for bla CTX-M . Further studies are needed to determine the optimal management when an isolate is phenotypically susceptible to ceftriaxone, but bla CTX-M is detected. IMPORTANCE There is limited literature on corresponding results obtained from rapid molecular diagnostics with the antibiotic susceptibility profile. We reported a correlation between the results obtained from ePlex and the antibiograms against a large collection of Gram-negative bacteria. We reported that there can be a discrepancy in a small number of cases, but the clinical significance of that is unknown.

Introduction: Scant data are available on the 30-day financial burden associated with incident complicated urinary tract infections (cUTIs) in a cohort of predominately elderly patients. This study sought to examine total and cUTI-related 30-day Medicare spending (MS), a proxy for healthcare costs, among Medicare fee-for-service (FFS) beneficiaries who resided in the community with newly diagnosed cUTIs., Methods: A retrospective multicenter cohort study of adult beneficiaries in the Medicare FFS database with a cUTI between 2017 and 2018 was performed. Patients were included if they were enrolled in Medicare FFS and Medicare Part D from 2016 to 2019, had a cUTI first diagnosis in 2017-2018, no evidence of any UTI diagnoses in 2016, and residence in the community between 2016 and 2018., Results: During the study period, 723,324 cases occurred in Medicare beneficiaries who met the study criteria. Overall and cUTI-related 30-day MS were $7.6 and $4.5 billion, respectively. The average overall and cUTI-related 30-day MS per beneficiary were $10,527 and $6181, respectively. The major driver of cUTI-related 30-day MS was acute care hospitalizations ($3.2 billion) and the average overall and cUTI-related 30-day MS per hospitalizations were $16,431 and $15,438, respectively., Conclusion: Overall 30-day MS for Medicare FSS patients who resided in the community with incident cUTIs was substantial, with cUTI-related MS accounting for 59%. As the major driver of cUTI-related 30-day MS was acute care hospitalizations, healthcare systems should develop well-defined criteria for hospital admissions that aim to avert hospitalizations in clinically stable patients and expedite the transition of patients to the outpatient setting to complete their care.

The coronavirus disease 2019 (COVID-19) pandemic has sparked the rapid development of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) diagnostics. However, emerging variants pose the risk for target dropout and false-negative results secondary to primer/probe binding site (PBS) mismatches. The Agena MassARRAY® SARS-CoV-2 Panel combines reverse-transcription polymerase chain reaction and matrix-assisted laser desorption/ionization time-of-flight mass-spectrometry to probe for five targets across N and ORF1ab genes, which provides a robust platform to accommodate PBS mismatches in divergent viruses. Herein, we utilize a deidentified data set of 1262 SARS-CoV-2-positive specimens from Mount Sinai Health System (New York City) from December 2020 to April 2021 to evaluate target results and corresponding sequencing data. Overall, the level of PBS mismatches was greater in specimens with target dropout. Of specimens with N3 target dropout, 57% harbored an A28095T substitution that is highly specific for the Alpha (B.1.1.7) variant of concern. These data highlight the benefit of redundancy in target design and the potential for target performance to illuminate the dynamics of circulating SARS-CoV-2 variants., (© 2021 Wiley Periodicals LLC.)

Background: Cladophialophora bantiana is a dematiaceous, saprophytic fungus and a rare but reported cause of intracranial abscesses due to its strong neurotropism. Although it predominantly affects immunocompetent individuals with environmental exposure, more recently, its significance as a highly lethal opportunistic infection in transplant recipients has been recognized. Successful treatment requires timely but often challenging diagnosis, followed by complete surgical excision. Next-generation sequencing of microbial cell-free DNA (cfDNA) from plasma is a novel diagnostic method with the potential to identify invasive fungal infections more rapidly and less invasively than conventional microbiological testing, including brain biopsy., Observations: The authors described the case of a recipient of a liver transplant who presented with seizures and was found to have innumerable ring-enhancing intracranial lesions. The Karius Test, a commercially available method of next-generation sequencing of cfDNA, was used to determine the causative organism. Samples from the patient's plasma identified C. bantiana 6 days before culture results of the surgical specimen, allowing optimization of the empirical antifungal regimen, which led to a reduction in the size of the abscesses., Lessons: The authors' findings suggest that microbial cfDNA sequencing may be particularly impactful in improving the management of brain abscesses in which the differential diagnosis is wide because of immunosuppression.

Equine mastication, as well as dental wear patterns, is highly important for the development of treatments in equine dentistry. During the last decades, the stress and strain distributions of equine teeth have been successfully simulated using finite element analysis. Yet, to date, there is no simulation available for dental tooth wear in equines. In this study, we developed a simplified two-dimensional computer simulation of dental wear. It provides a first tentative explanation for the development of the marked physiological inclination of the occlusal surface and for pathological conditions such as sharp enamel points in equine cheek teeth. The mechanical properties of the dental structures as well as the movement of the mandible during the equine chewing cycle were simulated according to previously published data. The simulation setup was optimized in preliminary test runs. Further simulations were conducted varying the lateral excursion of the mandible and the presence or absence of incisor contact during the chewing cycle. The results of simulations showed clear analogies to tooth wear patterns in living equids, including the formation of wear abnormalities. Our analysis indicates that small variations in the pattern of movement during the masticatory cycle, as well as incisor contacts, are leading to marked changes in the occlusal tooth wear patterns. This opens new research avenues to better understand the development of dental wear abnormalities in equines and might have serious implications on captive animal health, welfare, and longevity., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Sterkenburgh, Schulz-Kornas, Nowak and Staszyk.)

As severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) infections continue, there is a substantial need for cost-effective and large-scale testing that utilizes specimens that can be readily collected from both symptomatic and asymptomatic individuals in various community settings. Although multiple diagnostic methods utilize nasopharyngeal specimens, saliva specimens represent an attractive alternative as they can rapidly and safely be collected from different populations. While saliva has been described as an acceptable clinical matrix for the detection of SARS-CoV-2, evaluations of analytic performance across platforms for this specimen type are limited. Here, we used a novel sensitive RT-PCR/MALDI-TOF mass spectrometry-based assay (Agena MassARRAY®) to detect SARS-CoV-2 in saliva specimens. The platform demonstrated high diagnostic sensitivity and specificity when compared to matched patient upper respiratory specimens. We also evaluated the analytical sensitivity of the platform and determined the limit of detection of the assay to be 1562.5 copies/ml. Furthermore, across the five individual target components of this assay, there was a range in analytic sensitivities for each target with the N2 target being the most sensitive. Overall, this system also demonstrated comparable performance when compared to the detection of SARS-CoV-2 RNA in saliva by the cobas® 6800/8800 SARS-CoV-2 real-time RT-PCR Test (Roche). Together, we demonstrate that saliva represents an appropriate matrix for SARS-CoV-2 detection on the novel Agena system as well as on a conventional real-time RT-PCR assay. We conclude that the MassARRAY® system is a sensitive and reliable platform for SARS-CoV-2 detection in saliva, offering scalable throughput in a large variety of clinical laboratory settings., (© 2021 Wiley Periodicals LLC.)

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and its associated clinical syndrome COVID-19 are causing overwhelming morbidity and mortality around the globe and disproportionately affected New York City between March and May 2020. Here, we report on the first 100 COVID-19-positive autopsies performed at the Mount Sinai Hospital in New York City. Autopsies revealed large pulmonary emboli in six cases. Diffuse alveolar damage was present in over 90% of cases. We also report microthrombi in multiple organ systems including the brain, as well as hemophagocytosis. We additionally provide electron microscopic evidence of the presence of the virus in our samples. Laboratory results of our COVID-19 cohort disclose elevated inflammatory markers, abnormal coagulation values, and elevated cytokines IL-6, IL-8, and TNFα. Our autopsy series of COVID-19-positive patients reveals that this disease, often conceptualized as a primarily respiratory viral illness, has widespread effects in the body including hypercoagulability, a hyperinflammatory state, and endothelial dysfunction. Targeting of these multisystemic pathways could lead to new treatment avenues as well as combination therapies against SARS-CoV-2 infection., (© 2021. The Author(s), under exclusive licence to United States & Canadian Academy of Pathology.)

Bacillus cereus is a gram-positive, rod-shaped bacterium that is commonly implicated in foodborne illness but has also become increasingly recognized as a source of serious non-gastrointestinal infections, including sepsis, meningitis, and pneumonia. Non-gastrointestinal B. cereus infections have been identified in children, especially in neonates; however, there are no previously described cases of fetal demise associated with B. cereus placental infection. We present a case of acute chorioamnionitis-related intrauterine fetal demise of twin A at 17 weeks gestation, noted two days after selective termination of twin B. Histological examination revealed numerous gram-positive bacilli in placental tissue, as well as fetal vasculature, in the setting of severe acute necrotizing chorioamnionitis and subchorionitis, intervillous abscesses, acute villitis, and peripheral acute funisitis. Cultures of maternal blood and placental tissue both yielded growth of B. cereus . This case underscores the importance of B. cereus as a human pathogen, and specifically demonstrates its potential as an agent of severe intraamniotic and placental infection with poor outcomes for the fetus.

Numerous reports document the spread of SARS-CoV-2, but there is limited information on its introduction before the identification of a local case. This may lead to incorrect assumptions when modeling viral origins and transmission. Here, we utilize a sample pooling strategy to screen for previously undetected SARS-CoV-2 in de-identified, respiratory pathogen-negative nasopharyngeal specimens from 3,040 patients across the Mount Sinai Health System in New York. The patients had been previously evaluated for respiratory symptoms or influenza-like illness during the first 10 weeks of 2020. We identify SARS-CoV-2 RNA from specimens collected as early as 25 January 2020, and complete SARS-CoV-2 genome sequences from multiple pools of samples collected between late February and early March, documenting an increase prior to the later surge. Our results provide evidence of sporadic SARS-CoV-2 infections a full month before both the first officially documented case and emergence of New York as a COVID-19 epicenter in March 2020.

The Arctic is one of the most extreme terrestrial environments on the planet. Here, we present the first chromosome-scale genome assembly of a plant adapted to the high Arctic, Draba nivalis (Brassicaceae), an attractive model species for studying plant adaptation to the stresses imposed by this harsh environment. We used an iterative scaffolding strategy with data from short-reads, single-molecule long reads, proximity ligation data, and a genetic map to produce a 302 Mb assembly that is highly contiguous with 91.6% assembled into eight chromosomes (the base chromosome number). To identify candidate genes and gene families that may have facilitated adaptation to Arctic environmental stresses, we performed comparative genomic analyses with nine non-Arctic Brassicaceae species. We show that the D. nivalis genome contains expanded suites of genes associated with drought and cold stress (e.g., related to the maintenance of oxidation-reduction homeostasis, meiosis, and signaling pathways). The expansions of gene families associated with these functions appear to be driven in part by the activity of transposable elements. Tests of positive selection identify suites of candidate genes associated with meiosis and photoperiodism, as well as cold, drought, and oxidative stress responses. Our results reveal a multifaceted landscape of stress adaptation in the D. nivalis genome, offering avenues for the continued development of this species as an Arctic model plant., (© 2020 The Authors. Molecular Ecology Resources published by John Wiley & Sons Ltd.)

Introduction: Dalbavancin is a lipoglycopeptide antibiotic approved as a single- and two-dose regimen for adults with acute bacterial skin and skin structure infections (ABSSSI) caused by susceptible gram-positive organisms. We present nephrotoxicity rates for patients with ABSSSI who received dalbavancin in three pivotal clinical trials and compare the rates with vancomycin., Methods: In a phase 3b clinical trial (DUR001-303), patients were randomized to dalbavancin single-dose (1500 mg intravenous [IV]) or two-dose regimen (1000 mg IV on day 1, 500 mg IV on day 8). In two phase 3 clinical trials (DISCOVER 1 and DISCOVER 2), patients were randomized to dalbavancin (two-dose regimen) or vancomycin 1 g (or 15 mg/kg) IV every 12 h for at least 3 days with an option to switch to orally administered linezolid 600 mg every 12 h for 10-14 days. Patients on dalbavancin with a creatinine clearance below 30 mL/min not on regular dialysis received a reduced dose of 1000 mg (single-dose arm) or 750 mg IV on day 1, 375 mg IV on day 8 (two-dose arm). Nephrotoxicity was defined as a 50% increase from baseline serum creatinine (SCr) or an absolute increase in SCr of 0.5 mg/dL at any time point. P values were obtained using the Cochran-Mantel-Haenszel test., Results: In dalbavancin-treated patients, rates of nephrotoxicity were low. The safety population with available creatinine values included 1325/1347 patients on any regimen of dalbavancin, and 54/651 patients who received vancomycin intravenously for at least 10 days and were not switched to orally administered linezolid. Patients on any regimen of dalbavancin had a lower rate of nephrotoxicity compared with patients receiving vancomycin intravenously for at least 10 days (3.7% vs 9.3%, respectively; P = 0.039)., Conclusions: Nephrotoxicity rates were lower in patients on dalbavancin relative to vancomycin for at least 10 days. On the basis of this experience, dalbavancin may be less nephrotoxic than intravenously administered vancomycin.

Morphologically similar species, that is cryptic species, may be similar or quasi-similar owing to the deceleration of morphological evolution and stasis. While the factors underlying the deceleration of morphological evolution or stasis in cryptic species remain unknown, decades of research in the field of paleontology on punctuated equilibrium have originated clear hypotheses. Species are expected to remain morphologically identical in scenarios of shared genetic variation, such as hybridization and incomplete lineage sorting, or in scenarios where bottlenecks reduce genetic variation and constrain the evolution of morphology. Here, focusing on three morphologically similar Stygocapitella species, we employ a whole-genome amplification method (WGA) coupled with double-digestion restriction-site associated DNA sequencing (ddRAD) to reconstruct the evolutionary history of the species complex. We explore population structure, use population-level statistics to determine the degree of connectivity between populations and species, and determine the most likely demographic scenarios which generally reject for recent hybridization. We find that the combination of WGA and ddRAD allowed us to obtain genomic-level data from microscopic eukaryotes (∼1 millimetre) opening up opportunities for those working with population genomics and phylogenomics in such taxa. The three species share genetic variance, likely from incomplete lineage sorting and ancient admixture. We speculate that the degree of shared variation might underlie morphological similarity in the Atlantic species complex., Competing Interests: The authors declare there are no competing interests., (©2021 Cerca et al.)

In late 2019, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) was first detected in China and has since caused a pandemic of coronavirus disease 2019 (COVID-19). The first case of COVID-19 in New York City was officially confirmed on 1 March 2020 followed by a severe local epidemic 1 . Here, to understand seroprevalence dynamics, we conduct a retrospective, repeated cross-sectional analysis of anti-SARS-CoV-2 spike antibodies in weekly intervals from the beginning of February to July 2020 using more than 10,000 plasma samples from patients at Mount Sinai Hospital in New York City. We describe the dynamics of seroprevalence in an 'urgent care' group, which is enriched in cases of COVID-19 during the epidemic, and a 'routine care' group, which more closely represents the general population. Seroprevalence increased at different rates in both groups; seropositive samples were found as early as mid-February, and levelled out at slightly above 20% in both groups after the epidemic wave subsided by the end of May. From May to July, seroprevalence remained stable, suggesting lasting antibody levels in the population. Our data suggest that SARS-CoV-2 was introduced in New York City earlier than previously documented and describe the dynamics of seroconversion over the full course of the first wave of the pandemic in a major metropolitan area.

The complex nature of species boundaries has been a central topic in evolutionary biology ever since Darwin. Despite numerous separate studies on reproductive isolation and hybridization, their relationship remains underinvestigated. Are the strengths and asymmetries of reproductive barriers reflected in the extent and directionalities of interspecific genetic exchange? We combined field, experimental, and molecular data to quantify strengths and asymmetries of sympatric reproductive barriers and hybridization between florally heteromorphic primroses. We also assessed whether generalist pollinators discriminate between different floral cues and contribute to reproductive isolation, a long-debated topic. Sympatric reproductive isolation is high but incomplete, and most phenotypic intermediates are genetic F 1 hybrids, whereas backcrosses are rare, revealing low interspecific gene flow. Species integrity rests on multiple barriers, but ethological isolation is among the strongest, demonstrating that even generalist pollinators crucially contribute to the maintenance of species boundaries. Furthermore, reproductive barriers are weaker for Primula veris and short-styled plants, results corroborated by molecular data. Thus, in florally heteromorphic systems, both species- and morph-dependent asymmetries affect permeability of species boundaries. Our study illustrates how the interactions between complex floral syndromes and pollinators shape species boundaries in unique, previously undescribed ways., (© 2020 The Authors New Phytologist © 2020 New Phytologist Trust.)

Message Queuing Telemetry Transport (MQTT) is one of the dominating protocols for edge- and cloud-based Internet of Things (IoT) solutions. When a security vulnerability of an IoT device is known, it has to be fixed as soon as possible. This requires a firmware update procedure. In this paper, we propose a secure update protocol for MQTT-connected devices which ensures the freshness of the firmware, authenticates the new firmware and considers constrained devices. We show that the update protocol is easy to integrate in an MQTT-based IoT network using a semantic approach. The feasibility of our approach is demonstrated by a detailed performance analysis of our prototype implementation on a IoT device with 32 kB RAM. Thereby, we identify design issues in MQTT 5 which can help to improve the support of constrained devices.

The urgent need to implement and rapidly expand testing for severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) infection has led to the development of multiple assays. How these tests perform relative to one another is poorly understood. We evaluated the concordance between the Roche Diagnostics cobas 6800 SARS-CoV-2 test and a laboratory-developed test (LDT) real-time reverse transcription-polymerase chain reaction based on a modified Centers for Disease Control and Prevention protocol, for the detection of SARS-CoV-2 in samples submitted to the Clinical Laboratories of the Mount Sinai Health System. A total of 1006 nasopharyngeal swabs in universal transport medium from persons under investigation were tested for SARS-CoV-2 as part of routine clinical care using the cobas SARS-CoV-2 test with subsequent evaluation by the LDT. Cycle threshold values were analyzed and interpreted as either positive ("detected" or "presumptive positive"), negative (not detected), inconclusive, or invalid. Statistical analysis was performed using GraphPad Prism 8. The cobas SARS-CoV-2 test reported 706 positive and 300 negative results. The LDT reported 640 positive, 323 negative, 34 inconclusive, and 9 invalid results. When excluding inconclusive and invalid results, the overall percent agreement between the two platforms was 95.8%. Cohen's κ coefficient was 0.904 (95% confidence interval, 0.875-0.933), suggesting almost perfect agreement between both platforms. An overall discordance rate of 4.2% between the two systems may reflect differences in primer sequences, assay limit of detection, or other factors, highlighting the importance of comparing the performance of different testing platforms., (© 2020 Wiley Periodicals LLC.)

Extreme environments offer powerful opportunities to study how different organisms have adapted to similar selection pressures at the molecular level. Arctic plants have adapted to some of the coldest and driest biomes on Earth and typically possess suites of similar morphological and physiological adaptations to extremes in light and temperature. Here, we compare patterns of molecular evolution in three Brassicaceae species that have independently colonized the Arctic and present some of the first genetic evidence for plant adaptations to the Arctic environment. By testing for positive selection and identifying convergent substitutions in orthologous gene alignments for a total of 15 Brassicaceae species, we find that positive selection has been acting on different genes, but similar functional pathways in the three Arctic lineages. The positively selected gene sets identified in the three Arctic species showed convergent functional profiles associated with extreme abiotic stress characteristic of the Arctic. However, there was little evidence for independently fixed mutations at the same sites and for positive selection acting on the same genes. The three species appear to have evolved similar suites of adaptations by modifying different components in similar stress response pathways, implying that there could be many genetic trajectories for adaptation to the Arctic environment. By identifying candidate genes and functional pathways potentially involved in Arctic adaptation, our results provide a framework for future studies aimed at testing for the existence of a functional syndrome of Arctic adaptation in the Brassicaceae and perhaps flowering plants in general., (© The Author(s) 2020. Published by Oxford University Press on behalf of the Society for Molecular Biology and Evolution.)

Background: Little is known about the strains at the glenoid near the bone-implant interface in reverse shoulder arthroplasty. The purpose of the current study was to evaluate the strains on the glenoid bone under a compressive load after implantation of three different sizes of metal-backed baseplates in either inferior or superior position in combination with three different sizes of glenospheres and three different glenosphere designs., Methods: Three sizes of baseplates (small, medium, large) were implanted in thirty-six paired human cadaveric scapulae either inferior, flush with the glenoid neck, or with a 5 mm superior offset. Glenospheres were available in three sizes (36 mm, 39 mm, 42 mm) and designs (standard, 4 mm lateralized, 2.5 mm inferiorized). Specimens were mounted in a servo-hydraulic testing apparatus at a 60° angle between the glenoid and actuator holding the humeral component. Four strain-gauge rosettes were placed around the glenoid rim to measure strains transferred to the scapular bone under a compressive load (750 N) relative to the various baseplate-glenosphere combinations. Following repeated compression, a load-to-failure test was performed., Results: Mean overall registered strains were 161με (range: - 1165 to 2347) at the inferior sensor, -2με (range: - 213 to 90) at the superior sensor, -95με (range: - 381 to 254) at the anterior sensor, and 13με (range: - 298 to 128) at the posterior sensor. Measured bone strains did not show any significant differences across tested baseplate and glenosphere design, size, or positioning combinations (p > 0.05 for all sensors). Furthermore, linear regression analysis did not identify any of the evaluated parameters as an independent influential factor for strains (p > 0.05 for all sensors). Mean load-at-failure was significantly higher in the group of inferior (3347.0 N ± 704.4 N) compared to superior (2763.8 N ± 927.8 N) positioned baseplates (p = 0.046)., Conclusion: Different baseplate positions, baseplate sizes, glenosphere sizes, and glenosphere design or various combinations of these parameters did not significantly influence the measured bone strains at the glenoid near the bone-implant interface in a contemporary reverse shoulder arthroplasty system., Level of Evidence: Basic Science Study, Biomechanical Study.

We construct a graph representation for the topology and geometry of the vasculature presenting across the whole mouse brain (dataset: Knife-Edge Scanning Microscope Brain Atlas India Ink). We use our graph representation to calculate preliminary estimates of the average radius as 4:8 μm, total vascular volume as 1:1000 mm 3 , total vascular surface area as 6:5511 cm 2 , and total vascular length of 2866:6567 cm. We then isolate a posterior cerebral region, derive its graph representation, and then import that representation to a Neo4j graph database. We then detail how researchers can query this database online to isolate specific vascular networks for further analysis and reconstruction.

The island-like distribution of subalpine habitats across mountain ranges can trigger the parallel evolution of locally adapted ecotypes. Such naturally replicated scenarios allow testing hypotheses on how elevational differentiation structures genetic diversity within species. Nevertheless, the parallel colonization of subalpine habitats across different mountain ranges has only rarely been documented with molecular data. We chose Primula elatior (Primulaceae), naturally spanning entire elevation range in multiple mountain regions of central Europe, to test for the origin of its scattered subalpine populations. Nuclear microsatellite variation revealed three genetic groups corresponding with the distinct study regions. We found that genetic differentiation between foothill and subalpine populations within each region was relatively low, suggesting that the colonization of subalpine habitats occurred independently within each mountain range. Furthermore, the strongest differentiation was usually found between the subalpine populations suggesting that mountain ridges may act as migration barriers that can reduce gene flow more strongly than elevational differences between foothill and subalpine populations. Finally, we found that subalpine colonization did not result in a loss of genetic diversity relative to foothill populations in agreement with the high migration rates that we document here between the subalpine and the foothill populations. In summary, our study shows subalpine Primula elatior populations are genetically diverse and distinct results of parallel colonization events from multiple foothill gene pools.

Interactions between fungi and plants, including parasitism, mutualism, and saprotrophy, have been invoked as key to their respective macroevolutionary success. Here we evaluate the origins of plant-fungal symbioses and saprotrophy using a time-calibrated phylogenetic framework that reveals linked and drastic shifts in diversification rates of each kingdom. Fungal colonization of land was associated with at least two origins of terrestrial green algae and preceded embryophytes (as evidenced by losses of fungal flagellum, ca. 720 Ma), likely facilitating terrestriality through endomycorrhizal and possibly endophytic symbioses. The largest radiation of fungi (Leotiomyceta), the origin of arbuscular mycorrhizae, and the diversification of extant embryophytes occurred ca. 480 Ma. This was followed by the origin of extant lichens. Saprotrophic mushrooms diversified in the Late Paleozoic as forests of seed plants started to dominate the landscape. The subsequent diversification and explosive radiation of Agaricomycetes, and eventually of ectomycorrhizal mushrooms, were associated with the evolution of Pinaceae in the Mesozoic, and establishment of angiosperm-dominated biomes in the Cretaceous.

The increasing demand and popularity of bariatric surgery are not only due to the ever-increasing obesity epidemic but to tackle obesity-related comorbidities like diabetes and hypertension. However, bariatric surgery is not free of complications. One rare complication is intussusception, jejuno-jejunal intussusception being the most common. Intussusception has been defined both in pregnant and in non-pregnant women as well as men. We describe the case of a 40-year-old woman in the first trimester of pregnancy came to the hospital with worsening abdominal pain, was found to have intussusception of small bowel involving the jejunum, along with ischaemic perforation and necrosis of the fundus of the stomach. Postsurgery, the patient had a complete recovery and eventually, she successfully delivered an intact, viable fetus., Competing Interests: Competing interests: None declared., (© BMJ Publishing Group Limited 2018. No commercial re-use. See rights and permissions. Published by BMJ.)

Prostate carcinoma is the most common non-dermatological malignancy in men and only second to lung cancer as the leading cause of cancer related deaths in America. Moreover, metastasis to the skin is even more uncommon following apparently successful treatment of a preexisting prostate cancer. We present an 87-year-old man previously diagnosed with prostate cancer treated with brachytherapy, who presented with a metastatic prostate adenocarcinoma of the scalp mimicking a basal cell carcinoma. Herein, we present a patient with an NKX 3.1 positive cutaneous metastatic prostate carcinoma identified through NKX3.1 stain rather than the typical PSA/PSAP stain.

Self-organized robots may develop attracting states within the sensorimotor loop, that is within the phase space of neural activity, body and environmental variables. Fixpoints, limit cycles and chaotic attractors correspond in this setting to a non-moving robot, to directed, and to irregular locomotion respectively. Short higher-order control commands may hence be used to kick the system from one self-organized attractor robustly into the basin of attraction of a different attractor, a concept termed here as kick control. The individual sensorimotor states serve in this context as highly compliant motor primitives. We study different implementations of kick control for the case of simulated and real-world wheeled robots, for which the dynamics of the distinct wheels is generated independently by local feedback loops. The feedback loops are mediated by rate-encoding neurons disposing exclusively of propriosensoric inputs in terms of projections of the actual rotational angle of the wheel. The changes of the neural activity are then transmitted into a rotational motion by a simulated transmission rod akin to the transmission rods used for steam locomotives. We find that the self-organized attractor landscape may be morphed both by higher-level control signals, in the spirit of kick control, and by interacting with the environment. Bumping against a wall destroys the limit cycle corresponding to forward motion, with the consequence that the dynamical variables are then attracted in phase space by the limit cycle corresponding to backward moving. The robot, which does not dispose of any distance or contact sensors, hence reverses direction autonomously.

Background: Previous biomechanical studies regarding deltoid function during glenohumeral abduction have primarily used static testing protocols., Hypotheses: (1) Deltoid forces required for scapular plane abduction increase as simulated rotator cuff tears become larger, and (2) maximal abduction decreases despite increased deltoid forces., Study Design: Controlled laboratory study., Methods: Twelve fresh-frozen cadaveric shoulders with a mean age of 67 years (range, 64-74 years) were used. The supraspinatus and anterior, middle, and posterior deltoid tendons were attached to individual shoulder simulator actuators. Deltoid forces and maximum abduction were recorded for the following tear patterns: intact, isolated subscapularis (SSC), isolated supraspinatus (SSP), anterosuperior (SSP + SSC), posterosuperior (infraspinatus [ISP] + SSP), and massive (SSC + SSP + ISP). Optical triads tracked 3-dimensional motion during dynamic testing. Fluoroscopy and computed tomography were used to measure critical shoulder angle, acromial index, and superior humeral head migration with massive tears. Mean values for maximum glenohumeral abduction and deltoid forces were determined. Linear mixed-effects regression examined changes in motion and forces over time. Pearson product-moment correlation coefficients ( r) among deltoid forces, critical shoulder angles, and acromial indices were calculated., Results: Shoulders with an intact cuff required 193.8 N (95% CI, 125.5 to 262.1) total deltoid force to achieve 79.8° (95% CI, 66.4° to 93.2°) of maximum glenohumeral abduction. Compared with native shoulders, abduction decreased after simulated SSP (-27.2%; 95% CI, -43.3% to -11.1%, P = .04), anterosuperior (-51.5%; 95% CI, -70.2% to -32.8%, P < .01), and massive (-48.4%; 95% CI, -65.2% to -31.5%, P < .01) cuff tears. Increased total deltoid forces were required for simulated anterosuperior (+108.1%; 95% CI, 68.7% to 147.5%, P < .01) and massive (+57.2%; 95% CI, 19.6% to 94.7%, P = .05) cuff tears. Anterior deltoid forces were significantly greater in anterosuperior ( P < .01) and massive ( P = .03) tears. Middle deltoid forces were greater with anterosuperior tears ( P = .03). Posterior deltoid forces were greater with anterosuperior ( P = .02) and posterosuperior ( P = .04) tears. Anterior deltoid force was negatively correlated ( r = -0.89, P = .01) with critical shoulder angle (34.3°; 95% CI, 32.0° to 36.6°). Deltoid forces had no statistical correlation with acromial index (0.55; 95% CI, 0.48 to 0.61). Superior migration was 8.3 mm (95% CI, 5.5 to 11.1 mm) during testing of massive rotator cuff tears., Conclusion: Shoulders with rotator cuff tears require considerable compensatory deltoid function to prevent abduction motion loss. Anterosuperior tears resulted in the largest motion loss despite the greatest increase in deltoid force., Clinical Relevance: Rotator cuff tears place more strain on the deltoid to prevent abduction motion loss. Fatigue or injury to the deltoid may result in a precipitous decline in abduction, regardless of tear size.

The use of graphs to analyze cerebrovascular networks is quite common in studies of the microcirculation. While we have learned a lot from studies utilizing graphs as a tool for the analysis of microvessels, most methodologies for these procedures have only been described in brief and most are not publicly accessible. In this work, we introduce the foundation for an anticipated open-source framework that we hope will streamline the analysis of cerebrovascular structure. We believe that a standardized and accessible framework for the analysis vascular filaments is not only needed, but is necessary, for studies charting the microcirculation on image volumes spanning several grains of tissue. We set forth the foundations for a comprehensive and complete framework in our current work.

Background: AABB requires that red blood cells (RBCs) are maintained at 1 to 10°C during transport. Historically, blood banks used the 30-minute rule for returned RBCs transported outside of validated containers. The implications of this policy have not been previously reported in a real-life hospital setting., Study Design and Methods: A 2-year, retrospective review of RBC units returned outside of qualified containers was conducted. During the first year, the 30-minute rule was used to accept RBCs back into inventory. Sequentially, the following year, a temperature-based approach was implemented using a thermometer with an accuracy of ±1°C. Time out of the blood bank, temperature upon return, wastage, and transfusion reactions associated with the reissued RBCs were analyzed., Results: In our practice, the 30-minute rule would have accepted 15.2% of RBC units outside of the allowed temperature. Compared to the 30-minute rule, temperature-based acceptance was associated with a 13% increase in wastage (p < 0.001). During the 30-minute rule period, transfusion of returned and subsequently reissued RBCs was associated with a nonsignificant trend toward a higher transfusion reaction rate compared to the overall RBC transfusion reaction rate (1.4% vs. 0.6%, p = 0.084). During the temperature period, transfusion of returned and subsequently reissued RBCs had the same transfusion reaction rate compared to the overall RBC transfusion reaction rate (0.5% vs. 0.5%, p = 1.0)., Conclusion: Temperature-based acceptance of returned RBCs is associated with significantly higher wastage compared to the 30-minute rule. A temperature-based acceptance practice mitigates the risk of accepting RBCs with unacceptable temperatures returned within 30 minutes of issue., (© 2018 AABB.)

Cryptic species could represent a substantial fraction of biodiversity. However, inconsistent definitions and taxonomic treatment of cryptic species prevent informed estimates of their contribution to biodiversity and impede our understanding of their evolutionary and ecological significance. We propose a conceptual framework that recognizes cryptic species based on their low levels of phenotypic (morphological) disparity relative to their degree of genetic differentiation and divergence times as compared with non-cryptic species. We discuss how application of a more rigorous definition of cryptic species in taxonomic practice will lead to more accurate estimates of their prevalence in nature, better understanding of their distribution patterns on the tree of life, and increased abilities to resolve the processes underlying their evolution., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Plant mating systems have profound effects on levels and structuring of genetic variation and can affect the impact of natural selection. Although theory predicts that intermediate outcrossing rates may allow plants to prevent accumulation of deleterious alleles, few studies have empirically tested this prediction using genomic data. Here, we study the effect of mating system on purifying selection by conducting population-genomic analyses on whole-genome resequencing data from 38 European individuals of the arctic-alpine crucifer Arabis alpina We find that outcrossing and mixed-mating populations maintain genetic diversity at similar levels, whereas highly self-fertilizing Scandinavian A. alpina show a strong reduction in genetic diversity, most likely as a result of a postglacial colonization bottleneck. We further find evidence for accumulation of genetic load in highly self-fertilizing populations, whereas the genome-wide impact of purifying selection does not differ greatly between mixed-mating and outcrossing populations. Our results demonstrate that intermediate levels of outcrossing may allow efficient selection against harmful alleles, whereas demographic effects can be important for relaxed purifying selection in highly selfing populations. Thus, mating system and demography shape the impact of purifying selection on genomic variation in A. alpina These results are important for an improved understanding of the evolutionary consequences of mating system variation and the maintenance of mixed-mating strategies., Competing Interests: The authors declare no conflict of interest.

Purpose: Current techniques for anatomic repair of the dislocated acromioclavicular (AC) joint aim on reconstruction of the AC ligaments and utilize tunnels drilled through the acromion . This improves the stability of the reconstruction but might also increase the risk of fractures at the acromion. The purpose of this study was to evaluate the fracture risk for the acromion after transacromial tunnel placement for anatomic AC joint stabilization procedure. It was hypothesized that the risk of fracture of the acromion is correlated to size and orientation of bone tunnels commonly used for anatomic AC joint reconstruction., Methods: A finite element analysis was used to simulate multiple bone tunnels and incoming force vectors (lateral vs. superior). Different tunnels were analysed, horizontal meaning an anterior-posterior orientation versus a vertical inferior-superior orientation through the acromion. Two tunnel diameters were simulated (2.4 vs. 4.5 mm). Furthermore, the tunnel length and distance between tunnels were altered. Forty-five cadaveric specimens (median age: 64 years, range 33-71 years) were utilized for data acquisition. Out of these, 30 specimens were used to evaluate basic tunnel orientations and drill diameters using a MTS 858 servohydraulic test system., Results: With regard to the tunnel orientation and drill hole size, the loads to failure were limited. The acromion is at higher fracture risk, with a superior to inferior directed incoming force. Position, size and direction of bone tunnels influenced the loads to failure. Horizontal tunnels with a higher diameter (4.5 mm) had the most impact on load to failure reduction. A long horizontal tunnel with a diameter of 4.5 mm reduced the load to failure with medial direction of force to 25% of the native acromion. The identical tunnel with a diameter of 2.4 mm reduced the load to failure to 61%. Both 2.4-mm horizontal tunnels with a medium and short length did not reduce the load to failure., Conclusion: Tunnels placed at the acromion did not result in an increased risk of fracture. However, descriptive data showed a tendency for an increased fracture risk if tunnels are placed at the acromion, especially in horizontal direction with diameters of 4.5 mm. In addition, the pattern of fracture was dependent on the orientation of the bone tunnels and the size. However, the results indicate a "safe zone" for the placement of bone tunnels within the anterior half of the acromion, which does not affect the loads to failure at the acromion. Therefore, current techniques for anatomic AC joint reconstruction which utilize fixation of grafts or sutures at the acromion are safe within current ranges of tunnel placement and sizes.

Hansen disease, also known as leprosy, is a chronic granulomatous infectious disease that is caused by Mycobacterium leprae . We report an unusual case of a 65-year-old man who presented with multiple anesthetic, annular, erythematous, scaly plaques with a raised border without any known exposures to leprosy. Histologic examination revealed a perineural lymphohistiocytic infiltrate and rare bacilli demonstrated on Fite staining. After confirmation with polymerase chain reaction (PCR) and consultation with the National Hansen's Disease Program (Baton Rouge, Louisiana), the patient was placed on a regimen of rifampicin 600 mg once monthly and dapsone 100 mg once daily for 6 months, which showed considerable improvement. This case demonstrates the identification of leprosy in central Florida, a region that is not known to be endemic to the disease. Leprosy, however rare, must be part of a practitioner's differential diagnosis even without history of traditional exposures.

Defects in maintaining genome integrity are a hallmark of cancer. The DNA damage response kinase ATM is frequently mutated in human cancer, but the significance of these events to chemotherapeutic efficacy has not been examined deeply in whole organism models. Here we demonstrate that bi-allelic Atm deletion in mouse models of Kras -mutant lung adenocarcinoma does not affect cisplatin responses. In marked contrast, Atm -deficient tumors displayed an enhanced response to the topoisomerase-II poison etoposide. Moreover, Atm -deficient cells and tumors were sensitive to the PARP inhibitor olaparib. This actionable molecular addiction to functional PARP1 signaling was preserved in models that were proficient or deficient in p53, resembling standard or high-risk genetic constellations, respectively. Atm deficiency also markedly enhanced sensitivity to the ATR inhibitor VE-822. Taken together, our results provide a functional rationale to profile human tumors for disabling ATM mutations, particularly given their impact on PARP1 and ATR inhibitors. Cancer Res; 77(11); 3040-56. ©2017 AACR ., (©2017 American Association for Cancer Research.)

Purpose: To evaluate the number of connective tissue progenitor cells (CTPs) and nucleated cells obtained during bone marrow aspiration (BMA) from the proximal humerus using either a fenestrated or a nonfenestrated trocar and determine differences in varying amounts of aspiration volume. The first hypothesis was that the number of CTPs extracted with the fenestrated trocar would be greater due to its potential to extract more cells through its fenestrations. The second hypothesis was that using consecutive aspirations with either trocar would provide a consistent number of CTPs and nucleated cells throughout the aspiration with no significant decrease of cells at the end., Methods: Patients were eligible for inclusion if they underwent primary or revision arthroscopic rotator cuff surgery, were between 18 and 75 years of age, and signed the informed consent. Between January 2011 and September 2013, 24 patients underwent BMA from the proximal humerus during arthroscopic surgery. They were grouped according to which of 3 different trocars were used for aspiration: (1) nonfenestrated, (2) fenestrated trocar A, and (3) fenestrated trocar H. Four consecutive 12 mL double syringes were used for each aspiration: 1 (0-12 mL), 2 (12-24 mL), 3 (24-36 mL), and 4 (36-48 mL). One milliliter was removed from each syringe (nonconcentrated BMA). The remainder of the BMA was then spun using a centrifuge. BMA and concentrated BMA were brought to the laboratory, counted for nucleated cells (million cells/mL BMA) and cultured for 7 days to obtain colony-forming units (CTPs/million cells)., Results: No significant differences were observed in tubes 1 to 4 in the number of nucleated cells in the nonconcentrated and concentrated BMA using the nonfenestrated trocar compared with the fenestrated trocars A and H (all P > .05), except for concentrated BMA tube 3 (P = .014) and tube 4 (P = .003). Nonconcentrated and concentrated BMA from tubes 1 to 4 had a significantly higher CTP prevalence using the nonfenestrated trocar compared with the fenestrated trocars A and H (all P < .05). Most of the times the first tube of each aspiration showed a significantly greater amount of cells and a greater CTP prevalence compared with tubes 2, 3, and 4., Conclusions: Aspiration from the proximal humerus with the nonfenestrated trocar during BMA was associated with higher prevalence of CTPs, suggesting that more CTPs can be obtained using a nonfenestrated trocar. Furthermore, CTPs can be obtained through all consecutive aspirations with a greater amount in the first tubes., Level of Evidence: Level II, prospective comparative study., (Copyright © 2016 Arthroscopy Association of North America. Published by Elsevier Inc. All rights reserved.)