An integrated spatio-temporal view of riverine biodiversity using environmental DNA metabarcoding.

Anthropogenically forced changes in global freshwater biodiversity demand more efficient monitoring approaches. Consequently, environmental DNA (eDNA) analysis is enabling ecosystem-scale biodiversity assessment, yet the appropriate spatio-temporal resolution of robust biodiversity assessment remains ambiguous. Here, using intensive, spatio-temporal eDNA sampling across space (five rivers in Europe and North America, with an upper range of 20-35 km between samples), time (19 timepoints between 2017 and 2018) and environmental conditions (river flow, pH, conductivity, temperature and rainfall), we characterise the resolution at which information on diversity across the animal kingdom can be gathered from rivers using eDNA. In space, beta diversity was mainly dictated by turnover, on a scale of tens of kilometres, highlighting that diversity measures are not confounded by eDNA from upstream. Fish communities showed nested assemblages along some rivers, coinciding with habitat use. Across time, seasonal life history events, including salmon and eel migration, were detected. Finally, effects of environmental conditions were taxon-specific, reflecting habitat filtering of communities rather than effects on DNA molecules. We conclude that riverine eDNA metabarcoding can measure biodiversity at spatio-temporal scales relevant to species and community ecology, demonstrating its utility in delivering insights into river community ecology during a time of environmental change.

The Evolving Effectiveness of Biologics in Avoiding Surgery in Children With Ulcerative Colitis: At what Nutritional Cost?

BACKGROUND: Pediatric ulcerative colitis (UC) treatment has changed dramatically with the introduction of multiple biologics. The goal of this study was to determine the effectiveness of these new biologics on achieving remission, nutritional impact, and eventual need for surgery in children. METHODS: We retrospectively analyzed hospital records of UC patients (ages 1-19) seen at a pediatric gastroenterology clinic between January 2012 andAugust 2020. Patients were divided into groups: 1) medically without biologics or surgery; 2) patients treated with one biologic; and 3) patients treated with multiple biologics 4)patients that underwent colectomy. RESULTS: There were 115 UC patients with a mean follow-up of 5.9 ± 3.7 years (1 month-15.3 years). PUCAI score at diagnosis was mild in 52 patients (45%), moderate in 25 (21%), and severe in 5 (4.3%). PUCAI score for 33 patients (29%) could not be calculated. There were 48 (41.3%) in group 1 with 58% remission, 34 (29.6%) in group 2 with 71% remission, 24 (20.8%) in group 3 with 29% remission, and only 9 (7.8%) in group 4 with 100% remission. The majority (55%) of surgical patients had colectomy within the first year of diagnosis. BMI improved after surgery (P = 0.001). The change from one biologic to others did not improve nutrition over time. DISCUSSION: New biologics are changing the landscape in maintaining remission from UC. The current need for surgery is much lower than previously published studies. In medically refractive UC, nutritional status only improved after surgery. Addition of another biologic for medically refractory ulcerative colitis in order to avoid surgery must take into account the positive impact surgery has on nutrition and disease remission.

Neonatal sevoflurane exposure induces long-term changes in dendritic morphology in juvenile rats and mice.

General anesthetics are potent neurotoxins when given during early development, causing apoptotic deletion of substantial number of neurons and persistent neurocognitive and behavioral deficits in animals and humans. The period of intense synaptogenesis coincides with the peak of susceptibility to deleterious effects of anesthetics, a phenomenon particularly pronounced in vulnerable brain regions such as subiculum. With steadily accumulating evidence confirming that clinical doses and durations of anesthetics may permanently alter the physiological trajectory of brain development, we set out to investigate the long-term consequences on dendritic morphology of subicular pyramidal neurons and expression on genes regulating the complex neural processes such as neuronal connectivity, learning, and memory. Using a well-established model of anesthetic neurotoxicity in rats and mice neonatally exposed to sevoflurane, a volatile general anesthetic commonly used in pediatric anesthesia, we report that a single 6 h of continuous anesthesia administered at postnatal day (PND) 7 resulted in lasting dysregulation in subicular mRNA levels of cAMP responsive element modulator (Crem), cAMP responsive element-binding protein 1 (Creb1), and Protein phosphatase 3 catalytic subunit alpha, a subunit of calcineurin (Ppp3ca) (calcineurin) when examined during juvenile period at PND28. Given the critical role of these genes in synaptic development and neuronal plasticity, we deployed a set of histological measurements to investigate the implications of anesthesia-induced dysregulation of gene expression on morphology and complexity of surviving subicular pyramidal neurons. Our results indicate that neonatal exposure to sevoflurane induced lasting rearrangement of subicular dendrites, resulting in higher orders of complexity and increased branching with no significant effects on the soma of pyramidal neurons. Correspondingly, changes in dendritic complexity were paralleled by the increased spine density on apical dendrites, further highlighting the scope of anesthesia-induced dysregulation of synaptic development. We conclude that neonatal sevoflurane induced persistent genetic and morphological dysregulation in juvenile rodents, which could indicate heightened susceptibility toward cognitive and behavioral disorders we are beginning to recognize as sequelae of early-in-life anesthesia.

Vaping-Related Clotting Phenomena Presenting As Central Retinal Vein Occlusion.

Central retinal vein occlusion (CRVO) typically manifests as unilateral vision loss from thrombosis and occlusion of the central retinal vein in patients with thrombophilic risk factors. Here we report a case of a 23-year-old male with three weeks of intermittent left-sided eye pressure and vision loss, who was found to have decreased visual acuity, retinal hemorrhages, and an impending CRVO in his left eye. Upon further evaluation, infectious disease and autoimmune labs were normal, but he had mildly increased right heart pressures and hypercoagulable changes in the right middle cerebral artery. He denied any personal or family history of clotting disorders but noted a four-year history of vaping. He was started on anticoagulation and discharged. Outpatient genetic testing for Factor V Leiden, protein C, protein S, and prothrombin G20210 was normal. His visual acuity returned to normal in the left eye and the retinal hemorrhages resolved. After the exclusion of organic causes, significant vaping history was considered the likely etiology of his hypercoagulable state and resultant CRVO. Vaping-related clotting phenomena may explain the etiology of an otherwise unexplained CRVO, but further investigation of the long-term health consequences of electronic cigarette use is still needed.

A Scalable Approach Reveals Functional Responses of iPSC Cardiomyocyte 3D Spheroids.

Cardiomyocytes (CMs) derived from induced pluripotent stem cells (iPSCs) provide an in vitro model of the human myocardium. Complex 3D scaffolded culture methods improve the phenotypical maturity of iPSC-CMs, although typically at the expense of throughput. We have developed a novel, scalable approach that enables the use of iPSC-CM 3D spheroid models in a label-free readout system in a standard 96-well plate-based format. Spheroids were accurately positioned onto recording electrodes using a magnetic gold-iron oxide nanoparticle approach. Remarkably, both contractility (impedance) and extracellular field potentials (EFPs) could be detected from the actively beating spheroids over long durations and after automated dosing with pharmacological agents. The effects on these parameters of factors, such as co-culture (including human primary cardiac fibroblasts), extracellular buffer composition, and electrical pacing, were investigated. Beat amplitudes were increased greater than 15-fold by co-culture with fibroblasts. Optimization of extracellular Ca2+ fluxes and electrical pacing promoted the proper physiological response to positive inotropic agonists of increased beat amplitude (force) rather than the increased beat rate often observed in iPSC-CM studies. Mechanistically divergent repolarizations in different spheroid models were indicated by their responses to BaCl2 compared with E-4031. These studies demonstrate a new method that enables the pharmacological responses of 3D iPSC-CM spheroids to be determined in a label-free, standardized, 96-well plate-based system. This approach could have discovery applications across cardiovascular efficacy and safety, where parameters typically sought as readouts of iPSC-CM maturity or physiological relevance have the potential to improve assay predictivity.

Healthcare Disparities Correlated with In-Hospital Mortality in COVID-19 Patients.

INTRODUCTION: Increasing age, male gender, African American race, and medical comorbidities have been reported as risk factors for COVID-19 mortality. We aimed to identify health-care disparities associated with increased mortality in COVID-19 patients. METHODS: We performed an observational study of all hospitalized patients with SARS-CoV2 infection from within the largest multicenter healthcare system in Southeast Michigan, from February to December, 2020. RESULTS: From 11,304 hospitalized patients, 1295 died, representing an in-hospital mortality rate of 11.5%. The mean age of hospitalized patients was 63.77 years-old, with 49.96% being males. Older age (AOR = 1.05, p < 0.0001), male gender (AOR = 1.43, p < 0.0001), divorced status (AOR = 1.25, p = 0.0256), disabled status (AOR = 1.42, p = 0.0091), and homemakers (AOR = 1.96, p = 0.0216) were significantly associated with in-hospital mortality. CONCLUSION: Older age, male gender, divorced and disabled status and homemakers were significantly associated with in-hospital mortality if they developed COVID-19. Further research should aim to identify the underlying factors driving these disparities in COVID-19 in-hospital mortality.

Translation of national guidelines into local practice.

BACKGROUND: Addressing themes raised by the Royal College of Anaesthetists National Audit Project 4, we introduced new training programmes to improve the knowledge and skills necessary for the management of airway crises. A further large-scale multimodal training programme was introduced to implement guidelines published in 2015 by the Difficult Airway Society (DAS). METHODS: In 2014, questionnaires were used to assess the knowledge necessary to manage the unanticipated difficult airway before and after high-fidelity simulation sessions. In 2016, surveys assessed knowledge of new DAS guidelines before and 5 months after the implementation of a large-scale targeted teaching programme to educate staff. RESULTS: In 2014, 20 anaesthetic teams (75 delegates) attended high-fidelity simulations and demonstrated a significant immediate improvement in knowledge. In 2016, 185 participants attended a targeted teaching programme. Although after the teaching programme an increased number of anaesthetists reported having accessed training, there was a persistent knowledge gap, with some details retained by as few as 15% of participants. DISCUSSION: Whereas the knowledge of these important guidelines was improved in the immediate aftermath of high-fidelity simulation training, the knowledge retention was disappointing, raising questions as to why knowledge of highly important techniques used in crises was so poor. We ask whether training should be compulsory and how knowledge retention might be improved across all health care disciplines that rely on guidelines for the effective management of rarely occurring but safety-critical events.

Service improvement. In the driving seat.

Overseas manufacturers and healthcare providers can provide valuable lessons to the NHS in embedding a culture of service improvement. A key lesson is that empowering staff unleashes an organisation's most powerful asset. Service improvement has been proven to enhance a organisation's year-on-year fianancial position.

A retrospective evaluation of activity of gemcitabine/platinum regimens in the treatment of recurrent ovarian cancer.

BACKGROUND: While many of these agents have been compared in prospective clinical trials, the gemcitabine/platinumbased regimens have not been compared in a prospective, randomized clinical trial. While bothgemcitabine/carboplatin and gemcitabine/cisplatin have a similar ORR in separate clinical trials, the tworegimens have never been directly been compared. With overlapping dose-limiting toxicity of thrombocytopenia, the gemcitabine/carboplatin regimen has been challenging to employ in the clinical setting in previously treated ovarian cancer patients and is often associated with treatment delays and/or dose reductions. Gemcitabine/cisplatin can also be a challenge due to its dose limiting neuropathy and renal toxicity, especially in previously treated patients. In the absence of any prospective, head to head comparison this retrospective study was embarked upon to compare the response rate and toxicity profiles of gemcitabine/cisplatin verses gemcitabine/carboplatin for the treatment of platinum-sensitive verses platinum-resistant recurrent ovarian cancer. METHODS: This was a retrospective chart review study that identified patients that had received either gemcitabine/cisplatin or gemcitabine/carboplatin for treatment of recurrent ovarian cancer and compared documented hematological and non-hematological toxicity and response based on RECIST (v1.1). Data was evaluated based upon platinum sensitivity/resistance as well. RESULTS: A total of 93 patients were identified that had received a gemcitabine/platinum regimen with 48 with recurrent ovarian cancer that were included in the study. There were 21 patients in the gemcitabine/cisplatin arm and 27 patients identified in the gemcitabine/carboplatin arm. Objective response rate (ORR) was greater in platinum-sensitive patients that received gemcitabine/carboplatin compared to gemcitabine/cisplatin (8 (67%) vs 2 (25%), p < 0.05). Conversely, ORR was greater in platinum-resistant patients treated with gemcitabine/cisplatin (4 (57%) vs 1 (25%), NS). Mean time to progression was greater in gemcitabine/cisplatin patients (7.2 vs 5.1 months, p < 0.03). Patients treated with gemcitabine/carboplatin discontinued due to toxicity at a greater rate (8 (33%) vs 5 (24%)). Specifically gemcitabine/carboplatin had a greater incidence (85%) of grade 2 or greater leukopenia, thrombocytopenia, and neutropenia compared to gemcitabine/cisplatin (19%) However, there was no significant difference in dose reductions, treatment delays, or granulocyte-colony stimulating factor (G-CSF) administration between regimens. CONCLUSIONS: Gemcitabine/cisplatin appears to have greater efficacy in platinum-resistant patients, while gemcitabine/carboplatin seems to have greater efficacy in platinum-sensitive patients. Overall, gemcitabine/carboplatin was associated with a greater incidence of myelosuppression and discontinuation due to toxicity. Similar to findings in endometrial cancer, gemcitabine/cisplatin may have benefit specifically in platinum-resistant ovarian cancer.

Development of the System for Observing Student Movement in Academic Routines and Transitions (SOSMART).

National attention on whole-of-school approaches to decrease children's sedentary behavior and increase physical activity includes movement integration (MI) in classrooms. The purpose of this study was to describe instrument development, reliability, and validity of the System for Observing Student Movement in Academic Routines and Transitions (SOSMART), designed to assess MI in elementary classrooms. An a priori conceptual framework was developed based on existing literature. The framework was expanded/refined using videos from elementary classrooms and a Delphi survey. The survey, sent to 85 experts, yielded a 38% response rate. The final system includes 11 MI variables (three categories of teacher variables, two categories of student variables) and uses a 20-second continuous interval recording format. Reliability and validity data were collected in 12 classrooms across four elementary schools. Instrument reliability was tested using interval-by-interval percentage agreement for each category. Construct validity was tested by estimating multilevel random effects logistic regression models comparing student accelerometer derived activity with the presence/absence of each MI variable. Intraobserver reliability resulted in 97.5% agreement and exceeded 80% on all variables. Construct validity was supported for 8 out of 11 MI variables. SOSMART can provide valid, reliable, and objective data about MI in elementary schools.

Head and neck paragangliomas: A two-decade institutional experience and algorithm for management.

Objectives: Paragangliomas of the head and neck and cranial base are typically benign, slow-growing tumors arising within the jugular foramen, middle ear, carotid bifurcation, or vagus nerve proper. The objective of this study was to provide a comprehensive characterization of our institutional experience with clinical management of these tumors and posit an algorithm for diagnostic evaluation and treatment. Methods: This was a retrospective cohort study of patients undergoing treatment for paragangliomas of the head and neck and cranial base at our institution from 2000-2017. Data on tumor location, catecholamine levels, and specific imaging modalities employed in diagnostic work-up, pre-treatment cranial nerve palsy, treatment modality, utilization of preoperative angiographic embolization, complications of treatment, tumor control and recurrence, and hereditary status (ie, succinate dehydrogenase mutations) were collected and summarized. Results: The mean (SD) age of our cohort was 51.8 (±16.1) years with 123 (63.4%) female patients and 71 (36.6%) male patients. Catecholamine-secreting lesions were found in nine (4.6%) patients. Fifty-one patients underwent genetic testing, with mutations identified in 43 (20 SDHD, 13 SDHB, 7 SDHD, 1 SDHA, SDHAF2, and NF1). Observation with serial imaging, surgical extirpation, radiation, and stereotactic radiosurgery were variably employed as treatment approaches across anatomic subsites. Conclusion: An algorithmic approach to clinical management of these tumors, derived from our longitudinal institutional experience and current empiric evidence, may assist otolaryngologists, radiation oncologists, and geneticists in the care of these complex neoplasms. Level of Evidence: 4.

Developing non-technical ward-round skills.

BACKGROUND: Conducting clinical 'rounds' is one of the most onerous and important duties that every junior doctor is expected to perform. There is evidence that newly qualified doctors are not adequately prepared by their undergraduate experiences for this task. The aim of this study was to analyse the challenges pertaining to non-technical skills that students would face during ward rounds, and to create a model that facilitates the transition from medical student to doctor. METHODS: A total of 217 final-year medical students completed a simulated ward round. Free-text responses were analysed using template analysis applying an a priori template developed from the literature by the research team. This drew on the generic categories of non-technical skills suggested by Flin et al. RESULTS: Ninety-seven per cent of students agreed or strongly agreed that the simulated ward round improved their insight into the challenges of ward rounds and their perceived ability to work efficiently as an active member of the ward round. The responding students (206) submitted written feedback describing the learning that they planned to use: 800 learning points were recorded, and all could be categorised into one of seven non-technical skills. Conducting clinical 'rounds' is one of the most onerous and important duties that every junior doctor is expected to perform DISCUSSION: We believe that improved task efficiency and insight into the challenges of the ward round gained by medical students will lead to an enhancement in performance during clinical rounds, and will have a positive impact on patient safety. We would suggest that undergraduate medical schools consider this model in the preparation for the clinical practice element of the curriculum.