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West Nile virus (WNV) is a zoonotic disease that is endemic in North America and is known to cause a range of symptoms from mild to life threatening in humans. Culex quinquefasciatus is one of the most prominent vectors of WNV in Southern California. The goal of this study was to identify which animal species are most fed upon by these mosquitoes in various habitats in the West Valley area of San Bernardino County, California, and determine the relationship between blood-feeding patterns and WNV activity in the region. Culex quinquefasciatus specimens were collected by West Valley Mosquito and Vector Control District during 2011 from 32 different sites. The bloodmeals of 683 individuals (92.4% of those tested) were identified using the mitochondrial gene cytochrome c oxidase 1 (COI). These bloodmeals comprised 29 vertebrate species across four different habitats. Species richness (ranging from 10 to 17) was not significantly different between habitats when rarified to account for sample size. Across habitats, the highest percentage of avian bloodmeals were taken from house sparrows (18.8–39.1%) and house finches (2.6–31.5%). Bloodmeals were identified from five mammalian species, accounting for 5.1–59.2% of bloodmeals by habitat, including humans (0–4.1%). A seasonal shift towards increased mammalian bloodmeal prevalence, specifically for domestic dog and human bloodmeals, was observed in urban habitats. The WNV activity during 2011 in San Bernardino County occurred mostly in urban and suburban areas as indicated by minimum infection rate (MIR) in Culex quinquefasciatus, notable as all human bloodmeals were identified from these two habitats.

Supplemental Digital Content is available in the text., Background: Knowledge gaps remain in the epidemiology and clinical implications of myocardial injury in coronavirus disease 2019 (COVID-19). We aimed to determine the prevalence and outcomes of myocardial injury in severe COVID-19 compared with acute respiratory distress syndrome (ARDS) unrelated to COVID-19. Methods: We included intubated patients with COVID-19 from 5 hospitals between March 15 and June 11, 2020, with troponin levels assessed. We compared them with patients from a cohort study of myocardial injury in ARDS and performed survival analysis with primary outcome of in-hospital death associated with myocardial injury. In addition, we performed linear regression to identify clinical factors associated with myocardial injury in COVID-19. Results: Of 243 intubated patients with COVID-19, 51% had troponin levels above the upper limit of normal. Chronic kidney disease, lactate, ferritin, and fibrinogen were associated with myocardial injury. Mortality was 22.7% among patients with COVID-19 with troponin under the upper limit of normal and 61.5% for those with troponin levels >10 times the upper limit of normal (P

Organoid models of early tissue development have been produced for the intestine, brain, kidney and other organs, but similar approaches for the heart have been lacking. Here we generate complex, highly structured, three-dimensional heart-forming organoids (HFOs) by embedding human pluripotent stem cell aggregates in Matrigel followed by directed cardiac differentiation via biphasic WNT pathway modulation with small molecules. HFOs are composed of a myocardial layer lined by endocardial-like cells and surrounded by septum-transversum-like anlagen; they further contain spatially and molecularly distinct anterior versus posterior foregut endoderm tissues and a vascular network. The architecture of HFOs closely resembles aspects of early native heart anlagen before heart tube formation, which is known to require an interplay with foregut endoderm development. We apply HFOs to study genetic defects in vitro by demonstrating that NKX2.5-knockout HFOs show a phenotype reminiscent of cardiac malformations previously observed in transgenic mice., Heart-forming organoids model early cardiac development.

BACKGROUND: Home telemonitoring has been used with discharged patients in an attempt to reduce 30-day readmissions with mixed results. OBJECTIVE: To assess whether home 30-day telemonitoring after discharge for patients at high risk of readmission would reduce readmissions or mortality. DESIGN: Prospective, randomized controlled trial. PATIENTS: We compared 30-day readmission rates and mortality for patients at high risk for readmission who received home telemonitoring versus standard care between November 1, 2014, and November 30, 2018, in 2 tertiary care hospitals. INTERVENTIONS: The intervention group received home-installed equipment to measure blood pressure, heart rate, pulse oximetry, weight if heart failure was present, and glucose if diabetes was present. Results were transmitted daily and reviewed by a nurse. Both groups received standard care. MAIN MEASURES: The primary outcome was a composite end point of hospital readmission or death within 30 days after discharge. The secondary outcome was an emergency department visit within 30 days after discharge. KEY RESULTS: A total of 1380 participants (mean [SD] age, 66 [14] years; 722 [52.3%] men and 658 [47.7%] women) participated in this study. Using a modified intention-to-treat analysis, the risk of readmission or death within 30 days among patients at high readmission risk was 23.7% (137/578) in the control group and 18.2% (87/477) in the telemonitoring group (absolute risk difference, − 5.5% [95% CI, − 10.4 to − 0.6%]; relative risk, 0.77 [95% CI, 0.61 to 0.98]; P = .03). Emergency department visits occurred within 30 days after discharge in 14.2% (81/570) of patients in the control group and 8.6% (40/464) of patients in the telemonitoring group (absolute risk difference, − 5.6% [95% CI, − 9.4 to − 1.8%]; relative risk, 0.61 [95% CI, 0.42 to 0.87]; P = .005). CONCLUSIONS: Thirty days of postdischarge telemonitoring may reduce readmissions of high-risk patients. TRIAL REGISTRATION: ClinicalTrials.gov identifier: NCT02136186. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s11606-020-06589-1.

Background Risk factors for progression of coronavirus 2019 (COVID-19) to severe disease or death are underexplored in U.S. cohorts. Objective To determine the factors on hospital admission that are predictive of severe disease or death from COVID-19. Design Retrospective cohort analysis. Setting Five hospitals in the Maryland and Washington, DC, area. Patients 832 consecutive COVID-19 admissions from 4 March to 24 April 2020, with follow-up through 27 June 2020. Measurements Patient trajectories and outcomes, categorized by using the World Health Organization COVID-19 disease severity scale. Primary outcomes were death and a composite of severe disease or death. Results Median patient age was 64 years (range, 1 to 108 years); 47% were women, 40% were Black, 16% were Latinx, and 21% were nursing home residents. Among all patients, 131 (16%) died and 694 (83%) were discharged (523 [63%] had mild to moderate disease and 171 [20%] had severe disease). Of deaths, 66 (50%) were nursing home residents. Of 787 patients admitted with mild to moderate disease, 302 (38%) progressed to severe disease or death: 181 (60%) by day 2 and 238 (79%) by day 4. Patients had markedly different probabilities of disease progression on the basis of age, nursing home residence, comorbid conditions, obesity, respiratory symptoms, respiratory rate, fever, absolute lymphocyte count, hypoalbuminemia, troponin level, and C-reactive protein level and the interactions among these factors. Using only factors present on admission, a model to predict in-hospital disease progression had an area under the curve of 0.85, 0.79, and 0.79, at day 2, 4, and 7, respectively. Limitation The study was done in a single health care system. Conclusion A combination of demographic and clinical variables is strongly associated with severe COVID-19 disease or death and their early onset. The COVID-19 Inpatient Risk Calculator (CIRC), using factors present on admission, can inform clinical and resource allocation decisions. Primary funding source Hopkins inHealth and COVID-19 Administrative Supplement for the HHS Region 3 Treatment Center from the Office of the Assistant Secretary for Preparedness and Response.

Emergency care in Germany is in transition. Emergency departments (EDs) treat their patients based on symptoms and acuity. However, this perspective is not reflected in claims data. The aim of the AKTIN project was to establish an Emergency Department Data Registry as a data privacy-compliant infrastructure for the use of routine medical data.Data from the respective documentation systems are continuously transmitted to local data warehouses using a standardized interface. They are available for several applications such as internal reports but also multicentre studies, in compliance with data privacy regulations. Based on a 12-months period we evaluate the population with focus on acuity assessment (triage) and vital parameters in combination with presenting complaints.For the period April 2018 to March 2019, 436,149 cases from 15 EDs were available. A triage level is documented in 86.0% of cases, and 70.5% were triaged within 10 min of arrival. Ten EDs collected a presenting complaint regularly (82.3%). The respective documentation of vital signs shows plausible patterns.The AKTIN registry provides an almost real-time insight into German EDs, regardless of the primary documentation system and health insurance claims data. The Federal Joint Committee's requirements are largely met. Standardized presenting complaints allow for symptom-based analyses as well as health surveillance.HINTERGRUND: Die Notfallversorgung befindet sich im Umbruch. In Notaufnahmen werden Patienten ausgehend von Symptomen und Dringlichkeit versorgt; dies bildet sich jedoch in den etablierten Routinedaten der gesetzlichen Sozialversicherung nicht ab. Ziel des AKTIN-Projekts war der Aufbau einer datenschutzkonformen Registerinfrastruktur zur Nutzung von klinischen Routinedaten aus Notaufnahmen.Über eine standardisierte Schnittstelle werden aus den verschiedenen Dokumentationssystemen kontinuierlich Daten der Notaufnahmebehandlung in ein lokales Data Warehouse exportiert. Dort stehen sie sowohl für lokale Nutzungen, wie interne Berichte und Qualitätsmanagement, als auch gleichzeitig datenschutzkonform für multizentrische Auswertungen zur Verfügung. Anhand der Registerpopulation wird die Ersteinschätzung sowie die Erhebung von Vitalparametern in Abhängigkeit von Vorstellungsgründen für einen 12-Monats-Zeitraum analysiert.Für den Zeitraum 04/2018 bis 03/2019 wurden 436.149 gültige Fälle aus 15 Notaufnahmen übermittelt. In 86,0 % der Fälle ist eine Ersteinschätzung dokumentiert. Diese fand in 70,5 % innerhalb von 10 min nach Ankunft des Patienten statt. In 10 Kliniken wird regelhaft (82,3 %) ein Vorstellungsgrund erfasst. Die Erfassung von Vitalparametern variiert plausibel zwischen den Vorstellungsgründen.Das AKTIN-Notaufnahmeregister bietet einen zeitnahen Einblick in das Versorgungsgeschehen der Notaufnahmen ohne zusätzlichen Dokumentationsaufwand und unabhängig vom primären IT-System, Kostenträger, Fallart und Abrechnungsmodus. Die Vorgaben des Gemeinsamen Bundesausschusses zur Ersteinschätzung werden weitgehend umgesetzt. Durch die Etablierung von standardisierten Vorstellungsgründen werden symptombasierte Analysen und Gesundheitssurveillance ermöglicht.

BACKGROUND: B-cell affinity maturation in germinal center relies on regulated actin dynamics for cell migration and cell-to-cell communication. Activating mutations in the cytoskeletal regulator Wiskott-Aldrich syndrome protein (WASp) cause X-linked neutropenia (XLN) with reduced serum level of IgA. OBJECTIVE: We investigated the role of B cells in XLN pathogenesis. METHODS: We examined B cells from 6 XLN patients, 2 of whom had novel R268W and S271F mutations in WASp. By using immunized XLN mouse models that carry the corresponding patient mutations, WASp L272P or WASp I296T, we examined the B-cell response. RESULTS: XLN patients had normal naive B cells and plasmablasts, but reduced IgA+ B cells and memory B cells, and poor B-cell proliferation. On immunization, XLN mice had a 2-fold reduction in germinal center B cells in spleen, but with increased generation of plasmablasts and plasma cells. In vitro, XLN B cells showed reduced immunoglobulin class switching and aberrant cell division as well as increased production of immunoglobulin-switched plasma cells. CONCLUSIONS: Overactive WASp predisposes B cells for premature differentiation into plasma cells at the expense of cell proliferation and immunoglobulin class switching. ispartof: JOURNAL OF ALLERGY AND CLINICAL IMMUNOLOGY vol:149 issue:3 pages:1069-1084 ispartof: location:United States status: published

Summary Human pluripotent stem cell-derived cardiomyocytes (hPSC-CMs) represent an attractive model to investigate CM function and disease mechanisms. One characteristic marker of ventricular specificity of human CMs is expression of the ventricular, slow β-myosin heavy chain (MyHC), as opposed to the atrial, fast α-MyHC. The main aim of this study was to investigate at the single-cell level whether contraction kinetics and electrical activity of hESC-CMs are influenced by the relative expression of α-MyHC versus β-MyHC. For effective assignment of functional parameters to the expression of both MyHC isoforms at protein and mRNA levels in the very same hESC-CMs, we developed a single-cell mapping technique. Surprisingly, α- versus β-MyHC was not related to specific contractile or electrophysiological properties of the same cells. The multiparametric cell-by-cell analysis suggests that in hESC-CMs the expression of genes associated with electrical activity, contraction, calcium handling, and MyHCs is independently regulated., Highlights • Single-cell mapping technique allows multiparametric characterization of hESC-CMs • Twitch and action potential kinetics are independent of MyHC isoform in hESC-CMs • Myosin-mRNA levels are only weakly correlated with respective protein levels, In this article, Natalie Weber and colleagues developed a single-cell mapping technique for multiparametric analysis of hESC cardiomyocytes (hESC-CMs). Characterization of function (twitch and action potential) in direct relation to the expressed myosin heavy chain isoform by multiparametric analysis suggests that in hESC-CMs the expression of genes associated with electrical activity, contraction, calcium handling, and myosin heavy chains is independently regulated.

Barttin is the accessory subunit of the human ClC-K chloride channels, which are expressed in both the kidney and inner ear. Barttin promotes trafficking of the complex it forms with ClC-K to the plasma membrane and is involved in activating this channel. Barttin undergoes post-translational palmitoylation that is essential for its functions, but the enzyme(s) catalyzing this post-translational modification is unknown. Here, we identified zinc finger DHHC-type containing 7 (DHHC7) protein as an important barttin palmitoyl acyltransferase, whose depletion affected barttin palmitoylation and ClC-K-barttin channel activation. We investigated the functional role of barttin palmitoylation in vivo in Zdhhc7(−/−) mice. Although palmitoylation of barttin in kidneys of Zdhhc7(−/−) animals was significantly decreased, it did not pathologically alter kidney structure and functions under physiological conditions. However, when Zdhhc7(−/−) mice were fed a low-salt diet, they developed hyponatremia and mild metabolic alkalosis, symptoms characteristic of human Bartter syndrome (BS) type IV. Of note, we also observed decreased palmitoylation of the disease-causing R8L barttin variant associated with human BS type IV. Our results indicate that dysregulated DHHC7-mediated barttin palmitoylation appears to play an important role in chloride channel dysfunction in certain BS variants, suggesting that targeting DHHC7 activity may offer a potential therapeutic strategy for reducing hypertension.

Complete tracheal ring deformity (CTRD) is a rare abnormality of unknown etiology characterized by circumferentially continuous cartilaginous tracheal rings leading to variable degrees of tracheal stenosis with or without additional heart and lung malformations. Pleuropulmonary blastomas (PPB) are rare malignant mesenchymal tumors, which occur almost exclusively in young children. Pathogenic germlineKomplette Trachealringe (Complete tracheal ring deformity/CTRD) sind seltene angeborene Fehlbildungen unklarer Genese, welche zu Trachealstenosen verschiedener Schweregrade führen. CTRD können isoliert oder gemeinsam mit weiteren kardiopulmonalen Fehlbildungen auftreten. Pleuropulmonale Blastome (PBB) sind seltene maligne mesenchymale Tumore, die fast ausschließlich im frühen Kindesalter auftreten. Pathogene Keimbahnvarianten im DICER1-Gen sind mit PPB assoziiert, stehen aber auch in Zusammenhang mit anderen Tumoren wie Rhabdomyosarkomen oder syndromalen Erkrankungen wie dem GLOW (global developmental delay, lung cysts, overgrowth and Wilms tumor)-Syndrom. Wir berichten über einen Fall mit CTRD und rezidivierenden Pneumothoraces mit histologisch nachgewiesenem PPB und einer pathogenen Mutation im DICER1-Gen.

SARS-CoV-2 infection induces severe disease in a subpopulation of patients, but the underlying mechanisms remain unclear. We demonstrate robust IgM autoantibodies that recognize angiotensin converting enzyme-2 (ACE2) in 18/66 (27%) patients with severe COVID-19, which are rare (2/52; 3.8%) in hospitalized patients who are not ventilated. The antibodies do not undergo class-switching to IgG, suggesting a T-independent antibody response. Purified IgM from anti-ACE2 patients activates complement. Pathological analysis of lung obtained at autopsy shows endothelial cell staining for IgM in blood vessels in some patients. We propose that vascular endothelial ACE2 expression focuses the pathogenic effects of these autoantibodies on blood vessels, and contributes to the angiocentric pathology observed in some severe COVID-19 patients. These findings may have predictive and therapeutic implications., One-sentence summary: ACE2 autoantibodies in severe COVID-19 have features of a T-independent immune response, and may mediate vascular damage.

Progress in the systemic control of osteosarcoma has been limited over the past decades thus indicating the urgent clinical need for the development of novel treatment strategies. Therefore, we have recently developed new preclinical models to study promising novel agents for the treatment of pediatric osteosarcoma. The checkpoint kinase (chk) inhibitor prexasertib (LY2606368) and its salt form (LSN2940930) have recently been shown to be active in adult and pediatric malignancies, including sarcoma. We have now tested the potency of prexasertib in clonogenic survival assays in two new lines of primary patient‐derived osteosarcoma cells and in two established osteosarcoma cell lines as a single agent and in combination with cisplatin and the poly ADP‐ribose polymerase (PARP) inhibitor talazoparib. Prexasertib alone results in strongly reduced clonogenic survival at low nanomolar concentrations and acts by affecting cell cycle progression, induction of apoptosis and induction of double‐stranded DNA breakage at concentrations that are well below clinically tolerable and safe plasma concentrations. In combination with cisplatin and talazoparib, prexasertib acts in a synergistic fashion. Chk1 inhibition by prexasertib and its combination with the DNA damaging agent cisplatin and the PARP‐inhibitor talazoparib thus emerges as a potential new treatment option for pediatric osteosarcoma which will now have to be tested in preclinical primary patient derived in vivo models and clinical studies., What's new? Treatment options and outcomes for osteosarcoma have changed very little in decades. New treatment strategies are thus urgently needed. In this study, the authors found that the checkpoint inhibitor ‘prexasertib’ increased apoptosis and DNA damage in patient‐derived osteosarcoma cells, at very low concentrations. In addition, when combined with cisplatin and talazoparib, prexasertib acted in a synergistic fashion. This drug thus represents a promising therapeutic candidate for further preclinical and clinical development in osteosarcoma.

Free-living amoeba (FLA) group includes the potentially pathogenic genera Acanthamoeba, Naegleria, Balamuthia, Sappinia, and Vermamoeba, causative agents of human infections (encephalitis, keratitis, and disseminated diseases). In Brazil, the first report on pathogenic FLA was published in the 70s and showed meningoencephalitis caused by Naegleria spp. FLA studies are emerging, but no literature review is available to investigate this trend in Brazil critically. Thus, the present work aims to integrate and discuss these data. Scopus, PubMed, and Web of Science were searched, retrieving studies from 1974 to 2020. The screening process resulted in 178 papers, which were clustered into core and auxiliary classes and sorted into five categories: wet-bench studies, dry-bench studies, clinical reports, environmental identifications, and literature reviews. The papers dating from the last ten years account for 75% (134/178) of the total publications, indicating the FLA topic has gained Brazilian interest. Moreover, 81% (144/178) address Acanthamoeba-related matter, revealing this genus as the most prevalent in all categories. Brazil’s Southeast, South, and Midwest geographic regions accounted for 96% (171/178) of the publications studied in the present work. To the best of our knowledge, this review is the pioneer in summarising the FLA research history in Brazil.

Campylobacter is one of the leading bacterial foodborne pathogens worldwide. Poultry is the host species with this pathogen with the highest clinical impact. Flocks become colonised with Campylobacter, which leads to contamination of product entering the food-chain. Rapid and reliable Campylobacter detection methods could support controls to minimize the risks of contamination within the food-chain, which would easier enable the implementation of a logistical slaughter schedule or other control options. The present study evaluates current and emerging C. jejuni detection technologies on air samples in a unique study set-up of pre-defined C. jejuni prevalences. Both non-invasive detection technologies on air samples by subsequent measuring of volatile organic compounds (VOCs) or by qPCR detected the C. jejuni presence and could additionally distinguish between the number of present C. jejuni-positive birds in the study set-up. Nevertheless, electrostatic air samplers diagnosed fewer birds as C. jejuni-positive compared to the cultivation-based method. By measuring the VOCs, it was possible to detect the presence of two positive birds in the room. This apparent high sensitivity still needs to be verified in field studies. Techniques, such as these promising methods, that can facilitate C. jejuni surveillance in poultry flocks are desirable to reduce the risk of infection for humans.

Necrotic enteritis (NE) is an important intestinal disease of commercial poultry associated with poor performance, high mortality, and significant economic loss. In this case report, a novel presentation of NE is described in young broilers. Initially, affected farms presented with a moderate increase in mortality at or before 19 days of age. Farms experiencing the disease syndrome belonged to two complexes of the same company. However, all farms sourced chicks from the same hatchery. Farm postmortem examinations revealed moderate to severe enteritis of the upper small intestine characterized by multifocal, irregular, plaque-like, mucosal ulcerations. Additionally, thinning of the intestinal wall with consequential distension and ballooning and a necrotic, pseudomembranous layer covering the mucosa were observed in some birds. Clinically affected birds were submitted to the Poultry Research and Diagnostic Laboratory at Mississippi State University for further evaluation. Birds were between 5 and 11 days of age and presented with similar gross lesions. Anaerobic culture was performed, and

The objective of this study was to compare the combination of intramuscular (IM) droperidol/midazolam to haloperidol/lorazepam regarding time to sedation in patients with acute undifferentiated agitation in the emergency department (ED).This was a prospective, unblinded observational study in the ED of a university teaching hospital. Subjects with acute undifferentiated agitation refractory to verbal de-escalation were assigned to receive a combination of either haloperidol 5 mg/lorazepam 2 mg or droperidol 5 mg/midazolam 5 mg IM. The primary outcome was the proportion of patients adequately sedated at 10 min defined as ED Sedation Assessment Tool (SAT) score of 0 or less. Secondary outcomes included change in ED SAT score at 5, 15, 30, and 60 min, the need for oxygen supplementation, and the need for airway intervention.A total of 86 patients were enrolled in the study, with 43 patients receiving droperidol/midazolam and 43 patients receiving haloperidol/lorazepam. Ten minutes after receiving medication, 51.2% of patients in the droperidol/midazolam group were adequately sedated compared to 7% of patients in the haloperidol/lorazepam group (OR: 14; 95% CI: 3.7, 52.1). Median time to adequate sedation was 10 min for the droperidol/midazolam group and 30 min for the haloperidol/lorazepam group. Eleven patients (25.6%) in the droperidol/midazolam group received oxygen supplementation compared to four patients (9.3%) in the haloperidol/lorazepam group. No study patients experienced extrapyramidal symptoms or required endotracheal intubation.Intramuscular droperidol/midazolam was superior to intramuscular haloperidol/lorazepam in achieving adequate sedation at 10 min. Patients in the droperidol/midazolam arm may be more likely to receive oxygen supplementation than those in the haloperidol/lorazepam arm.

Ecosystem services such as food provisioning, climate regulation, nutrient cycling, or recreation in open landscapes underpin human wellbeing. They are highly dependent on land use, land cover and utilization pattern as well as environmental factors like climate, topography and soil. In consequence, ecosystem services supply shows a high spatial variability. However, it is less clear if the perception of the importance of ecosystem services is similarly heterogeneous in space and amongst societal actors. The aim of this large-scale study was to explore whether land cover and climate gradients as well as socio-cultural factors influence the perceptions of ecosystem services of four groups of societal actors: citizens, farmers, foresters and nature managers. Spatially explicit survey data of 3018 respondents allowed to gain insight into the distribution of perceived importance of 21 ecosystem services in the federal state of Bavaria, Germany together with the respondents’ socio-cultural characterisation (e.g. gender, education and hobbies in nature). Responses were analysed through descriptive statistics, redundancy analysis, and Generalized Linear Models. Results reveal that the perceived importance of many ecosystem services was consistently high across groups, although perception differed for some ecosystem services (e.g. production of energy plants and timber as well as recreation in urban green space). Compared to other actor groups, farmers attributed slightly lower importance to all ES except provisioning services. Socio-cultural factors better explained variability in perceived importance of ecosystem services than land cover and climate gradients. This might be either explained by the fact that the environmental gradients vary not strong enough in our case study or that they do not shape the perceptions of respondents. A limitation of the study is that the sample of respondents obtained is not representative for the population, but biased towards persons interested in the topics of the survey. Still the consensus indicated by the overall positive perception of ecosystem services among respondents highlights the integrative potential of ecosystem services when included in decision-making.

Task-unrelated thoughts (TUTs) are frequent distractions from our everyday tasks, which can reduce productivity and safety during task performance. This necessitates the examination of factors that modulate TUT occurrence in daily life. One factor that has previously been implicated as a source of TUT is personally salient concerns. External factors such as news media serve as cues for these concerns, potentially increasing TUT occurrence. However, this has not been thoroughly examined in everyday life settings. We thus utilized Ecological Momentary Assessment to survey participants throughout the day for ten days, on their TUTs and news consumption in two separate studies. Study 1 focused on the impact of news related to the Coronavirus Disease (COVID-19) pandemic, as a common and global concern. We found that TUT occurrence was significantly predicted by COVID-19 news consumption, such that TUT occurrence increased with COVID-19 news consumption. To extend these findings, we implemented Study 2 using similar methods, but focusing on the consumption of news media in general. TUT occurrence was predicted by general news consumption, with an increase in reports of TUT following consumption of news media in general. We thus extended the association found between TUT and COVID-19-related news in Study 1, to any news topic in Study 2. We speculate that the increase in TUTs was due to heightened salience of current concerns, cued by the news. These findings highlight the importance of considering when we choose to consume news media and the value of examining contextual factors when studying TUTs in daily life.

Patient: Female, 19-year-old Final Diagnosis: Poisoning by bupropion Symptoms: Altered mental status • cardiogenic shock • seizure Medication: — Clinical Procedure: — Specialty: Toxicology Objective: Management of emergency care Background: Bupropion is a norepinephrine/dopamine-reuptake inhibitor (NDRI) that has been reported to increase the risk of suicide attempts in some patients. This report is of a case of a 19-year-old woman with a history of depression who suffered fatal cardiorespiratory failure following an overdose of prescribed bupropion. Case Report: A 19-year-old woman presented to the Emergency Department with an estimated bupropion overdose of 28.2 g and possible oxcarbazepine co-ingestion. This serum level was estimated based on the patient’s history of medication reconciliation and number of pills remaining in the prescription bottle at presentation. The patient was unresponsive on arrival to the Emergency Department and was treated for intermittent seizures and shock. Despite aggressive medical interventions, her condition progressed to cardiogenic shock and eventually cardiac arrest, from which she could not be resuscitated. Several existing reports regarding bupropion overdose describe sinus tachycardia and seizures corrected by symptomatic treatment. This case may document the highest reported ingestion of bupropion recorded thus far in the literature and demonstrates the rapid onset of cardiac dysfunction and cardiogenic shock. Conclusions: In the context of this case, we discuss the clinical manifestations of bupropion overdose and the rapid progression to cardiogenic shock. By examining the pathophysiology of overdose in an adolescent who consumed an extremely high dose of bupropion, we hope this information can be helpful to clinicians who are managing similarly challenging critical cases.

Contradictory reports of play strengths and weaknesses for children with autism spectrum disorders (ASD) persist in the literature. We compared the play of 19 children with ASD to 19 typically developing (TD) children matched on language and cognitive skills. All children were verbal. Results revealed no differences in indiscriminate actions, functional play, and object interest. The children with ASD showed less symbolic play and a significantly fewer number of children met criteria for emerging or mastered symbolic play. A specific deficit was observed for “doll as agent” symbolic play. Outcomes suggest that compared to children without disabilities, children with ASD may have comparable functional play skills and struggle with the transition to some, but not all types of symbolic play.

With the appearance of publicly available, high-resolution, physiological datasets in neurocritical care, like Collaborative European NeuroTrauma Effectiveness Research in Traumatic Brain Injury (CENTER-TBI), there is a growing need for tools that could be used by clinical researchers to interrogate this information-rich data. The ICM+ software is widely used for processing data acquired from bedside monitors. Considering the growing popularity of scripting simple-syntax programming languages like Python, particularly among clinical researchers, we have developed an interface in ICM+ that provides a streamlined way of adding Python scripting functionality to the ICM+ calculation engine. The new interface imposes certain requirements on the scripts and needs an accompanying descriptor file that tells ICM+ about the functions implemented, so that they become available to the end user in the same way as native ICM+ functions. ICM+ also now includes a tool that eases the creation of Python functions to be imported. The Python extension works very efficiently, and any user with some degree of experience in scripting can use it to enrich capabilities of ICM+. Depending on the data analysed and calculations performed, Python functions are 15-60% slower than built-in ICM+ functions, which is a more-than-acceptable trade-off for empowering ICM+ with the unlimited analytical freedom offered by extensive Python libraries.

With the appearance of publicly available, high-resolution, physiological datasets in neurocritical care, like Collaborative European NeuroTrauma Effectiveness Research in Traumatic Brain Injury (CENTER-TBI), there is a growing need for tools that could be used by clinical researchers to interrogate this information-rich data. The ICM+ software is widely used for processing data acquired from bedside monitors. Considering the growing popularity of scripting simple-syntax programming languages like Python, particularly among clinical researchers, we have developed an interface in ICM+ that provides a streamlined way of adding Python scripting functionality to the ICM+ calculation engine. The new interface imposes certain requirements on the scripts and needs an accompanying descriptor file that tells ICM+ about the functions implemented, so that they become available to the end user in the same way as native ICM+ functions. ICM+ also now includes a tool that eases the creation of Python functions to be imported. The Python extension works very efficiently, and any user with some degree of experience in scripting can use it to enrich capabilities of ICM+. Depending on the data analysed and calculations performed, Python functions are 15-60% slower than built-in ICM+ functions, which is a more-than-acceptable trade-off for empowering ICM+ with the unlimited analytical freedom offered by extensive Python libraries.

Osteoid osteomas belong to the benign osseous neoplasia and can occur on the entire skeleton. The prolonged diagnosis of an osteoid osteoma of the great toe prompted us to prepare a case report with a literature review to identify possible sources of error and to contribute to a more expeditious diagnosis and therapy.Currently, only two case reports on osteoid osteomas of the toes are known in the German literature. Internationally, however, a large number of cases have been described, which, when viewed as a whole, present a homogeneous picture. A very clear accumulation of osteoid osteomas at the distal phalanx of the great toe suggests a possible post-traumatic aetiology. The typical NSAID-sensitive night pain is clinically unambiguous and should make one think of an osteoid osteoma at any site on the body. The symptom of macrodactyly, which is unique to the toes, is striking and may provide a helpful clue to establishing the diagnosis. The median time from symptom onset to correct diagnosis was 12 months. One possible reason for this prolonged diagnosis is the often atypical and ambiguous imaging. Surgical therapy is the method of choice for osteoid osteomas of the toes and should always be supplemented by histopathologic examination.HINTERGRUND: Osteoidosteome zählen zu den benignen ossären Neoplasien und können am gesamten Skelett auftreten. Die verlängerte Diagnosestellung eines Osteoidosteoms der Großzehe veranlasste uns, einen Fallbericht mit Review der Literatur zu verfassen, um mögliche Fehlerquellen zu identifizieren und einen Beitrag zur zügigeren Diagnosefindung und Therapie zu leisten.In der deutschen Literatur sind aktuell nur zwei Fallberichte zu Osteoidosteomen der Zehen bekannt. International wurde allerdings eine Vielzahl an Fällen beschrieben, die in der Zusammenschau ein homogenes Bild ergeben. Eine sehr deutliche Häufung von Osteoidosteomen am Endglied der Großzehe lassen eine posttraumatische Ätiologie möglich erscheinen. Der typische NSAR-sensible Nachtschmerz ist klinisch eindeutig und sollte an jeder Lokalität des Körpers an ein Osteoidosteom denken lassen. Auffällig ist das an den Zehen einzigartige Symptom der Makrodaktylie, was ein dankbarer Hinweis zur Diagnosefindung sein kann. Die mittlere Zeit vom Symptombeginn bis zur korrekten Diagnosestellung betrug 12 Monate. Häufig wurden Fehldiagnosen und Fehltherapien unter einer anderen Verdachtsdiagnose beschrieben. Eine mögliche Ursache ist die häufig atypische und uneindeutige Bildgebung. Die operative Therapie stellt bei Osteoidosteomen der Zehen die Methode der Wahl dar und sollte stets durch eine histopathologische Untersuchung ergänzt werden.

Protein tyrosine phosphatases (PTPs) are key virulence factors in pathogenic bacteria, consequently, they have become important targets for new approaches against these pathogens, especially in the fight against antibiotic resistance. Among these targets of interest YopH (Yersinia outer protein H) from virulent species of Yersinia is an example. PTPs can be reversibly inhibited by nitric oxide (NO) since the oxidative modification of cysteine residues may influence the protein structure and catalytic activity. We therefore investigated the effects of NO on the structure and enzymatic activity of Yersinia enterocolitica YopH in vitro. Through phosphatase activity assays, we observe that in the presence of NO YopH activity was inhibited by 50%, and that this oxidative modification is partially reversible in the presence of DTT. Furthermore, YopH S-nitrosylation was clearly confirmed by a biotin switch assay, high resolution mass spectrometry (MS) and X-ray crystallography approaches. The crystal structure confirmed the S-nitrosylation of the catalytic cysteine residue, Cys403, while the MS data provide evidence that Cys221 and Cys234 might also be modified by NO. Interestingly, circular dichroism spectroscopy shows that the S-nitrosylation affects secondary structure of wild type YopH, though to a lesser extent on the catalytic cysteine to serine YopH mutant. The data obtained demonstrate that S-nitrosylation inhibits the catalytic activity of YopH, with effects beyond the catalytic cysteine. These findings are helpful for designing effective YopH inhibitors and potential therapeutic strategies to fight this pathogen or others that use similar mechanisms to interfere in the signal transduction pathways of their hosts.

CD81 plays a role in a variety of physiological and pathological processes. Recent structural analysis of CD81 indicates that it contains an intramembrane cholesterol-binding pocket and that interaction with cholesterol may regulate a conformational switch in the extracellular domain of CD81. Therefore, CD81 possesses a potential cholesterol sensing mechanism; however, its relevance for protein function is thus far unknown. In this study we investigate CD81 cholesterol sensing in the context of its activity as a receptor for hepatitis C virus. Structure-led mutagenesis of the cholesterol-binding pocket reduced CD81-cholesterol association, but had disparate effects on HCV, both reducing and enhancing CD81 receptor activity. We reasoned that this could be explained by alterations in the consequences of cholesterol binding. To investigate this further we performed molecular dynamic simulations of CD81 with and without cholesterol; this identified an allosteric mechanism by which cholesterol binding regulates the conformation of CD81. To test this, we designed further mutations to force CD81 into either the open (cholesterol unbound) or closed (cholesterol bound) conformation. The open mutant of CD81 exhibited reduced receptor activity whereas the closed mutant was enhanced. These data are consistent with cholesterol switching CD81 between a receptor active and inactive state. CD81 interactome analysis also suggests that conformational switching may modulate the assembly of CD81-partner networks. This work furthers our understanding of the molecular mechanism of CD81 cholesterol sensing, how this relates to HCV entry and CD81's function as a molecular scaffold; these insights are relevant to CD81's varied roles in health and disease.

Modern research projects in healthcare research and medical research are oftentimes multi-centered, multi-disciplinary, and conducted by a consortium of multiple collaborators. Increasingly, the resulting data emanating emerging from different primary and secondary sources is linked on a personal level. The General Data Protection Regulation regulates many fundamental processes in such research projects. Despite the regulatory framework given, it can happen that a collaborator does not handle data properly. We propose a systematic risk management for the handling of data as well as a systematic error management.

Background Symptoms of depression and anxiety experienced by undergraduate medical students have become a prominent concern. Evidence about students’ depression and anxiety including prevalence, trajectory during medical education, gender differences and comparisons with age-matched peers is conflicting. However few studies of medical students’ mental health specify the precise time of assessment. Proximity to examinations may be relevant. Precise identification of the time of data collection might help explain contradictory findings and facilitate provision of more timely support. Methods This study addressed whether: Proximity of final examinations affected students’ depression and anxiety symptomsMales and females differed in this respect. We analysed data provided by 446 final year students from 6 UK medical schools. These students were a subset of data provided by 14 UK medical schools which participated in an online survey comparing first and final year students and in which final year response rates exceeded 30%. We used the Hospital Anxiety and Depression Scale to assess symptoms of depression and anxiety and the norms to indicate potentially clinically relevant cases. We grouped students into those for whom final exams were imminent i.e. within 2 months of completing the survey (n = 164) and those for whom exams were more distant or had been taken (n = 282). We used parametric and non-parametric tests to compare both groups and gender differences in respect of depression and anxiety sum scores and cases rates. Results For both depression and anxiety male and female students facing imminent final exams recorded greater prevalence and significantly higher mean scores. The effect size of differences for anxiety were large. No substantial gender differences were found for depression. Regardless of the timing of final exams female students recorded both significantly higher mean scores and clinically relevant rates for anxiety. Conclusions Proximity to final exams negatively affected the mental health of both male and female final year students. The study suggests that there may be times in the undergraduate medical curriculum when additional or targeted support is needed. It also highlights the need for research to provide a greater specificity of context when investigating medical students’ mental health.

EXECUTIVE SUMMARY Quality improvement, regulatory, and payer organizations use various definitions of hospital mortality as clinical outcome measures. In this prospective study, the authors evaluated a multicomponent intervention aimed at reducing inpatient mortality in a multistate healthcare delivery system. The project was initiated because of a statistically nonsignificant upward trend in mortality suggested by a six-quarter rise in the observed/expected mortality ratio generated by the Vizient Clinical Data Base and Resource Manager. The design of the mortality reduction plan was influenced by the known limitations of using hospital-wide mortality as a quality improvement measure. The primary objective was to reduce mortality through focused care redesign. The project leadership team attempted to implement standardized system-wide improvements while allowing individual hospitals to simultaneously pursue site-specific practice redesign opportunities. Between Q3, 2015, and Q4, 2017, system-wide mortality reduced from 1.78 to 1.53 (per 100 admissions; p = .01). The actual plan implemented in Mayo Clinic's hospitals is included as Appendix A to this article, published online as Supplemental Digital Content. The authors included it to allow comparison with similar efforts at other healthcare systems, as well as to stimulate criticism and discussion by readers.

Several methods aid with reconstructing features of the human nose, including angle, projection and width, but only one study by Davy-Jow et al. (2012) has focused on nose tip shape. The main finding was that the shape of the nasal bridge is consistent with the shape of the nose tip. The study also theorised that the method would not be suitable for snub (upturned) noses. Although promising, further investigation with a larger sample of different origin would be of benefit. In addition, grouping samples into upturned, horizontal and downturned nose tips could reveal the need for a difference in the applied method. The approach has been recreated with a larger sample size (N = 103 versus N = 25) derived from a modern German population. Based on soft tissue models, the individuals were firstly grouped into three categories; upturned, horizontal, and downturned noses. Computed Tomography (CT) data allowed the simultaneous visualisation of both skull and (semi-transparent) facial surfaces. Each head was viewed frontally in the Frankfurt Horizontal Plane (FHP), and then tilted back until the nasal tip superimposed the nasal bridge, with the angle of tilt measured from the FHP. The results show that the angle of tilt is significantly different for upturned, horizontal, and downturned noses, but that it can be equally applied to all three groups. The mean angle was 44° for upturned noses, 51° for horizontal, and 56° for downturned. Error studies suggest a very high accuracy and repeatability with intra-class correlation coefficients of 0.991 (inter-observer error) and 0.972 (intra-observer error) respectively.

The alveolar ridge splitting technique (ARST) offers an alternative to classic ridge augmentation techniques for successful insertion of dental implants. However, the buccal lamella is at risk of fracturing during ARST distraction. To better understand the fracture mechanisms and displacement limits of the split lamella, this study conducted biomechanical tests on human cadaveric maxilla specimens having extremely atrophied alveolar ridges treated with ARST. A total of 12 standardized alveolar splits were prepared on the maxillae of 3 elderly female donors using an oscillating piezoelectric saw. Mimicking the surgical distraction process of the lamella, each split was tested to failure using a dental osteotome attached to the crosshead of an electromechanical testing system. All specimens were scanned by means of high-resolution peripheral quantitative computed tomography prior to and post testing to evaluate split geometries and failure modes. Split stiffness, failure force, and displacement were 27.4 ± 18.7 N/mm, 12.0 ± 8.4 N, and 0.97 ± 0.31 mm, with no significant differences between anatomical sides and split locations ( p ≥ 0.17 ). Stiffness correlated significantly with failure force ( R 2 = 0.71 , p < 0.01 ). None of the alveolar split widths correlated significantly with the outcomes from biomechanical testing ( p ≥ 0.10 ). The results suggest that simple geometrical measures do not predict the allowed extent of lamella distraction prior to failure. More sophisticated methods are required for surgical planning to optimize the ARST outcomes. Still, the present study may advocate a clinical protocol for the maxilla where the implant site is prepared directly after osteotomy setting and immediately before full lamella dislocation, when the lamella is still stable, resistant to mechanical stress, and bone loss caused by the abrasion of the burr is minimized.

CD40 ligand (TNFSF5/CD154/CD40L), a member of the tumor necrosis factor (TNF) superfamily is a key regulator of the immune system. The cognate receptor CD40 (TNFRSF5) is expressed broadly on antigen-presenting cells and many tumor types, and has emerged as an attractive target for immunologic cancer treatment. Most of the CD40 targeting drugs in clinical development are antibodies which display some disadvantages: their activity typically depends on Fcγ receptor-mediated crosslinking, and depletion of CD40-expressing immune cells by antibody-dependent cellular cytotoxicity compromises an efficient antitumor response. To overcome the inadequacies of antibodies, we have developed the hexavalent receptor agonist (HERA) Technology. HERA compounds are fusion proteins composed of 3 receptor binding domains in a single chain arrangement, linked to an Fc-silenced human IgG1 thereby generating a hexavalent molecule. HERA-CD40L provides efficient receptor agonism on CD40-expressing cells and, importantly, does not require FcγR-mediated crosslinking. Strong activation of NFκB signaling was observed upon treatment of B cells with HERA-CD40L. Monocyte treatment with HERA-CD40L promoted differentiation towards the M1 spectrum and repolarization of M2 spectrum macrophages towards the M1 spectrum phenotype. Treatment of in vitro co-cultures of T and B cells with HERA-CD40L-triggered robust antitumor activation of T cells, which depended upon direct interaction with B cells. In contrast, bivalent anti-CD40 antibodies and trivalent soluble CD40L displayed weak activity which critically depended on crosslinking. In vivo, a murine surrogate of HERA-CD40L-stimulated clonal expansion of OT-I-specific murine CD8 T cells and showed single agent antitumor activity in the CD40 syngeneic MC38-CEA mouse model of colorectal cancer, suggesting an involvement of the immune system in controlling tumor growth. We conclude that HERA-CD40L is able to establish robust antitumor immune responses both in vitro and in vivo.

Morphine delays oral P2Y12 platelet inhibitor absorption and is associated with adverse outcomes after myocardial infarction. Consequently, many physicians and first responders are now considering fentanyl as an alternative. We conducted a single-centre trial randomizing cardiac patients undergoing coronary angiography to intravenous fentanyl or not. All participants received local anaesthetic and intravenous midazolam. Those requiring percutaneous coronary intervention (PCI) with stenting received 180 mg oral ticagrelor intra-procedurally. The primary outcome was area under the ticagrelor plasma concentration–time curve (AUC0–24 hours). The secondary outcomes were platelet function assessed at 2 hours after loading, measured by P2Y12 reaction units (PRUs) and light transmission platelet aggregometry. Troponin-I was measured post-PCI using a high-sensitivity troponin-I assay (hs-TnI). All participants completed a survey of pain and anxiety. Of the 212 randomized, 70 patients required coronary stenting and were loaded with ticagrelor. Two participants in the no-fentanyl arm crossed over to receive fentanyl for pain. In as-treated analyses, ticagrelor concentrations were higher in the no-fentanyl arm (AUC0–24 hours 70% larger, p = 0.03). Platelets were more inhibited by 2 hours in the no-fentanyl arm (71 vs. 113 by PRU, p = 0.03, and 25% vs. 41% for adenosine diphosphate response by platelet aggregation, p Clinical Trial Registration: https://clinicaltrials.gov/ct2/show/NCT02683707 (NCT02683707).

Naegleria fowleri is a pathogenic amoeboflagellate most prominently known for its role as the etiological agent of the Primary Amoebic Meningoencephalitis (PAM), a disease that afflicts the central nervous system and is fatal in more than 95% of the reported cases. Although being fatal and with potential risks for an increase in the occurrence of the pathogen in populated areas, the organism receives little public health attention. A great underestimation in the number of PAM cases reported is assumed, taking into account the difficulty in obtaining an accurate diagnosis. In this review, we summarize different techniques and methods used in the identification of the protozoan in clinical and environmental samples. Since it remains unclear whether the protozoan infection can be successfully treated with the currently available drugs, we proceed to discuss the current PAM therapeutic strategies and its effectiveness. Finally, novel compounds for potential treatments are discussed as well as research on vaccine development against PAM.

Purpose This study examined effects of a peer-mediated intervention that provided training on the use of a speech-generating device for preschoolers with severe autism spectrum disorder (ASD) and peer partners. Method Effects were examined using a multiple probe design across 3 children with ASD and limited to no verbal skills. Three peers without disabilities were taught to Stay, Play, and Talk using a GoTalk 4+ (Attainment Company) and were then paired up with a classmate with ASD in classroom social activities. Measures included rates of communication acts, communication mode and function, reciprocity, and engagement with peers. Results Following peer training, intervention effects were replicated across 3 peers, who all demonstrated an increased level and upward trend in communication acts to their classmates with ASD. Outcomes also revealed moderate intervention effects and increased levels of peer-directed communication for 3 children with ASD in classroom centers. Additional analyses revealed higher rates of communication in the added context of preferred toys and snack. The children with ASD also demonstrated improved communication reciprocity and peer engagement. Conclusions Results provide preliminary evidence on the benefits of combining peer-mediated and speech-generating device interventions to improve children's communication. Furthermore, it appears that preferred contexts are likely to facilitate greater communication and social engagement with peers.

Cancer-related fatigue is the most prevalent and distressing symptom experienced by adolescents and young adults (AYAs). An electronic survey was undertaken to ascertain current fatigue management and perceptions of its effectiveness. Eighty-five percent of responders (68/80) experienced fatigue, and it was worse more than 1 year after cancer treatment ended, compared to

Forensic facial reconstruction may be the final option available to draw public attention in cases where the identity of an individual cannot be established by standard identification methods. Two fundamental components of all forensic facial reconstruction techniques are cranial morphology and soft tissue depth databases. The purpose of this study was to extend such databases by providing a complete set of accurate facial soft tissue thickness measurements, acquired from a contemporary adult population from Germany. The material for the study consisted of 320 (160 male, 160 female) anonymised multi-slice computerised tomography (MSCT) scans of individuals drawn from a German population. In Amira®, 3D models of the surfaces of the skull and the facial skin were semi-automatically segmented using calculated thresholds and surface extraction algorithms. Facial soft tissue depth was measured at 10 midline and 28 bilateral anatomical landmarks. The analysis of facial soft tissue thickness versus BMI, sex and age indicated that, at a number of the landmarks, facial soft tissue depth is significantly (p

Fentanyl is a potent opiate commonly administered during cardiac catheterization procedures in North America.1 The question of whether fentanyl could have adverse consequences in patients undergoing percutaneous coronary intervention (PCI) is raised by recent research demonstrating that intravenous morphine significantly delays the absorption of oral P2Y12 platelet inhibitors.2 The presumed mechanism is slowed gastric emptying. The single-center PACIFY trial (Platelet Aggregation With Ticagrelor Inhibition and Fentanyl; ClinicalTrials.gov. Unique identifier: NCT02683707) randomized adults undergoing clinically indicated elective coronary angiography to receive the procedure with or without intravenous fentanyl.3 The study was approved by the Johns Hopkins Medicine Institutional Review Board, and all participants provided written informed consent. Eligible adults had not received P2Y12 inhibitors for 14 days before enrollment. Other exclusion criteria included preprocedural treatment with oral anticoagulants or opiates, platelet count

Each year, millions of rescue operations occur in Germany. Despite rising numbers, there is a shortfall of systematic quality management (QM). Focusing on finding the required quality parameters, this work aims at linking heterogeneous and distributed data sources on the patient-level. Presented here is necessary IT infrastructure for linking patients' data records properly and efficiently. The focus of the IT infrastructure is to provide a lightweight and easy applicable solution with an utmost generic approach.

Background Glucocorticoid-induced TNFR-related protein (TNFRSF18, GITR, CD357), expressed by T cells, and its ligand (TNFSF18, GITRL), expressed by myeloid populations, provide co-stimulatory signals that boost T cell activity. Due to the important role that GITR plays in regulating immune functions, agonistic stimulation of GITR is a promising therapeutic concept. Multiple strategies to induce GITR signaling have been investigated. The limited clinical efficacy of antibody-based GITR agonists results from structural and functional characteristics of antibodies that are unsuitable for stimulating the well-defined trimeric members of the TNFRSF. Methods To overcome limitations of antibody-based TNFRSF agonists, we have developed HERA-GITRL, a fully human hexavalent TNF receptor agonist (HERA) targeting GITR and mimicking the natural signaling concept. HERA-GITRL is composed of a trivalent but single-chain GITRL-receptor-binding-domain (scGITRL-RBD) unit fused to an IgG1 derived silenced Fc-domain serving as dimerization scaffold. A specific mouse surrogate, mmHERA-GITRL, was also generated to examine in vivo activity in respective mouse tumor models. Results For functional characterization of HERA-GITRL in vitro, human immune cells were isolated from healthy-donor blood and stimulated with anti-CD3 antibody in the presence of HERA-GITRL. Consistently, HERA-GITRL increased the activity of T cells, including proliferation and differentiation, even in the presence of regulatory T cells. In line with these findings, mmHERA-GITRL enhanced antigen-specific clonal expansion of both CD4+ (OT-II) and CD8+ (OT-I) T cells in vivo while having no effect on non-specific T cells. In addition, mmHERA-GITRL showed single-agent anti-tumor activity in two subcutaneous syngeneic colon cancer models (CT26wt and MC38-CEA). Importantly, this activity is independent of its FcγR-binding functionality, as both mmHERA-GITRL with a functional Fc- and a silenced Fc-domain showed similar tumor growth inhibition. Finally, in a direct in vitro comparison to a bivalent clinical benchmark anti-GITR antibody and a trivalent GITRL, only the hexavalent HERA-GITRL showed full biological activity independent of additional crosslinking. Conclusion In this manuscript, we describe the development of HERA-GITRL, a true GITR agonist with a clearly defined mechanism of action. By clustering six receptor chains in a spatially well-defined manner, HERA-GITRL induces potent agonistic activity without being dependent on additional FcγR-mediated crosslinking. Electronic supplementary material The online version of this article (10.1186/s40425-019-0671-4) contains supplementary material, which is available to authorized users.

ALG3-CDG is one of the very rare types of congenital disorder of glycosylation (CDG) caused by variants in the ER-mannosyltransferase ALG3. Here, we summarize the clinical, biochemical, and genetic data of four new ALG3-CDG patients, who were identified by a type I pattern of serum transferrin and the accumulation of Man5 GlcNAc2 -PP-dolichol in LLO analysis. Additional clinical symptoms observed in our patients comprise sensorineural hearing loss, right-descending aorta, obstructive cardiomyopathy, macroglossia, and muscular hypertonia. We add four new biochemically confirmed variants to the list of ALG3-CDG inducing variants: c.350G>C (p.R117P), c.1263G>A (p.W421*), c.1037A>G (p.N346S), and the intron variant c.296+4A>G. Furthermore, in Patient 1 an additional open-reading frame of 141 bp (AAGRP) in the coding region of ALG3 was identified. Additionally, we show that control cells synthesize, to a minor degree, a hybrid protein composed of the polypeptide AAGRP and ALG3 (AAGRP-ALG3), while in Patient 1 expression of this hybrid protein is significantly increased due to the homozygous variant c.160_196del (g.165C>T). By reviewing the literature and combining our findings with previously published data, we further expand the knowledge of this rare glycosylation defect.

Loss-of-function mutations of the SCN5A gene encoding for the sodium channel α-subunit NaV1.5 result in the autosomal dominant hereditary disease Brugada Syndrome (BrS) with a high risk of sudden cardiac death in the adult. We here engineered human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) carrying the CRISPR/Cas9 introduced BrS-mutation p.A735V-NaV1.5 (g.2204C > T in exon 14 of SCN5A) as a novel model independent of patient´s genetic background. Recent studies raised concern regarding the use of hiPSC-CMs for studying adult-onset hereditary diseases due to cells’ immature phenotype. To tackle this concern, long-term cultivation of hiPSC-CMs on a stiff matrix (27–42 days) was applied to promote maturation. Patch clamp recordings of A735V mutated hiPSC-CMs revealed a substantially reduced upstroke velocity and sodium current density, a prominent rightward shift of the steady state activation curve and decelerated recovery from inactivation as compared to isogenic hiPSC-CMs controls. These observations were substantiated by a comparative study on mutant A735V-NaV1.5 channels heterologously expressed in HEK293T cells. In contrast to mutated hiPSC-CMs, a leftward shift of sodium channel inactivation was not observed in HEK293T, emphasizing the importance of investigating mechanisms of BrS in independent systems. Overall, our approach supports hiPSC-CMs’ relevance for investigating channelopathies in a dish.

Purpose The purpose of this study was to measure changes in communication of preschoolers with autism using the Communication Complexity Scale (CCS; Brady et al., 2012 ) and to examine the utility of the CCS in measuring pretreatment and posttreatment changes within peer and adult assessment contexts. Method The CCS was used to code preassessment and postassessment for 23 children with autism randomly assigned to a treatment that incorporated a peer-mediated approach and a speech-generating device and 22 assigned to a business-as-usual condition with untrained peers. Children were assessed in 2 structured 30-min contexts—1 with an adult examiner and 1 with a peer partner coached by an adult. Results Children in both groups showed significant changes in communication complexity CCS scores from pretreatment to posttreatment in the adult and peer contexts. At both occasions, CCS scores were higher with adult partners yet showed greater improvements over time with peer partners. Conclusions Results showed that the CCS was sensitive to change over time but did not discriminate changes in communication complexity associated with maturation versus treatment. It did show some differences based on interactions with peer versus adult partners. Outcomes provide preliminary support for using this scale to measure communication changes in different contexts. Supplemental Material https://doi.org/10.23641/asha.7408856

Neural regions important for smell are proximal and closely connected to cortical areas that have been strongly implicated in higher order functions of value-based decision making and emotional memory. The integrity of these neural regions are affected in aging and neurodegenerative conditions. Two specific predictions follow from these neuroanatomical arrangements-namely, that olfaction would be associated with value-based decision making and with emotional memory.To test these predictions, we measured these different capacities in participants with presumed varying degrees of integrity of the relevant brain structures: specifically, 13 patients with Alzheimer's disease, 8 patients with mild cognitive impairment, and 20 healthy older adults. The participants completed detailed tests of olfaction, value-based decision making, emotional memory, and general cognitive ability.Olfactory functioning was significantly associated with emotional and nonemotional memory. The association was especially strong and consistent for memory recall with olfaction, explaining as much as 10% additional variance over and above general cognition. Olfactory functioning was not strongly or consistently associated with decision making over and above general cognition.Olfaction is a strong predictor of memory recall. These findings may contribute to a better understanding of olfaction and specific cognitive domains known to be affected by aging and implicated in neurodegenerative disease.

Objective To extend available dosing options in the treatment of growth hormone deficiency, a comparative pharmacokinetic and pharmacodynamic phase-1 clinical study involving subcutaneous administration of growth hormone was conducted. Design The test formulation (biosimilar recombinant human growth hormone; r-hGH; Somatotropin) and reference formulation (Genotropin®) were tested in 38 adult healthy subjects after their subcutaneous administration of 12.8 IU in an open label, single dose, randomized, two period cross over study separated with a washout period of 11 days. Endogenous growth hormone release was suppressed by a continuous Octreotide infusion up to 24 h after r-hGH administration. All the subjects were evaluated for local tolerance using Wong-Baker Faces pain rating scale and an injection site reaction (ISR) score. Detection of serum levels of r-hGH, insulin-like growth factor-1 (IGF-1) and insulin-like growth factor binding protein-3 (IGFBP-3) was done by suitable validated bio-analytical methods. Assessment of bioequivalence for pharmacokinetic parameters was done using log-transformed area under the curve (AUC) and maximum concentration (Cmax) for r-hGH. The pharmacodynamic assessment was done by comparing the area under the effect-time curve (AUEClast) and maximum measured effect concentration (Emax) of IGF-1 and IGFBP-3. Results The biosimilar formulation of recombinant human growth hormone fulfilled the predefined bioequivalence criteria for pharmacokinetic and pharmacodynamic parameters. Conclusion The new biosimilar recombinant human growth hormone bears the potential to become an alternative option for the treatment of growth hormone deficiency.

Few studies of oral glucose-lowering drugs exist in newly diagnosed type 2 diabetes (T2D) patients with marked hyperglycemia, and insulin is often proposed as initial treatment. We evaluated the oral initial combination of metformin and linagliptin, a dipeptidyl peptidase-4 inhibitor, in this population.We performed a pre-specified subgroup analysis of a randomized study in which newly diagnosed T2D patients with glycated hemoglobin A1c (HbA1c) 8.5%-12.0% received linagliptin/metformin or linagliptin monotherapy. Subgroups of baseline HbA1c, age, body-mass index (BMI), renal function, race, and ethnicity were evaluated, with efficacy measured by HbA1c change from baseline after 24 weeks.HbA1c reductions from baseline (mean 9.7%) at week 24 in the overall population were an adjusted mean -2.81% ± 0.12% with linagliptin/metformin (n = 132) and -2.02% ± 0.13% with linagliptin (n = 113); treatment difference -0.79% (95% CI -1.13 to -0.46, P 0.0001). In patients with baseline HbA1c ≥9.5%, HbA1c reduction was -3.37% with linagliptin/metformin (n = 76) and -2.53% with linagliptin (n = 61); difference -0.84% (95% CI -1.32 to -0.35). In those with baseline HbA1c9.5%, HbA1c reduction was -2.08% with linagliptin/metformin (n = 56) and -1.39% with linagliptin (n = 52); difference -0.69% (95% CI -1.23 to -0.15). Changes in HbA1c and treatment differences between the linagliptin/metformin and linagliptin groups were of similar magnitudes to the overall population across patient subgroups based on age, BMI, renal function, and race. Drug-related adverse events occurred in 8.8% and 5.7% of linagliptin/metformin and linagliptin patients, respectively; no severe hypoglycemia occurred.Linagliptin/metformin combination in newly diagnosed T2D patients with marked hyperglycemia was well tolerated and elicited substantial improvements in glycemic control regardless of baseline HbA1c, age, BMI, renal function, or race. Thus, newly diagnosed, markedly hyperglycemic patients may be effectively treated by combinations of oral agents.www.clinicaltrials.gov identifier is NCT01512979.

Galectins are a family of mammalian carbohydrate-binding proteins expressed by many cell types. Galectins can function intracellularly and can also be secreted to bind to cell surface glycoconjugate counterreceptors. Some galectins are made by immune cells, whereas other galectins are secreted by different cell types, such as endothelial or epithelial cells, and bind to immune cells to regulate immune responses. Galectin binding to a single glycan ligand is a low-affinity interaction, but the multivalency of galectins and the glycan ligands presented on cell surface glycoproteins results in high-avidity binding that can reversibly scaffold or cluster these glycoproteins. Galectin binding to a specific glycoprotein counterreceptor is regulated in part by the repertoire of glycosyltransferase enzymes (which make the glycan ligands) expressed by that cell, and the effect of galectin binding results from clustering or retention of specific glycoprotein counterreceptors bearing these specific ligands.