Bi-allelic Loss-of-Function CACNA1B Mutations in Progressive Epilepsy-Dyskinesia.

The occurrence of non-epileptic hyperkinetic movements in the context of developmental epileptic encephalopathies is an increasingly recognized phenomenon. Identification of causative mutations provides an important insight into common pathogenic mechanisms that cause both seizures and abnormal motor control. We report bi-allelic loss-of-function CACNA1B variants in six children from three unrelated families whose affected members present with a complex and progressive neurological syndrome. All affected individuals presented with epileptic encephalopathy, severe neurodevelopmental delay (often with regression), and a hyperkinetic movement disorder. Additional neurological features included postnatal microcephaly and hypotonia. Five children died in childhood or adolescence (mean age of death: 9 years), mainly as a result of secondary respiratory complications. CACNA1B encodes the pore-forming subunit of the pre-synaptic neuronal voltage-gated calcium channel Cav2.2/N-type, crucial for SNARE-mediated neurotransmission, particularly in the early postnatal period. Bi-allelic loss-of-function variants in CACNA1B are predicted to cause disruption of Ca2+ influx, leading to impaired synaptic neurotransmission. The resultant effect on neuronal function is likely to be important in the development of involuntary movements and epilepsy. Overall, our findings provide further evidence for the key role of Cav2.2 in normal human neurodevelopment.MAK is funded by an NIHR Research Professorship and receives funding from the Wellcome Trust, Great Ormond Street Children's Hospital Charity, and Rosetrees Trust. E.M. received funding from the Rosetrees Trust (CD-A53) and Great Ormond Street Hospital Children's Charity. K.G. received funding from Temple Street Foundation. A.M. is funded by Great Ormond Street Hospital, the National Institute for Health Research (NIHR), and Biomedical Research Centre. F.L.R. and D.G. are funded by Cambridge Biomedical Research Centre. K.C. and A.S.J. are funded by NIHR Bioresource for Rare Diseases. The DDD Study presents independent research commissioned by the Health Innovation Challenge Fund (grant number HICF-1009-003), a parallel funding partnership between the Wellcome Trust and the Department of Health, and the Wellcome Trust Sanger Institute (grant number WT098051). We acknowledge support from the UK Department of Health via the NIHR comprehensive Biomedical Research Centre award to Guy's and St. Thomas' National Health Service (NHS) Foundation Trust in partnership with King's College London. This research was also supported by the NIHR Great Ormond Street Hospital Biomedical Research Centre. J.H.C. is in receipt of an NIHR Senior Investigator Award. The research team acknowledges the support of the NIHR through the Comprehensive Clinical Research Network. The views expressed are those of the author(s) and not necessarily those of the NHS, the NIHR, Department of Health, or Wellcome Trust. E.R.M. acknowledges support from NIHR Cambridge Biomedical Research Centre, an NIHR Senior Investigator Award, and the University of Cambridge has received salary support in respect of E.R.M. from the NHS in the East of England through the Clinical Academic Reserve. I.E.S. is supported by the National Health and Medical Research Council of Australia (Program Grant and Practitioner Fellowship)

Retrospective evaluation of whole exome and genome mutation calls in 746 cancer samples

Funder: NCI U24CA211006Abstract: The Cancer Genome Atlas (TCGA) and International Cancer Genome Consortium (ICGC) curated consensus somatic mutation calls using whole exome sequencing (WES) and whole genome sequencing (WGS), respectively. Here, as part of the ICGC/TCGA Pan-Cancer Analysis of Whole Genomes (PCAWG) Consortium, which aggregated whole genome sequencing data from 2,658 cancers across 38 tumour types, we compare WES and WGS side-by-side from 746 TCGA samples, finding that ~80% of mutations overlap in covered exonic regions. We estimate that low variant allele fraction (VAF < 15%) and clonal heterogeneity contribute up to 68% of private WGS mutations and 71% of private WES mutations. We observe that ~30% of private WGS mutations trace to mutations identified by a single variant caller in WES consensus efforts. WGS captures both ~50% more variation in exonic regions and un-observed mutations in loci with variable GC-content. Together, our analysis highlights technological divergences between two reproducible somatic variant detection efforts

Effect of angiotensin-converting enzyme inhibitor and angiotensin receptor blocker initiation on organ support-free days in patients hospitalized with COVID-19

IMPORTANCE Overactivation of the renin-angiotensin system (RAS) may contribute to poor clinical outcomes in patients with COVID-19. Objective To determine whether angiotensin-converting enzyme (ACE) inhibitor or angiotensin receptor blocker (ARB) initiation improves outcomes in patients hospitalized for COVID-19. DESIGN, SETTING, AND PARTICIPANTS In an ongoing, adaptive platform randomized clinical trial, 721 critically ill and 58 non–critically ill hospitalized adults were randomized to receive an RAS inhibitor or control between March 16, 2021, and February 25, 2022, at 69 sites in 7 countries (final follow-up on June 1, 2022). INTERVENTIONS Patients were randomized to receive open-label initiation of an ACE inhibitor (n = 257), ARB (n = 248), ARB in combination with DMX-200 (a chemokine receptor-2 inhibitor; n = 10), or no RAS inhibitor (control; n = 264) for up to 10 days. MAIN OUTCOMES AND MEASURES The primary outcome was organ support–free days, a composite of hospital survival and days alive without cardiovascular or respiratory organ support through 21 days. The primary analysis was a bayesian cumulative logistic model. Odds ratios (ORs) greater than 1 represent improved outcomes. RESULTS On February 25, 2022, enrollment was discontinued due to safety concerns. Among 679 critically ill patients with available primary outcome data, the median age was 56 years and 239 participants (35.2%) were women. Median (IQR) organ support–free days among critically ill patients was 10 (–1 to 16) in the ACE inhibitor group (n = 231), 8 (–1 to 17) in the ARB group (n = 217), and 12 (0 to 17) in the control group (n = 231) (median adjusted odds ratios of 0.77 [95% bayesian credible interval, 0.58-1.06] for improvement for ACE inhibitor and 0.76 [95% credible interval, 0.56-1.05] for ARB compared with control). The posterior probabilities that ACE inhibitors and ARBs worsened organ support–free days compared with control were 94.9% and 95.4%, respectively. Hospital survival occurred in 166 of 231 critically ill participants (71.9%) in the ACE inhibitor group, 152 of 217 (70.0%) in the ARB group, and 182 of 231 (78.8%) in the control group (posterior probabilities that ACE inhibitor and ARB worsened hospital survival compared with control were 95.3% and 98.1%, respectively). CONCLUSIONS AND RELEVANCE In this trial, among critically ill adults with COVID-19, initiation of an ACE inhibitor or ARB did not improve, and likely worsened, clinical outcomes. TRIAL REGISTRATION ClinicalTrials.gov Identifier: NCT0273570

Where families and healthcare meet

Recent developments in professional healthcare pose moral problems that standard bioethics cannot even identify as problems, but that are fully visible when redefined as problems in the ethics of families. Here, we add to the growing body of work that began in the 1990s by demonstrating the need for a distinctive ethics of families. First, we discuss what 'family' means and why families can matter so deeply to the lives of those within them. Then, we briefly sketch how, according to an ethics of families, responsibilities must be negotiated against the backdrop of family relationships, treatment decisions must be made in the light of these negotiated responsibilities and justice must be served, both between families and society more generally and within families themselves

Vitamin D Status and Indices of Bone Turnover in Older European Adults

International audienceAn increased rate of bone turnover increases risk of osteoporotic fracture later in life. The concentration of 25-hydroxyvitamin D that contributes to an elevated rate of bone turnover in older adults is unclear. The objective of this study was to investigate the associations between 25-hydroxyvitamin D and biochemical markers of bone turnover in an older, pan-European cohort. 25-hydroxyvitamin D and serum markers of bone-formation (osteocalcin and bone-specific alkaline phosphatase) were assessed by ELISA, while urinary markers of bone-resorption (pyridinoline and deoxypyridinoline) were assessed by HPLC. Six percent, 36%, and 64% of subjects had 25-hydroxyvitamin D concentrations 85.8 [T-3] nmol/L), showed that urinary pyridinoline and deoxypyridinoline were significantly lower in subjects in the 2nd and 3rd compared to the 1st tertile (p < 0.015). Low vitamin D status (<50 nmol/L) was associated with an increased rate of bone turnover in this older pan-European cohort

Validating cognitive screening in young people with first‐episode psychosis:The CogScreen protocol

Aim: Cognitive impairments are a core feature of first‐episode psychosis (FEP) and one of the strongest predictors of long‐term psychosocial functioning. Cognition should be assessed and treated as part of routine clinical care for FEP. Cognitive screening offers the opportunity to rapidly identify and triage those in most need of cognitive support. However, there are currently no validated screening measures for young people with FEP. CogScreen is a hybrid effectiveness‐implementation study which aims to evaluate the classification accuracy (relative to a neuropsychological assessment as a reference standard), test–retest reliability and acceptability of two cognitive screening tools in young people with FEP. Methods: Participants will be 350 young people (aged 12–25) attending primary and specialist FEP treatment centres in three large metropolitan cities (Adelaide, Sydney, and Melbourne) in Australia. All participants will complete a cross‐sectional assessment over two sessions including two cognitive screening tools (Screen for Cognitive Impairment in Psychiatry and Montreal Cognitive Assessment), a comprehensive neuropsychological assessment battery, psychiatric and neurodevelopmental assessments, and other supplementary clinical measures. To determine the test–retest reliability of the cognitive screening tools, a subset of 120 participants will repeat the screening measures two weeks later. Results: The protocol, rationale, and hypotheses for CogScreen are presented. Conclusions: CogScreen will provide empirical evidence for the validity and reliability of two cognitive screening tools when compared to a comprehensive neuropsychological assessment. The screening measures may later be incorporated into clinical practice to assist with rapid identification and treatment of cognitive deficits commonly experienced by young people with FEP

Data from: Flight capacity and behavior of Ephestia kuehniella Zeller (Lepidoptera: Pyralidae) in response to kairomonal and pheromonal stimuli

Kairomonal and pheromonal stimuli The pheromone stimuli consisted of a proprietary blend (IL-103, Insects Limited, Westfield, IN, USA) of the sex pheromone for E. kuehniella, including less than 5% concentration by weight of (Z, E)-9,12-tetradecadienyl acetate (e.g., ZETA). The kairomonal stimuli consisted of 20 g of a food cue mixture comprised of cracked wheat, wheat shorts, of wheat germ, and of brewer’s yeast in a 23:7:2:1 ratio. These were the dry components of the E. kuehniella diet. Flight Mill Apparatus Flight mills were assembled after Smith et al. (2012) and Ruiz et al. (2022) (Figure 1). The base assembly consisted of a Delrin rod (95.25 mm length × 38.1 mm diameter) mounted to a 6.35-mm thick equilateral triangular Plexiglass base (each side 13.6 cm long), with screws used as levels. At the top of the rod, a 1.5 cm length (0.72 mm, 22 ga) hypodermic needle was mounted into a hole, and a 5 mm circular magnet was glued to each end of dual 35 mm long metal #1 insect pins at the top of the rod around the axle pin. A unipolar digital Hall effect sensor (OHN3120U, Optek, Inc., Carrollton, TX) was wired to a coupler with one wire for the 5V DC, one ground, and one output signal. This sensor assembly was attached to the top of the Delrin rod with a Plexiglass assembly and thumbscrews. The rotation flight arm assembly consisted of a Teflon bar (2.0 cm length × 1.1 cm diameter) with circular magnets (polarity conserved) glued to the bottom and a hole bored into the bottom to accept the other end of the axle pin that was mounted in the Delrin rod. A hypodermic needle (28 cm length × 0.72 mm diameter) was threaded through a hole in the flight arm assembly, with the ends bent downward at a 95° angle. As a result, the rotation flight arm assembly fit onto the base assembly axle pin, and the circular magnets on the base and bottom of the flight arm assemblies configured to the same polarity kept the arm suspended over the axle, allowing it to spin on the Teflon bearing with very little friction. When the small magnet facing down from the flight arm assembly rotated over the Hall effect sensor, a signal was sent through the output signal wire. The six flight mills were situated on a 71.1 cm × 91.4 cm piece of plexiglass to minimize vibrations. Each mill was spaced 39 cm apart in tandem (e.g., three in a row) with 25.4 cm between rows. Each mill contained three wires connected to the main connector board (#777101-01, National Instruments, Austin, TX, USA). A DC Power Supply (QW-MS3010D, QW, Tampa, FL) was set to the site and connected to the main wiring block to provide power. A 50-pin ribbon cable (180524-10, National Instruments) ran from the main wiring connector block into a specially made PC port (#77690-01, National Instruments), where a computer with the software Labview 2017 (version 17.0.1f3, National Instruments) automatically recorded the data from the mills. Data intervals for Hall effect sensors were set to 1 ms. Hall sensors were triggered when a magnet from the flight arm passed over, and this was recorded by the software as a voltage change. Schematics, photographs, wiring diagrams, ordering information, and data acquisition programs for this flight mill setup are all available online (Jones et al. 2010)(Flight Mill Studies n.d.). Flight Capacity Experiments Six adults blocked by sex were run simultaneously on the six flight mills described above (15-FMASM SDP Unit, Crist Instrument Co., Hagerstown, MD) to test flight capacity. Freshly eclosed moths were used for the experiments to standardize age, and were likely unmated. A 14-gauge copper wire was stripped into individual threads and cut into 4 cm segments. The copper wire thread was wrapped around an insect pin (#6 insect pin, BioQuip Products, Rancho Dominguez, CA) embedded in modelling clay to form a loop. The loop was pinched with standard metal forceps, the excess wire was ablated on the shorter end with scissors, and the loop was flattened relative to the plane of gravity. The moths were placed singly on a metal mason jar lid suspended on ice and restrained with a 2 oz plastic lid until sessile. Subsequently, the pronotum of E. kuehniella was descaled lightly with an artist’s paintbrush. The copper loop was dipped in instant adhesive (#347908, Evo Stik Multi-Purpose Impact Adhesive, Bostik, Ltd., Leicester, United Kingdom) and affixed onto the pronotum of the adult, ensuring not to impair proper wing functioning in addition to avoiding the eyes and antennae of the adults. The individual was tethered by inserting the point of the copper wire into the end of the hypodermic needle attached to the rotation arm assembly of each flight mill. Each trial was started between 15:00-18:00 by gently blowing by mouth over the back of the insects to initiate flight to provide the opportunity for flight (per the established guidance for tethered flight mill systems: Naranjo 2019) and flight was recorded over initiate the 24 h observation period. Any and all flight bouts were recorded during this time. A flight bout was defined as consecutive rotations of the flight arm of the flight mill by E. kuehniella separated by a pause of 3 s in rotation. Flight mills were housed in a dedicated space in the laboratory under constant conditions (at 21.6 ± 0.01°C, 43 ± 0.2% RH, 14:10 L:D photoperiod). Air flow was gently vented at a mean velocity of 4.57 m per min. Treatments included an unbaited control, kairomone (e.g., food cue), and pheromone (e.g., all as above). Treatment stimuli sources were placed on a platform located directly in the center of the set of flight mills. There was a total of n = 18 reps per treatment combination of stimulus. At the end of a trial, insects were detached from the apparatus and weighed on a balance. Data were streamed in real-time to a computer containing the software, which was used to automatically record the flight parameters, namely distance flown, as well as the number of tandem flight bouts over the sampling interval (flights lasting more than 1 s), and their average length, and average distance flown per bout. To conservatively estimate flight distance, all flight distances consisting of a singleton flight bout were eliminated from the datasets. In addition, to support data quality and avoid the false accumulation of data, revolutions were eliminated if the sum of revolutions on the flight mill divided by the total time spent flying was larger than 3.6 s (the conservatively slowest time E. kuehniella could make a revolution and still be actively flying) and the number of revolutions was greater than 30. Data were also parsed by time of day to determine the time of maximum dispersal. Data were analyzed with R software (v. 2022.02.1 Build 461) (R Core Team 2022). Ggplot2 was used for some of the figures (Wickham et al. 2016). Cage Assay To test flight initiation patterns of E. kuehniella, a cage assay was utilized. Cohorts of 20 adult moths blocked by sex were collected in a plastic bag from colony jars, then briefly anesthetized for no longer than 30 s with N2 gas. Moths were gently placed in a petri dish (100 × 15 mm) lined with polytetrafluoroethylene (PTFE) (MilliporeSigma, Burlington, MA, USA), which was then situated in a randomly chosen corner of a mesh cage (28 × 28 × 28 cm, Bug Dorm, MegaView Science Co. Ltd., Taiwan) and covered with a funnel (height: 12.5 cm; large opening: 15 cm D; small opening: 2.6 cm D). The semiochemical treatments listed above were placed randomly in one of the three remaining corners. Cages were placed in a walk-in environmental chamber under constant conditions (at 25 ℃, 65% RH, 14:10 L:D photoperiod) and given 24 h to respond to the stimuli. The proportion of adults leaving the funnel was measured at the end of the trial period. In total, n = 8 replications were performed with each treatment, translating to 480 total E. kuehniella individuals tested. </p