Pathways of Enzymic Cholesterol Syntheses

The purpose of this paper is to summarize the results of my research on the enzymic steps of cholesterol biosynthesis (Dempsey, 1962; 1964; 1965; Dempsey, Seaton, and Trockmon,1963; Dempsey, Seaton, Sanford, and Trockmon,19640; Dempsey, Seaton, Schroepfer, and Trackman, 19646.2 These data were obtained by use of an enzyme system isolated from rat liver homogenates. The findings have elucidated the pathways of and cofactor requirements for cholesterol synthesis from naturally occurring sterol precursors

Geostatistical evaluation of integrated marsh management impact on mosquito vectors using before-after-control-impact (BACI) design

<p>Abstract</p> <p>Background</p> <p>In many parts of the world, salt marshes play a key ecological role as the interface between the marine and the terrestrial environments. Salt marshes are also exceedingly important for public health as larval habitat for mosquitoes that are vectors of disease and significant biting pests. Although grid ditching and pesticides have been effective in salt marsh mosquito control, marsh degradation and other environmental considerations compel a different approach. Targeted habitat modification and biological control methods known as Open Marsh Water Management (OMWM) had been proposed as a viable alternative to marsh-wide physical alterations and chemical control. However, traditional larval sampling techniques may not adequately assess the impacts of marsh management on mosquito larvae. To assess the effectiveness of integrated OMWM and marsh restoration techniques for mosquito control, we analyzed the results of a 5-year OMWM/marsh restoration project to determine changes in mosquito larval production using GIS and geostatistical methods.</p> <p>Methods</p> <p>The following parameters were evaluated using "Before-After-Control-Impact" (BACI) design: frequency and geographic extent of larval production, intensity of larval production, changes in larval habitat, and number of larvicide applications. The analyses were performed using Moran's I, Getis-Ord, and Spatial Scan statistics on aggregated before and after data as well as data collected over time. This allowed comparison of control and treatment areas to identify changes attributable to the OMWM/marsh restoration modifications.</p> <p>Results</p> <p>The frequency of finding mosquito larvae in the treatment areas was reduced by 70% resulting in a loss of spatial larval clusters compared to those found in the control areas. This effect was observed directly following OMWM treatment and remained significant throughout the study period. The greatly reduced frequency of finding larvae in the treatment areas led to a significant decrease (~44%) in the number of times when the larviciding threshold was reached. This reduction, in turn, resulted in a significant decrease (~74%) in the number of larvicide applications in the treatment areas post-project. The remaining larval habitat in the treatment areas had a different geographic distribution and was largely confined to the restored marsh surface (i.e. filled-in mosquito ditches); however only ~21% of the restored marsh surface supported mosquito production.</p> <p>Conclusion</p> <p>The geostatistical analysis showed that OMWM demonstrated considerable potential for effective mosquito control and compatibility with other natural resource management goals such as restoration, wildlife habitat enhancement, and invasive species abatement. GPS and GIS tools are invaluable for large scale project design, data collection, and data analysis, with geostatistical methods serving as an alternative or a supplement to the conventional inference statistics in evaluating the project outcome.</p

Cytotoxic G-rich oligodeoxynucleotides: putative protein targets and required sequence motif

It has recently been shown that certain oligodeoxynucleotides (ODNs) designed as catalytic DNA molecules (DNAzymes) exhibit potent cytotoxicity independent of RNA-cleavage activity in a number of cell lines. These cytotoxic ODNs all featured a 5′ G-rich sequence and induced cell death by a TLR9-independent mechanism. In this study, we examined the sequence and length dependence of ODNs for cytotoxicity. A G-rich sequence at the 5′ terminus of the molecule was necessary for cytotoxicity and the potency of ODNs with active 5′ sequences was length dependent. Cytotoxicity appeared to be generally independent of 3′ sequence composition, although 3′ sequences totally lacking G-nucleotides were mostly inactive. Nucleolin, elongation factor 1-alpha (eEF1A) and vimentin were identified as binding to a cytotoxic ODN (Dz13) using protein pull-down assays and LC-MS/MS. Although these proteins have previously been described to bind G-rich ODNs, the binding of eEF1A correlated with cytotoxicity, whereas binding of nucleolin and vimentin did not. Quiescent non-proliferating cells were resistant to cytotoxicity, indicating cytotoxicity may be cell cycle dependent. Although the exact mechanism of cytotoxicity remains unknown, marked potency of the longer (⩾25 nt) ODNs in particular, indicates the potential of these molecules for treatment of diseases associated with abnormal cell proliferation

Hyperoxemia and excess oxygen use in early acute respiratory distress syndrome : Insights from the LUNG SAFE study

Publisher Copyright: © 2020 The Author(s). Copyright: Copyright 2020 Elsevier B.V., All rights reserved.Background: Concerns exist regarding the prevalence and impact of unnecessary oxygen use in patients with acute respiratory distress syndrome (ARDS). We examined this issue in patients with ARDS enrolled in the Large observational study to UNderstand the Global impact of Severe Acute respiratory FailurE (LUNG SAFE) study. Methods: In this secondary analysis of the LUNG SAFE study, we wished to determine the prevalence and the outcomes associated with hyperoxemia on day 1, sustained hyperoxemia, and excessive oxygen use in patients with early ARDS. Patients who fulfilled criteria of ARDS on day 1 and day 2 of acute hypoxemic respiratory failure were categorized based on the presence of hyperoxemia (PaO2 > 100 mmHg) on day 1, sustained (i.e., present on day 1 and day 2) hyperoxemia, or excessive oxygen use (FIO2 ≥ 0.60 during hyperoxemia). Results: Of 2005 patients that met the inclusion criteria, 131 (6.5%) were hypoxemic (PaO2 < 55 mmHg), 607 (30%) had hyperoxemia on day 1, and 250 (12%) had sustained hyperoxemia. Excess FIO2 use occurred in 400 (66%) out of 607 patients with hyperoxemia. Excess FIO2 use decreased from day 1 to day 2 of ARDS, with most hyperoxemic patients on day 2 receiving relatively low FIO2. Multivariate analyses found no independent relationship between day 1 hyperoxemia, sustained hyperoxemia, or excess FIO2 use and adverse clinical outcomes. Mortality was 42% in patients with excess FIO2 use, compared to 39% in a propensity-matched sample of normoxemic (PaO2 55-100 mmHg) patients (P = 0.47). Conclusions: Hyperoxemia and excess oxygen use are both prevalent in early ARDS but are most often non-sustained. No relationship was found between hyperoxemia or excessive oxygen use and patient outcome in this cohort. Trial registration: LUNG-SAFE is registered with ClinicalTrials.gov, NCT02010073publishersversionPeer reviewe

AI is a viable alternative to high throughput screening: a 318-target study

: High throughput screening (HTS) is routinely used to identify bioactive small molecules. This requires physical compounds, which limits coverage of accessible chemical space. Computational approaches combined with vast on-demand chemical libraries can access far greater chemical space, provided that the predictive accuracy is sufficient to identify useful molecules. Through the largest and most diverse virtual HTS campaign reported to date, comprising 318 individual projects, we demonstrate that our AtomNet® convolutional neural network successfully finds novel hits across every major therapeutic area and protein class. We address historical limitations of computational screening by demonstrating success for target proteins without known binders, high-quality X-ray crystal structures, or manual cherry-picking of compounds. We show that the molecules selected by the AtomNet® model are novel drug-like scaffolds rather than minor modifications to known bioactive compounds. Our empirical results suggest that computational methods can substantially replace HTS as the first step of small-molecule drug discovery

Diagnosis and management of Cornelia de Lange syndrome:first international consensus statement

Cornelia de Lange syndrome (CdLS) is an archetypical genetic syndrome that is characterized by intellectual disability, well-defined facial features, upper limb anomalies and atypical growth, among numerous other signs and symptoms. It is caused by variants in any one of seven genes, all of which have a structural or regulatory function in the cohesin complex. Although recent advances in next-generation sequencing have improved molecular diagnostics, marked heterogeneity exists in clinical and molecular diagnostic approaches and care practices worldwide. Here, we outline a series of recommendations that document the consensus of a group of international experts on clinical diagnostic criteria, both for classic CdLS and non-classic CdLS phenotypes, molecular investigations, long-term management and care planning

Screening of healthcare workers for SARS-CoV-2 highlights the role of asymptomatic carriage in COVID-19 transmission

Funder: Addenbrooke's Charitable Trust, Cambridge University Hospitals; FundRef: http://dx.doi.org/10.13039/501100002927Significant differences exist in the availability of healthcare worker (HCW) SARS-CoV-2 testing between countries, and existing programmes focus on screening symptomatic rather than asymptomatic staff. Over a 3 week period (April 2020), 1032 asymptomatic HCWs were screened for SARS-CoV-2 in a large UK teaching hospital. Symptomatic staff and symptomatic household contacts were additionally tested. Real-time RT-PCR was used to detect viral RNA from a throat+nose self-swab. 3% of HCWs in the asymptomatic screening group tested positive for SARS-CoV-2. 17/30 (57%) were truly asymptomatic/pauci-symptomatic. 12/30 (40%) had experienced symptoms compatible with coronavirus disease 2019 (COVID-19)>7 days prior to testing, most self-isolating, returning well. Clusters of HCW infection were discovered on two independent wards. Viral genome sequencing showed that the majority of HCWs had the dominant lineage B∙1. Our data demonstrates the utility of comprehensive screening of HCWs with minimal or no symptoms. This approach will be critical for protecting patients and hospital staff

Complement lectin pathway activation is associated with COVID-19 disease severity, independent of MBL2 genotype subgroups

IntroductionWhile complement is a contributor to disease severity in severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infections, all three complement pathways might be activated by the virus. Lectin pathway activation occurs through different pattern recognition molecules, including mannan binding lectin (MBL), a protein shown to interact with SARS-CoV-2 proteins. However, the exact role of lectin pathway activation and its key pattern recognition molecule MBL in COVID-19 is still not fully understood.MethodsWe therefore investigated activation of the lectin pathway in two independent cohorts of SARS-CoV-2 infected patients, while also analysing MBL protein levels and potential effects of the six major single nucleotide polymorphisms (SNPs) found in the MBL2 gene on COVID-19 severity and outcome.ResultsWe show that the lectin pathway is activated in acute COVID-19, indicated by the correlation between complement activation product levels of the MASP-1/C1-INH complex (p=0.0011) and C4d (p&lt;0.0001) and COVID-19 severity. Despite this, genetic variations in MBL2 are not associated with susceptibility to SARS-CoV-2 infection or disease outcomes such as mortality and the development of Long COVID.ConclusionIn conclusion, activation of the MBL-LP only plays a minor role in COVID-19 pathogenesis, since no clinically meaningful, consistent associations with disease outcomes were noted