Rab27a controls HIV-1 assembly by regulating plasma membrane levels of phosphatidylinositol 4,5-bisphosphate

During the late stages of the HIV-1 replication cycle, the viral polyprotein Pr55(Gag) is recruited to the plasma membrane (PM), where it binds phosphatidylinositol 4,5-bisphosphate (PI(4,5)P2) and directs HIV-1 assembly. We show that Rab27a controls the trafficking of late endosomes carrying phosphatidylinositol 4-kinase type 2 α (PI4KIIα) toward the PM of CD4(+) T cells. Hence, Rab27a promotes high levels of PM phosphatidylinositol 4-phosphate and the localized production of PI(4,5)P2, therefore controlling Pr55(Gag) membrane association. Rab27a also controls PI(4,5)P2 levels at the virus-containing compartments of macrophages. By screening Rab27a effectors, we identified that Slp2a, Slp3, and Slac2b are required for the association of Pr55(Gag) with the PM and that Slp2a cooperates with Rab27a in the recruitment of PI4KIIα to the PM. We conclude that by directing the trafficking of PI4KIIα-positive endosomes toward the PM, Rab27a controls PI(4,5)P2 production and, consequently, HIV-1 replication.Universidad de Buenos Aires and CONICET doctoral fellowships, Agencia Nacional de Pro- moción Científica y Tecnológica (Argentina) grants: (2010-1681, 2012-00353), Creative and Novel Ideas in HIV Research Program, University of Alabama at Birmingham Center for AIDS Research funding grant P30 AI027767-24

The management of acute venous thromboembolism in clinical practice. Results from the European PREFER in VTE Registry

Venous thromboembolism (VTE) is a significant cause of morbidity and mortality in Europe. Data from real-world registries are necessary, as clinical trials do not represent the full spectrum of VTE patients seen in clinical practice. We aimed to document the epidemiology, management and outcomes of VTE using data from a large, observational database. PREFER in VTE was an international, non-interventional disease registry conducted between January 2013 and July 2015 in primary and secondary care across seven European countries. Consecutive patients with acute VTE were documented and followed up over 12 months. PREFER in VTE included 3,455 patients with a mean age of 60.8 ± 17.0 years. Overall, 53.0 % were male. The majority of patients were assessed in the hospital setting as inpatients or outpatients (78.5 %). The diagnosis was deep-vein thrombosis (DVT) in 59.5 % and pulmonary embolism (PE) in 40.5 %. The most common comorbidities were the various types of cardiovascular disease (excluding hypertension; 45.5 %), hypertension (42.3 %) and dyslipidaemia (21.1 %). Following the index VTE, a large proportion of patients received initial therapy with heparin (73.2 %), almost half received a vitamin K antagonist (48.7 %) and nearly a quarter received a DOAC (24.5 %). Almost a quarter of all presentations were for recurrent VTE, with >80 % of previous episodes having occurred more than 12 months prior to baseline. In conclusion, PREFER in VTE has provided contemporary insights into VTE patients and their real-world management, including their baseline characteristics, risk factors, disease history, symptoms and signs, initial therapy and outcomes

Reproducibility and reliability of the ankle-brachial index as assessed by vascular experts, family physicians and nurses

The reliability of ankle—brachial index (ABI) measurements performed by different observer groups in primary care has not yet been determined. The aims of the study were to provide precise estimates for all effects influencing the variability of the ABI (patients' individual variability, intra- and inter-observer variability), with particular focus on the performance of different observer groups. Using a partially balanced incomplete block design, 144 unselected individuals aged 65 years underwent double ABI measurements by one vascular surgeon or vascular physician, one family physician and one nurse with training in Doppler sonography. Three groups comprising a total of 108 individuals were analyzed (only two with ABI < 0.90). Errors for two repeated measurements for all three observer groups did not differ (experts 8.5%, family physicians 7.7%, and nurses 7.5%, p = 0.39). There was no relevant bias among observer groups. Intra-observer variability expressed as standard deviation divided by the mean was 8%, and inter-observer variability was 9%. In conclusion, reproducibility of the ABI measurement was good in this cohort of elderly patients who almost all had values in the normal range. The mean error of 8—9% within or between observers is smaller than with established screening measures. Since there were no differences among observers with different training backgrounds, our study confirms the appropriateness of ABI assessment for screening peripheral arterial disease (PAD) and generalized atherosclerosis in the primary case setting. Given the importance of the early detection and management of PAD, this diagnostic tool should be used routinely as a standard for PAD screening. Additional studies will be required to confirm our observations in patients with PAD of various severities

Alternative Splicing of G9a Regulates Neuronal Differentiation

Chromatin modifications are critical for the establishment and maintenance of differentiation programs. G9a, the enzyme responsible for histone H3 lysine 9 dimethylation in mammalian euchromatin, exists as two isoforms with differential inclusion of exon 10 (E10) through alternative splicing. We find that the G9a methyltransferase is required for differentiation of the mouse neuronal cell line N2a and that E10 inclusion increases during neuronal differentiation of cultured cells, as well as in the developing mouse brain. Although E10 inclusion greatly stimulates overall H3K9me2 levels, it does not affect G9a catalytic activity. Instead, E10 increases G9a nuclear localization. We show that the G9a E10+ isoform is necessary for neuron differentiation and regulates the alternative splicing pattern of its own pre-mRNA, enhancing E10 inclusion. Overall, our findings indicate that by regulating its own alternative splicing, G9a promotes neuron differentiation and creates a positive feedback loop that reinforces cellular commitment to differentiation