Combining molecular dynamics and docking simulations to develop targeted protocols for performing optimized virtual screening campaigns on the HTRPM8 channel

Background: There is an increasing interest in TRPM8 ligands of medicinal interest, the rational design of which can be nowadays supported by structure-based in silico studies based on the recently resolved TRPM8 structures. Methods: The study involves the generation of a reliable hTRPM8 homology model, the reliability of which was assessed by a 1.0 \u3bcs MD simulation which was also used to generate multiple receptor conformations for the following structure-based virtual screening (VS) campaigns; docking simulations utilized different programs and involved all monomers of the selected frames; the so computed docking scores were combined by consensus approaches based on the EFO algorithm. Results: The obtained models revealed very satisfactory performances; LiGen\u2122 provided the best results among the tested docking programs; the combination of docking results from the four monomers elicited a markedly beneficial effect on the computed consensus models. Conclusions: The generated hTRPM8 model appears to be amenable for successful structure-based VS studies; cross-talk modulating effects between interacting monomers on the binding sites can be accounted for by combining docking simulations as performed on all the monomers; this strategy can have general applicability for docking simulations involving quaternary protein structures with multiple identical binding pockets

Multidrug-resistant HIV viral rebound during early syphilis: a case report

Background Syphilis has been associated with an increase in HIV RNA and a temporary decline in CD4 T cell counts in people living with HIV who are not receiving antiretroviral treatment (ART), and may be associated with a transient HIV RNA rebound in those who are receiving ART. Our case is the first to highlight the risk of a multidrug-resistant HIV viral rebound during the course of early syphilis even if antiretroviral drug concentrations are within the therapeutic range. Case presentation This 50-year-old HIV-1-positive male patient with concomitant early syphilis presented with an HIV RNA rebound (8908 copies/mL) during a scheduled visit to our clinic. He was receiving a stable ART regimen consisting of darunavir/cobicistat plus dolutegravir, and had a 15-year history of viral suppression. Good short-term drug adherence could be inferred as liquid chromatography tandem mass spectrometry showed that his trough antiretroviral drug concentrations were within the therapeutic range: darunavir 2353 ng/mL (minimum effective concentration > 500 ng/mL) and dolutegravir 986 ng/mL (minimum effective concentration > 100 ng/mL). A plasma RNA genotype resistance test revealed wild-type virus in the integrase region and protease region (PR), but extensive resistance in the reverse transcriptase (RT) region (M41L, E44D, D67N, K70R, M184V, L210W and T215Y). Phylogenetic analysis of next-generation sequences (used to investigate the presence of minor viral variants), the PR and RT sequences from plasma HIV RNA and pro-viral DNA extracted from peripheral blood mononuclear cells during the viral rebound, and a Sanger sequence obtained during a previous virological failure suggested clonal viral expression because the previous PR resistance mutations had been lost or had not been archived in pro-viral DNA. Conclusions This case shows that early syphilis may cause an HIV RNA rebound in patients under stable virological control with the potential of transmitting an extensively drug-resistant virus

Is a wearable sensor-based characterisation of gait robust enough to overcome differences between measurement protocols? A multi-centric pragmatic study in patients with multiple sclerosis

Inertial measurement units (IMUs) allow accurate quantification of gait impairment of people with multiple sclerosis (pwMS). Nonetheless, it is not clear how IMU-based metrics might be influenced by pragmatic aspects associated with clinical translation of this approach, such as data collection settings and gait protocols. In this study, we hypothesised that these aspects do not significantly alter those characteristics of gait that are more related to quality and energetic efficiency and are quantifiable via acceleration related metrics, such as intensity, smoothness, stability, symmetry, and regularity. To test this hypothesis, we compared 33 IMU-based metrics extracted from data, retrospectively collected by two independent centres on two matched cohorts of pwMS. As a worst-case scenario, a walking test was performed in the two centres at a different speed along corridors of different lengths, using different IMU systems, which were also positioned differently. The results showed that the majority of the temporal metrics (9 out of 12) exhibited significant between-centre differences. Conversely, the between-centre differences in the gait quality metrics were small and comparable to those associated with a test-retest analysis under equivalent conditions. Therefore, the gait quality metrics are promising candidates for reliable multi-centric studies aiming at assessing rehabilitation interventions within a routine clinical context

Proven intra and interobserver reliability in the echographic assessments of body fat changes related to HIV associated Adipose Redistribution Syndrome (HARS)

OBJECTIVE: To prove intra- and inter-observer's reliability of ultrasound (US) in the assessment of lipoatrophic findings related to the HIV associated Adipose Redistribution Syndrome (HARS). PATIENTS AND METHODS: In two separated sessions, 2 consecutive measurements of subcutaneous fat thickness (SFT) were performed by each observer at the deepest point of Bichat pad, the dorsal face of arm and the mid thigh for the assessment of facial, brachial and crural lipoatrophy, respectively. We enrolled 20 HIV patients, rotating an experienced and untrained sonologist. The assessments were performed avoiding any stand off pads in the skin and excluding artefacts due to the too abundant quantity of gel to obtaining, with minimal transducer pressure, the best resolution of the reference points. RESULTS: Means of facial, brachial and crural SFT showed no significant differences between the workers. Coefficients of variability (SD/mean x100) were similar for facial (ranges: 4.7-5.2% vs 4.9-5.6%, respectively), brachial (ranges: 5.8-8.4% vs 9.7-11.2%) and crural SFTs (ranges: 5.9-6% vs 6.2-8.7%). There was greater consistency in the measurements performed by the experienced vs the untrained worker. Inter-observer agreement, assessed through kappa statistic (k) analysis, confirmed increased measurement's agreement in the facial (k ranged from 0.40 to 0.60), brachial (k: 0.23-0.63) and crural SFT assessments (k: 0.58-0.70) from the 1(st) to 2(nd) session. CONCLUSIONS: US shows low intra observer variability and good inter observer reliability in the assessment of body fat changes related to the HARS. The different degree of consistency by the workers and the improvement of interobserver agreement, suggest to stating a well defined period of training to obtain better US reliabilit

Implante de Desfibrilador: a Propósito de um Limiar de Desfibrilaçao Elevado

Este artigo tem por finalidade orientar a conduta nos casos de pacientes com taquicardia ventricular sustentada e disfunçao ventricular grave, encaminhados para implante de desfibrilador, situaçao cada vez mais freqüente. Discute-se a possibilidade de utilizar alternativas cirúrgicas para solucionar situaçoes em que os limiares de desfibrilaçao permanecem altos durante os testes relizados no momento do implante

Implante de Desfibrilador: a Propósito de um Limiar de Desfibrilaçao Elevado

Este artigo tem por finalidade orientar a conduta nos casos de pacientes com taquicardia ventricular sustentada e disfunçao ventricular grave, encaminhados para implante de desfibrilador, situaçao cada vez mais freqüente. Discute-se a possibilidade de utilizar alternativas cirúrgicas para solucionar situaçoes em que os limiares de desfibrilaçao permanecem altos durante os testes relizados no momento do implante