Detection of Chemical Engagement of Solute Carrier Proteins by a Cellular Thermal Shift Assay

Solute carriers (SLCs) are transmembrane proteins that transport various nutrients, metabolites, and drugs across cellular membranes. Despite the relevance of SLCs to cell homeostasis, metabolism, and disease states, for the majority of SLCs we lack experimental evidence regarding the nature of the cognate ligands, whether endobiotic or xenobiotic. Moreover, even for the roughly 20 SLCs for which inhibitors have been characterized, engagement assays in cells are limited to the accessibility of radiolabeled or fluorescent probes. The cellular thermal shift assay (CETSA) has been introduced as a powerful method to assess target engagement by monitoring ligand-induced changes in the thermal stability of cellular proteins. We addressed the question of whether CETSA could be modified to become routinely applicable to membrane transporters such as SLCs. We used SLC16A1 (MCT1) and SLC1A2 (EAAT2) as targets to establish robust conditions by which chemical engagement of SLCs can be detected. Using immunoblotting, we demonstrate that treatment with the SLC16A1 inhibitors AZD3965 and AR-C155858 stabilized endogenous SLC16A1 in HEK293 cell lysates as well as intact cells. In addition, the high-affinity ligand of SLC16A1, l-lactate, and the low-affinity ligand, formate, resulted in strong and weak stabilization of SLC16A1, respectively. Moreover, we observed stabilization of SLC1A2 upon treatment with the selective inhibitor WAY-213613. We propose that the experimental approach presented here should be generally and easily applicable for monitoring the engagement of chemical ligands by SLCs in cellular settings and thus assisting in their deorphanization

Tuberculosis Transmission from Healthcare Workers to Patients and Co-workers: A Systematic Literature Review and Meta-Analysis

<div><p>Healthcare workers (HCWs) are at risk of becoming infected with tuberculosis (TB), and potentially of being infectious themselves when they are ill. To assess the magnitude of healthcare-associated TB (HCA-TB) transmission from HCWs to patients and colleagues, we searched three electronic databases up to February 2014 to select primary studies on HCA-TB incidents in which a HCW was the index case and possibly exposed patients and co-workers were screened.We identified 34 studies out of 2,714 citations. In 29 individual investigations, active TB was diagnosed in 3/6,080 (0.05%) infants, 18/3,167 (0.57%) children, 1/3,600 (0.03%) adult patients and 0/2,407 HCWs. The quantitative analysis of 28 individual reports showed that combined proportions of active TB among exposed individuals were: 0.11% (95% CI 0.04–0.21) for infants, 0.38% (95% CI 0.01–1.60) for children, 0.09% (95% CI 0.02–0.22) for adults and 0.00% (95% CI 0.00–0.38) for HCWs. Combined proportions of individuals who acquired TB infection were: 0.57% (95% CI 7.28E-03 – 2.02) for infants, 0.9% (95% CI 0.40–1.60) for children, 4.32% (95% CI 1.43–8.67) for adults and 2.62% (95% CI 1.05–4.88) for HCWs. The risk of TB transmission from HCWs appears to be lower than that recorded in other settings or in the healthcare setting when the index case is not a HCW. To provide a firm evidence base for the screening strategies, more and better information is needed on the infectivity of the source cases, the actual exposure level of screened contacts, and the environmental characteristics of the healthcare setting.</p></div

Characteristics of 117 Health Care Workers index cases of pulmonary Tuberculosis in 34 included studies.

<p>Characteristics of 117 Health Care Workers index cases of pulmonary Tuberculosis in 34 included studies.</p

Proportion meta-analysis (random effects).

<p>Forest plots for: A. Proportion of active TB cases among infants; B. Proportion of active TB cases among children; C. Proportion of active TB cases among adult patients; D. Proportion of active TB cases among HCWs. (A B C D elements are ordered from top to bottom and left to right).</p

Response to selected RD1 peptides in Ugandan HIV-infected patients with smear positive pulmonary tuberculosis: a pilot study-1

Ming cells (SFCs) per million PBMC for each individual at the time of diagnosis and 6 months after therapy are reported. For 9/12 individuals data are reported also after 3 months of therapy. The p value denotes the difference between the responders in each group.<p><b>Copyright information:</b></p><p>Taken from "Response to selected RD1 peptides in Ugandan HIV-infected patients with smear positive pulmonary tuberculosis: a pilot study"</p><p>http://www.biomedcentral.com/1471-2334/8/11</p><p>BMC Infectious Diseases 2008;8():11-11.</p><p>Published online 28 Jan 2008</p><p>PMCID:PMC2267196.</p><p></p

Response to selected RD1 peptides in Ugandan HIV-infected patients with smear positive pulmonary tuberculosis: a pilot study-0

Cate individuals with tuberculosis (TB patients) and unshaded symbols indicate individuals without tuberculosis (No active TB). Horizontal lines indicate median values. The TB patient group had a significantly higher response to the sum of ESAT-6 and CFP-10 peptides compared with the group without active TB (p = 0.02).<p><b>Copyright information:</b></p><p>Taken from "Response to selected RD1 peptides in Ugandan HIV-infected patients with smear positive pulmonary tuberculosis: a pilot study"</p><p>http://www.biomedcentral.com/1471-2334/8/11</p><p>BMC Infectious Diseases 2008;8():11-11.</p><p>Published online 28 Jan 2008</p><p>PMCID:PMC2267196.</p><p></p

Table_1_B-cell-depleted patients with persistent SARS-CoV-2 infection: combination therapy or monotherapy? A real-world experience.DOCX

ObjectivesThe aim of the study was to describe a cohort of B-cell-depleted immunocompromised (IC) patients with prolonged or relapsing COVID-19 treated with monotherapy or combination therapy.MethodsThis is a multicenter observational retrospective study conducted on IC patients consecutively hospitalized with a prolonged or relapsing SARS-CoV-2 infection from November 2020 to January 2023. IC COVID-19 subjects were stratified according to the monotherapy or combination anti-SARS-CoV-2 therapy received.ResultsEighty-eight patients were enrolled, 19 under monotherapy and 69 under combination therapy. The study population had a history of immunosuppression (median of 2 B-cells/mm3, IQR 1–24 cells), and residual hypogammaglobulinemia was observed in 55 patients. A reduced length of hospitalization and time to negative SARS-CoV-2 molecular nasopharyngeal swab (NPS) in the combination versus monotherapy group was observed. In the univariable and multivariable analyses, the percentage change in the rate of days to NPS negativity showed a significant reduction in patients receiving combination therapy compared to those receiving monotherapy.ConclusionIn IC persistent COVID-19 patients, it is essential to explore new therapeutic strategies such as combination multi-target therapy (antiviral or double antiviral plus antibody-based therapies) to avoid persistent viral shedding and/or severe SARS-CoV-2 infection.</p

Table_2_B-cell-depleted patients with persistent SARS-CoV-2 infection: combination therapy or monotherapy? A real-world experience.DOCX

ObjectivesThe aim of the study was to describe a cohort of B-cell-depleted immunocompromised (IC) patients with prolonged or relapsing COVID-19 treated with monotherapy or combination therapy.MethodsThis is a multicenter observational retrospective study conducted on IC patients consecutively hospitalized with a prolonged or relapsing SARS-CoV-2 infection from November 2020 to January 2023. IC COVID-19 subjects were stratified according to the monotherapy or combination anti-SARS-CoV-2 therapy received.ResultsEighty-eight patients were enrolled, 19 under monotherapy and 69 under combination therapy. The study population had a history of immunosuppression (median of 2 B-cells/mm3, IQR 1–24 cells), and residual hypogammaglobulinemia was observed in 55 patients. A reduced length of hospitalization and time to negative SARS-CoV-2 molecular nasopharyngeal swab (NPS) in the combination versus monotherapy group was observed. In the univariable and multivariable analyses, the percentage change in the rate of days to NPS negativity showed a significant reduction in patients receiving combination therapy compared to those receiving monotherapy.ConclusionIn IC persistent COVID-19 patients, it is essential to explore new therapeutic strategies such as combination multi-target therapy (antiviral or double antiviral plus antibody-based therapies) to avoid persistent viral shedding and/or severe SARS-CoV-2 infection.</p

Additional file 1 of A machine learning approach for early identification of patients with severe imported malaria

Additional file 1: Fig. S1. A visualization of the clusters related to the severe malaria patients, obtained using K-means

Layout of TB laboratory.

<p>Layout of TB laboratory.</p