Changes in ROS production induced by pentose phosphate pathway inhibition.

<p>Mean fluorescence intensity (MFI—arbitrary units) by pancreatic islets from fed (CT) or 48-hour fasted (48h) rats, treated with dihydroethidium (DHE), and incubated for 60 minutes in the presence of 2.8 mM glucose with or without addition of dehydroepiandrosterone (DHEA—75 μM)–experimental protocol 5. The results are presented as mean ± SEM of five different cell preparations for each group. * p <0.05 compared to fed control (CT) under the same conditions as indicated by the Student t-test.</p

ROS production and insulin secretion by pancreatic islets from fed and fasted rats in the presence of 20 mM leucine and 2.8 or 16.7 mM glucose.

<p><b>(A)</b> Mean fluorescence intensity (MFI—arbitrary units) and <b>(B)</b> insulin secretion by pancreatic islets from fed (CT) or 48-hour fasted (48h) rats, treated with dihydroethidium (DHE), and incubated for 60 minutes in the presence of 2.8 mM glucose with or without addition of VAS2870 (20 μM) or DPI (5 μM), associated or not with leucine (LEU—20 mM)–experimental protocol 4. The results are presented as mean ± SEM of four different cell preparations for each group * p <0.05 compared to fed control (CT) under the same conditions as indicated by the Student t-test. <b>(C)</b> Inverse correlation between MFI and insulin secretion in the presence of 2.8 mM glucose with or without addition of VAS2870 (20 μM) or DPI (5 μM) in pancreatic islets isolated from fed or 48-hour fasted rats as indicated by the correlation test using the GraphPad Prism 5.</p

Metabolic pathways associated to NADPH production from glucose and leucine oxidation in pancreatic islets.

<p>The sites indicated are: the pentose phosphate pathway and reactions involving intermediates of the Krebs cycle. ACO, aconitase; CYT, cytosol; DHEA, dehydroepiandrosterone; G3P, glyceraldehyde-3-phosphate dehydrogenase; GDH, glutamate dehydrogenase; GPx, glutathione peroxidase; GR, glutathione reductase; GSH, reduced glutathione; GSSG, oxidized glutathione; IDH, isocitrate dehydrogenase; ME, malic enzyme; MIT, mitochondria; PDH, pyruvate dehydrogenase; PPP, pentose phosphate pathway; SOD1, superoxide dismutase 1; 2OG, 2-oxoglutarate; α-KIC, α-ketoisocaproic acid.</p

Changes of NADPH/NADP+ ratio in INS-1E cells cultivated in the presence of 20 mM leucine and 2.8 or 16.7 mM glucose.

<p>NADPH/NADP⁺ ratio in INS-1E cells incubated for 60 minutes in the presence of 2.8 or 16.7 mM glucose with or without addition of leucine (LEU—20 mM)–experimental protocol 6. The results are presented as mean ± SEM of three different cell preparations for each group. *p <0.05 compared to 2.8 mM glucose as indicated by the Student t-test.</p

Kinetic measurement of net cytosolic ROS production.

<p><b>(A)</b> Mean fluorescence intensity (MFI—arbitrary units) by pancreatic islets isolated from fed rats, treated with dihydroethidium (DHE), and incubated with 2.8 mM glucose for 120 minutes or for 60 minutes with 2.8 mM glucose that was replaced by 16.7 mM glucose and then maintained for another 60-minute period incubation—experimental protocol 2. <b>(B)</b> The corresponding areas under the curves were then calculated. The results are presented as mean ± SEM of five cell preparations for each group. *p <0.05 as compared to 2.8 mM glucose and indicated by the Student’s t-test.</p

Kinetic measurement of net cytosolic ROS production.

<p><b>(A)</b> Mean fluorescence intensity (MFI—arbitrary units) by pancreatic islets isolated from fed rats, treated with dihydroethidium (DHE), and measured every 5 minutes during 120-minute incubation in the presence of 2.8 or 16.7 mM glucose—experimental protocol 1. <b>(B)</b> The corresponding areas under the curves were then calculated. The results are presented as mean ± SEM of four cell preparations for each group. * p <0.05 as compared to 2.8 mM glucose and indicated by the Student’s t-test.</p

Mitochondrial ROS measurement.

<p>Mean fluorescence intensity (MFI—arbitrary units) by pancreatic islets from fed (CT) or 48-hour fasted (48h) rats. The islets were treated with MitoSOX Red reagent and incubated for 60 minutes in the presence of 2.8 or 16.7 mM glucose with or without addition of VAS2870 (20 μM) or DPI (5 μM) associated or not with leucine (LEU—20 mM)—experimental protocol 4. The results are presented as mean ± SEM of four cell preparations for each group as indicated by one-way ANOVA and Dunnett's post test.</p

Data_Sheet_1_Determinants of HIV late presentation among men who have sex with men in Portugal (2014–2019): who’s being left behind?.docx

IntroductionHIV late presentation (LP) remains excessive in Europe. We aimed to analyze the factors associated with late presentation in the MSM population newly diagnosed with HIV in Portugal between 2014 and 2019.MethodsWe included 391 newly HIV-1 diagnosed Men who have Sex with Men (MSM), from the BESTHOPE project, in 17 countrywide Portuguese hospitals. The data included clinical and socio-behavioral questionnaires and the viral genomic sequence obtained in the drug resistance test before starting antiretrovirals (ARVs). HIV-1 subtypes and epidemiological surveillance mutations were determined using different bioinformatics tools. Logistic regression was used to estimate the association between predictor variables and late presentation (LP).ResultsThe median age was 31 years, 51% had a current income between 501–1,000 euros, 28% were migrants. 21% had never been tested for HIV before diagnosis, with 42.3% of MSM presenting LP. 60% were infected with subtype B strains. In the multivariate regression, increased age at diagnosis, higher income, lower frequency of screening, STI ever diagnosed and higher viral load were associated with LP.ConclusionOur study suggests that specific subgroups of the MSM population, such older MSM, with higher income and lower HIV testing frequency, are not being targeted by community and clinical screening services. Overall, targeted public health measures should be strengthened toward these subgroups, through strengthened primary care testing, expanded access to PrEP, information and promotion of HIV self-testing and more inclusive and accessible health services.</p

Data_Sheet_1_Determinants of HIV late presentation among men who have sex with men in Portugal (2014–2019): who’s being left behind?.docx

IntroductionHIV late presentation (LP) remains excessive in Europe. We aimed to analyze the factors associated with late presentation in the MSM population newly diagnosed with HIV in Portugal between 2014 and 2019.MethodsWe included 391 newly HIV-1 diagnosed Men who have Sex with Men (MSM), from the BESTHOPE project, in 17 countrywide Portuguese hospitals. The data included clinical and socio-behavioral questionnaires and the viral genomic sequence obtained in the drug resistance test before starting antiretrovirals (ARVs). HIV-1 subtypes and epidemiological surveillance mutations were determined using different bioinformatics tools. Logistic regression was used to estimate the association between predictor variables and late presentation (LP).ResultsThe median age was 31 years, 51% had a current income between 501–1,000 euros, 28% were migrants. 21% had never been tested for HIV before diagnosis, with 42.3% of MSM presenting LP. 60% were infected with subtype B strains. In the multivariate regression, increased age at diagnosis, higher income, lower frequency of screening, STI ever diagnosed and higher viral load were associated with LP.ConclusionOur study suggests that specific subgroups of the MSM population, such older MSM, with higher income and lower HIV testing frequency, are not being targeted by community and clinical screening services. Overall, targeted public health measures should be strengthened toward these subgroups, through strengthened primary care testing, expanded access to PrEP, information and promotion of HIV self-testing and more inclusive and accessible health services.</p

Data_Sheet_1_Determinants of HIV late presentation among men who have sex with men in Portugal (2014–2019): who’s being left behind?.docx

IntroductionHIV late presentation (LP) remains excessive in Europe. We aimed to analyze the factors associated with late presentation in the MSM population newly diagnosed with HIV in Portugal between 2014 and 2019.MethodsWe included 391 newly HIV-1 diagnosed Men who have Sex with Men (MSM), from the BESTHOPE project, in 17 countrywide Portuguese hospitals. The data included clinical and socio-behavioral questionnaires and the viral genomic sequence obtained in the drug resistance test before starting antiretrovirals (ARVs). HIV-1 subtypes and epidemiological surveillance mutations were determined using different bioinformatics tools. Logistic regression was used to estimate the association between predictor variables and late presentation (LP).ResultsThe median age was 31 years, 51% had a current income between 501–1,000 euros, 28% were migrants. 21% had never been tested for HIV before diagnosis, with 42.3% of MSM presenting LP. 60% were infected with subtype B strains. In the multivariate regression, increased age at diagnosis, higher income, lower frequency of screening, STI ever diagnosed and higher viral load were associated with LP.ConclusionOur study suggests that specific subgroups of the MSM population, such older MSM, with higher income and lower HIV testing frequency, are not being targeted by community and clinical screening services. Overall, targeted public health measures should be strengthened toward these subgroups, through strengthened primary care testing, expanded access to PrEP, information and promotion of HIV self-testing and more inclusive and accessible health services.</p