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Aims: This study aimed to evaluate the role of the 24-Hour Ambulatory Blood Pressure Monitoring (ABPM) as a possible predictor of vascular outcomes in office normotensive people with type 1 diabetes mellitus (T1DM)., Methods: This is a systematic review including cohort studies from the Embase, PubMed/Medline, and Web of Science databases on people with T1DM undergoing ABPM and subsequent evaluation of vascular complications. Measurements of difference (MD) were obtained using random effect model meta-analysis., Results: We found 364 articles and 49 duplicates. Seven studies were included, comprising 635 participants aged 25.8 ± 6.2 years. Most (57.5 %) were men, mean duration of diabetes was 11.8 ± 5.3 years, mean glycated hemoglobin level among participants was 8.5 % ± 1.6 %, and mean follow-up time was 4.2 years. Lower night systolic blood pressure MD - 4.37 mmHg (p = 0.0009) and night diastolic blood pressure MD - 3.97 mmHg (p < 0.0001) were associated with lower incidence of albuminuria. People withT1DM who presented no beginning or progression of retinopathy were those with lower night diastolic blood pressure MD - 3.62 mmHg (p = 0.042), diurnal diastolic blood pressure MD - 2.69 mmHg (p = 0.0138), and 24-hour diastolic blood pressure MD - 3.65 mmHg (p = 0.037)., Conclusion: Small mean differences in blood pressure parameters, as measured by ABPM, between people with T1DM are associated with a lower incidence or risk of progression of nephropathy and retinopathy., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Streptococcus pneumoniae is an important human pathogen that can colonize the respiratory tract of healthy individuals. The respiratory tract mucosa is thus the first barrier for this pathogen. In this study, we have tested three models of the respiratory epithelium with immune cells: (i) monolayer of A549 human lung epithelial cells, (ii) A549 + macrophages differentiated from the human monocytic THP-1 cell line (dMφ) and (iii) A549 + dMφ + dendritic cells differentiated from THP-1 (dDC) using a two-chamber system. Pneumococcal strains Rx1 (non-encapsulated) and BHN418 (serotype 6B) were incubated with the cells and secretion of IL-6, IL-8, IL-1β, TNF-α and IL-10 was evaluated. Overall, the models using co-cultures of A549 + dMφ and A549 + dMφ + dDC elicited higher levels of pro-inflammatory cytokines and the non-encapsulated strain elicited an earlier cytokine response. BHN418 pspA (pneumococcal surface protein A) and pspC (pneumococcal surface protein C) knockouts elicited similar cytokine secretion in the co-culture models, whereas BHN18 ply (pneumolysin) knockout induced much lower levels. The results are in accordance with the activation of the inflammasome by Ply. Finally, we evaluated pneumococcal extracellular vesicles (pEVs) in the co-culture models and observed secretion of pro-inflammatory cytokines in the absence of cytotoxicity. Since pEVs are being studied as vaccine candidate against pneumococcal infections, the co-cultures of A549 + dMφ and A549 + dMφ + dDC are simple models that could be used to evaluate pEV vaccine batches., (© 2024. The Author(s) under exclusive licence to Sociedade Brasileira de Microbiologia.)

Streptococcus pneumoniae can cause diseases with high mortality and morbidity. The licensed vaccines are based on capsular polysaccharides and induce antibodies with low cross reactivity, leading to restricted coverage of serotypes. For surpassing this limitation, new pneumococcal vaccines are needed for induction of broader protection. One important candidate is the pneumococcal surface protein A (PspA), which can be classified in 6 clades and 3 families. We have reported an efficient process for production and purification of untagged recombinant PspA from clade 4 (PspA4Pro). We now aim to obtain a highly pure recombinant PspA from clade 1 (PspA1) to be included, together with PspA4Pro, in a vaccine formulation to broaden response against pneumococci. The vector pET28a-pspA1 was constructed and used to transform Escherichia coli BL21(DE3) strain. One clone with high production of PspA1 was selected and adapted to high-density fermentation (HDF) medium. After biomass production in 6 L HDF using a bioreactor, the purification was defined after testing 3 protocols. During the batch bioreactor cultivation, plasmid stability remained above 90% and acetate formation was not detected. The final protein purification process included treatment with a cationic detergent after lysis, anion exchange chromatography, cryoprecipitation, cation exchange chromatography, and multimodal chromatography. The final purification process showed PspA1 purity of 93% with low endotoxin content and an overall recovery above 20%. The novel established process can be easily scaled-up and proved to be efficient to obtain a highly pure untagged PspA1 for inclusion in vaccine formulations. KEY POINTS: • Purification strategy for recombinant PspA1 from Streptococcus pneumoniae • Downstream processing for untagged protein antigens, the case of PspA1 • Purification strategy for PspA variants relies on buried amino acids in their sequences., (© 2024. The Author(s).)

Aim: To assess the relationship of the presence of sarcopenia and malnutrition with unfavorable clinical outcomes: prolonged length of hospital stay (LOS), readmission, and one-year mortality in older patients with type 2 diabetes (T2D)., Methods: Were included 319 patients with ≥ 60 years of age with T2D hospitalized at a university hospital in Southern Brazil. Sarcopenia was diagnosed according to handgrip strength (HGS), calf circumference (CC), and the timed up and go (TUG) walking test, and malnutrition according to the subjective global assessment (SGA) and the mini nutritional assessment long form (MNA-LF). Multivariate analyses, adjusted for confounders, were performed to assess the association of sarcopenia and malnutrition with clinical outcomes. One-year survival was compared using Kaplan-Meier analysis., Results: The association between sarcopenia and malnutrition increased by 2.42 times (95 %CI 1.35-4.36) the probability of LOS ≥ 14 days and by 2.01 times (95 %CI 1.09-3.72) the risk of one-year mortality. Older patients with malnutrition and sarcopenia have a higher risk of one-year mortality (log-rank p < 0.05) compared with well-nourished patients without sarcopenia., Conclusion: In older patients with type 2 diabetes, those with sarcopenia, and malnutrition have higher odds of prolonged hospitalization and risk of mortality within one year after hospital discharge., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Objective: Body composition changes are associated with adverse effects such as increased insulin resistance (IR) in individuals with diabetes mellitus. This study aims to evaluate the association between different body adiposity markers and IR in adults with type 1 diabetes (T1D)., Subjects and Methods: The cross-sectional study included outpatient adults with T1D from a university public hospital in southern Brazil. The body adiposity markers studied were waist circumference (WC), waist-height ratio (WHtR), body mass index (BMI), conicity index (CI), lipid accumulation product (LAP) and body adiposity index (BAI). IR was calculated using an Estimated Glucose Disposal Rate (EGDR) equation (analyzed in tertiles), considering an inverse relation between EGDR and IR. Poisson regression models were used to estimate the odds ratio (OR) and 95% CIs of association of adiposity markers with IR., Results: A total of 128 patients were enrolled (51% women), with a median EGDR of 7.2 (4.4-8.7) mg.kg -1 .min -1 . EGDR was negatively correlated with WC ( r = -0.36, p < 0.01), WHtR ( r = -0.39, p < 0.01), CI ( r = -0.44, p < 0.01), LAP ( r = -0.41, p < 0.01) and BMI ( r = -0.24, p < 0.01). After regression analyses, WC (OR = 2.07; CIs: 1.12-3.337; p = 0.003), WHtR (OR = 2.77; CIs: 1.59-4.79; p < 0.001), CI (OR = 2.59; CIs: 1.43-4.66; p = 0.002), LAP (OR = 2.27; CIs: 1.25-4.11; p = 0.007) and BMI (OR = 1.78; CIs: 1.09-2.91; p = 0.019) remained associated with IR., Conclusion: The authors suggest using the studied adiposity markers as a routine since they were shown to be suitable parameters in association with IR.

Objective: The objective of this study was to verify the impact of carbohydrate counting (CC) on glycemic control and body weight variation (primary and secondary outcomes, respectively) between consultations in patients with diabetes mellitus (T1D) followed at a tertiary hospital in southern Brazil in a public health system environment. We also sought to investigate CC adherence., Materials and Methods: This retrospective cohort study included 232 patients with T1D who underwent nutritional monitoring at a referral hospital for diabetes care between 2014 and 2018. To assess primary and secondary outcomes, data from 229 patients, 49 of whom underwent CC during this period and 180 individuals who used fixed doses of insulin, were analyzed. The impact of CC on glycemic control was assessed with the mean glycated hemoglobin (HbA1c) level at all consultations during the follow-up period., Results: In the model adjusted for the most confounders (except pregnancy), the mean HbA1c was better in the CC group (8.66 ± 0.4% vs. 9.36 ± 0.39%; p = 0.016), and body weight variation was lower (0.13 ± 0.28 kg vs. 0.53 ± 0.24 kg; p = 0.024). Adherence to CC was reported in 69.2% of consultations., Conclusion: CC optimized the glycemic control of individuals with T1D, resulting in less weight variation than in the fixed insulin dose group, which indicates that CC is an important care strategy for these patients.

Introduction: Drug scheduling in older adults can be a challenge, especially considering polypharmacy, physical dependency, and possible drug interactions. Properly testing alternative treatment regimens could therefore help to overcome treatment barriers. Hypothyroidism is a prevalent condition in older adults, however, studies evaluating L-thyroxine treatment effectiveness in this specific age group are still lacking. Most studies testing an evening administration of levothyroxine were mainly composed of younger adults. Therefore, this trial is aimed to assess if evening levothyroxine (LT4) administration can effectively control hypothyroidism in older patients., Materials and Methods: A randomized crossover clinical trial was conducted between June 2018 and March 2020 at the Hospital de Clínicas de Porto Alegre, a teaching hospital in Brazil, to compare the efficacy of morning and evening administration of LT4 for hypothyroidism control in older patients. The study protocol is published elsewhere. A total of 201 participants, ≥60 years old, with primary hypothyroidism treated with LT4 for at least 6 months and on stable doses for at least 3 months were included. Participants were randomly assigned to a starting group of morning LT4 intake (60 min before breakfast) or bedtime LT4 intake (60 min after the last meal). After ≥12 weeks of follow-up, a crossover between strategies was performed. The primary outcome was the change in serum thyrotropin (Thyroid-Stimulating Hormone; TSH) levels after 12 weeks of each LT4 administration regimen., Results: A total of 201 participants with mean age of 72.4 ± 7.2 years were included, out of which 84.1% were women; baseline characteristics and frequency of controlled hypothyroidism were similar between groups. Mean baseline TSH was 3.43 ± 0.25 mUI/L. In total, 118 participants attended three meetings, allowing 135 comparisons by crossover analytic strategy. Mean TSH levels after follow-up were 2.95 ± 2.86 in the morning group and 3.64 ± 2.86 in the bedtime group, p = 0.107., Discussion: Thyroid-Stimulating Hormone levels and frequency of controlled hypothyroidism were similar during the follow-up period regardless of the treatment regimen (morning or bedtime)., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 de Mello, Giassi, Stahl, Machado Assis, Flores, de Lima, Piccoli and da Costa Rodrigues.)

Objective: To assess the prevalence of psychiatric disorders in patients with type 1 diabetes mellitus (T1D) and to compare patients with and without psychiatric disorder., Methods: We made a cross-sectional study including patients with T1D assisted in the outpatient clinics of the Brazilian National Health System. To assess depression and anxiety, we used the PHQ-9 questionnaire and the DSM-5th edition criteria, respectively. B-PAID evaluated the level of emotional distress associated with diabetes; EAT-26, eating disorders; SCI-R, adherence to the proposed clinical treatment., Results: We analyzed 166 patients aged 33 (22-45.2) years, 53.6% female. The prevalence of depression and anxiety was 20.5% and 40.4%, respectively. HbA1c was worse in the depressed (9.0% vs. 8.4%, p = 0.008), in the anxious ones (9.0% vs. 8.3%, p = 0.012) and in the patients with high levels of B-PAID (8.8 % vs. 8.3 %, p = 0.009). There was no difference in the prevalence of complications related to diabetes., Conclusion: The prevalence of psychiatric disorders and emotional distress related to diabetes was high in our population of T1D patients, and depression and high levels of B-PAID were associated with the worse glycemic control.

Background: Obesity is associated with increased general mortality and comorbidities, it is multifactorial and some evidence has shown that sleep duration and shift work may be implicated in its pathogenesis., Objectives: The aim of this study was to evaluate the association between shift work, quality of life and obesity among healthcare workers of a Brazilian University Hospital., Methods: A cross-sectional study was performed from April 2013 to December 2014 with 200 workers of a University Hospital. Sociodemographic data were evaluated and BREF WHOQOL was used for quality of life. The physical activity was evaluated using the International Physical Activity Questionnaire (IPAQ), Chronotypes and daily sleep preference were investigated using Munich Chronotype Questionnaire (MCTQ). Venous blood was collected after 12-h of fasting for laboratory tests., Results: In this sample, the night shift workers had higher income and were older compared to day shift workers. Night shift workers sleep less hours, had higher weight, body mass index and abdominal circumference when compared to the day shift workers. Night shift workers had almost 3 times higher association with abdominal obesity independent of age and gender, than day shift workers. MCTQ parameters showed that night shift workers had lower sleep duration during working days and also during free days, associated with a higher level of social jetlag. Social jetlag had an association with obesity. We found no difference for quality of life between shifts., Conclusions: Night work was a risk factor for abdominal obesity, social jetlag was higher in night shift workers and it was associated with presence of obesity., Competing Interests: Competing interestsThe authors declare that they have no competing interests., (© The Author(s) 2020.)

Background: The aging population is associated with increased multimorbidity and polypharmacy. Older adults are at a higher risk of adverse events and reduced therapeutic response. This phenomenon is partially explained by drug interactions and treatment adherence. Most randomized clinical trials have found no significant differences between morning and evening levothyroxine (LT 4 ) administration in young adults, but there is little evidence regarding alternative LT 4 regimens in older populations. Thus, the MONIALE trial aims to test an alternative schedule for LT 4 administration in older adults., Methods/design: This randomized crossover clinical trial will include participants aged 60 years or older with primary hypothyroidism. The trial groups will consist of morning LT 4 intake (60 min before breakfast) or evening LT 4 intake (60 min after supper). The primary outcome will be variation in serum thyrotropin (TSH) levels after 24 weeks of the LT 4 protocol. The secondary outcomes will be the prevalence of drugs that potentially interact with LT 4 and hypothyroidism control according to interaction status. The sample size was calculated to detect a minimum mean difference of 1 mUI/L in serum TSH level between the groups with 80% power and a 5% probability of type I error, resulting in 91 patients per group. The project was approved by the Hospital de Clínicas de Porto Alegre Ethics Committee., Discussion: Considering the aging population, the increased prevalence of multimorbidity and polypharmacy, as well as potential drug interactions and treatment adherence difficulties, an alternative LT 4 protocol could be useful for hypothyroidism treatment in the elderly. Prior studies comparing alternative LT4 administration protocols have mainly included young adult populations and have not addressed potential drug interactions., Trial Registration: ClinicalTrials.gov, NCT03614988. Registered 30 July 2018.

Background: To investigate the association between dietary components and development of chronic diabetic complications, the dietary evaluation should include a long period, months or years. The present manuscript aims to develop a quantitative food frequency questionnaire (FFQ) and a portfolio with food photos to assess the usual intake pattern of Brazilian patients with type 2 diabetes to be used in future studies., Methods: Dietary data using 3-day weighed diet records (WDR) from 188 outpatients with type 2 diabetes were used to construct the list of usually consumed foods. Foods were initially clustered into eight groups: "cereals, tubers, roots, and derivatives"; "vegetables and legumes"; "fruits"; "beans"; "meat and eggs"; "milk and dairy products"; "oils and fats", and "sugars and sweets". The frequency of food intake and the relative contribution of each food item to the total energy and nutrient intakes were calculated. Portion sizes were determined according to the 25th, 50th, 75th, and 95th percentiles of intake for each food item., Results: A total of 62 food items were selected based on the 3-day WDR and another 27 foods or how they are prepared and nine beverages were included after the expert examination. Also, a portfolio with food photos of each included food item and portion sizes was made to assist the patients in identifying the consumed portion., Conclusions: We developed a practical quantitative FFQ and portfolio with photos of 98 food items covering those most commonly consumed in the past 12 months, to assess the usual diet pattern of patients with type 2 diabetes in Southern Brazil.

To investigate the relationship between diabetic retinopathy (DR) and 24-h ambulatory blood pressure (ABP) in a cohort of normotensive, normoalbuminuric type 1 diabetic patients. This is a 6.1+/-3.3 year prospective study of 44 normotensive, normoalbuminuric type 1 diabetic patients. ABP was measured at the beginning and at the end of the study. Measurements of urinary albumin excretion rate (UAER) and direct and indirect ophthalmoscopy after mydriasis were performed at the start and end of the study and at least once a year. DR was observed in 12 patients at baseline. At the end of the study, eight of these patients had progressed to more advanced stages of retinopathy. Four patients developed retinopathy after the study began. These patients were grouped and classified as progressors. At baseline, progressors were older, had longer duration of diabetes, higher levels of UAER, and higher 24-h diastolic (P=0.03) and diurnal diastolic blood pressure (P=0.03). UAER and diastolic blood pressure (24h or day) remained significantly associated with development and progression of DR after multivariate analysis. High normal ABP was associated with the development or progression of DR in this cohort of normotensive, normoalbuminuric type 1 diabetic patients. Abnormalities in blood pressure homeostasis could indicate higher susceptibility to DR.