Brazilian Journal of Medical and Biological Research最新文献

Diabetic osteoporosis (DOP) is a complication of prolonged hyperglycemia. Hydrogen sulfide (H2S) has been identified as a protective factor in bone development. However, the mechanism by which H2S antagonizes the effects of high glucose (HG) on osteoblasts remains unclear. The effects of HG and H2S on osteoblasts were assessed through transcriptomic and metabolomic sequencing to identify key changes in gene expression and metabolism. Reactive oxygen species (ROS) levels, mitochondrial membrane potential (MMP), alkaline phosphatase (ALP) activity, mineralization, iron ion levels, malondialdehyde (MDA) levels, cell proliferation, and protein expression were evaluated. Transcriptomic analysis revealed significant upregulation of the ferroptosis pathway in HG-treated osteoblasts. Fer-1 and H2S antagonized the HG-induced decrease in osteoblast cell proliferation, increase in ROS production, decrease in MMP, decrease in ALP, decrease in mineralized nodules, and increase in iron ions and MDA. Transcriptome analysis showed Fer-1 was involved in upregulating the synthesis, secretion, and action of parathyroid hormone and estrogen synthesis, while downregulating the mitogen-activated protein kinases (MAPK) pathway. Metabolomic analysis showed H2S restored glutathione metabolism, reducing pyroglutamic acid and L-5-oxoproline levels. Transcriptome sequencing identified downregulated genes (hmox1, ncoa4) and an upregulated gene (slc40a1) related to ferroptosis in the H2S + HG group compared with the HG group. Western blot analysis indicated H2S increased GPX4 and SLC7A11 levels while reducing ACSL4 expression compared with the HG group. Ferroptosis may be involved in the pathogenesis of DOP and H2S can effectively alleviate osteoblast injury by inhibiting ferroptosis in DOP.

This study aimed to investigate the impact of targeted nursing care combined with nutritional support on the clinical outcomes of diabetic nephropathy (DN) patients undergoing maintenance hemodialysis (HD). Clinical indicators such as serum creatinine (SCr), blood urea nitrogen (BUN), fasting blood glucose (FBG), hemoglobin A1c (HbA1c), and K (urea clearance) × t (dialysis time) / V (volume of urea distribution) (Kt/V), as well as inflammatory indicators such as high-sensitivity C-reactive protein (hs-CRP), interleukin (IL)-6, and tumor necrosis factor-α (TNF-α), and nutritional indicators such as transferrin (TRF), albumin (ALB), and prealbumin (PA) were measured. SF-36 quality of life scale scores were assessed, and adverse events and patient satisfaction with care were recorded. Post-intervention, the experimental group exhibited lower SCr, BUN, FBG, HbA1c, hs-CRP, IL-6, and TNF-α, and higher body mass index, Kt/V, TRF, ALB, and PA than the control group (all P<0.05). Additionally, the experimental group demonstrated higher nursing satisfaction scores, and lower total incidence of adverse events compared to the control group (all P<0.05). Targeted nursing care combined with nutritional support applied to DN patients during HD helped improve residual renal function, reduce the body's inflammatory response, improve nutritional status and the quality of life, reduce adverse events, and at the same time, improve nursing satisfaction.

After SARS-CoV-2 infection, severe COVID-19 may develop with persistent sequelae, even after hospital discharge. This condition may result from tissue damage or immune alterations caused by the virus, including immune dysregulation, hyperinflammation, loss of immune tolerance, excessive neutrophil extracellular trap (NET) production, and antibody cross-reactivity (molecular mimicry), which can promote autoantibody development. This study evaluated autoantibody expression in patients with long COVID-19 and its potential relationship with symptoms. Conducted in Baixada Santista, São Paulo, Brazil, the study involved 55 participants aged 21-85 years who had tested positive for SARS-CoV-2. Blood samples were collected two years post-discharge, and serum was analyzed for inflammatory and autoimmune markers, including antinuclear antibody (ANA), rheumatoid factor (RF), anti-cyclic citrullinated peptide (anti-CCP), procalcitonin (PCT), Venereal Disease Research Laboratory test (VDRL), and C-reactive protein (CRP). Results were compared to a control group of 21 individuals who never tested positive for COVID-19. Among severe COVID-19 patients, 26 reacted to ANA, 16 to VDRL, 2 had elevated RF, 12 had increased PCT, and 11 had high CRP, whereas the control group showed no reactive results. Anti-CCP values were not significant. Findings suggest that hyperinflammation may contribute to autoimmunity, particularly in cases of reactive ANA levels, linking COVID-19 symptoms to autoimmune responses.

A hallmark of COVID-19 patients is the reduction of the lymphocyte population accompanied by activation, senescence, and exhaustion markers. We investigated patients admitted to hospital wards who either recovered after a short hospitalization or progressed to critical illness. Patients (n=48) were recruited between May and September 2020; 19 healthy volunteers were enrolled as controls. Blood samples were collected on days 0, 3, and 7 of hospitalization and around 30 days after discharge (convalescence sample, CS30). Lymphocyte counts and extended immunophenotyping were performed by flow cytometry and analyzed using conventional and stochastic methods. At D0 and D7, total lymphocytes, natural killer cells, T cells, TCD4 cells, and TCD8 cells were lower in patients than in volunteers but were restored at CS30. The stochastic analysis identified 11 distinct clusters of lymphocytes, nine of them with significant differences between patients and healthy controls. Clusters of TCD8+ memory cells showing activation, senescence, and exhaustion were increased in patients during hospitalization and in the convalescence samples. In contrast, clusters 5 (TCD4+ Central Memory exhausted activated) and 7 (TCD4+ Central Memory exhausted) were decreased in patients during the disease compared to healthy controls. Overall, the conventional flow cytometry analyses corroborated the findings from the stochastic analysis, showing that effector memory (EM) and TEMRA subsets exhibited sustained markers of exhaustion and senescence, particularly in TCD8+ cells. Our findings reinforce lymphopenia, T cell activation, senescence, and exhaustion as essential immunological features of COVID-19; while cell counts fully recovered, lymphocytes remained dysfunctional in convalescent samples.

Sacha inchi oil (SIO) is characterized by its high content of polyunsaturated fatty acids (PUFAs), metabolites with beneficial properties on health. The objective was to evaluate the fatty acid (FA) profile of wild SIO and its effect on biochemical parameters of lipid metabolism under a high-fat diet. Twenty-four albino rats were grouped into groups I, II, III, and IV, which ingested ad libitum the following diets: conventional diet without supplementation (CDOS), conventional diet supplemented with SIO (CDWS), hyperlipidic diet without supplementation (HDOS), and hyperlipidic diet supplemented with SIO (HDWS) for 6 weeks. The FA content of SIO was assessed by gas chromatography-mass spectrometry. The lipid profile was analyzed by the enzymatic-spectrophotometric method, and cytokines and lipid mediator levels were measured via enzyme-linked immunosorbent assay (ELISA). Α-linolenic acid (ALA) and linoleic acid (LA) constitute 82% of this oil. Two-way ANOVA showed interaction effects between diet and supplement on interleukin (IL)-10 levels, and SIO-supplemented diet significantly decreased triglycerides (TG), very low-density lipoprotein cholesterol (VLDL-C), and the TG/HDL-C ratio levels. Wild SIO is high in ALA and LA. SIO supplementation reduced TG, VLDL-C, and the TG/HDL-C ratio, modulated IL-10, and slightly improved leptin, resolvin-D1 (RvD1), and IL-6 levels.

Inflammatory bowel disease (IBD), encompassing Crohn's disease (CD) and ulcerative colitis (UC), is characterized by chronic inflammation, which may be associated with hepatic and biliary manifestations such as non-alcoholic fatty liver disease (NAFLD). Despite the high risk of hepatic manifestations among patients with IBD, few studies in Brazil have assessed the frequency of these diseases. Therefore, this study aimed to analyze the prevalence of liver disease by ultrasound in patients with IBD. This was a single-center, cross-sectional study that included patients with IBD who were followed up at an outpatient clinic. The clinical and sociodemographic data, disease activity, biochemical test results, and Doppler liver ultrasonography results were assessed. Descriptive and association tests were used for statistical analyses. A total of 138 patients were included: 64.49% females, mean age 45.55±14.17 years, and body mass index of 26.92±5.07 kg/m2. In total, 63 (45.65%) patients had CD and 75 (54.35%) had UC. Most patients were in either clinical (58.39%) or endoscopic remission (52.55%). Liver ultrasound revealed NAFLD in 58 patients (42.03%), which was classified as mild (36.21%), moderate (46.55%), or severe (17.24%). Seven patients had choledocholithiasis and three had chronic liver disease. Liver disease was associated with changes in aspartate aminotransferase (AST), alanine aminotransferase (ALT), hematocrit, hemoglobin, and fasting glucose levels. Liver disease is frequent in IBD patients, with NAFLD being the most prevalent. Screening for liver disease in patients with IBD is recommended for early detection and immediate treatment of the alterations, in order to prevent complications and progression to cirrhosis.

The aim of this study was to establish reference intervals for lymphocyte subpopulations in the peripheral blood of Brazilian adults, and to assess potential variations by gender and age groups. The study assessed 351 healthy participants of the Brazilian Longitudinal Study of Adult Health (ELSA-Brasil). Lymphocyte subpopulations were analyzed by dual platform using automated hematological analyzer Sysmex XN-3000 and the four-color flow cytometer on the FACSCalibur. Reference intervals were established using the 2.5th and 97.5th percentiles. Z-score was employed to ascertain the need for distinct reference intervals across gender and age groups. The Mann-Whitney test, with a significance level set at P<0.05, was conducted to identify differences among population groups. The absolute and relative reference intervals were: total lymphocytes: 1.0-2.9 (×103/µL); CD3+: 721.0-2311.5 cells/μL, 59.1-84.5%; CD4+: 421.4-1523.8 cells/μL, 32.5-61.6%; CD8+: 175.3-879.8 cells/μL, 12.3-39.1%; CD4+CD8+: 0.8-4.1 cells/μL; CD19+: 85.2-501.6 cells/μL, 5.6-21.0%; NK: 83.9-444.4 cells/μL, 4.3-23.5%. Significant gender and age differences were observed in both the relative and absolute values of most lymphocyte subpopulations. There are variations in lymphocyte subsets across the global population, underscoring the need to establish tailored reference intervals for distinct populations, particularly for Helper T lymphocytes, B lymphocytes, and NK cells.

Plastination is a technique for preserving biological tissues, in which body fluids are replaced by a curable polymer. Epoxy resin is used for 2-5 mm sections of anatomical segments, with the German-made Biodur® E12 being the best known and most widely used resin. A few alternative epoxies can be used in the technique, but research should be developed to identify options that are cheaper and less bureaucratic to acquire. This study aimed to find, adapt, and apply an alternative epoxy resin formulation and its curing system for the plastination process as a potential substitute for Biodur® E12. The methodology was divided into the search of a resin for national commercialization in Brazil, the development of the final formulation, the testing of its use in plastination, and the evaluation of the resin and final specimens. From market research, E48 epoxy (brand not disclosed) was selected, and its formulation was changed with the addition of a plasticizer for use in impregnation. A total of 150 Wistar rat cross-sections were plastinated with the control polymer (E12) and with the developed test resin (E48). Based on the positive results of the shrinkage analysis (no statistical difference) and confocal and stereoscopic microscopy, it was concluded that the modified E48 is a great alternative to E12.

Minimally invasive autopsy (MIA) is used to understand the pathophysiological mechanisms and clinical outcome of diseases. One of its many advantages is the lower risk of contamination in case of infectious diseases and the quick tissue collection procedure compared to conventional autopsies, which reduces cold ischemia time. Here we investigated the potential role of MIA for molecular pathology studies by comparing RNA yield and quality after RNA extraction from frozen lung tissue samples collected from different methods: MIA, lobectomy, and transbronchial biopsy (TBB). Our results revealed that RNA yield was significantly higher (P<0.05) in the TBB group compared to the lobectomy and MIA groups. However, 93% of MIA samples were non-degraded, showing similar results to TBB, where all had a DV200≥70%. Therefore, MIA proves to be a novel tool for molecular pulmonary pathology in diagnostic and/or research settings.

Escherichia coli is a common intestinal microorganism that can cause a variety of diseases in humans and animals. The aim of this study was to evaluate the therapeutic effects of fish oil rich in omega-3 against intestinal infection caused by enteroaggregative E. coli (EAEC). Minimum inhibitory and bactericidal concentrations were determined, along with toxicity assays using HT-29 intestinal cells and Tenebrio molitor larvae. Swiss mice infected with EAEC 042 were used to assess the in vivo therapeutic potential of fish oil. Histological analyses of the liver, kidney, and colon were conducted to identify tissue alterations such as inflammation and necrosis. Fish oil exhibited a bacteriostatic effect on E. coli and was non-toxic to HT-29 cells at concentrations up to 50 mg/mL. It also enhanced survival in treated larvae. In infected mice, bacterial colony counts were significantly lower in the fish oil-treated group. Histological evaluation showed reduced inflammatory infiltrates in the liver and colon, and no progression of hepatic hydropic degeneration was observed in treated animals, unlike in the untreated infected group. These findings indicated that fish oil rich in omega-3 possesses antimicrobial activity against E. coli, is non-toxic to both cells and animal models, and effectively reduces intestinal infection and associated tissue damage in mice. This suggests its potential as a supportive therapeutic agent for infections caused by pathogenic E. coli.