Objective: To investigate clinicopathological correlations, cell proliferation, and immortalization during induced oral carcinogenesis. Methods: Forty-three Wistar rats were divided into a control group (n=10) or a 4-Nitroquinoline 1-oxide (4NQO) group (n=33). Control animals were euthanized after 20 weeks, and 4NQO-treated animals after 4 (n=10), 12 (n=10), or 20 weeks (n=13). Oral lesions were classified macroscopically and histologically, with Ki-67 and BMI-1 immunolabeling used to assess cell proliferation and immortalization. Results: Histological alterations, including hyperplasia/hyperkeratosis (n=4) and severe dysplasia (n=2), were observed in clinically normal mucosa. Leukoplakic lesions exhibited varying severity, ranging from hyperplasia/hyperkeratosis (n=3) to squamous cell carcinoma (SCC, n=2). Most SCCs appeared as ulcers (n=3) or nodules (n=4). Ki-67 expression increased progressively with histopathological changes, while BMI-1 levels rose significantly in later stages. A positive correlation was found between Ki-67 and BMI-1 (R=0.33, p=0.03). Conclusion: Cellular alterations often precede visible clinical lesions. Clinical appearances, particularly of leukoplakic lesions, frequently did not align with histopathological findings. Proliferation and immortalization were interconnected but occurred at distinct stages of carcinogenesis.

Sudden cardiac death (SCD) is an unpredictable and common mode of death in patients with heart failure (HF). Alterations in calcium handling may lead to malignant arrhythmias, resulting in SCD, and variants in calcium signaling-related genes have a significant association with SCD. Therefore, the aim of the present retrospective cohort study was to investigate the association of Ser96Ala [histidine-rich calcium-binding protein (HRC)], Ser49Gly [β1-adrenergic receptor (ADRB1)], Arg389Gly (ADRB1) and Gly1886Ser [ryanodine receptor 2 (RYR2)] polymorphisms with serious arrhythmic events and overall mortality in patients with HF with reduced left ventricular ejection fraction of non-ischemic etiology. In total, 136 patients with HF underwent physical examination, routine laboratory tests, non-invasive assessment of cardiac function and an invasive electrophysiological study. The primary outcome was the occurrence of serious arrhythmic events, set as either SCD or appropriate implantable cardioverter-defibrillator (ICD) therapy, and the secondary outcome was all-cause death. During a median follow-up of 37 months, arrhythmic events occurred in 26 patients (19%) and 41 patients (30%) died. Patients carrying the Ser allele of the Ser96Ala polymorphism in HRC had worse survival than those with the Ala/Ala genotype (log-rank P=0.043). Despite the difference in survival time, the Ala/Ala genotype was not associated with all-cause death in the regression analysis [unadjusted hazard ratio (HR)=0.17; 95% CI, 0.02-1.21]. Regarding the Ser49Gly and Arg389Gly polymorphisms in ADRB1, homozygosity for the major alleles at both sites (Ser49Ser and Arg389Arg) was associated with a two-fold increased risk of all-cause death compared with the other genotype combinations (unadjusted HR=1.98; 95% CI, 1.02-3.82). However, this association was lost after controlling for clinical covariates. No association was observed for the Gly1886Ser polymorphism in RYR2. Overall, the present findings are concurrent with the hypothesis that the Ser96Ala (HRC), Ser49Gly (ADRB1) and Arg389Gly (ADRB1) polymorphisms may be associated with HF prognosis. In particular, the Ser96Ala polymorphism might aid in risk stratification and patient selection for ICD implantation.

Trastuzumab is a monoclonal antibody used in oncotherapy for HER2-positive tumors. However, as an adverse effect, trastuzumab elevates the risk of heart failure, implying the involvement of energy production and mitochondrial processes. Past studies with transcriptome analysis have offered insights on pathways related to trastuzumab safety and toxicity but limited study sizes hinder conclusive findings. Therefore, we meta-analyzed mitochondria-related gene expression data in trastuzumab-treated cardiomyocytes. We searched the transcriptome databases for trastuzumab-treated cardiomyocytes in the ArrayExpress, DDBJ Omics Archive, Gene Expression Omnibus, Google Scholar, PubMed, and Web of Science repositories. A subset of 1270 genes related to mitochondrial functions (biogenesis, organization, mitophagy, and autophagy) was selected from the Kyoto Encyclopedia of Genes and Genomes and Gene Ontology Resource databases to conduct the present meta-analysis using the Metagen package (Study register at PROSPERO: CRD42021270645). Three datasets met the inclusion criteria and 1243 genes were meta-analyzed. We observed 69 upregulated genes after trastuzumab treatment which were related mainly to autophagy (28 genes) and mitochondrial organization (28 genes). We also found 37 downregulated genes which were related mainly to mitochondrial biogenesis (11 genes) and mitochondrial organization (24 genes). The present meta-analysis indicates that trastuzumab therapy causes an unbalance in mitochondrial functions, which could, in part, help explain the development of heart failure and yields a list of potential molecular targets. These findings contribute to our understanding of the molecular mechanisms underlying the cardiotoxic effects of trastuzumab and may have implications for the development of targeted therapies to mitigate such effects.

Background: Cardiovascular diseases (CVD) are the leading cause of death globally. Oxidative stress plays a significant role in cell damage, inflammation, and progression of CVD. The use of N-acetylcysteine (NAC), a well-known antioxidant and a precursor to glutathione (GSH), has shown potential in preventing different types of CVD. Yet, there is limited data assessing NAC’s impact on GSH levels in animal models of CVD. Objective: To evaluate reduced glutathione (GSH) and total glutathione levels in cardiac tissue or serum/plasma from animals with established heart disease treated with NAC. Method: The literature search included PubMed, Embase, WoS, Scielo, Lilacs/BINACIS, and rXiv databases. The search in the databases was completed in December 2022. Results: We identified 619 articles, fully reviewed 35, and included 7 in the meta-analysis. The included studies varied in cardiovascular disease models, NAC doses, and outcomes. Meta-analysis revealed that NAC supplementation significantly increased total glutathione levels (SMD = 1.23, 95% CI = 0.56–1.89; p = 0.0003) with low heterogeneity (I² = 37%). GSH levels showed a marginal increase (SMD = 2.49, 95% CI = -0.20–5.18; p = 0.07), with high heterogeneity (I² = 93%). Conclusion: NAC supplementation effectively increases total glutathione in animal models of heart disease. The impact on GSH levels was less clear, possibly due to dose variability and limited study numbers. Standardization of NAC dosing and improved methodological reporting are needed for more consistent and clinically relevant findings.

Dear Dr. Nunamaker and Dr. ReynoldsWe have read with interest your recent paper published in PlosOne where you demonstrated how poor the reporting is in oncology preclinical studies [1]. After selecting 400 articles published during 2020 in 20 oncology-related journals that explicitly endorsed the ARRIVE guidelines - a 20-item checklist to improve animal studies reporting - you found just 23% of compliance with five key domains (ethical oversight assurance, animal signalment, animal husbandry, welfare, and euthanasia) [2]. That’s unfortunate, way below acceptable, and not novel. Still, it’s highly important to show how bad the portrait is today. The ARRIVE guidelines were first launched in 2010 [3] but they didn’t stick as they should and poor reporting performance of preclinical studies has persisted in several fields [4–6]. Among the worst reported items is euthanasia and, as pointed out by you, only 14% of the articles fully described the procedure. Before you, Abbas et. al also found only 14% of the articles with tissue engineering approaches for urethral repair properly described the euthanasia methodology [4] and Reynolds et. al showed that 52% of the articles published in Shock journal did it properly [7]. A detailed description of the euthanasia methods is often underestimated by researchers but has huge consequences for the validity and interpretation of the outcomes. Ignoring the variation in the chosen shift, duration, and most importantly, anesthetics employed for euthanasia can simply make the results ungeneralizable. We have investigated the impact of anesthetics in a systematic review and meta-analysis [8] and we noticed that:i) the interference of a given anesthetic is not constant across several biological pathways;ii) the degree of interference in a given pathway differs according to the anesthetic employed;iii) the anesthetic dose has a wide variation across studies;iv) the confirmatory euthanasia method (i.e. overdose, decapitation, bleeding after organ harvesting) may be a confounding factor. It’s noteworthy that these interferences do not justify their abrogation. We must bear in mind that animal killing methods must comply with the ability to cause a rapid loss of consciousness and death with minimum distress, fear, or pain in the animals. Thus, using anesthetics right before euthanasia is meant to achieve these goals. Instead of abrogation, we should develop standards and, while these standards are lacking, we should, at least, describe the procedure in detail. That’s the researcher's burden. However, we cannot forget the publisher's part in this lack of adherence to good standards. Nunamaker and Reynolds chose just journals that endorse the ARRIVE guidelines. Why didn’t the editors and reviewers enforce the journal policies? It seems to be another demonstration that scientific publishers have not been working just for the advancement of science.

Introduction: Cardiac biomarkers have been proposed as a new tool to improve risk stratification of serious arrhythmic events in patients with heart failure (HF) beyond estimates of left ventricular ejection fraction. Growth differentiation factor (GDF)-15, a stress-induced cytokine, has been found to correlate with markers of myocardial fibrosis and adverse clinical outcomes, but its role as a predictor of arrhythmic events in patients with nonischemic HF is uncertain. Methods and Results: A prospective observational study was conducted in 148 nonischemic patients with HF who underwent comprehensive clinical and laboratory evaluation, including measurement of serum GDF-15. The study endpoints were serious arrhythmic events (which included appropriate implantable cardioverter-defibrillator therapy and sudden cardiac death) and all-cause mortality. Mean age of the cohort was 54.8±12.7 years, and mean left ventricular ejection fraction (LVEF) was 27.4±7.5. During a mean follow-up time of 42 months, arrhythmic events occurred in 28 patients (19%), and 40 patients (27%) died. An increase in serum GDF-15 (log-transformed) correlated linearly with a higher risk of serious arrhythmic events (HR 1.14, 95% CI 1.01-1.28, p=0.03) even after adjustment for other potential clinical predictors (HR 1.16, 95% CI 1.02-1.32, p=0.02). GDF-15 was also strongly and independently associated with all-cause mortality (HR 1.17, 1.05-1.31, p=0.004). Conclusion: In this cohort of nonischemic HF patients on optimized medical treatment, serum GDF-15 levels were independently associated with major arrhythmic events and overall mortality. This biomarker may add prognostic information beyond LVEF to better stratify the risk of sudden death in this particular population.

Grape juice consumption may influence the early occurrence of ductal constriction during pregnancy, since the consumption of foods rich in polyphenols can be linked to the premature constriction of the ductus arteriosus. This study aimed to evaluate the effect of purple grape juice consumption during gestation on fetal ductus arteriosus closure, prostaglandin levels, and oxidative stress markers in Wistar rats. We divided 18 pregnant rats into four groups: a control group (C), a single-dose grape juice group (SDGJ), a two-dose grape juice group (TDGJ) of 7 μl/g body weight per day, and an indomethacin group (I). Blood was collected on gestational day (GD) 0, 14, and 20. Prostaglandin levels were measured, and the livers and hearts were removed from the mothers and fetuses for oxidative stress analysis; histology of the fetal ductus arteriosus was performed. Prostaglandin levels (pg/ml) at GD 20 were (C:1462.10 ± 314.61); (SDGJ:987.66 ± 86.25); (TDGJ:1290.00 ± 221.57), and (I:584.75 ± 46.77). Fetal ductus arteriosus closure occurred only in the indomethacin group. Lipid peroxidation evaluated through thiobarbituric acid reactive substances (nmol/mg protein) in maternal livers was lower in the grape juice groups (C: 4.11 ± 0.76 nmol/mg protein), (SDGJ: 2.34 ± 0.36), (TDGJ: 1.52 ± 0.18), and (I: 4.20 ± 0.76). Sulfhydryls (nmol/mg protein) were lower in the TDGJ group (C:763.59 ± 61.38 nmol/mg protein), (SDGJ:978.88 ± 158.81), (TDGJ:385.32 ± 86.78), and (I:727.72 ± 49.12). Also, superoxide dismutase activity (USOD/mg protein) was higher in fetal hearts in this group: (C:5.29 ± 0.33), (SDGJ:4.48 ± 0.47), (TDGJ:7.35 ± 0.43), and (I:6.00 ± 0.18). We conclude that grape juice consumption in pregnancy does not induce ductus arteriosus closure in the fetus and presented potential antioxidant effects.

Abstract Background Interleukin-6 (IL-6) is an inflammation-related cytokine associated with an elevated risk of cardiovascular events. In a previous study, we demonstrated that increased IL-6 was predictive of sub-clinical atherosclerotic coronary disease in intermediate-risk patients undergoing coronary angiography. In the present study, we investigated whether increased serum IL-6 is predictive of cardiovascular events in high-risk patients. Methods In this observational study, consecutive patients referred for elective coronary angiography due to stable chest pain/myocardial ischemia had IL-6 measured immediately before the procedure. Long-term follow-up was performed by phone call or e-mail, and their clinical registries were revised. The primary outcome was a composite of new myocardial infarction, new ischemic stroke, hospitalization due to heart failure, new coronary revascularization, cardiovascular death, and death due to all causes. Results From 141 patients submitted to coronary angiography and IL-6 analysis, 100 had complete follow-up data for a mean of 5.7 years. The median age was 61.1 years, 44% were men, and 61% had type-2 diabetes. The median overall time-to-event for the primary outcome was 297 weeks (95% confidence interval [CI] 266.95–327.16). A receiver operator characteristic curve defined the best cut-off value of baseline serum IL-6 (0.44 pg/mL) with sensitivity (84.37%) and specificity (38.24%) to define two groups. High (> 0.44 pg/mL) IL-6 levels were predictive of cardiovascular events. (p for interaction = 0.015) (hazard ratio = 2.81; 95% CI 1.38–5.72, p = 0.01). Subgroup analysis did not find interactions between patients with or without diabetes, obesity, or hypertension. Conclusion In conclusion, an interleukin-6 level higher than 0.44 pg/mL, obtained just before elective coronary angiography, was associated with a poorer prognosis after a mean of 5,7-year. A pre-procedure IL-6 below 0.44 pg/mL, on the other hand, has a very good negative predictive value, suggesting a good prognosis, and may be useful to better indicate coronary angiography in high-risk patients. .

Aim: To summarize the knowledge on the effect of anesthetics employed right before euthanasia on biological outcomes. Data source: A systematic review of the literature to find studies with isoflurane, ketamine, halothane, pentobarbital, or thiopental just before euthanasia of laboratory rats or mice. Study selection: Controlled studies with quantitative data available. Data extraction: The search, data extraction, and risk of bias (RoB) were performed independently by two reviewers using a structured form. For each outcome, an effect size (ES) was calculated relative to the control group. Meta-analysis was performed using robust variance meta-regression for hierarchical data structures, with adjustment for small samples. Data synthesis: We included 20 studies with 407 biological outcomes (110 unique). RoB analysis indicated that 87.5% of the domains evaluated showed unclear risk, 2% high risk, and 10.5% low risk. The effect size for all anesthetics considered together was 0.99 (CI95% = 0.75-1.23;p < 0.0001). Sub-analyses indicate high effect sizes for pentobarbital (1.14; CI95% = 0.75-1.52; p < 0.0001), and isoflurane (1.01; CI95% = 0.58-1.44; p = 0.0005) but not for ketamine (1.49; CI95% = -7.95-10.9; p = 0.295). Conclusion: We showed that anesthetics interfere differently with the majority of the outcomes assessed. However, our data did not support the use of one anesthetic over others or even the killing without anesthetics. We conclude that outcomes cannot be compared among studies without considering the killing method. This protocol was registered at Prospero (CRD42019119520).

Pulmonary arterial hypertension (PAH) is a disease characterized by increased pulmonary vascular resistance and right ventricle (RV) failure. In this context, oxidative stress is an essential element contributing to PAH’s pathophysiology. Thus, blueberry (BB), which has a high antioxidant capacity, emerges as a natural therapeutic approach in PAH. This work evaluated the effect of BB extract on redox balance in RV in a PAH’s animal model. Male Wistar rats (200 ± 20 g) (n = 72) were randomized into eight groups: control (CTR); monocrotaline (MCT); CTR and MCT treated at doses of 50, 100, and 200 mg/kg BB. PAH was induced by administration of MCT (60 mg/kg, intraperitoneal). Rats were treated with BB orally for 5 weeks (2 weeks before monocrotaline and 3 weeks after monocrotaline injection). On day 35, rats were submitted to echocardiography and catheterization, then euthanasia and RV harvesting for biochemical analyses. RV hypertrophy, observed in the MCT groups, was reduced with BB treatment. MCT elevated RV systolic pressure and pressure/time derivatives, while the intervention with BB decreased these parameters. PAH decreased RV output and pulmonary artery outflow acceleration/ejection time ratio, while increased RV diameters, parameters restored by BB treatment. Animals from the MCT group showed elevated lipid peroxidation and NADPH oxidase activity, outcomes attenuated in animals treated with BB, which also led to increased catalase activity. Treatment with BB partially mitigated PAH, which could be associated with improvement of RV redox state. Such findings constitute an advance in the investigation of the role of BB extract in chronic progressive cardiovascular diseases that involve the redox balance, such as PAH.

Resumo Fundamento: A estratificação de risco continua sendo clinicamente desafiadora em pacientes com insuficiência cardíaca (IC) de etiologia não isquêmica. A galectina-3 é um marcador sérico de fibrose que pode ajudar no prognóstico. Objetivo: Determinar o papel da galectina-3 como preditora de eventos arrítmicos graves e mortalidade total. Métodos: Este é um estudo de coorte prospectivo que incluiu 148 pacientes com IC não isquêmica. Todos os pacientes foram submetidos a uma avaliação clínica e laboratorial abrangente para coleta de dados de referência, incluindo níveis de galectina-3 sérica. O desfecho primário foi a ocorrência de síncope arrítmica, intervenções apropriadas do cardioversor desfibrilador implantável, taquicardia ventricular sustentada ou morte súbita cardíaca. O desfecho secundário foi a morte por todas as causas. Para todos os testes estatísticos, considerou-se significativo o valor p<0,05 (bicaudal). Resultados: Em seguimento mediano de 941 dias, os desfechos primário e secundário ocorreram em 26 (17,5%) e 30 (20%) pacientes, respectivamente. A galectina-3 sérica>22,5 ng/mL (quartil mais alto) não foi preditora de eventos arrítmicos graves (HR: 1,98; p=0,152). Os preditores independentes do desfecho primário foram diâmetro diastólico final do ventrículo esquerdo (DDFVE)>73 mm (HR: 3,70; p=0,001), ventilação periódica durante o exercício (VPE) no teste de esforço cardiopulmonar (HR: 2,67; p=0,01) e taquicardia ventricular não sustentada (TVNS)>8 batimentos na monitorização por Holter (HR: 3,47; p=0,027). Os preditores de morte por todas as causas foram: galectina-3>22,5 ng/mL (HR: 3,69; p=0,001), DDFVE>73 mm (HR: 3,35; p=0,003), VPE (HR: 3,06; p=0,006) e TVNS>8 batimentos (HR: 3,95; p=0,007). A ausência de todos os preditores de risco foi associada a um valor preditivo negativo de 91,1% para o desfecho primário e 96,6% para a mortalidade total. Conclusões: Em pacientes com IC não isquêmica, níveis elevados de galectina-3 não foram preditores de eventos arrítmicos graves, mas foram associados à mortalidade total. A ausência de preditores de risco revelou um subgrupo prevalente de pacientes com IC com excelente prognóstico.

Background: Risk stratification remains clinically challenging in patients with heart failure (HF) of non-ischemic etiology. Galectin-3 is a serum marker of fibrosis that might help in prognostication.Objective: To determine the role of galectin-3 as a predictor of major arrhythmic events and overall mortality.Methods: We conducted a prospective cohort study that enrolled 148 non-ischemic HF patients. All patients underwent a comprehensive baseline clinical and laboratory assessment, including levels of serum galectin-3. The primary outcome was the occurrence of arrhythmic syncope, appropriate implantable cardioverter defibrillator therapy sustained ventricular tachycardia, or sudden cardiac death. The secondary outcome was all-cause death. for all statistical tests, a tvvo-tailed p-value< 0.05 was considered significant.Results: In a median follow-up of 941 days, the primary and secondary outcomes occured in 26(17.5%) and 30 (20%) patients, respectively Serum galectin-3> 22.5 ng/mL (highest quartile) did not predict serious arrhythmic events (HR: 1.98, p=0.152). Independent predictors of the primary outcome were left ventricular end-diastolic diameter (LVEDD)>73mm (HR 3.70, p=0.001), exercise periodic breathing (EPB) on cardiopulmonary exercise testing (HR: 2.67, p=0.01), and non-sustained ventricular tachycardia (NSVD>8 beats on Hotter monitoring (HR 3.47, p=0.027). Predictors of all-cause death were galectin-3> 22.5 ng/mL (HR 3.69, p=0.001), LVEDD> 73mm (HR: 3.35, p=0.003), EPB (HR: 3.06, p=0.006), and NSVT> 8 beats (HR: 3.95, p=0.007). The absence of all risk predictors was associated with a 91.1% negative predictive value for the primary outcome and 96.6% for total mortality.Conclusions: In non-ischemic HF patients, elevated piectin-3 levels did not predict major arrhythmic events but were associated with total mortality. Absence of risk predictors revealed a prevalent subgroup of HF patients with an excellent prognosis.

The tissue response to acute myocardial infarction (AMI) is key to avoiding heart complications due to inflammation, mitochondrial dysfunction, and oxidative stress. Antioxidant and anti-inflammatory agents can minimize the effects of AMI. This study investigated the role of 2-methoxy-isobutyl-isonitrile (MIBI)-associated gold nanoparticles (AuNP) on reperfusion injury after ischemia and its effect on cardiac remodeling in an experimental AMI model. Three-month-old Wistar rats were subjected to a temporary blockade of the anterior descending artery for 30 min followed by reperfusion after 24 h and 7 days by intraventricularly administering 0.4, 1.3, and 3 mg/kg AuNP-MIBI. The cardiac toxicity and renal and hepatic function levels were determined, and the infarct and peri-infarct regions were surgically removed for histopathology, analysis of inflammation from oxidative stress, and echocardiography. MIBI-conjugated AuNP promoted changes in oxidative stress and inflammation depending on the concentrations used, suggesting promising applicability for therapeutic purposes.

The effects of non-nutritive sweeteners (NNS) on the gut microbiota are an area of increasing research interest due to their potential influence on weight gain, insulin resistance, and inflammation. Studies have shown that mice and rats fed saccharin develop weight gain and metabolic alterations, possibly related to changes in gut microbiota. Here, we hypothesized that chronic exposure to a commercial NNS would change the gut microbiota composition in Wistar rats when compared to sucrose exposure. To test this hypothesis, Wistar rats were fed either NNS- or sucrose-supplemented yogurt for 17 weeks alongside standard chow (ad libitum). The gut microbiome was assessed by 16S rDNA deep sequencing. Assembly and quantification were conducted using the Brazilian Microbiome Project pipeline for Ion Torrent data with modifications. Statistical analyses were performed in the R software environment. We found that chronic feeding of a commercial NNS-sweetened yogurt to Wistar rats, within the recommended dose range, did not significantly modify gut microbiota composition in comparison to sucrose-sweetened yogurt. Our findings do not support the hypothesis that moderate exposure to NNS is associated with changes in gut microbiota pattern compared to sucrose, at least in this experimental model.

Objectives We explored the effects of mucoadhesive formulation with Curcuma longa L. extract (MCL) on oral mucositis induced by 5-fluorouracil in hamsters. Study Design Seventy-two golden Syrian hamsters separated into 4 groups: control group, placebo group (no active substance), positive control group (AdMuc topical chamomile), and MCL group (topical use of mucoadhesive Curcuma longa). The animals received an intraperitoneal injection of 5-fluorouracil on days 0 and 2. On days 3 and 4, the buccal mucosa was scratched, and therapy was initiated on day 5. The animals received 2 daily applications of the product according to the experimental group. Wound area was calculated and histologic sections of 3 μm were stained by hematoxylin and eosin for semiquantitative analysis of re-epithelization and degree of tissue inflammation. Immunohistochemistry was used for transforming growth factor β1 (TGF-β1) and CD31 analysis. Results The MCL group showed a greater clinical reduction of the mucositis lesions at day 8, a higher degree of re-epithelization, and a lower inflammatory process, with a lower angiogenesis and TGF-β1 epithelial labeling compared with the placebo and control groups (P Conclusions MCL Curcuma longa L. has a therapeutic effect accelerating the healing of oral mucositis reducing the inflammatory response and increasing re-epithelization.

Objective We aimed to evaluate the effect of photobiomodulation therapy (PBMT) on acetyl-histone H3 (acH3) expression during oral ulcer wound healing. Study Design Forty-eight male Wistar rats were divided into Control Group (CG) and PBMT Group. Traumatic ulcers were caused in the dorsum of the tongue with punch. Irradiation with InGaAlP laser, 660 nm, 40 mW, 0.04 cm2 spot size, 4 J/cm2, 4 seconds, and 0.16 J per point was performed once a day in close contact for 10 consecutive days. The CG received only daily handling. Rats were euthanized at days 3, 5, and 10 (n = 8) and were monitored daily to determine wound status. Immunohistochemical analysis for the detection of acH3 was performed. One thousand epithelial cells were counted and the mean of acH3 was calculated and compared between groups. Results PBMT accelerated the repair of oral ulcers. At day 3, PBMT showed a significantly higher mean of acH3 than GC (P = .04). On day 5, no difference was observed between the groups. On day 10, PBMT presented a lower mean of acH3 than the CG (P = .05). Conclusions PBMT stimulates the oral mucosa wound healing by activating epigenetic mechanisms such as histone acetylation in the early stages of the process. Fipe HCPA: 14-0572 We aimed to evaluate the effect of photobiomodulation therapy (PBMT) on acetyl-histone H3 (acH3) expression during oral ulcer wound healing. Forty-eight male Wistar rats were divided into Control Group (CG) and PBMT Group. Traumatic ulcers were caused in the dorsum of the tongue with punch. Irradiation with InGaAlP laser, 660 nm, 40 mW, 0.04 cm2 spot size, 4 J/cm2, 4 seconds, and 0.16 J per point was performed once a day in close contact for 10 consecutive days. The CG received only daily handling. Rats were euthanized at days 3, 5, and 10 (n = 8) and were monitored daily to determine wound status. Immunohistochemical analysis for the detection of acH3 was performed. One thousand epithelial cells were counted and the mean of acH3 was calculated and compared between groups. PBMT accelerated the repair of oral ulcers. At day 3, PBMT showed a significantly higher mean of acH3 than GC (P = .04). On day 5, no difference was observed between the groups. On day 10, PBMT presented a lower mean of acH3 than the CG (P = .05). PBMT stimulates the oral mucosa wound healing by activating epigenetic mechanisms such as histone acetylation in the early stages of the process. Fipe HCPA: 14-0572

Circulating advanced glycation end products (AGE) and their receptor, RAGE, are increased after a myocardial infarction (MI) episode and seem to be associated with worse prognosis in patients. Despite the increasing importance of these molecules in the course of cardiac diseases, they have never been characterized in an animal model of MI. Thus, the aim of this study was to characterize AGE formation and RAGE expression in plasma and cardiac tissue during cardiac remodeling after MI in rats. Adult male Wistar rats were randomized to receive sham surgery (n = 15) or MI induction (n = 14) by left anterior descending coronary artery ligation. The MI group was stratified into two subgroups based on postoperative left ventricular ejection fraction: low (MIlowEF) and intermediate (MIintermEF). Echocardiography findings and plasma levels of AGEs, protein carbonyl, and free amines were assessed at baseline and 2, 30, and 120 days postoperatively. At the end of follow-up, the heart was harvested for AGE and RAGE evaluation. No differences were observed in AGE formation in plasma, except for a decrease in absorbance in MIlowEF at the end of follow-up. A decrease in yellowish-brown AGEs in heart homogenate was found, which was confirmed by immunodetection of N-ε-carboxymethyl-lysine. No differences could be seen in plasma RAGE levels among the groups, despite an increase in MI groups over the time. However, MI animals presented an increase of 50% in heart RAGE at the end of the follow-up. Despite the inflammatory and oxidative profile of experimental MI in rats, there was no increase in plasma AGE or RAGE levels. However, AGE levels in cardiac tissue declined. Thus, we suggest that the rat MI model should be employed with caution when studying the AGE-RAGE signaling axis or anti-AGE drugs for not reflecting previous clinical findings.

Background: Obesity is a risk factor for cardiovascular diseases; however, in obese heart failure (HF) patients live longer than lean HF patients, this observation is known as obesity paradox. MicroRNAs (miRs) regulate processes involved in both cardiac remodeling and obesity. Objective: We investigated whether the levels of circulating miRs in HF patients are influenced by obesity. Methods: In this case–control study, twenty HF patients (10 obese and 10 lean) and 10 healthy control individuals were analyzed using Affymetrix GeneChip miRNA 4.0 Array following manufacturer's instruction. Raw data was normalized using the Robust Multiarray Average method, batch effect was adjusted with Surrogate Variable Analysis, pairwise differential expression analysis was carry-out with Limma R package, and miRPath v3.0 was used to interrogate pathways enriched for the dysregulated miRNAs based on validated targets from Tarbase v7.0 and KEGG pathways annotation database. Bioinformatics and statistical analysis were performed using R and a p-value <0.01 was considered significant. Results and conclusions: We discovered a set of 36 and 48 miRNAs that were differentially expressed in obese HF and lean HF, respectively, in relation to controls. Thirteen miRNAs were commonly dysregulated in both HF groups. In addition, we found hsa-miR-451a to be up-regulated in obese HF in relation to controls, as well as to lean HF patients, suggesting that obesity may accentuate its dysregulation. On the other hand, hsa-miR-4738-5p, hsa-miR-1260a, and hsa-miR-98-5p were differentially expressed in lean HF in relation to both remaining groups. Pathways enrichment analysis suggested that hsa-miR-451a regulates genes from the mTOR signaling pathway (p=0.002), whereas hsa-miR-1260a modulates Hippo (p=0.02) and AMPK (p=0.03) signaling pathways, all of which have been previously related to cardiac hypertrophy. Further investigation and validation of their targets may contribute to a better understanding of the obesity paradox.

The aim of this study was to evaluate the acute effect of aerobic (AER) and eccentric (ECC) exercise on glucose variability, correlating it with circulating markers of inflammation and oxidative stress in healthy subjects. Sixteen healthy subjects (32 ± 12 years old) wore a continuous glucose monitoring system for three days. Participants randomly performed single AER and ECC exercise sessions. Glucose variability was evaluated by glucose variance (VAR), glucose coefficient of variation (CV%) and glucose standard deviation (SD). Blood samples were collected to evaluate inflammatory and oxidative stress markers. When compared with the pre-exercise period of 0-6 h, all the indices of glucose variability presented comparable reductions 12-18 h after both exercises (∆AER: VAR= 151.5, ∆CV% = 0.55 and ∆SD = 3.1 and ECC: ∆VAR = 221.2 , ∆CV% = 3.7 and ∆SD = 6.5). Increased interleukin-6 (IL-6) levels after AER (68.5%) and ECC (30.8%) (P<0.001) were observed, with no differences between sessions (P = 0.459). Uric acid levels were increased after exercise sessions (3% in AER and 4% in ECC, P = 0.001). In conclusion, both AER and ECC exercise sessions reduced glucose variability in healthy individuals. Inflammatory cytokines, such as IL-6, and stress oxidative markers might play a role in underlying mechanisms modulating the glucose variability responses to exercise (clinicalTrials.gov NCT02262208).