High-intensity interval training mitigates the progression of periodontitis and improves behavioural aspects in rats.

AIM: To investigate the effects of high-intensity interval training (HIIT) on periodontitis (PD) progression and behavioural outcomes. MATERIALS AND METHODS: Forty-eight Wistar rats were divided into four groups: non-trained (NT); non-trained with PD; HIIT with PD; and HIIT. The HIIT protocol, involving daily treadmill sessions, spanned 8 weeks, with PD induced by ligature after the 6th week. Behavioural tests were conducted to assess anxiety and memory. Post euthanasia, we evaluated the systemic inflammatory profile and oxidative stress markers in the hippocampus and amygdala. A morphological evaluation and elemental composition analysis of the mandibular alveolar bone were performed. RESULTS: PD exacerbated alveolar bone level, bone surface damage and alterations in calcium and phosphorus percentages on the bone surface (p < .05), while HIIT attenuated these changes (p < .05). HIIT improved systemic inflammatory markers altered by PD (tumour necrosis factor [TNF]-α, interleukin [IL]-10, TNF-α/IL-10 and IL-1ß/IL-10 ratios, p < .05). PD animals exhibited lower total antioxidant capacity and levels of thiobarbituric acid reactive substances in the amygdala and hippocampus, respectively (p < .05). HIIT maintained these parameters at levels similar to those in NT animals. HIIT improved anxiety and memory outcomes altered by PD (p < .05). CONCLUSIONS: HIIT attenuates systemic inflammation, anxiety and memory outcomes promoted by PD.

The ASSIST trial: Acute effects of manipulating strength exercise volume on insulin sensitivity in obese adults: A protocol for a randomized controlled, crossover, clinical trial.

Type 2 diabetes mellitus is a disease in which insulin action is impaired, and an acute bout of strength exercise can improve insulin sensitivity. Current guidelines for strength exercise prescription suggest that 8 to 30 sets could be performed, although it is not known how variations in exercise volume impact insulin sensitivity. Additionally, this means an almost 4-fold difference in time commitment, which might directly impact an individual's motivation and perceived capacity to exercise. This study will assess the acute effects of high- and low-volume strength exercise sessions on insulin sensitivity. After being thoroughly familiarized, 14 obese individuals of both sexes (>40 year old) will undergo 3 random experimental sessions, with a minimum 4-day washout period between them: a high-volume session (7 exercises, 3 sets per exercise, 21 total sets); a low-volume session (7 exercises, 1 set per exercise, 7 total sets); and a control session, where no exercise will be performed. Psychological assessments (feeling, enjoyment, and self-efficacy) will be performed after the sessions. All sessions will be held at night, and the next morning, an oral glucose tolerance test will be performed in a local laboratory, from which indexes of insulin sensitivity will be derived. We believe this study will aid in strength exercise prescription for individuals who claim not to have time to exercise or who perceive high-volume strength exercise intimidating to adhere to. This trial was prospectively registered (ReBEC #RBR-3vj5dc5 https://ensaiosclinicos.gov.br/rg/RBR-3vj5dc5).

Effects of Branched-Chain Amino Acids on the Inflammatory Response Induced by LPS in Caco-2 Cells.

Branched-chain amino acids (BCAA) are essential for maintaining intestinal mucosal integrity. However, only a few studies have explored the role of BCAA in the modulation of intestinal inflammation. In this study, we investigated in vitro effects of BCAA on the inflammatory response induced by lipopolysaccharide (LPS) (1 µg/mL) in Caco-2 cells. Caco-2 cells were assigned to six groups: control without BCAA (CTL0), normal BCAA (CTL; 0.8 mM leucine, 0.8 mM isoleucine, and 0.8 mM valine); leucine (LEU; 2 mM leucine), isoleucine (ISO; 2 mM isoleucine), valine (VAL; 2 mM valine), and high BCAA (LIV; 2 mM leucine, 2 mM isoleucine, and 2 mM valine). BCAA was added to the culture medium 24 h before LPS stimulation. Our results indicated that BCAA supplementation did not impair cell viability. The amino acids leucine and isoleucine attenuated the synthesis of IL-8 and JNK and NF-kB phosphorylation induced by LPS. Furthermore, neither BCAA supplementation nor LPS treatment modulated the activity of glutathione peroxidase or the intracellular reduced glutathione/oxidized glutathione ratio. Therefore, leucine and isoleucine exert anti-inflammatory effects in Caco-2 cells exposed to LPS by modulating JNK and NF-kB phosphorylation and IL-8 production. Further in vivo studies are required to validate these findings and gather valuable information for potential therapeutic or dietary interventions.

Randomized Controlled Trial to Study the Acute Effects of Strength Exercise on Insulin Sensitivity in Obese Adults.

An acute session of strength exercise (SE) ameliorates insulin sensitivity (IS) for several hours; however, the effects of SE volume (i.e., number of sets) have not been studied thoroughly. Although it is intuitive that some SE is better than none, and more is better than some for the improvement of IS, high-volume sessions might be challenging for diseased populations to complete, especially obese adults, for whom even a brisk walk can be challenging. This protocol details a randomized clinical trial to assess the acute effects of SE on IS in obese adults. The inclusion criteria are body mass index >30 kg/m2, central obesity (waist circumference >88 cm and >102 cm for women and men, respectively), and age >40 years. Participants will be familiarized with the SE (7 exercises targeting major muscle groups) and then will perform three sessions in a randomized order: session 1 - high-volume session (3 sets/exercise); session 2 - low-volume session (1 set/exercise); session 3 - control session (no exercise). Diet will be controlled the day before and on the day of the sessions. Sessions will be completed at night, and an oral glucose tolerance test will be performed the next morning, from which several indexes of IS will be derived, such as the area under the curve (AUC) of glucose and insulin, the Matsuda index, the Cederholm index, the muscle IS index, and the Gutt index. Based on pilot studies, we expect ~15% improvement in IS (insulin AUC, and Matsuda and Cederholm indexes) after the high-volume session, and ~8% improvement after the low-volume session compared to the control session. This study will benefit individuals who find high-volume SE sessions challenging but still aim to improve their IS by investing 1/3 of their time and effort.

Post-translational modification by the Pgf glycosylation machinery modulates Streptococcus mutans OMZ175 physiology and virulence.

Streptococcus mutans is commonly associated with dental caries and the ability to form biofilms is essential for its pathogenicity. We recently identified the Pgf glycosylation machinery of S. mutans, responsible for the post-translational modification of the surface-associated adhesins Cnm and WapA. Since the four-gene pgf operon (pgfS-pgfM1-pgfE-pgfM2) is part of the S. mutans core genome, we hypothesized that the scope of the Pgf system goes beyond Cnm and WapA glycosylation. In silico analyses and tunicamycin sensitivity assays suggested a functional overlap between the Pgf machinery and the rhamnose-glucose polysaccharide synthesis pathway. Phenotypic characterization of pgf mutants (ΔpgfS, ΔpgfE, ΔpgfM1, ΔpgfM2, and Δpgf) revealed that the Pgf system is important for biofilm formation, surface charge, membrane stability, and survival in human saliva. Moreover, deletion of the entire pgf operon (Δpgf strain) resulted in significantly impaired colonization in a rat oral colonization model. Using Cnm as a model, we showed that Cnm is heavily modified with N-acetyl hexosamines but it becomes heavily phosphorylated with the inactivation of the PgfS glycosyltransferase, suggesting a crosstalk between these two post-translational modification mechanisms. Our results revealed that the Pgf machinery contributes to multiple aspects of S. mutans pathobiology that may go beyond Cnm and WapA glycosylation.

SARS-CoV-2 genome incidence on the inanimate surface of the material used in the flow of biological samples from the collection point to the testing unit.

Despite the undeniable effect of vaccination against COVID-19 in reducing disease severity, there is still a need to monitor and limit SARS-CoV-2 circulation and transmission. Thus, this study evaluated the presence of the SARS-CoV-2 genome on the surfaces of highly touched objects manipulated in the biological sample collection point and at the reception unit of the diagnostic laboratory. Surfaces were sampled once a week, for 6 weeks, between September 18th and October 23rd, 2020. RT-qPCR was used for SARS-CoV-2 detection. The coolers for biological sample transportation and the envelope containing the patient form were the objects with the highest occurrence of viral genome detection, although it was detected in each object in only two of the 6 evaluations. And the SARS-CoV-2 genome was detected just once on the vehicle steering wheel, computer keyboard, bathroom door handle and disinfection bench. The virus genome was not detected in any object on three of the six evaluations. And eight was the largest number of surfaces contaminated by the virus genome on one occasion. The reduced incidence of object contamination by the SARS-CoV-2 genome can be explained by the exposure of the objects to environmental conditions and the adoption of virus-spread containment measures. It can also reflect the low incidence of SARS-CoV-2 during the study's development period. Despite the low frequency of SARS-CoV-2 genome detection, our findings show that the virus was present in the environment at some point. This highlights the importance of adopting personal preventive measures to reduce respiratory virus spread, especially during epidemics and outbreaks.

Intense Caloric Restriction from Birth Prevents Cardiovascular Aging in Rats.

We previously demonstrated that a 50% caloric restriction (CR) from birth improves several cardiometabolic risk factors in young rats. In this study, we investigated in middle-aged rats the consequences of a 50% CR from birth on cardiometabolic risk factors, heart function/morphology, ventricular arrhythmia, and fibrillation incidence, and cardiac intracellular proteins involved with redox status and cell survival. From birth to the age of 18 months, rats were divided into an Ad Libitum (AL18) group, which had free access to food, and a CR18 group, which had food limited to 50% of that consumed by the AL18. Resting metabolic rate, blood pressure, and heart rate were recorded, and oral glucose and intraperitoneal insulin tolerance tests were performed. Blood was collected for biochemical analyses, and visceral fat and liver were harvested and weighed. Hearts were harvested for cardiac function, histological, redox status, and western blot analyses. The 50% CR from birth potentially reduced several cardiometabolic risk factors in 18-month-old rats. Moreover, compared with AL18, the CR18 group showed a ∼50% increase in cardiac contractility and relaxation, nearly three to five times less incidence of ventricular arrhythmia and fibrillation, ∼18% lower cardiomyocyte diameter, and ∼60% lower cardiac fibrosis. CR18 hearts also improved biomarkers of antioxidant defense and cell survival. Collectively, these results reveal several metabolic and cardiac antiaging effects of a 50% CR from birth in middle-aged rats.

Don't face writer's block alone.

Creating a writing club allowed a Brazilian PhD student to confront her fears, improve her English and, ultimately, change the way she sees research.

Catalase produced by Candida albicans protects Streptococcus mutans from H2O2 stress-one more piece in the cross-kingdom synergism puzzle.

Co-infection with Streptococcus mutans and Candida albicans is associated with dental caries, and their co-cultivation results in enhanced biofilm matrix production that contributes to increased virulence and caries risk. Moreover, the catalase-negative S. mutans demonstrates increased oxidative stress tolerance when co-cultivated in biofilms with C. albicans, a catalase-producing yeast. Here, we sought to obtain mechanistic insights into the increased H2O2 tolerance of S. mutans when co-cultivated with clinical isolates of Candida glabrata, Candida tropicalis, and C. albicans. Additionally, the C. albicans SC5314 laboratory strain, its catalase mutant (SC5314Δcat1), and S. mutans UA159 and its glucosyltransferase B/C mutant (UA159ΔgtfB/C) were grown as single- and dual-species biofilms. Time-kill assays revealed that upon acute H2O2 challenge, the survival rates of S. mutans in dual-species biofilms with the clinical isolates and C. albicans SC5314 were greater than when paired with SC5314Δcat1 or as a single-species biofilm. Importantly, this protection was independent of glucan production through S. mutans GtfB/C. Transwell assays and treatment with H2O2-pre-stimulated C. albicans SC5314 supernatant revealed that this protection is contact-dependent. Biofilm stability assays with sublethal H2O2 or peroxigenic Streptococcus A12 challenge resulted in biomass reduction of single-species S. mutans UA159 and dual-species with SC5314Δcat1 biofilms compared to UA159 biofilms co-cultured with C. albicans SC5314. S. mutans oxidative stress genes were upregulated in single-species biofilms when exposed to H2O2, but not when S. mutans was co-cultivated with C. albicans SC5314. Here, we uncovered a novel, contact-dependent, synergistic interaction in which the catalase of C. albicans protects S. mutans against H2O2. IMPORTANCE It is well established that co-infection with the gram-positive caries-associated bacterium Streptococcus mutans and the yeast pathobiont Candida albicans results in aggressive forms of caries in humans and animal models. Together, these microorganisms form robust biofilms through enhanced production of extracellular polysaccharide matrix. Further, co-habitation in a biofilm community appears to enhance these microbes' tolerance to environmental stressors. Here, we show that catalase produced by C. albicans protects S. mutans from H2O2 stress in a biofilm matrix-independent manner. Our findings uncovered a novel synergistic trait between these two microorganisms that could be further exploited for dental caries prevention and control.

Seroprevalence of Dengue, Chikungunya, and Zika viruses antibodies in a cohort of asymptomatic pregnant women in a low-income region of Minas Gerais, Brazil, 2018-2019.

Dengue, Chikungunya, and Zika viruses are arthropod-borne viruses (arboviruses) that infect millions of individuals in tropical and subtropical regions. In the Americas, arboviruses represent a major public health problem, especially among vulnerable groups such as the elderly, children, and pregnant women. In this study, the seroprevalence of IgM or IgG against these arboviruses in pregnant, young women in the city of Diamantina, Minas Gerais, Brazil, and the influence of sociodemographic factors on the incidence/prevalence of infection in this group were investigated. A cross-sectional investigation was conducted on a total of 135 pregnant women for Dengue and Chikungunya IgM and 88 pregnant women for Zika IgG. Dengue IgM was found on the serum of twenty participants (14.8%) and only one woman (0.7%) tested positive for Chikungunya IgM. Zika IgG was found in three (3.4%) participants and 2 women who tested positive for Zika virus were also positive for Dengue virus IgM. Although the arboviruses seroprevalence was higher frequency among young (20-25 years old), brown and high school women, with a monthly income of 1-3 minimum wages, no association between these sociodemographic factors and arboviruses seroprevalence was found.

Early Six-Minute Walk Test May Predict Midterm Outcomes Following Coronary Artery Bypass Grafting.

OBJECTIVE: This study aims to investigate the ability of the six-minute walk distance (6MWD) as a prognostic marker for midterm clinical outcomes three months after coronary artery bypass grafting (CABG), to identify possible predictors of fall in 6MWD in the early postoperative period, and to establish the percentage fall in early postoperative 6MWD, considering the preoperative baseline as 100%. METHODS: A prospective cohort of patients undergoing elective CABG were included. The percentage fall in 6MWD was assessed by the difference between preoperative and postoperative day (POD) five. Clinical outcomes were evaluated three months after hospital discharge. RESULTS: There was a significant decrease in 6MWD on POD5 compared with preoperative baseline values (percentage fall of 32.5±16.5%, P<0.0001). Linear regression analysis showed an independent association of the percentage fall of 6MWD with cardiopulmonary bypass (CPB) and preoperative inspiratory muscle strength. Receiver operating characteristic curve analysis revealed that the best cutoff value of percentage fall in 6MWD to predict poorer clinical outcomes at three months was 34.6% (area under the curve = 0.82, sensitivity = 78.95%, specificity = 76.19%, P=0.0001). CONCLUSION: This study indicates that a cutoff value of 34.6% in percentage fall of 6MWD on POD5 was able to predict poorer clinical outcomes at three months of follow-up after CABG. Use of CPB and preoperative inspiratory muscle strength were independent predictors of percentage fall of 6MWD in the postoperative period. These findings further support the clinical application of 6MWD and propose an inpatient preventive strategy to guide clinical management over time.

Mitochondria are midfield players in steroid synthesis.

Steroids are important membrane components and signaling metabolites and thus are required for cellular homeostasis. All mammalian cells retain the ability to uptake and synthesize steroids. Dysregulation of steroid levels leads to profound effects on cellular function and organismal health. Hence it comes as no surprise that steroid synthesis is tightly regulated. It is well established that the main site for steroid synthesis and regulation is the endoplasmic reticulum. However, mitochondria are essential for: (1) cholesterol production (the precursor of all steroids) by exporting citrate and; (2) the products of steroidogenesis (such as mineralocorticoids and glucocorticoids). In this review, we describe the midfield player role of mitochondria in steroid synthesis and bring the idea of mitochondria actively participating in steroid synthesis regulation. A better understanding of the mitochondrial regulatory roles in steroid synthesis would open new avenues to targeted approaches aiming to control steroid levels.

Antimicrobial Photodynamic Therapy as a Technique for Decontamination of Acrylic Resin Devices Provided by Different Dental Laboratories.

Introduction: Dentures, occlusal splints, surgical guides and orthodontic appliances are examples of acrylic resin devices made in dental laboratories, which must be disinfected and even sterilized before insertion into the oral cavity. This study evaluated the antimicrobial effect of photodynamic therapy (PDT) applied to acrylic resin specimens received from different laboratories. Methods: Three hundred standardized specimens were ordered from six randomly selected laboratories registered in the Council of Dentistry of Ceará (n=50). The PDT consisted in the association of 22 µM erythrosine, as a photosensitizer (P), and a 520-nm LED at 38 J/cm2 (L). The specimens of each laboratory were randomly distributed into five groups: positive control, sterilized with ethylene oxide; negative control, untreated (P-L-); erythrosine control, only stained (P+L-); LED control, only irradiated (P-L+); PDT (P+L+). Then, the specimens were individually sonicated in saline solution; the suspension was diluted, plated on culture mediums (blood agar, sabouraud dextrose agar and a non-selective chromogenic agar), and incubated for 48 hours at 37°C. Colony-forming-unit (CFU) counts were done and statistical tests of Kruskal-Wallis/Dunn were carried out. Results: The specimens from all laboratories were contaminated with bacteria and yeasts. Staphylococcus aureus, Staphylococcus saprophyticus, Escherichia coli, Enterococcus spp., Klebsiella and Pseudomonas spp. were identified. The PDT significantly reduced CFU counts (P<0.0001), compared to P-L-. Conclusion: PDT was able to effectively decontaminate the acrylic resin specimens provided from dental laboratories.

Association between Anti-DENV IgM Serum Prevalence and CD11b Expression by Classical Monocytes in Obesity.

Dengue and obesity are currently highly prevalent conditions worldwide and the association between these two conditions may result in greater risk for DENV infection and disease severity. In this study the association between obesity and recent, inapparent dengue was investigated. Serum DENV IgM and NS1 were evaluated in 49 adult volunteers (15 lean and 34 individuals with obesity, according to body mass index), between September 2017 and June 2018. Adiposity, endocrine, metabolic, and immune data of the participants were also obtained. None of the study participants tested positive for the DENV NS1 antigen. DENV IgM was detected in 33.3% of the lean individuals, and in 44.1% of those with obesity; the presence of DENV IgM was not associated with body mass index (OR = 1.32, 95% CI = 0.59-2.98, p = 0.48). However, body fat index was higher in obese individuals who had recent inapparent dengue (14.7 ± 3.1 versus 12.7 ± 2.1 kg/m2, p = 0.04), as was the expression of CD11b by classical (CD14++CD16-) monocytes (1103.0 ± 311.3 versus 720.3 ± 281.1 mean fluoresce intensity). Our findings suggest an association between adiposity and recent inapparent dengue and the involvement of classical monocytes in this association.

Early Six-Minute Walk Test May Predict Midterm Outcomes Following Coronary Artery Bypass Grafting

ABSTRACT Objective: This study aims to investigate the ability of the six-minute walk distance (6MWD) as a prognostic marker for midterm clinical outcomes three months after coronary artery bypass grafting (CABG), to identify possible predictors of fall in 6MWD in the early postoperative period, and to establish the percentage fall in early postoperative 6MWD, considering the preoperative baseline as 100%. Methods: A prospective cohort of patients undergoing elective CABG were included. The percentage fall in 6MWD was assessed by the difference between preoperative and postoperative day (POD) five. Clinical outcomes were evaluated three months after hospital discharge. Results: There was a significant decrease in 6MWD on POD5 compared with preoperative baseline values (percentage fall of 32.5±16.5%, P<0.0001). Linear regression analysis showed an independent association of the percentage fall of 6MWD with cardiopulmonary bypass (CPB) and preoperative inspiratory muscle strength. Receiver operating characteristic curve analysis revealed that the best cutoff value of percentage fall in 6MWD to predict poorer clinical outcomes at three months was 34.6% (area under the curve = 0.82, sensitivity = 78.95%, specificity = 76.19%, P=0.0001). Conclusion: This study indicates that a cutoff value of 34.6% in percentage fall of 6MWD on POD5 was able to predict poorer clinical outcomes at three months of follow-up after CABG. Use of CPB and preoperative inspiratory muscle strength were independent predictors of percentage fall of 6MWD in the postoperative period. These findings further support the clinical application of 6MWD and propose an inpatient preventive strategy to guide clinical management over time.

Efeito dos aminoácidos de cadeia ramificada na modulação da resposta inflamatória induzida por LPS em células intestinais Caco-2 / Effect of branched-chain amino acids on the inflammatory response modulation induced by LPS in Caco-2 intestinal cells

Introdução: A inflamação é um fator presente na fisiopatologia de doenças, e, o intestino, órgão susceptível a disfunções e à disbiose, que está em constante exposição a microrganismos, pode estar associado a este processo. Por outro lado, aminoácidos de cadeia ramificada (ACR) podem apresentar efeitos anti-inflamatórios em linhagem celular intestinal. Objetivo: Investigar os efeitos da suplementação de ACR, in vitro, na modulação da resposta inflamatória e do estresse oxidativo induzida por LPS em modelo de células intestinais Caco-2. Métodos: As culturas de células foram distribuídas em seis grupos, sendo: dois grupos controle - controle com meio Dulbecco's Modified Eagle's Medium sem ACR (CTL0) e com ACR (CTL) - e quatro grupos suplementados - grupo leucina (LEU), grupo isoleucina (ISO), grupo valina (VAL) e associação de ACR (LIV). Foi adotado um protocolo de pré-tratamento, com os ACR e o LPS. A viabilidade celular foi avaliada pelo ensaio de redução do MTT (brometo de [3-(4,5-dimetiltiazol-2yl 2,5-difenil tetrazolium]). A análise de proteínas (mTOR, AKT, AMPK, NF-kB, TAK-1 e JNK) foi feita por Western Blotting. A dosagem de citocina IL-8 no sobrenadante de cultura celular foi realizada por meio do kit Multiplex para imunoensaio. Foram avaliados componentes do sistema antioxidante relacionado à glutationa (GSH, GSSG e GPx) por meio de kit comercial. Os resultados foram expressos como média ± erro padrão. Resultados: No teste de viabilidade celular por MTT, verificou-se que os grupos tratados com ACR e LPS não apresentaram comprometimento da viabilidade celular em relação ao grupo CTL sem LPS. Os grupos CTL0, CTL, VAL e LIV, quando estimulados com LPS, apresentaram maior capacidade de síntese, in vitro, de IL-8, bem como maior fosforilação das proteínas JNK e NF-kB em relação aos seus respectivos grupos sem LPS. A suplementação. Todavia, as células estimuladas com LPS e suplementadas com leucina e isoleucina apresentaram capacidade de síntese, in vitro, de IL-8 e fosforilação das proteínas JNK e NF-kB que não diferiram significativamente das células suplementadas com esses aminoácidos e não estimuladas com LPS. A ausência de ACR (grupo CTL0) e a suplementação desses aminoácidos, in vitro, em células intestinais Caco-2, tratadas com ou sem LPS, não induziram alteração da atividade da enzima GPx e da razão intracelular GSH/GSSG. Conclusões: Os aminoácidos leucina e isoleucina apresentaram potencial efeito anti-inflamatório nas células Caco-2 por meio da modulação da fosforilação das proteínas JNK e NF-kB, cujo fato está associado à modulação da síntese, in vitro, de IL-8, a qual está envolvida na resposta inflamatória no intestino.

Introduction: Inflammation is a factor present in the pathophysiology of diseases, and the intestine, an organ susceptible to dysfunction and dysbiosis, which is constantly exposed to microorganisms, may be associated with this process. On the other hand, branched-chain amino acids (BCAAs) may have anti-inflammatory effects on intestinal cell lines. Objective: To investigate the effects of ACR supplementation, in vitro, on the modulation of inflammatory response and oxidative stress induced by LPS in a model of intestinal Caco-2 cells. Methods: The cell cultures were divided into six groups, as follows: two control groups - control with Dulbecco's Modified Eagle's Medium without ACR (CTL0) and with ACR (CTL) - and four supplemented groups - leucine group (LEU), isoleucine group (ISO), valine group (VAL) and ACR association (LIV). A pre-treatment protocol was adopted, with ACR and LPS. Cell viability was assessed by MTT reduction assay ([3-(4,5-dimethylthiazol-2yl 2,5-diphenyl tetrazolium] bromide)) Protein analysis (mTOR, AKT, AMPK, NF-kB, TAK -1 and JNK) was performed by Western Blotting. The dosage of cytokine IL-8 in the cell culture supernatant was performed using the Multiplex kit for immunoassay. Components of the antioxidant system related to glutathione (GSH, GSSG, and GPx) were evaluated by commercial kit. The results were expressed as mean ± standard error. Results: In the cell viability test by MTT, it was verified that the groups treated with ACR and LPS did not show impairment of cell viability compared to the CTL group without LPS. The CTL0, CTL, VAL, and LIV groups, when stimulated with LPS, showed a greater capacity for in vitro synthesis of IL-8, as well as greater phosphorylation of JNK and NF-kB proteins in relation to their respective groups without LPS. However, cells stimulated with LPS and supplemented with leucine and isoleucine showed in vitro synthesis capacity of IL-8 and phosphorylation of JNK and NF-kB proteins that did not differ significantly from cells supplemented with these amino acids and not stimulated with LPS. The absence of ACR (CTL0 group) and the supplementation of these amino acids, in vitro, in intestinal Caco-2 cells, treated with or without LPS, did not induce changes in the activity of the GPx enzyme and the intracellular GSH/GSSG ratio. Conclusions: The amino acids leucine and isoleucine had a potential anti-inflammatory effect on Caco-2 cells by modulating the phosphorylation of JNK and NF-kB proteins, which fact is associated with the modulation of the in vitro synthesis of IL-8, which is involved in the inflammatory response in the gut.

Corrigendum: Modulation of leukocyte subsets mobilization in response to exercise by water immersion recovery.

[This corrects the article DOI: 10.3389/fphys.2022.867362.].

Modulation of Leukocyte Subsets Mobilization in Response to Exercise by Water Immersion Recovery.

Purpose: To investigate the effect of different water immersion temperatures on the kinetics of blood markers of skeletal muscle damage and the main leukocyte subpopulations. Methods: Eleven recreationally trained young men participated in four experimental sessions consisting of unilateral eccentric knee flexion and 90 min of treadmill running at 70% of peak oxygen uptake, followed by 15 min of water immersion recovery at 15, 28 or 38°C. In the control condition participants remained seated at room temperature. Four hours after exercise recovery, participants completed a performance test. Blood samples were obtained before and immediately after exercise, after immersion, immediately before and after the performance test and 24 h after exercise. The number of leukocyte populations and the percentage of lymphocyte and monocytes subsets, as well as the serum activity of creatine kinase and aspartate aminotransferase were determined. Results: Leukocytosis and increase in blood markers of skeletal muscle damage were observed after the exercise. Magnitude effect analysis indicated that post-exercise hot-water immersion likely reduced the exercise-induced lymphocytosis and monocytosis. Despite reduced monocyte count, recovery by 38°C immersion, as well as 28°C, likely increased the percentage of non-classical monocytes in the blood. The percentage of CD25+ cells in the CD4 T cell subpopulation was possibly lower after immersion in water at 28 and 15°C. No effect of recovery by water immersion was observed for serum levels of creatine kinase and aspartate aminotransferase. Conclusions: Recovery by hot-water immersion likely attenuated the leukocytosis and increased the mobilization of non-classical monocytes induced by a single session of exercise combining resistance and endurance exercises, despite no effect of water immersion on markers of skeletal muscle damage. The monocyte response mediated by hot water immersion may lead to the improvement of the inflammatory response evoked by exercise in the skeletal muscle.

Clinical study of the biofilm of implant-supported complete dentures in healthy patients.

OBJECTIVE: The purpose of this study was to quantify the area covered by biofilm and identify bacteria and yeasts present in mandibular acrylic resin full-arch implant-supported fixed prostheses. BACKGROUND: Biofilm control of implant-supported fixed prosthesis is hampered by their design, and it can cause oral and systemic problems, mainly in immunocompromised patients like the elder. Knowledge about microbiota reinforces the awareness about the need for periodic professional cleaning maintenance. MATERIALS AND METHODS: Twenty prostheses were unscrewed, washed in 0.89% sodium chloride, stained with eosin 1% and photographed. The area covered by biofilm was digitally delimited and quantified. Biofilm samples were collected, diluted up to 1:107 , seeded in chromogenic agar media and incubated for 48 hours, at 37°C, for counting of colony-forming units (CFU/mL). DNA hybridization was performed to complement the identification and quantification of microorganisms. Data were analyzed using Mann-Whitney test, Spearman correlation and Fisher's exact test (α = .05). RESULTS: An average of 62% of the gingival surface of the prostheses was covered by biofilm. Enterococcus spp. (5.82 ± 1.38 log10 CFU/mL) and Staphylococcus aureus (5.75 ± 2.02 log10 CFU/mL) showed higher prevalence in cultures. Patients with five implants had less biofilm compared to those with four implants (P = .031) but had higher Escherichia coli counts (P = .039). In DNA hybridization, Streptococcus pneumoniae, Veillonella parvula and Fusobacterium nucleatum presented higher quantification and were present in all the samples; patients over 65 years old contained more Candida tropicalis (P = .049); prostheses on five implants presented lower quantification for several species. CONCLUSION: Biofilm was present on all prostheses, containing potentially pathogenic microorganisms. The number of implants may play a role in quantification of biofilm and in microorganism counts.

Storage Duration Affects the Quantification of Oxidative Stress Markers in the Gastrocnemius, Heart, and Brain of Mice Submitted to a Maximum Exercise.

This study investigated the effect of sample storage duration on the quantification of oxidative stress markers in the gastrocnemius, heart, and brain of mice submitted to a maximum swimming exercise. Thiobarbituric acid reactive substances (TBARSs), protein carbonyl derivatives, total antioxidant capacity (TAC), and the activity of superoxide dismutase (SOD) and catalase (CAT) were quantified in fresh tissues and in samples stored at -80°C for 1, 3, or 6 months, from exercised (n = 13) and nonexercised mice (n = 13). Except for protein carbonyl derivatives in the heart, the exercise resulted in the modification of all markers in all fresh-evaluated samples (p < 0.001). The storage duration did not modify the effect of exercise on protein carbonyl derivatives and TAC. TBARS was stable for 3 months in the gastrocnemius and for 1 month in frozen heart and brain. Accordingly, the exercise effect on TBARS levels observed in fresh samples was absent in the gastrocnemius frozen for 6 months (p = 0.98) and in the heart and brain frozen for 3 months (p = 0.07 and 0.28, respectively) or more (p = 0.21 for heart and p > 0.99 for brain). In addition, CAT and SOD activities were reduced by storage duration in all tissues evaluated (p < 0.05). Our findings show that sample storage duration alters the quantification of oxidative stress markers in mice submitted to maximum exercise, and its effect is tissue and marker dependent. Some recommendations to achieve more accurate and reproducible data in the exercise physiology and oxidative stress markers field are presented.