Alteration in the Expression of Circular Rnas and its association with the Development and Progression of Osteosarcoma, an Integrative Review with High Sensitivity Research.

BACKGROUND: Osteosarcoma is the most common primary malignant bone tumor, mainly affecting children, young adults, and the elderly. It is an aggressive cancer with a poor prognosis, exhibiting low survival rates even with standard treatment. Recently, circular RNA molecules capable of influencing gene expression through various functions, with their main role being acting as microRNA sponges and reducing their intracellular expression, have been identified. Recent studies have linked circular RNAs to osteosarcoma development and progression. Therefore, the present study aimed to investigate the alteration in circular RNA expression during osteosarcoma development and progression. METHODS: An integrative literature review was conducted from September 10th to November 12th, 2021, using the following databases: PubMed/MEDLINE, SCOPUS, Web of Science, OVID, and EMBASE. 129 full articles were included in the review. The obtained data were organized using a standardized data collection instrument, which included the following information: altered expression profile of circular RNAs, associated cancer hallmarks, clinical-pathological relationships of circular RNAs, and perspectives on the studied circular RNAs. RESULTS: A total of 94 distinct circular RNAs were identified, predominantly showing an increased expression pattern. Approximately 91% of the studies that aimed to identify the mechanisms of action of circular RNAs highlighted the function of circular RNAs as microRNA sponges. The most associated cancer hallmarks with the identified circular RNAs were proliferative signaling induction, invasion and metastasis, and resistance to cell death. The altered expression of these circular RNAs generally correlated with a worse prognosis for patients, as evidenced by clinical features such as shorter survival, advanced Enneking and/or TNM stage, higher incidence of metastasis, larger tumor size, and increased chemoresistance. CONSLUSION: These findings indicate the significance of circular RNA molecules in osteosarcoma carcinogenesis, suggesting their potential as new prognostic and/or diagnostic biomarkers, as well as alternative therapeutic targets in the fight against osteosarcoma.

Feasibility and effect of high-intensity training on the progression of motor symptoms in adult individuals with Parkinson's disease: A systematic review and meta-analysis.

BACKGROUND: To determine the feasibility and effect of high-intensity interval training (HIIT) in individuals with Parkinson's and their effect on symptom modification and progression. METHODS: We conducted this systematic review following the Preferred Reporting Items for systematic review and meta-analysis (PRISMA). All studies were searched in seven databases: MEDLINE (PubMed), Cochrane Central Register of Controlled Trials, Web of Science, EMBASE, SPORTDiscus, Virtual Health Library (VHL) and SCOPUS in September 2020 and updated in June 2023. The risk of bias was assessed by the Cochrane Collaboration tool and Grading of Recommendations Assessment, Development and Evaluation (GRADE) tool. We used standardized mean difference (SMD) with a 95% confidence interval (CI) and random effects models, as well as the non-parametric Cochran's Q test and I2 inconsistency test to assess heterogeneity. RESULTS: A total of 15 randomized clinical trials with 654 participants (mean age, 65.4 years). The majority of studies included high intensity training interventions versus moderate intensity, usual care, or control group. The meta-analysis comparing high-intensity exercise versus control group showed an improvement in the disease severity (MD = -4.80 [95%CI, -6.38; -3.21 high evidence certainty); maximum oxygen consumption (MD = 1.81 [95%CI, 0.36; 3.27] very low evidence certainty) and quality of life (MD = -0.54 [95%CI, -0.94; -0.13] moderate evidence certainty). The results showed that high-intensity exercise compared with moderate intensity exercise group showed a improve motor function and functional mobility measured by the TUG test (MD = -0.38 [95%CI, -0.91; 0.16] moderate evidence certainty) with moderate heterogeneity between studies. CONCLUSION: High-intensity exercise performed in both continuous and interval modes when compared with control groups may provide motor function benefits for individuals with Parkinson's disease. HIIT may be feasible, but the intensity of the exercise may influence individuals with Parkinson's disease. However, there was a lack of evidence comparing high intensity and moderate intensity for this population, as the results showed heterogeneity.

O papel dos RNAs longos não codificantes na insuficiência cardíaca: uma revisão de literatura / The role of non-coding long RNAs in heart failure: a literature review / El papel de las RNAs largas no codificantes en la insuficiencia cardiaca: una revisión de la literatura

A Insuficiência Cardíaca (IC) é uma das principais causas de internação hospitalar no mundo e tem um elevado grau de morbidade e mortalidade, sendo um grave problema de saúde pública. Os lncRNAs (RNAs longo não codificantes), têm funções regulatórias transcricionais e/ou pós transcricionais bem complexas e que ainda não são totalmente claras, mas que podem exercer influência sobre as doenças cardiovasculares, dentre elas a IC. Assim o estudo teve como objetivo identificar na literatura o papel dos lncRNAs na patogênese da IC por meio de uma revisão integrativa com busca sistemática. Foram considerados elegíveis para leitura e composição do estudo 33 artigos e os principais papéis dos lncRNA na IC foram relatados como possíveis marcadores biológicos para diagnóstico e prognóstico da doença devido a sua expressividade na corrente sanguínea. Além disso, os lncRNAs podem estar relacionados à capacidade funcional uma vez que o aumento ou diminuição de sua expressão promove redução da apoptose de células endoteliais, melhora a disfunção cardíaca, distúrbios de contratilidade e dos canais de cálcio em pacientes com IC. Portanto, os lncRNAs parecem estar envolvidos na patogênese e/ou fisiopatologia da IC, podendo ser utilizados como biomarcadores genéticos com sensibilidade e especificidade semelhantes ou superiores aos empregados atualmente no diagnóstico e prognóstico da IC.

Heart Failure (HF) is one of the main causes of hospitalization worldwide and has a high degree of morbidity and mortality being considered a public health pro- blem. lncRNAs (non-coding long RNAs) have very complex transcriptional and/or post- transcriptional regulatory functions that are still not entirely clear but may influence car- diovascular diseases, including HF. Thus, the study aimed to identify in the literature the role of lncRNAs in the pathogenesis of HF through an integrative review with a systema- tic search. A total of 33 articles were considered eligible for reading and composition of the study. The roles of lncRNA in HF were reported as possible biological markers for the diagnosis and prognosis of the disease due to its expressiveness in the bloodstream. In addition, lncRNAs may be related to functional capacity since the increase or decrease in their expression promotes a reduction in endothelial cell apoptosis, and improves car- diac dysfunction, contractility, and calcium channel disorders in patients with HF. The- refore, lncRNAs seem to be involved in the pathogenesis and/or pathophysiology of HF and can be used as genetic biomarkers with sensitivity and specificity similar or superior to those currently employed in the diagnosis and prognosis of HF.

La Insuficiencia Cardiaca (IC) es una de las principales causas de hospita- lización en el mundo y tiene un alto grado de morbimortalidad considerándose un pro- blema de salud pública. Los lncRNAs (ARN largos no codificantes) tienen funciones re- guladoras transcripcionales y/o post-transcripcionales muy complejas que aún no están del todo claras pero que pueden influir en las enfermedades cardiovasculares, incluida la IC. Así pues, el estudio se propuso identificar en la literatura el papel de los lncRNAs en la patogénesis de la IC mediante una revisión integradora con una búsqueda sistemática. Un total de 33 artículos fueron considerados elegibles para su lectura y composición del estudio. Las funciones de los lncRNA en la IC se señalaron como posibles marcadores biológicos para el diagnóstico y pronóstico de la enfermedad debido a su expresividad en el torrente sanguíneo. Además, los lncRNAs pueden estar relacionados con la capacidad funcional, ya que el aumento o disminución de su expresión promueve una reducción de la apoptosis de las células endoteliales y mejora la disfunción cardiaca, la contractilidad y los trastornos de los canales de calcio en pacientes con IC. Por tanto, los lncRNAs parecen estar implicados en la patogénesis y/o fisiopatología de la IC y pueden ser utili- zados como biomarcadores genéticos con sensibilidad y spe-cificidad similares o superi- ores a los empleados actualmente en el diagnóstico y pronóstico de la IC.

The impact of the ABO/Rh blood group on susceptibility and severity among COVID-19 patients in Luanda, Angola

SARS-CoV-2 is a public health concern worldwide. Identification of biological factors that could influence transmission and worsen the disease has been the subject of extensive investigation. Herein, we investigate the impact of the ABO/Rh blood group on susceptibility and severity among COVID-19 patients in Luanda, Angola. This was a multicentric cohort study conducted with 101 COVID-19 patients. Chi-square and logistic regression were calculated to check factors related to the worsening of the disease and deemed significant when p<0.05. Blood type O (51.5%) and Rh-positive (93.1%) were the most frequent. Patients from blood type O had a high risk to severe disease [OR: 1.33 (95% CI: 0.42 - 4.18), p=0.630] and hospitalization [OR: 2.59 (95% CI: 0.84 - 8.00), p=0.099]. Also, Rh-positive blood type presented a high risk for severe disease (OR: 10.6, p=0.007) and hospitalization (OR: 6.04, p=0.026). We find a high susceptibility, severity, hospitalization, and mortality, respectively, among blood group O and Rh-positive patients, while blood group AB presented a low susceptibility, severity, hospitalization, and mortality, respectively. Our findings add to the body of evidence suggesting that ABO/Rh blood groups play an important role in the course of SARS-CoV-2 infection.

Patient-Reported Outcomes after Subpectoral Breast Augmentation with Microtextured or Macrotextured Implants Using the BREAST-Q.

Background Breast augmentation with implants is the most commonly performed cosmetic plastic surgery in Brazil and worldwide. The aim of this study was to assess patient satisfaction and quality of life following subpectoral breast augmentation with either microtextured or macrotextured implants, using the BREAST-Q. Methods A prospective study was conducted with 40 women with hypomastia undergoing subpectoral breast augmentation. The patients were randomly allocated to two groups to receive either microtextured or macrotextured breast implants. All participants were assessed preoperatively (baseline) and after 2 and 4 months of surgery for quality of life and patient satisfaction with the surgical results, using the BREAST-Q augmentation module, a patient-reported outcome measure. Results The patients had a mean age of 28.9 ± 6.45 years. The microtextured ( n = 20) and macrotextured ( n = 20) groups were homogeneous for sex, age, education level, marital status, and number of children ( p > 0.05). Both groups showed significant improvement in satisfaction with breasts ( p < 0.001), psychosocial well-being ( p < 0.001), and sexual well-being ( p < 0.001) at the 2- and 4-month follow-up visits compared with baseline. The observed improvements were associated with high effect size values of 5.09, 3.44, and 3.90, respectively. In contrast, significant decreases from baseline in physical well-being scores ( p = 0.001) were found 2 and 4 weeks after surgery in both groups. Conclusion Subpectoral breast augmentation with either microtextured or macrotextured breast implants improved satisfaction with breasts and quality of life in patients with hypomastia.

A canthin-6-one derivative induces cell death by apoptosis/necroptosis-like with DNA damage in acute myeloid cells.

Natural products have long been considered a relevant source of new antitumor agents. Despite advances in the treatment of younger patients with acute myeloid leukemia (AML), the prognosis of elderly patients remains poor, with a high frequency of relapse. The cytotoxicity of canthin-6-one alkaloids has been extensively studied in different cell types, including leukemic strains. Among the canthin-6-one analogs tested, 10-methoxycanthin-6-one (Mtx-C) showed the highest cytotoxicity in the malignant AML cells Kasumi-1 and KG-1. Thus, we evaluated the cytotoxicity and cell death mechanisms related to Mtx-C using the EC50 (80 µM for Kasumi-1 and 36 µM for KG-1) treatment for 24 h. Our results identify reactive oxygen species production, mitochondrial depolarization, annexin V-FITC/7-AAD double staining, caspase cleave and upregulation of mitochondria-dependent apoptosis proteins (Bax, Bim, Bik, Puma and phosphorylation of p53) for both cell lineages. However, downregulation of Bcl-2 and the simultaneous execution of the apoptotic and necroptotic programs associated with the phosphorylation of the proteins receptor-interacting serine/threonine-protein kinase 3 and mixed lineage kinase domain-like pseudokinase occurred only in Kasumi-1 cells. About the lasted events, Kasumi-1 cell death was inhibited by pharmacological agents such as Zvad-FMK and necrostatin-1. The underlying molecular mechanisms of Mtx-C still include participation in the DNA damage and stress-signaling pathways involving p38 and c-Jun N-terminal mitogen-activated protein kinases and interaction with DNA. Thus, Mtx-C represents a promising tool for the development of new antileukemic molecules.

Reproducibility of an endurance test for master swimmers

Abstract Aim: This study aimed to verify the reproducibility of an endurance test for master swimmers. Methods: Seventeen male swimmers, aged from 30 to 50 years (35.06 ± 5.36), weight (78.71 ± 13.06 kg), height (174.35 ± 0.07 cm), participated in this study. The participants underwent the Progressive Swim Test (PSwT), which corresponds to swimming progressively to exhaustion in a 25-meter pool, guided by the recording of sound signals indicating the swimming rhythm. We verified the following variables: heart rate before the test (BHR) and right after the end of the test (AHR), followed by the rate of perceived exertion (RPE) and a total of laps performed (SN - swimming number). For comparison between the three days of tests, the one-way repeated measures ANOVA with Tukey's Post-Hoc was performed. Relative reproducibility was performed by the r test and the intraclass correlation coefficient (ICC). Absolute reproducibility was determined using the coefficient of variation (CV) and the confidence interval (CI). Results: The results showed high reproducibility in (PSwT) in master swimmers in the analyzed variables of intergroup and intragroup with relative reproducibility of r > 0.60 and ICC > 0.70 and absolute reproducibility with CI 95% (lower −1.10 and upper 0.99) and CV < 10%. Most of the cases were within the limits of agreement of the Bland-Altman tests, all variables were considered a level of significance with (p <0.05). Conclusion: The Progressive Swim Test (PSwT) is a reproducible instrument in the assessment of endurance in master swimmers.

Reality and Virtual: Hemodynamic Variables and Rating of Perceived Exertion Between Dance Sessions and Active Videogame Just Dance in Women.

Objective: To analyze the hemodynamic variables and the rating of perceived exertion (RPE) between an active videogame (AVG) session and a dance session. Materials and Methods: The sample consisted of 30 university women between 18 and 30 years of age, physically active and having some kind of experience with dancing. The volunteers participated in the two sessions held on different days, the first with an active dance videogame and the other with a dance instructor, both lasting 50 minutes, with a sequence of songs and moderate intensity. For the evaluation of the two sessions, heart rate (HR), blood pressure (BP), and RPE measurements were taken before and every 10 minutes, until the time for each modality (AVG or dance) was completed. The Shapiro-Wilk and Levene test were used for normal data. For the comparison of two sessions, the two-way ANOVA test was used for repeated measures, and were considered to have a significance level of P < 0.05. Results: There were significant differences in HR [F(6, 96) = 2.606, P = 0.02, η2 P = 0.140], from 10 to 50 minutes (P < 0.01), and diastolic BP [F(6, 84) = 1.858, P = 0.10, η2 P = 0.117], as well as, in the 20th minute (P = 0.02) between Just Dance 2016® and a dance session. All the variables analyzed showed an effect size as large in the classification of the degree of magnitude. Conclusion: There are similarities between the virtual and real dance sessions regarding hemodynamic variables and RPE. Thus, AVGs, especially Just Dance®, may be a favorable option for individuals who wish to exercise at home.

The search for assisted reproduction: profile of patients seen in the fertility outpatient clinic of a public hospital.

OBJECTIVE: To analyze the epidemiological profile of patients treated at the Fertility Outpatient Clinic of a tertiary public hospital in Juiz de Fora. METHODS: This cross-sectional study analyzed the medical records of 448 patients who sought fertility treatment at a tertiary public hospital. The data collected from the medical records were used to assess the main causes of infertility, find the most frequently performed procedures, and the cases eligible to therapeutic or prophylactic intervention. RESULTS: Of the 448 patients included in the study, 385 (86%) sought fertility consultation, 49 (10%) came in for repeated miscarriages, and 14 (3%) for other reasons. Of the 438 infertile patients, 280 (63.9%) had primary and 158 (36.1%) had secondary infertility. The top-three conditions of the 295 patients with established diagnoses were chronic anovulation (n=98; 33%); tubal factor infertility (n=86; 29%); and male factor infertility (n=59; 20%). CONCLUSIONS: Improving care in reproductive health requires a more profound comprehension of the epidemiological profile of patients seeking treatment. There are alternative cost-effective means to contain the development of infertility. Additional expenditure in public healthcare is needed to accommodate the growing number of individuals seeking fertility treatment in Brazil.

Involvement of P2 receptors in hematopoiesis and hematopoietic disorders, and as pharmacological targets.

Several reports have shown the presence of P2 receptors in hematopoietic stem cells (HSCs). These receptors are activated by extracellular nucleotides released from different sources. In the hematopoietic niche, the release of purines and pyrimidines in the milieu by lytic and nonlytic mechanisms has been described. The expression of P2 receptors from HSCs until maturity is still intriguing scientists. Several reports have shown the participation of P2 receptors in events associated with modulation of the immune system, but their participation in other physiological processes is under investigation. The presence of P2 receptors in HSCs and their ability to modulate this population have awakened interest in exploring the involvement of P2 receptors in hematopoiesis and their participation in hematopoietic disorders. Among the P2 receptors, the receptor P2X7 is of particular interest, because of its different roles in hematopoietic cells (e.g., infection, inflammation, cell death and survival, leukemias and lymphomas), making the P2X7 receptor a promising pharmacological target. Additionally, the role of P2Y12 receptor in platelet activation has been well-documented and is the main example of the importance of the pharmacological modulation of P2 receptor activity. In this review, we focus on the role of P2 receptors in the hematopoietic system, addressing these receptors as potential pharmacological targets.

Strength training improves body composition, muscle strength and increases CD4+ T lymphocyte levels in people living with HIV/AIDS.

The establishment of physical training programs for people living with HIV/AIDS (PLWHA) has several benefits. The study aimed to analyze the effect of resistance training using prediction of intensity by subjective perception of effort (SPE) on body composition, muscle strength, and TCD4+ lymphocyte levels in PLWHA. This a randomized controlled trial study. Participants (11 men and 8 women), were divided in two groups: exercise group (EG) and control group (CG). The EG was submitted to 12 weeks of a resistance-training program based in the prediction of intensity by SPE. Body mass percentages were evaluated using tetrapolar bioelectrical impedance and skinfold methods. We used Flow Cytometry to quantify CD4+ T lymphocytes. Patients showed significant changes in Body Fat Percentage (Δ%=-6.23%), Lean Body Mass (Δ%=2.45%), and CD4T lymphocytes levels (Δ%=15.77%). They also showed significant increase in muscular strength presented in the test for one repetition maximum in all the evaluated exercises. Our data suggest that exercising program prescribed by SPE is capable of improving immune function, body composition, and muscular strength in PLWHA.

Reconstrução de parede torácica em extensas ressecções oncológicas / Reconstruction of the chest wall in extensive oncological resections

Introdução: Apesar do avanço no diagnóstico e no tratamento das pacientes com neoplasia mamária, ainda há casos que se apresentam como doença localmente avançada. Nesse contexto, o controle oncológico requer ressecções extensas e complexos procedimentos reparadores. A participação do cirurgião plástico nas equipes multidisciplinares de tratamento oncológico tem apresentado importância crescente tanto nas etapas de planejamento como de execução. O objetivo é analisar prospectivamente casos de pacientes com extensas ressecções no tórax e reconstrução de parede torácica quanto aos seus resultados e complicações no período de 2014-2016. Métodos: Seguimento prospectivo por 6 meses de 15 casos de pacientes submetidas a extensas ressecções oncológicas no tórax e reconstrução por um mesmo cirurgião plástico em hospitais da rede pública de São Paulo-SP. Foram coletados os dados: idade, diagnóstico, queixa pré-operatória, extensão da área de ressecção e incidência de complicações locais. Resultados: As pacientes tinham idade média de 49,2 anos, a queixa mais comum era massa papável, o diagnóstico mais comum era neoplasia da mama (80%), a maioria das ressecções preservou o plano muscular e a área média de ressecção foi de 259,2 cm2. O retalho miocutâneo do grande dorsal em V-Y foi o mais utilizado, e o toracoabdominal foi o segundo. Epiteliólise foi detectada em 13,3% das pacientes e necrose parcial menor que 5% do retalho em 13,3%. Conclusão: Na presente casuística, as pacientes portadoras de neoplasias extensas no tórax representaram um desafio para a reconstrução local, necessitando de planejamento pré-operatório minucioso e com múltiplas opções. A participação do cirurgião plástico no tratamento desses casos contribuiu para redução de morbidade e demonstrou baixo índice de complicações.

Introduction: In spite of advances in the diagnosis and treatment of patients with breast cancer, there are still cases that present with locally advanced disease. In this context, cancer control requires extensive resections and complex repair procedures. The participation of the plastic surgeon in the multidisciplinary cancer treatment teams has been increasingly important in both the planning and execution stages. The aim of this study is to prospectively analyze patients who underwent extensive chest resections and thoracic wall reconstruction with regard to their results and complications in the period 2014-2016. Methods: Prospective 6-month follow-up of 15 patients who underwent extensive oncological resections in the chest and reconstruction by the same plastic surgeon in public hospitals of São Paulo was done. The following data were collected: age, diagnosis, preoperative complaint, extent of resection area, and incidence of local complications. Results: Patients had a mean age of 49.2 years, the most common complaint was a papillary mass, the most common diagnosis was breast cancer (80%), the most resection area preserved was the muscular plane, and the mean resection area was 259.2 cm2. The V-Y latissimus dorsi myocutaneous flap was the most used, and the thoracoabdominal flap was the second. Epitheliolysis was detected in 13.3% of the patients, and partial necrosis less than 5% of the flap in 13.3%. Conclusion: In the present study, patients with extensive cancers in the thorax presented a challenge for local reconstruction, requiring detailed preoperative planning and multiple options. The participation of the plastic surgeon in the treatment of these patients contributed to the reduction of morbidity rate and low rate of complications.

Physiological adaptations induced by swimming in mice fed a high fat diet.

This study examined physiological variables of animals fed with a high-fat diet (HFD) or with a normal diet (ND) subjected to swimming at low and moderate level. Over 16 weeks, a group of animals was fed with HFD or ND, and at the 8 weeks, they started swimming with 50% or 80% of the maximum load achieved in the progressive work test. Weekly, body weight and the amount of ingested food were registered. The glycemic level was measured at the beginning, middle and at the end of the experiment. Adipose tissue, gastrocnemius muscles and hearts were collected for morphometry. The results showed that the animals fed an HFD had a minor caloric intake; however, the HFD increased body weight and adiposity, likely causing cardiac hypertrophy and an increase in the glycemic level. In this context, swimming with an 80% load contributed positively to weight control, adiposity, glycemic level, to control cardiac hypertrophy and induce hypertrophy in the gastrocnemius muscle. All parameters assessed showed better results for the ND animals. Therefore, the importance of fat consumption was emphasized in relation to obesity onset. The practice of swimming with an 80% load produced greater benefits than swimming with a 50% load for overweight treatment.

Functional and molecular evidence for heteromeric association of P2Y1 receptor with P2Y2 and P2Y4 receptors in mouse granulocytes.

BACKGROUND: All hematopoietic cells express P2 receptors, however pharmacological characteristics such as expression and affinity in granulocytes are unknown. METHODS: Pharmacological characteristics of P2 receptors were evaluated by Ca(2+) measurements using Fura-2 fluorophore. P2 receptors expression were analyzed by flow cytometry and RT-PCR. P2 interaction were shown by coimmunoprecipitation, western blotting and FRET. RESULTS: Granulocytes were responsive to P2Y agonists, whereas P2X agonists were ineffective. Ca(2+) increase, elicited by ADP and UTP was dependent on intracellular stocks and sensitive to G-coupled receptor inhibition. Moreover, MRS2179, a specific antagonist of the P2Y1 receptor, abolished ADP response. Interestingly, ADP and UTP exhibited full heterologous desensitization, suggesting that these agonists interact with the same receptor. The heteromeric association between P2Y1 receptor and the P2Y2 and P2Y4 receptors was shown by immunoprecipitation and FRET analysis. CONCLUSION: Clear evidence of heteromeric association of P2Y receptors was found during the evaluation of P2 receptors present in mice granulocytes, which could impact in the classical pharmacology of P2Y receptors in granulocytes.

Post-exercise cold water immersion does not alter high intensity interval training-induced exercise performance and Hsp72 responses, but enhances mitochondrial markers.

This study aims to evaluate the effect of regular post-exercise cold water immersion (CWI) on intramuscular markers of cellular stress response and signaling molecules related to mitochondria biogenesis and exercise performance after 4 weeks of high intensity interval training (HIIT). Seventeen healthy subjects were allocated into two groups: control (CON, n = 9) or CWI (n = 8). Each HIIT session consisted of 8-12 cycling exercise stimuli (90-110 % of peak power) for 60 s followed by 75 s of active recovery three times per week, for 4 weeks (12 HIIT sessions). After each HIIT session, the CWI had their lower limbs immersed in cold water (10 °C) for 15 min and the CON recovered at room temperature. Exercise performance was evaluated before and after HIIT by a 15-km cycling time trial. Vastus lateralis biopsies were obtained pre and 72 h post training. Samples were analyzed for heat shock protein 72 kDa (Hsp72), adenosine monophosphate-activated protein kinase (AMPK), and phosphorylated p38 mitogen-activated protein kinase (p-p38 MAPK) assessed by western blot. In addition, the mRNA expression of heat shock factor-1 (HSF-1), peroxisome proliferator-activated receptor gamma coactivator-1α (PGC-1α), nuclear respiratory factor 1 and 2 (NRF1 and 2), mitochondrial transcription factor A (Tfam), calcium calmodulin-dependent protein kinase 2 (CaMK2) and enzymes citrate synthase (CS), carnitine palmitoyltransferase I (CPT1), and pyruvate dehydrogenase kinase (PDK4) were assessed by real-time PCR. Time to complete the 15-km cycling time trial was reduced with training (p < 0.001), but was not different between groups (p = 0.33). The Hsp72 (p = 0.01), p38 MAPK, and AMPK (p = 0.04) contents increased with training, but were not different between groups (p > 0.05). No differences were observed with training or condition for mRNA expression of PGC-1α (p = 0.31), CPT1 (p = 0.14), CS (p = 0.44), and NRF-2 (p = 0.82). However, HFS-1 (p = 0.007), PDK4 (p = 0.03), and Tfam (p = 0.03) mRNA were higher in CWI. NRF-1 decrease in both groups after training (p = 0.006). CaMK2 decreased with HIIT (p = 0.003) but it was not affected by CWI (p = 0.99). Cold water immersion does not alter HIIT-induced Hsp72, AMPK, p38 MAPK, and exercise performance but was able to increase some markers of cellular stress response and signaling molecules related to mitochondria biogenesis.

Dual role of P2 receptors during osteoblast differentiation.

UNLABELLED: P2 receptors activated by ATP are expressed in the skeletal system. However, the role of P2 receptors in osteoblast differentiation remains unclear. METHODS: Participation of P2 receptors in differentiation was investigated in the preosteoblast MC3T3-M1 cell line. Preosteoblasts were stimulated for 7 or 14 days in the presence of osteogenic medium containing ATP and its analogs, and then alkaline phosphatase (ALP) activity, gene expression analyses, and protein expression were assessed. RESULTS: We observed that ATP and its analogs promoted increased ALP activity after 7 days of treatment. In contrast, these agonists promoted reductions in ALP activity after 14 days. Some antagonists, such as PPADS (P2 antagonist), MRS2179 (P2Y1 antagonist), MRS2578 (P2Y6 antagonist), and AZ11645373 (P2X7 antagonist) reduced the increases in ALP activity after 7 days. However, only AZ11645373 inhibited the reduction in ALP activity after 14 days. The expression of the P2Y2, P2Y6, P2X4, and P2X7 receptors was observed. Furthermore, treatment with ATP modulated the expression of P2 receptors, increasing P2X4 expression and reducing P2Y6 and P2X7 expression. Similar results were observed after 14 days. In addition, ATP treatment for 7 days increased the expression of transcription factors associated with osteoblast differentiation, such as Runx2, SP7, and Dix5, whereas SP7 and Dix5 expression was reduced at 14 days. These results suggest that P2 receptor activation modulates the differentiation of osteoblasts and is dependent upon the stage of differentiation. These results also suggest that several P2 receptors are involved in this process.

γ-Rays-generated ROS induce apoptosis via mitochondrial and cell cycle alteration in smooth muscle cells.

PURPOSE: γ-rays (IR) cause an increase in intracellular calcium [Ca(2+)], alters contractility and triggers apoptosis via the activation of protein kinase C in intestinal guinea pig smooth muscle cells. The present study investigated the role of the mitochondria in these processes and characterized proteins involved in IR-induced apoptosis. MATERIALS AND METHODS: Intestinal smooth muscle cells were exposed to 10-50 Gy from a (60)Co γ-source. Reactive oxygen species (ROS) levels were measured by colourimetry with a fluorescente probe. Protein expression was analyzed by immunoblotting and immunofluorescence. RESULTS: Apoptosis was inhibited by glutathione, possible by inhibiting the generation or scavenging ROS. Apoptosis was mediated by the mitochondria releasing cytochrome c leading to caspase 3 activation. IR increased the expression of the cyclins A, B2 and E and led to unbalanced cellular growth in an absorption dose-dependent manner. However, radiation did not induce alterations in the mitochondrial ultrastructure or in transmembrane electric potential. In contrast, IR increased the nuclear expression of cytoplasmic proteins and cyclins A and E. CONCLUSION: Smooth muscle cells subjected to IR undergo mitochondrial-mediated apoptosis that involves oncoproteins activation and preserves mitochondrial structure. IR also cause alterations in the expression and localization of both pro- and anti-apoptotic proteins.

Transplantation of bone marrow mesenchymal stem cells decreases oxidative stress, apoptosis, and hippocampal damage in brain of a spontaneous stroke model.

Stroke is the most common cause of motor disabilities and is a major cause of mortality worldwide. Adult stem cells have been shown to be effective against neuronal degeneration through mechanisms that include both the recovery of neurotransmitter activity and a decrease in apoptosis and oxidative stress. We chose the lineage stroke-prone spontaneously hypertensive rat (SHRSP) as a model for stem cell therapy. SHRSP rats can develop such severe hypertension that they generally suffer a stroke at approximately 1 year of age. The aim of this study was to evaluate whether mesenchymal stem cells (MSCs) decrease apoptotic death and oxidative stress in existing SHRSP brain tissue. The results of qRT-PCR assays showed higher levels of the antiapoptotic Bcl-2 gene in the MSC-treated animals, compared with untreated. Our study also showed that superoxide, apoptotic cells, and by-products of lipid peroxidation decreased in MSC-treated SHRSP to levels similar those found in the animal controls, Wistar Kyoto rats. In addition, we saw a repair of morphological damage at the hippocampal region after MSC transplantation. These data suggest that MSCs have neuroprotective and antioxidant potential in stroke-prone spontaneously hypertensive rats.

Hydrogen peroxide (H2O2) induces leukemic but not normal hematopoietic cell death in a dose-dependent manner.

Over the last few years, studies have suggested that oxidative stress plays a role in the regulation of hematopoietic cell homeostasis. In particular, the effects of hydrogen peroxide (H2O2) range from hematopoietic cell proliferation to cell death, depending on its concentration in the intracellular milieu. In this work, we evaluated the effects of an oxidative environment on normal and leukemic hematopoietic cells by stimulating normal human (umbilical cord blood) and murine (bone marrow) hematopoietic cells, as well as human myeloid leukemic cells (HL-60 lineage), upon H2O2 stimulus. Total cell populations and primitive subsets were evaluated for each cell type. H2O2 stimulus induces HL-60 cell death, whereas the viability of human and murine normal cells was not affected. The effects of H2O2 stimulus on hematopoietic stem/progenitor cell subsets were examined and the normal primitive cells were found to be unaffected; however, the percentage of leukemic stem cells (LSC) increased in response to H2O2, while clonogenic ability of these cells to generate myeloid clones was inhibited. In addition, H2O2 stimulus caused a decrease in the levels of p-AKT in HL-60 cells, which most likely mediates the observed decrease of viability. In summary, we found that at low concentrations, H2O2 preferentially affects both the LSC subset and total HL-60 cells without damage normal cells.

Intoxicação por ivermectina em cães / Intoxicatin ivermectin in dogs

As ivermectinas são substâncias obtidas originalmente da fermentação de amostras de solo, contendo o fungo Streptomyces avermitilis (LYNN et al., 1999). É um dos antiparasitários mais utilizados mundialmente, sendo aprovado para uso em bovinos, suínos, ovinos e equinos como endectocida (SINDAN et al., 2001). Rotineiramente, o produto à base de ivermectina de uso bovino é empregado em clínica de pequenos animais, provocando intoxicações (SAKATE et al., 2002). O medicamento vem sendo usado para o tratamento da sarna sarcóptica, demodécica, notoédrica e otodécica de pequenos animais (SAKATE et al., 2002). Segundo a literatura, em caninos, a toxidade está relacionada às altas dosagens e à predisposição racial. Cães das raças Collie, Old English Sheepdog, Pastor de Shetland, Pastor Alemão, Afgan Hounds, ou seus mestiços são particularmente sensíveis, o que torna a barreira hematoencefálica desses cães mais permeável a ivermectina, ocasionando depressão do sistema nervoso central (MEALEY et al., 2006). Os sinais clínicos de intoxicação incluem ataxia, hipertermia, desorientação, sialorréia, midríase, hiperestesia, tremores, depressão, paralisia, ausência dos reflexos pupilares, cegueira, bradicardia, pulso fraco e em casos graves, coma, hipotermia e morte (SAKATE et al., 2002). Com esse trabalho, objetivou-se relatar um caso de intoxicação por ivermectina em dois cães sensíveis ao princípio ativo, desencadeando um quadro tóxico.