Rotenone inhibits embryonic chick myogenesis in a ROS-dependent mechanism.

Skeletal muscle function is highly dependent on the energy supply provided by mitochondria. Besides ATP production, mitochondria have several other roles, such as calcium storage, heat production, cell death signaling, autophagy regulation and redox state modulation. Mitochondrial function is crucial for skeletal muscle fiber formation. Disorders that affect mitochondria have a major impact in muscle development and function. Here we studied the role of mitochondria during chick skeletal myogenesis. We analyzed the intracellular distribution of mitochondria in myoblasts, fibroblasts and myotubes using Mitotracker labeling. Mitochondrial respiration was investigated in chick muscle cells. Our results show that (i) myoblasts and myotubes have more mitochondria than muscle fibroblasts; (ii) mitochondria are organized in long lines within the whole cytoplasm and around the nuclei of myotubes, while in myoblasts they are dispersed in the cytoplasm; (iii) the area of mitochondria in myotubes increases during myogenesis, while in myoblasts and fibroblasts there is a slight decrease; (iv) mitochondrial length increases in the three cell types (myoblasts, fibroblasts and myotubes) during myogenesis; (v) the distance of mitochondria to the nucleus increases in myoblasts and myotubes during myogenesis; (vi) Rotenone inhibits muscle fiber formation, while FCCP increases the size of myotubes; (vii) N-acetyl cysteine (NAC), an inhibitor of ROS formation, rescues the effects of Rotenone on muscle fiber size; and (viii) Rotenone induces the production of ROS in chick myogenic cells. The collection of our results suggests a role of ROS signaling in mitochondrial function during chick myogenesis.

Localization and function of humanized F508del-CFTR in mouse intestine following activation of serum glucocorticoid kinase 1 and Trikafta.

The CFTR modulator Trikafta has markedly improved lung disease for Cystic Fibrosis (CF) patients carrying the common delta F508 (F508del-CFTR) CFTR mutation. F508del-CFTR results in an apical trafficking defect and loss of function in CFTR-expressing epithelial cells. However, Trikafta has not resulted in improved gastrointestinal function in CF patients. A humanized mouse model of F508del-CFTR was recently generated to evaluate CFTR modulators and other compounds to treat human F508del-CFTR CF intestinal disease. Short-term (4 h) treatment of rats with Dexamethasone (Dex) potently activates serum glucocorticoid kinase 1 (SGK1) and increases CFTR apical traffic and ion transport in the native intestine. This study examined CFTR localization and ion transport in intestinal segments from humanized F508del-CFTR mice following treatment with Dex in the presence/absence of Trikafta. Dex treatment improved apical CFTR localization and function but was inconsistent along intestinal segments. Combined treatment with Dex and Trikafta was superior to Dex alone but inconsistently improved CFTR localization and function. These data suggest further optimization of humanized CF mouse models will be necessary to test the efficacy of compounds to treat human CF intestinal disease.

Ozone therapy improves early stages of osseointegration in ovariectomized rats.

OBJECTIVE: the aim of this study was to analyze the influence of ozone therapy (OZN) on peri-implant bone repair in critical bones by installing osseointegrated implants in the tibia of ovariectomized rats. METHODOLOGY: ovariectomy was performed on 30 Wistar rats, aged six months (Rattus novergicus), and, after 90 days, osseointegrated implants were installed in each tibial metaphysis. The study groups were divided into the animals that received intraperitoneal ozone at a concentration of 700 mcg/kg - OZ Group (n=15) - and a control group that received an intraperitoneal saline solution and, for this reason, was named the SAL group (n=15). The applications for both groups occurred during the immediate post-operative period on the 2nd, 4th, 6th, 8th, 10th, and 12th day post-surgery. At various stages (14, 42, and 60 days), the animals were euthanized, and tests were performed on their tibiae. These tests include histomorphometric and immunohistochemical analyses, computerized microtomography, sampling in light-cured resin for calcified sections, and confocal microscopy. The obtained data were then analyzed using One-way ANOVA and the Shapiro-Wilk, Kruskal-Wallis, and student t-tests (P<0.05). RESULTS: our findings indicate that the OZ group (3.26±0.20 mm) showed better cellular organization and bone neoformation at 14 days (SAL group, 0.90±1.42 mm) (P=0.001). Immunohistochemistry revealed that osteocalcin labeling was moderate in the OZ group and mild in the SAL group at 14 and 42 days post-surgery. The data from the analysis of calcified tissues (microtomography, histometric, and bone dynamism analysis) at 60 days showed no statistically significant differences between the groups (P=0.32). CONCLUSION: it was concluded that ozone therapy anticipated the initial phases of the peri-implant bone repair process.

Antimanic-like effect of dipyridamole in the methylphenidate-induced hyperlocomotion.

BACKGROUND: Adenosinergic system has been implicated in the pathophysiology of bipolar disorder and drugs that affect adenosine neurotransmission have shown some efficacy as add-on therapy in manic patients. OBJECTIVE: Thus, the aim of the present study was to screen adenosinergic drugs for antimanic-like effect in methylphenidate (MPH)-induced hyperlocomotion in mice. METHODS: Male and female Swiss mice received a single allopurinol (50 and 200 mg/kg, ip), dipyridamole (20 mg/kg, ip), or inosine (50 mg/kg, ip) administration before an acute MPH challenge (5 mg/kg, sc). In experiments with repeated treatment, male mice received a daily administration of allopurinol (25 and 50 mg/kg, ip), dipyridamole (20 mg/kg, ip), or inosine (50 mg/kg, ip) for 14 days. Finally, pretreatment with aminophylline (2 mg/kg, sc), an unspecific adenosine receptor antagonist, was used to evaluate a putative adenosinergic mediation. Locomotor activity was measured in the automated activity chamber for 20 min. RESULTS: Acute and repeated dipyridamole reduced the increase in locomotor activity induced by MPH, while allopurinol and inosine had no effect. Aminophylline blocked the effect of dipyridamole in MPH-induced hyperlocomotion. CONCLUSION: The present results suggest that dipyridamole may have an antimanic-like effect through adenosine receptors and reinforce the proposal that the adenosine system may be an interesting target for new antimanic drugs.

A new hydroxyapatite-alginate-gelatin biocomposite favor bone regeneration in a critical-sized calvarial defect model

Abstract This study aimed to evaluate the osteogenic potential of hydroxyapatite (HA), Alginate (Alg), and Gelatine (Gel) composite in a critical-size defect model in rats. Twenty-four male rats were divided into three groups: a negative control with no treatment (Control group), a positive control treated with deproteinized bovine bone mineral (DBBM group), and the experimental group treated with the new HA-Alg-Gel composite (HA-Alg-Gel group). A critical size defect (8.5mm) was made in the rat's calvaria, and the bone formation was evaluated by in vivo microcomputed tomography analysis (µCT) after 1, 15, 45, and 90 days. After 90 days, the animals were euthanized and histological and histomorphometric analyses were performed. A higher proportion of mineralized tissue/biomaterial was observed in the DBBM group when compared to the HA-Alg-Gel and Control groups in the µCT analysis during all analysis periods. However, no differences were observed in the mineralized tissue/biomaterial proportion observed on day 1 (immediate postoperative) in comparison to later periods of analysis in all groups. In the histomorphometric analysis, the HA-Alg-Gel and Control groups showed higher bone formation than the DBBM group. Moreover, in histological analysis, five samples of the HA-Alg-Gal group exhibited formed bone spicules adjacent to the graft granules against only two of eight samples in the DBBM group. Both graft materials ensured the maintenance of defect bone thickness, while a tissue thickness reduction was observed in the control group. In conclusion, this study demonstrated the osteoconductive potential of HA-Alg-Gel bone graft by supporting new bone formation around its particles.

Resumo Este estudo teve como objetivo avaliar o potencial osteogênico de um compósito de hidroxiapatita (HA), alginato (Alg) e gelatina (Gel) em um modelo de defeito de tamanho crítico em ratos. Vinte e quatro ratos machos foram divididos em três grupos: um controle negativo sem tratamento (grupo controle), um controle positivo tratado com osso bovino desproteinizado (grupo DBBM) e o grupo experimental tratado com o novo compósito HA-Alg-Gel (grupo HA-Alg-Gel). Um defeito de tamanho crítico (8,5mm) foi feito na calvária dos ratos, e a formação óssea foi avaliada por análise de microtomografia computadorizada in vivo (µCT) após 1, 15, 45 e 90 dias. Após 90 dias, os animais foram eutanasiados e análises histológicas e histomorfométricas foram realizadas. Uma maior proporção de tecido mineralizado/biomaterial foi observada no grupo DBBM quando comparado aos grupos HA-Alg-Gel e controle na análise de µCT durante todos os períodos de análise. Entretanto, não foram observadas diferenças na proporção tecido mineralizado/biomaterial no dia 1 (pós-operatório imediato) em relação aos períodos posteriores de análise em todos os grupos. Na análise histomorfométrica, os grupos HA-Alg-Gel e controle apresentaram maior formação óssea do que o grupo DBBM. Além disso, na análise histológica, cinco amostras do grupo HA-Alg-Gal exibiram espículas ósseas formadas adjacentes aos grânulos do enxerto contra apenas duas das oito amostras do grupo DBBM. Ambos os materiais de enxerto garantiram a manutenção da espessura óssea do defeito, enquanto uma redução da espessura do tecido foi observada no grupo controle. Em conclusão, este estudo demonstrou o potencial osteocondutor do enxerto ósseo de HA-Alg-Gel, promovendo a formação de osso novo ao redor das suas partículas.

Conhecimento dos profissionais intra-hospitalares acerca do suporte básico de vida em uma parada cardiorrespiratória / In-hospital professionals' knowledge about basic life support in cardiac arrest / Conocimiento de los profesionales intrahospitalarios sobre el soporte vital básico en la parada cardíaca

Objetivo: avaliar o efeito da intervenção educativa no conhecimento da equipe de enfermagem sobre o suporte básico de vida para o atendimento à parada cardiorrespiratória de adultos no ambiente intra-hospitalar. Método: estudo transversal com abordagem quantitativa, realizado com 25 profissionais de enfermagem em dois hospitais de região oeste de Santa Catarina - Brasil. Avaliou-se por meio da aplicação de um pré-teste, intervenção educativa e pós-teste. Resultados: houve aumento significativo no conhecimento dos profissionais. O hospital A obteve a média de acertos de 7,23 no pré-teste, elevando para 11,33 no pós-teste, com valor de p ≤ 0,0001. Já o hospital B pontuou 6,07 no pré-teste, progredindo para 11,15 no pós-teste, valor de p ≤ 0,0006. Conclusão: a intervenção realizada demonstrou ser uma estratégia eficaz, visto que os resultados pré-teste demostravam déficit significativo de conhecimento, e após a intervenção educativa, mostraram melhoria na maioria dos itens avaliados em relação ao atendimento específico.

Objective: to evaluate the effect of an educational intervention on the nursing team's knowledge about basic life support for adult cardiac arrest care in the in-hospital environment. Method: cross-sectional study with a quantitative approach, carried out with 25 nursing professionals in two hospitals in the western region of Santa Catarina - Brazil. A pre-test, educational intervention and post-test were applied. Results: there was a significant increase in the professionals' knowledge. Hospital A had a mean score of 7.23 in the pre-test, increasing to 11.33 in the post-test, with p-value ≤ 0.0001. Hospital B scored 6.07 in the pre-test, increasing to 11.15 in the post-test, p-value ≤ 0.0006. Conclusion: the intervention proved to be an effective strategy, since the pre-test results showed significant knowledge deficit, and after the educational intervention, showed improvement in most of the items evaluated in relation to specific care.

Objetivos:evaluar el efecto de una intervención educativa en el conocimiento del equipo de enfermería sobre el soporte vital básico para la atención del paro cardíaco del adulto en el ambiente intrahospitalario. Método: estudio transversal con abordaje cuantitativo, realizado con 25 profesionales de enfermería en dos hospitales de la región oeste de Santa Catarina - Brasil. Se aplicó un pre-test, una intervención educativa y un post-test. Resultados: hubo un aumento significativo de los conocimientos de los profesionales. El Hospital A obtuvo una puntuación media de 7,23 en el pre-test, aumentando a 11,33 en el post-test, con valor p ≤ 0,0001. El Hospital B obtuvo una puntuación de 6,07 en el pre-test, aumentando a 11,15 en el post-test, con valor p ≤ 0,0006. Conclusión: una intervención realizada demostró ser una estrategia eficaz, visto que os resultados previos demostraron un déficit significativo de conhecimento, y después de una intervención educativa, mostraron una mejoría na maioria dos itens avaliados em relação ao atendimento específico.

Ozone therapy improves early stages of osseointegration in ovariectomized rats

Abstract Objective the aim of this study was to analyze the influence of ozone therapy (OZN) on peri-implant bone repair in critical bones by installing osseointegrated implants in the tibia of ovariectomized rats. Methodology ovariectomy was performed on 30 Wistar rats, aged six months (Rattus novergicus), and, after 90 days, osseointegrated implants were installed in each tibial metaphysis. The study groups were divided into the animals that received intraperitoneal ozone at a concentration of 700 mcg/kg — OZ Group (n=15) — and a control group that received an intraperitoneal saline solution and, for this reason, was named the SAL group (n=15). The applications for both groups occurred during the immediate post-operative period on the 2nd, 4th, 6th, 8th, 10th, and 12th day post-surgery. At various stages (14, 42, and 60 days), the animals were euthanized, and tests were performed on their tibiae. These tests include histomorphometric and immunohistochemical analyses, computerized microtomography, sampling in light-cured resin for calcified sections, and confocal microscopy. The obtained data were then analyzed using One-way ANOVA and the Shapiro-Wilk, Kruskal-Wallis, and student t-tests (P<0.05). Results our findings indicate that the OZ group (3.26±0.20 mm) showed better cellular organization and bone neoformation at 14 days (SAL group, 0.90±1.42 mm) (P=0.001). Immunohistochemistry revealed that osteocalcin labeling was moderate in the OZ group and mild in the SAL group at 14 and 42 days post-surgery. The data from the analysis of calcified tissues (microtomography, histometric, and bone dynamism analysis) at 60 days showed no statistically significant differences between the groups (P=0.32). Conclusion it was concluded that ozone therapy anticipated the initial phases of the peri-implant bone repair process.

Delivery of Plasmid DNA by Ionizable Lipid Nanoparticles to Induce CAR Expression in T Cells.

Introduction: Chimeric antigen receptor (CAR) cell therapy represents a hallmark in cancer immunotherapy, with significant clinical results in the treatment of hematological tumors. However, current approved methods to engineer T cells to express CAR use viral vectors, which are integrative and have been associated with severe adverse effects due to constitutive expression of CAR. In this context, non-viral vectors such as ionizable lipid nanoparticles (LNPs) arise as an alternative to engineer CAR T cells with transient expression of CAR. Methods: Here, we formulated a mini-library of LNPs to deliver pDNA to T cells by varying the molar ratios of excipient lipids in each formulation. LNPs were characterized and screened in vitro using a T cell line (Jurkat). The optimized formulation was used ex vivo to engineer T cells derived from human peripheral blood mononuclear cells (PBMCs) for the expression of an anti-CD19 CAR (CAR-CD19BBz). The effectiveness of these CAR T cells was assessed in vitro against Raji (CD19+) cells. Results: LNPs formulated with different molar ratios of excipient lipids efficiently delivered pDNA to Jurkat cells with low cytotoxicity compared to conventional transfection methods, such as electroporation and lipofectamine. We show that CAR-CD19BBz expression in T cells was transient after transfection with LNPs. Jurkat cells transfected with our top-performing LNPs underwent activation when exposed to CD19+ target cells. Using our top-performing LNP-9-CAR, we were able to engineer human primary T cells to express CAR-CD19BBz, which elicited significant specific killing of CD19+ target cells in vitro. Conclusion: Collectively, our results show that LNP-mediated delivery of pDNA is a suitable method to engineer human T cells to express CAR, which holds promise for improving the production methods and broader application of this therapy in the future.

SOFT TISSUE SARCOMA - SANTA CASA DE SÃO PAULO EXPERIENCE FROM 2006 TO 2019.

Objective: To conduct an epidemiologic review, analyzing treatment, evolution, and survival of soft tissue sarcomas. Methods: Retrospective study based on medical records of patient with STS treated by the Orthopedic Oncology Group at the Santa Casa de São Paulo, from 2006 to 2019. Data from 121 patients were analyzed according to age, sex, histological type, tumor location, treatment, previous surgery in a non-specialized service, local recurrences, lung metastases, and survival analysis. Results: The most frequent location was the thigh. Patients who underwent surgery with a non-specialized group had higher rates of local recurrence and those with pulmonary metastasis had a lower survival rate. Conclusion: STS can occur at any age and the prevalence of the histological type depends on the patients' age group. Level of Evidence II, Prognostic Study.

Objetivo: Conduzir uma avaliação epidemiológica analisando tratamento, evolução e sobrevida dos sarcomas de partes moles (SPMs). Métodos: Estudo retrospectivo de prontuários de pacientes com SPM tratados pelo Grupo de Oncologia Ortopédica da Santa Casa de Misericórdia de São Paulo, no período de 2006 a 2019. Foram analisados os dados de 121 pacientes referentes a idade, sexo, tipo histológico, localização do tumor, tratamento, cirurgia prévia em serviço não especializado, presença de recidivas, metástases pulmonares e análise de sobrevida. Resultados: A localização mais frequente foi a coxa. Verificou-se que pacientes que realizaram cirurgia com grupo não especializado tiveram maiores índices de recidiva local, e aqueles com metástase pulmonar tiveram menor sobrevida. Conclusão: Os SPMs podem ocorrer em qualquer idade, e a prevalência do tipo histológico depende da faixa etária dos pacientes. Nível de Evidência II, Estudo Prognóstico.

Comparison of Manual (2D) and Digital (3D) Methods in the Assessment of Simulated Facial Edema.

BACKGROUND: An edema assessment following dental surgeries is essential to improving the dental surgeon's technique and, consequentially, patient comfort. PURPOSE: Two-dimensional (2D) methods are limited in analyzing 3-dimensional (3D) surfaces. Currently, 3D methods are used to investigate postoperative swelling. However, there are no studies that directly compare 2D and 3D methods. The goal of this study is to directly compare 2D and 3D methods used in the assessment of postoperative edema. STUDY DESIGN AND SAMPLE: The investigators implemented a prospective, cross-sectional study with each subject serving as its own control. The sample was composed of dental student volunteers without facial deformities. PREDICTOR VARIABLE: The predictor variable is the method used to measure edema. After simulating edema, manual (2D) and digital (3D) techniques were used to measure edema. The manual method used direct facial perimeter measurements. The two digital methods were photogrammetry using a smartphone (iPhone 11, Apple Inc, Cupertino, California), and facial scanning with a smartphone application (Bellus3D FaceApp, Bellus3D Inc, Campbell, California) [3D measurements] MAIN OUTCOME VARIABLE: The coefficient of variation (CV) (CV = standard deviation /mean) was used to assess homogeneity of edema measurements. ANALYSIS: The Shapiro-Wilk and equal variance tests were applied to assess data homogeneity. Next, one-way analysis of variance was performed, followed by a correlation analysis. Finally, the data were submitted to Tukey's test. The statistical significance threshold was set at 5% (P < .05). RESULTS: The sample was composed of 20 subjects aged 18-38 years. The CV showed higher values using the manual (2D) method (47%; 4.88% ± 2.99), compared with the photogrammetry method (18%; 8.55 mm ± 1.52) and the smartphone application (21%; 8.97 mm ± 1.93). A statistically significant difference was observed between the manual method values and those of the other two groups (P < .001). There was no difference between the facial scanning and photogrammetry groups (3D methods) (P = .778) CONCLUSION AND RELEVANCE: Both digital measuring methods (3D) demonstrated greater homogeneity than the manual method in analyzing facial distortions caused by the same swelling simulation. Therefore, it can be affirmed that digital methods may be more reliable that manual methods for assessing facial edema.

How is smoking distributed in relation to socioeconomic status? Evidence from Brazil in the years 2013 and 2019.

The present study aimed to analyze income-related inequality in tobacco consumption in Brazil using data from the National Health Survey at two points in time (2013 and 2019). This study contributes to the growing literature analyzing socioeconomic inequalities in tobacco use by investigating income-related inequalities in the consumption of different tobacco products in Brazil. The inequality measure is the concentration index with an Erreygers correction (EI), and the analysis of its decomposition allows the identification of the factors that determine such inequality. There is inequality in smoking concentrated in the poorest persons, and this pattern also occurs for manufactured cigarettes and roll-your-own cigarettes (RYO), while inequality in smoking cessation is concentrated among the wealthiest. Smoking inequalities were greater in men, older age groups, and RYO. In terms of evolution, the overall results indicated a small decline in smoking inequality. For the decomposition analysis, the results show that the main factors that affect tobacco inequality in terms of concentration in the poorest are education, income, and having private health insurance. The region variable, by contrast, has a positive contribution, since the wealthiest regions have individuals who are more likely to smoke. These results have important implications that serve as a basis for formulating public health policies. For example, greater inequalities for men and older individuals can be targeted by public policies with a special focus on these cases.

Single intraoperative infrared laser optimized bone repair in rat femoral osteotomies with experimentally induced osteoporosis.

This study aimed to evaluate the effect of infrared laser (IRL) on bone repair in ovariectomized rats subjected to femoral osteotomies. Of 32 rats, half underwent bilateral ovariectomy (OVX) and the other half underwent sham ovariectomy (SHAM). A period of 3 months was defined to observe the presence of osteoporosis. The rats were subjected to osteotomies in the femurs and then fixed with a miniplate and 1.5-mm system screws. Thereafter, half of the rats from both SHAM and OVX groups were not irradiated, and the other half were irradiated by IRL using the following parameters: wavelength, 808 nm; power, 100 mW; 60 s for each point; 6 J/point; and a total of 5 points of bone gap. All animals were euthanized 60 days after surgery. The femur gap was scanned using micro-computed tomography (micro-CT). The samples were then examined under a confocal laser microscope to determine the amounts of calcein and alizarin red. The slides were stained with alizarin red and Stevenel's blue for histometric analysis. In the micro-CT analysis, the OVX groups had the lowest bone volume (P < 0.05). When the laser was applied to the OVX groups, bone turnover increased (P < 0.05). New bone formation (NBF) was comparable between SHAM and OVX/IR (P > 0.05) groups; however, it was less in the OVX groups (P < 0.05). In conclusion, the results encourage the use of IRL intraoperatively as it optimizes bone repair, mainly in animals with low bone mineral density.

CFTR High Expresser Cells in cystic fibrosis and intestinal diseases.

Cystic Fibrosis Transmembrane Conductance Regulator (CFTR), the Cl-/HCO3 - channel implicated in Cystic Fibrosis, is critical to the pathophysiology of many gastrointestinal diseases. Defects in CFTR lead to intestinal dysfunction, malabsorption, obstruction, infection, inflammation, and cancer that increases morbidity and reduces quality of life. This review will focus on CFTR in the intestine and the implications of the subpopulation of CFTR High Expresser Cells (CHEs) in Cystic Fibrosis (CF), intestinal physiology and pathophysiology of intestinal diseases.

Soft tissue sarcoma - santa casa de são paulo experience from 2006 to 2019 / Sarcoma de partes moles - experiência da santa casa de são paulo de 2006 a 2019

ABSTRACT Objective: To conduct an epidemiologic review, analyzing treatment, evolution, and survival of soft tissue sarcomas. Methods: Retrospective study based on medical records of patient with STS treated by the Orthopedic Oncology Group at the Santa Casa de São Paulo, from 2006 to 2019. Data from 121 patients were analyzed according to age, sex, histological type, tumor location, treatment, previous surgery in a non-specialized service, local recurrences, lung metastases, and survival analysis. Results: The most frequent location was the thigh. Patients who underwent surgery with a non-specialized group had higher rates of local recurrence and those with pulmonary metastasis had a lower survival rate. Conclusion: STS can occur at any age and the prevalence of the histological type depends on the patients' age group. Level of Evidence II, Prognostic Study.

RESUMO Objetivo: Conduzir uma avaliação epidemiológica analisando tratamento, evolução e sobrevida dos sarcomas de partes moles (SPMs). Métodos: Estudo retrospectivo de prontuários de pacientes com SPM tratados pelo Grupo de Oncologia Ortopédica da Santa Casa de Misericórdia de São Paulo, no período de 2006 a 2019. Foram analisados os dados de 121 pacientes referentes a idade, sexo, tipo histológico, localização do tumor, tratamento, cirurgia prévia em serviço não especializado, presença de recidivas, metástases pulmonares e análise de sobrevida. Resultados: A localização mais frequente foi a coxa. Verificou-se que pacientes que realizaram cirurgia com grupo não especializado tiveram maiores índices de recidiva local, e aqueles com metástase pulmonar tiveram menor sobrevida. Conclusão: Os SPMs podem ocorrer em qualquer idade, e a prevalência do tipo histológico depende da faixa etária dos pacientes. Nível de Evidência II, Estudo Prognóstico.

Voltammetric Determination of 3-Methylmorphine Using Glassy Carbon Electrode Modified with rGO and Bismuth Film.

This work reports the development and application of a simple, rapid and low-cost voltammetric method for the determination of 3-methylmorphine at nanomolar levels in clinical and environmental samples. The proposed method involves the combined application of a glassy carbon electrode modified with reduced graphene oxide, chitosan and bismuth film (Bi-rGO-CTS/GCE) via square-wave voltammetry using 0.04 mol L-1 Britton-Robinson buffer solution (pH 4.0). The application of the technique yielded low limit of detection of 24 × 10-9 mol L-1 and linear concentration range of 2.5 × 10-7 to 8.2 × 10-6 mol L-1. The Bi-rGO-CTS/GCE sensor was successfully applied for the detection of 3-methylmorphine in the presence of other compounds, including paracetamol and caffeine. The results obtained also showed that the application of the sensor for 3-methylmorphine detection did not experience any significant interference in the presence of silicon dioxide, povidone, cellulose, magnesium stearate, urea, ascorbic acid, humic acid and croscarmellose. The applicability of the Bi-rGO-CTS/GCE sensor for the detection of 3-methylmorphine was evaluated using synthetic urine, serum, and river water samples through addition and recovery tests, and the results obtained were found to be similar to those obtained for the high-performance liquid chromatography method (HPLC)-used as a reference method. The findings of this study show that the proposed voltammetric method is a simple, fast and highly efficient alternative technique for the detection of 3-methylmorphine in both biological and environmental samples.

Trypanosoma cruzi iron superoxide dismutases: insights from phylogenetics to chemotherapeutic target assessment.

BACKGROUND: Components of the antioxidant defense system in Trypanosoma cruzi are potential targets for new drug development. Superoxide dismutases (SODs) constitute key components of antioxidant defense systems, removing excess superoxide anions by converting them into oxygen and hydrogen peroxide. The main goal of the present study was to investigate the genes coding for iron superoxide dismutase (FeSOD) in T. cruzi strains from an evolutionary perspective. METHODS: In this study, molecular biology methods and phylogenetic studies were combined with drug assays. The FeSOD-A and FeSOD-B genes of 35 T. cruzi strains, belonging to six discrete typing units (Tcl-TcVI), from different hosts and geographical regions were amplified by PCR and sequenced using the Sanger method. Evolutionary trees were reconstructed based on Bayesian inference and maximum likelihood methods. Drugs that potentially interacted with T. cruzi FeSODs were identified and tested against the parasites. RESULTS: Our results suggest that T. cruzi FeSOD types are members of distinct families. Gene copies of FeSOD-A (n = 2), FeSOD-B (n = 4) and FeSOD-C (n = 4) were identified in the genome of the T. cruzi reference clone CL Brener. Phylogenetic inference supported the presence of two functional variants of each FeSOD type across the T. cruzi strains. Phylogenetic trees revealed a monophyletic group of FeSOD genes of T. cruzi TcIV strains in both distinct genes. Altogether, our results support the hypothesis that gene duplication followed by divergence shaped the evolution of T. cruzi FeSODs. Two drugs, mangafodipir and polaprezinc, that potentially interact with T. cruzi FeSODs were identified and tested in vitro against amastigotes and trypomastigotes: mangafodipir had a low trypanocidal effect and polaprezinc was inactive. CONCLUSIONS: Our study contributes to a better understanding of the molecular biodiversity of T. cruzi FeSODs. Herein we provide a successful approach to the study of gene/protein families as potential drug targets.

BioAutoML: automated feature engineering and metalearning to predict noncoding RNAs in bacteria.

Recent technological advances have led to an exponential expansion of biological sequence data and extraction of meaningful information through Machine Learning (ML) algorithms. This knowledge has improved the understanding of mechanisms related to several fatal diseases, e.g. Cancer and coronavirus disease 2019, helping to develop innovative solutions, such as CRISPR-based gene editing, coronavirus vaccine and precision medicine. These advances benefit our society and economy, directly impacting people's lives in various areas, such as health care, drug discovery, forensic analysis and food processing. Nevertheless, ML-based approaches to biological data require representative, quantitative and informative features. Many ML algorithms can handle only numerical data, and therefore sequences need to be translated into a numerical feature vector. This process, known as feature extraction, is a fundamental step for developing high-quality ML-based models in bioinformatics, by allowing the feature engineering stage, with design and selection of suitable features. Feature engineering, ML algorithm selection and hyperparameter tuning are often manual and time-consuming processes, requiring extensive domain knowledge. To deal with this problem, we present a new package: BioAutoML. BioAutoML automatically runs an end-to-end ML pipeline, extracting numerical and informative features from biological sequence databases, using the MathFeature package, and automating the feature selection, ML algorithm(s) recommendation and tuning of the selected algorithm(s) hyperparameters, using Automated ML (AutoML). BioAutoML has two components, divided into four modules: (1) automated feature engineering (feature extraction and selection modules) and (2) Metalearning (algorithm recommendation and hyper-parameter tuning modules). We experimentally evaluate BioAutoML in two different scenarios: (i) prediction of the three main classes of noncoding RNAs (ncRNAs) and (ii) prediction of the eight categories of ncRNAs in bacteria, including housekeeping and regulatory types. To assess BioAutoML predictive performance, it is experimentally compared with two other AutoML tools (RECIPE and TPOT). According to the experimental results, BioAutoML can accelerate new studies, reducing the cost of feature engineering processing and either keeping or improving predictive performance. BioAutoML is freely available at https://github.com/Bonidia/BioAutoML.

Influência de indicadores sociais na incidência e mortalidade da COVID-19 no Brasil em junho de 2020 / Influence of social indicators on the occurrence and mortality of COVID-19 in Brazil in june 2020

O estudo tem por objetivo analisar a influência de indicadores sociais das regiões e verificar sua correlação com a incidência e mortalidade causada pela COVID-19 no Brasil no período de junho de 2020. Trata-se de um estudo observacional, ecológico e analítico na qual foram incluídas nas análises todas Unidades Federativas brasileiras de 10/06/2020 à 13/06/2020. Considerou-se como variáveis relacionadas à COVID-19: taxa de incidência e mortalidade acumulada por 100 mil habitantes e taxa de letalidade acumulada. Como variáveis independentes, determinou-se: índice de desenvolvimento humano, percentual de pobreza e renda per capita. Verificou-se que regiões mais pobres (Norte e Nordeste) possuem maior incidência, mortalidade e letalidade por COVID-19 se comparado com as regiões mais ricas do (Sudeste, Sul e Centro-oeste). Esses resultados demonstram que os indicadores sociais possuem influência nas taxas de incidência e óbitos por 100 mil habitantes devido a COVID-19, além de apresentar correlações significativas entre si. Os achados do estudo podem ser usados para embasar e dar direcionamento a ações de combate ao COVID-19 no país, levando à criação de políticas públicas bem estruturadas e garantindo eficácia para o enfrentamento da doença em áreas com vulnerabilidade.

Current paper analyzes the influence of social indicators in Brazilian regions and verifies their correlation with the occurrence and mortality caused by COVID-19 in Brazil. All the Brazilian Federative States were included in current observational, ecological and analytical study from 06/10/2020 to 06/13/2020. Variables were related to COVID-19´s occurrence rate, lethality and accumulated mortality per 100,000 inhabitants. Independent variables comprised human development index, poverty percentage and per capita income. Results show that poorer regions have higher incidence, mortality and lethality by COVID-19 when compared to richer ones. These results demonstrate that social indicators influence the incidence and death rates per 100,000 inhabitants due to COVID-19, in addition to showing significant correlations with each other. The study findings may be used to support and guide actions to combat COVID-19 in the country.

Diminazene Aceturate, an angiotensin converting enzyme 2 (ACE2) activator, promotes cardioprotection in ischemia/reperfusion-induced cardiac injury.

This study aimed to investigate whether the Diminazene Aceturate (DIZE), an angiotensin-converting enzyme 2 (ACE2) activator, can revert cardiac dysfunction in ischemia reperfusion-induced (I/R) injury in animals and examine the mechanism underlying this effect. Wistar rats systemically received DIZE (1 mg/kg) for thirty days. Cardiac function in isolated rat hearts was evaluated using the Langendorff technique. After I/R, ventricular non-I/R and I/R samples were used to evaluate ATP levels. Mitochondrial function was assessed using cardiac permeabilized fibers and isolated cardiac mitochondria. Cardiac cellular electrophysiology was evaluated using the patch clamp technique. DIZE protected the heart after I/R from arrhythmia and cardiac dysfunction by preserving ATP levels, independently of any change in coronary flow and heart rate. DIZE improved mitochondrial function, increasing the capacity for generating ATP and reducing proton leak without changing the specific citrate synthase activity. The activation of the ACE2 remodeled cardiac electrical profiles, shortening the cardiac action potential duration at 90 % repolarization. Additionally, cardiomyocytes from DIZE-treated animals exhibited reduced sensibility to diazoxide (KATP agonist) and a higher KATP current compared to the controls. DIZE was able to improve mitochondrial function and modulate cardiac electrical variables with a cardio-protective profile, resulting in direct myocardial cell protection from I/R injury.