Immunolocalization of melatonin receptors in bovine ovarian follicles and in vitro effects of melatonin on growth, viability and gene expression in secondary follicles.

This study aims to investigate the (1) expression of melatonin receptors types 1A/B (MTNR1A/B) in bovine ovaries and (2) the in vitro effects of melatonin on secondary follicle development, antrum formation, viability, and expression of messenger ribonucleic acid (mRNA) for superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase-1 (GPX1) and peroxiredoxin 6 (PRDX6). The expression of MTNR1A/B in bovine ovarian follicles was demonstrated by immunohistochemistry. To choose the most effective concentration of melatonin on follicular growth and viability, isolated secondary follicles were cultured individually at 38.5°C, with 5% CO2 in air, for 18 d in TCM-199+ alone or supplemented with 10-11, 10-9, 10-7 or 10-5 M melatonin. Then, melatonin receptor antagonist, luzindole, was tested to further evaluate the mechanisms of actions of melatonin, that is, the follicles were cultured in control medium alone or supplemented with 10-7 M melatonin, 10 µM luzindole and both 10-7 M melatonin and 10 µM luzindole. Follicular growth, morphology and antrum formation were evaluated at days 6, 12 and 18. At the end of culture, viability of secondary follicles was analyzed by calcein-AM and ethidium homodimer-1, and the relative levels of mRNA for SOD, CAT, GPX1 and PRDX6 were evaluated by real time polymerase chain reaction. Immunohistochemistry results showed expression of MTNR1A/B in oocyte and granulosa cells of primordial, primary, secondary and antral follicles. Secondary follicles cultured in medium supplemented with melatonin at different concentrations had well preserved follicles after 18 d of culture. Furthermore, follicles cultured in presence of 10-7 M melatonin presented significantly higher diameters than those cultured in other treatments. The presence of melatonin receptor antagonist, luzindole, blocked the effects of melatonin on follicular growth and viability. In addition, follicles cultured in medium containing only melatonin had significantly higher rates of antrum formation. Follicles cultured in medium containing only melatonin had higher relative levels of mRNA for CAT, SOD and PRDX-6 than those cultured with both melatonin and luzindole. Follicles cultured with luzindole only or both melatonin and luzindole had lower relative levels of mRNA for PRDX6 and GPX1 than those cultured control medium. In conclusion, melatonin promotes growth of bovine secondary follicles through its membrane-coupled receptors, while luzindole blocks the effects of melatonin on follicle growth and reduces the expression of antioxidant enzymes in cultured follicles.

Nanohydroxyapatite-Blasted Bioactive Surface Drives Shear-Stressed Endothelial Cell Growth and Angiogenesis.

Nanosized crystalline hydroxyapatite coating (HAnano®) accelerates the osteointegration of dental implants which is hypothesized to drive angiogenesis. In order to test this hypothesis, we have subjected shear-stressed human umbilical vein endothelial cells (HUVECs) to a HAnano®-enriched medium, as well as to surface presenting dual acid etching (DAE) as a control. To note, the titanium implants were coated with 10 nm in diameter HA particles using the Promimic HAnano method. Our data reveals that HAnano® modulates higher expression of genes related with endothelial cell performance and viability, such as VEGF, eNOS, and AKT, and further angiogenesis in vitro by promoting endothelial cell migration. Additionally, the data shows a significant extracellular matrix (ECM) remodeling, and this finding seems developing a dual role in promoting the expression of VEGF and control endothelial cell growth during angiogenesis. Altogether, these data prompted us to further validate this phenomenon by exploring genes related with the control of cell cycle and in fact our data shows that HAnano® promotes higher expression of CDK4 gene, while p21 and p15 genes (suppressor genes) were significantly lower. In conjunction, our data shows for the first time that HAnano®-coated surfaces drive angiogenesis by stimulating a proliferative and migration phenotype of endothelial cells, and this finding opens novel comprehension about osseointegration mechanism considering nanosized hydroxyapatite coating dental implants.

A 60 MeV proton beam-line dedicated to research and development programs.

A new proton beam-line dedicated to R&D programs has been developed at CentreAntoine Lacassagne (CAL), in Nice (France), in collaboration with the Centrenational d'études spatiales (CNES). This is the second beam-line of the MEDICYC 65 MeV cyclotron that is currently in operation, the first being the clinical 'eye-line' used for ocular proton therapy. The R&D beam-line is proposed with two configurations, the first producing a Gaussian narrow beam of a few mm width, the second a 100 mm diameter flat beam with a homogeneity better than ±3%. The energy range is (20 - ∼60) MeV, where the exact upper limit depends on the beam configuration being used. The energy spread of the non-degraded beam is (0.3 ± 0.1) MeV. A beam current between 10 pA and 10 µA can be produced with a stability better than 0.2% above 100 pA, and 2% below. The beam can be monitored online at a precision better than 5% in the flux range 1E5 (1E6) - 1E9 (1E10) p/cm2/s for a flat (Gaussian) configuration, although work is in progress to extend this range. Targeted applications for the R&D beam-line are instrumentation research, radiation tolerance tests of components and radiobiology.

Evaluation of vectorcardiography parameters matrixed synthesized

The form of analysis of the cardiac signal most used today is the electrocardiogram (ECG). However, in addition to this form of data visualization, there is the vectorcardiogram (VCG), that allows a visualization of the signal in 3 dimensions. This study aims to compare the different ECG to VCG transformation matrices Kors and Inverse Dower (iDower), by analyzing some known parameters taken from VCG's mathematically synthesized from 12-lead ECG's of amyloidosis patients. The idea is also to compare that similarity for patients with different types of amyloidosis. The study was done through the analysis of electrocardiograms taken from a sample of 12 humans who have amyloidosis, either mutant or wild-type. The results indicated that there is not much similarity between the signals, although the similarity was higher for patients with mutant amyloidosis than for those with wild-type amyloidosis.

Phytochemical characterization and biological activities of Plectranthus barbatus Andrews / Caracterização fitoquímica e atividades biológicas de Plectranthus barbatus Andrews

Plectranthus barbatus Andrews (Lamiaceae) is widely distributed in the world and has a range of popular therapeutic indications. This work aimed to evaluate the phytochemical characterization of two leaf extracts of P. barbatus, and their antimicrobial, antineoplastic and immunomodulatory potential. After collection, herborization and obtainment of the P. barbatus aqueous extract (PBA) and acetone:water 7:3 P. barbatus organic extract (PBO), the phytochemical characterization was performed by high-performance liquid chromatography (HPLC). The antimicrobial activity was performed to determine the minimum inhibitory concentration (MIC) against eight bacterial strains using the microdilution test and the fungus Trichophyton rubrum by disc diffusion assay and microdilution test. Cytotoxicity was assessed by MTT and trypan blue methods in normal peripheral blood mononuclear cells (PBMCs) at concentrations ranged between 0.1 to 100 µg.mL-1 and in neoplastic cell lines Toledo, K562, DU-145 and PANC-1 at 1, 10 and 100 µg.mL-1 . Immunomodulatory activity, was evaluated by sandwich ELISA of proinflammatory cytokines at BALB/c mice splenocytes cultures supernatant. Both extracts presented flavonoids, cinnamic derivatives, steroids and ellagic acid. PBO showed bacteriostatic activity against Acinetobacter baumannii (MIC = 250 µg.mL-1) clinical isolate and PBA fungistatic activity against Trichophyton rubrum (MIC = 800 µg.mL-1). The extracts did not exhibit toxicity to PBMCs and neoplastic cells (IC50 > 100 µg.mL-1). Additionally, PBO at 100 µg.mL-1 significantly inhibited IFN-γ and IL-17A cytokines (p = 0.03). Plectranthus barbatus is a potential candidate for therapeutic use due to its low toxicity in healthy human cells and exhibits biological activities of medical interest as bacteriostatic, fungistatic and immunomodulatory.

Plectranthus barbatus Andrews (Lamiaceae) é amplamente distribuída no mundo e com uma série de indicações terapêuticas populares. Este trabalho teve como objetivo avaliar a caracterização fitoquímica de dois extratos da folha de P. barbatus e seu potencial antimicrobiano, antineoplásico e imunomodulador. Após coleta, herborização e obtenção do extrato aquoso (PBA) e acetona: água 7: 3 (orgânico) (PBO) de P. barbatus, a caracterização fitoquímica foi realizada por cromatografia líquida de alta eficiência (CLAE). A atividade antimicrobiana foi realizada para determinar a concentração inibitória mínima (CIM) contra oito cepas bacterianas usando o teste de microdiluição e o fungo Trichophyton rubrum por ensaio de difusão em disco e teste de microdiluição. A citotoxicidade foi avaliada por métodos MTT e azul de tripan em células normais mononucleares do sangue periférico (CMSP) em concentrações variadas entre 0,1 a 100 µg.mL-1 e nas linhagens celulares neoplásicas Toledo, K562, DU-145 e PANC-1 em 1, 10 e 100 µg.mL-1 . A atividade imunomoduladora foi avaliada por ELISA sanduíche de citocinas pró-inflamatórias em sobrenadante de culturas de esplenócitos de camundongos BALB/c. Ambos os extratos apresentaram flavonoides, derivados cinâmicos, esteróides e ácido elágico. O PBO mostrou atividade bacteriostática contra Acinetobacter baumannii (CIM = 250 µg.mL-1) e atividade fungistática do PBA contra Trichophyton rubrum (CIM = 800 µg.mL-1). Os extratos não apresentaram toxicidade para CMSP e células neoplásicas (IC50 > 100 µg.mL-1). Além disso, o PBO a 100 µg.mL-1 inibiu significativamente as citocinas IFN-γ e IL-17A (p = 0,03). Plectranthus barbatus é um candidato potencial para uso terapêutico devido à sua baixa toxicidade em células humanas saudáveis e exibe atividade de interesse médico como bacteriostática, fungistática e imunomoduladora.

Phytochemical characterization and biological activities of Plectranthus barbatus Andrews / Caracterização fitoquímica e atividades biológicas de Plectranthus barbatus Andrews

Plectranthus barbatus Andrews (Lamiaceae) is widely distributed in the world and has a range of popular therapeutic indications. This work aimed to evaluate the phytochemical characterization of two leaf extracts of P. barbatus, and their antimicrobial, antineoplastic and immunomodulatory potential. After collection, herborization and obtainment of the P. barbatus aqueous extract (PBA) and acetone:water 7:3 P. barbatus organic extract (PBO), the phytochemical characterization was performed by high-performance liquid chromatography (HPLC). The antimicrobial activity was performed to determine the minimum inhibitory concentration (MIC) against eight bacterial strains using the microdilution test and the fungus Trichophyton rubrum by disc diffusion assay and microdilution test. Cytotoxicity was assessed by MTT and trypan blue methods in normal peripheral blood mononuclear cells (PBMCs) at concentrations ranged between 0.1 to 100 µg.mL-¹ and in neoplastic cell lines Toledo, K562, DU-145 and PANC-1 at 1, 10 and 100 µg.mL-¹ . Immunomodulatory activity, was evaluated by sandwich ELISA of proinflammatory cytokines at BALB/c mice splenocytes cultures supernatant. Both extracts presented flavonoids, cinnamic derivatives, steroids and ellagic acid. PBO showed bacteriostatic activity against Acinetobacter baumannii (MIC = 250 µg.mL-¹) clinical isolate and PBA fungistatic activity against Trichophyton rubrum (MIC = 800 µg.mL-¹). The extracts did not exhibit toxicity to PBMCs and neoplastic cells (IC50 > 100 µg.mL-¹). Additionally, PBO at 100 µg.mL-1 significantly inhibited IFN-γ and IL-17A cytokines (p = 0.03). Plectranthus barbatus is a potential candidate for therapeutic use due to its low toxicity in healthy human cells and exhibits biological activities of medical interest as bacteriostatic, fungistatic and immunomodulatory.

Plectranthus barbatus Andrews (Lamiaceae) é amplamente distribuída no mundo e com uma série de indicações terapêuticas populares. Este trabalho teve como objetivo avaliar a caracterização fitoquímica de dois extratos da folha de P. barbatus e seu potencial antimicrobiano, antineoplásico e imunomodulador. Após coleta, herborização e obtenção do extrato aquoso (PBA) e acetona: água 7: 3 (orgânico) (PBO) de P. barbatus, a caracterização fitoquímica foi realizada por cromatografia líquida de alta eficiência (CLAE). A atividade antimicrobiana foi realizada para determinar a concentração inibitória mínima (CIM) contra oito cepas bacterianas usando o teste de microdiluição e o fungo Trichophyton rubrum por ensaio de difusão em disco e teste de microdiluição. A citotoxicidade foi avaliada por métodos MTT e azul de tripan em células normais mononucleares do sangue periférico (CMSP) em concentrações variadas entre 0,1 a 100 µg.mL-¹ e nas linhagens celulares neoplásicas Toledo, K562, DU-145 e PANC-¹ em 1, 10 e 100 µg.mL-¹ . A atividade imunomoduladora foi avaliada por ELISA sanduíche de citocinas pró-inflamatórias em sobrenadante de culturas de esplenócitos de camundongos BALB/c. Ambos os extratos apresentaram flavonoides, derivados cinâmicos, esteróides e ácido elágico. O PBO mostrou atividade bacteriostática contra Acinetobacter baumannii (CIM = 250 µg.mL-¹) e atividade fungistática do PBA contra Trichophyton rubrum (CIM = 800 µg.mL-¹). Os extratos não apresentaram toxicidade para CMSP e células neoplásicas (IC50 > 100 µg.mL-¹). Além disso, o PBO a 100 µg.mL-¹ inibiu significativamente as citocinas IFN-γ e IL-17A (p = 0,03). Plectranthus barbatus é um candidato potencial para uso terapêutico devido à sua baixa toxicidade em células humanas saudáveis e exibe atividade de interesse médico como bacteriostática, fungistática e imunomoduladora.

Phytochemical characterization and biological activities of Plectranthus barbatus Andrews

Abstract Plectranthus barbatus Andrews (Lamiaceae) is widely distributed in the world and has a range of popular therapeutic indications. This work aimed to evaluate the phytochemical characterization of two leaf extracts of P. barbatus, and their antimicrobial, antineoplastic and immunomodulatory potential. After collection, herborization and obtainment of the P. barbatus aqueous extract (PBA) and acetone:water 7:3 P. barbatus organic extract (PBO), the phytochemical characterization was performed by high-performance liquid chromatography (HPLC). The antimicrobial activity was performed to determine the minimum inhibitory concentration (MIC) against eight bacterial strains using the microdilution test and the fungus Trichophyton rubrum by disc diffusion assay and microdilution test. Cytotoxicity was assessed by MTT and trypan blue methods in normal peripheral blood mononuclear cells (PBMCs) at concentrations ranged between 0.1 to 100 µg.mL-1 and in neoplastic cell lines Toledo, K562, DU-145 and PANC-1 at 1, 10 and 100 µg.mL-1 . Immunomodulatory activity, was evaluated by sandwich ELISA of proinflammatory cytokines at BALB/c mice splenocytes cultures supernatant. Both extracts presented flavonoids, cinnamic derivatives, steroids and ellagic acid. PBO showed bacteriostatic activity against Acinetobacter baumannii (MIC = 250 µg.mL-1) clinical isolate and PBA fungistatic activity against Trichophyton rubrum (MIC = 800 µg.mL-1). The extracts did not exhibit toxicity to PBMCs and neoplastic cells (IC50 > 100 µg.mL-1). Additionally, PBO at 100 µg.mL-1 significantly inhibited IFN- and IL-17A cytokines (p = 0.03). Plectranthus barbatus is a potential candidate for therapeutic use due to its low toxicity in healthy human cells and exhibits biological activities of medical interest as bacteriostatic, fungistatic and immunomodulatory.

Resumo Plectranthus barbatus Andrews (Lamiaceae) é amplamente distribuída no mundo e com uma série de indicações terapêuticas populares. Este trabalho teve como objetivo avaliar a caracterização fitoquímica de dois extratos da folha de P. barbatus e seu potencial antimicrobiano, antineoplásico e imunomodulador. Após coleta, herborização e obtenção do extrato aquoso (PBA) e acetona: água 7: 3 (orgânico) (PBO) de P. barbatus, a caracterização fitoquímica foi realizada por cromatografia líquida de alta eficiência (CLAE). A atividade antimicrobiana foi realizada para determinar a concentração inibitória mínima (CIM) contra oito cepas bacterianas usando o teste de microdiluição e o fungo Trichophyton rubrum por ensaio de difusão em disco e teste de microdiluição. A citotoxicidade foi avaliada por métodos MTT e azul de tripan em células normais mononucleares do sangue periférico (CMSP) em concentrações variadas entre 0,1 a 100 µg.mL-1 e nas linhagens celulares neoplásicas Toledo, K562, DU-145 e PANC-1 em 1, 10 e 100 µg.mL-1 . A atividade imunomoduladora foi avaliada por ELISA sanduíche de citocinas pró-inflamatórias em sobrenadante de culturas de esplenócitos de camundongos BALB/c. Ambos os extratos apresentaram flavonoides, derivados cinâmicos, esteróides e ácido elágico. O PBO mostrou atividade bacteriostática contra Acinetobacter baumannii (CIM = 250 µg.mL-1) e atividade fungistática do PBA contra Trichophyton rubrum (CIM = 800 µg.mL-1). Os extratos não apresentaram toxicidade para CMSP e células neoplásicas (IC50 > 100 µg.mL-1). Além disso, o PBO a 100 µg.mL-1 inibiu significativamente as citocinas IFN- e IL-17A (p = 0,03). Plectranthus barbatus é um candidato potencial para uso terapêutico devido à sua baixa toxicidade em células humanas saudáveis e exibe atividade de interesse médico como bacteriostática, fungistática e imunomoduladora.

Association between visceral/subcutaneous adipose tissue ratio and plasma inflammatory markers and score for cardiovascular risk prediction in a Brazilian cohort: Pró-Saúde Study.

Visceral adipose tissue (VAT) is associated with various metabolic disorders, and adipokines, secreted by adipose tissue, are involved in their pathogenesis. This study investigated associations between VAT/subcutaneous adipose tissue (SAT) ratio, inflammatory markers, and cardiovascular (CV) risk-score in adults. Plasma levels of adipokines, plasma lipid profile, blood pressure, and body composition (using dual-emission x-ray absorptiometry) were determined. CV risk-score based on the American College of Cardiology and the American Heart Association (ACC/AHA) score was calculated in a sample of 309 Brazilian civil servants aged <60 years. Participants' VAT/SAT ratio were categorized into quartiles. Among males, plasma leptin (2.8 ng/mL) and C reactive protein (CRP) (0.2 mg/dL) (P<0.05) levels were higher at P75 and P50 than P5, and the highest calculated CV risk-score was observed at P75 (7.1%). Among females, higher plasma adiponectin levels were observed at P25 (54.3 ng/mL) compared with P75 (36 ng/mL) (P<0.05). Higher plasma CRP levels were observed at P75 (0.4 mg/dL) compared with P5 (0.1 mg/dL) (P<0.05). Higher CV risk-score was observed at P75 (2.0%) compared with P5 (0.7%). In both sexes, VAT and VAT/SAT ratio were directly associated with plasma leptin, CRP, and CV risk-score, and inversely associated with adiponectin; SAT was directly associated with plasma leptin and CRP (P<0.01); interleukin (IL)-10 and CRP were directly associated with adiponectin and leptin, respectively (P<0.05). Among men only, IL-10 (inversely) and CRP (directly) were associated with CV risk-score (P=0.02). Our results strengthened the relevance of the VAT/SAT ratio in cardiovascular risk.

A deep learning based surrogate model for the parameter identification problem in probabilistic cellular automaton epidemic models.

BACKGROUND AND OBJECTIVE: an accurate estimation of the epidemiological model coefficients helps understand the basic principles of disease spreading. Some studies showed that dozens of hours are needed to simulate the traditional probabilistic cellular automaton (PCA) model, and dozens of hours are spent for a fine-tuning of the system. Here, we propose a deep learning-based surrogate model to mimic a PCA model to reduce the simulations' computational time, maintaining an equivalent precision in the estimates. METHOD: we consider PCA models based on regular lattices of different sizes to generate training data sets varying the parameters related to individuals' movement in the lattice and the disease infectivity. These parameters are the input variables for training the surrogate model, and the outputs parameters to be fitted are the percentages of susceptible and infected individuals at the steady-state, the basic reproduction number R0, the peak value and the peak instant of infected individuals, I(τ) and τ, respectively. RESULTS: The proposed surrogate model can predict all the output variables with a low relative error. The surrogate model's training time is independent of the size of the lattice, and the time for evaluating a solution by the surrogate model is low and independent of the lattice size. CONCLUSIONS: The surrogate model provides a fast simulation time for a generic Susceptible-Infected-Removed (SIR) model in a PCA, which is helpful for tuning the model before final simulations, supporting the initial search for inverse problems of parameters estimation in SIR models and providing a satisfactory estimation of the output variables for large populations.

Análise de parâmetros do VCG obtidos a partir de matrizes de transformação do ECG em pacientes com hipertrofia cardíaca / Analysis of VCG parameters obtained from ECG transformation matrices in patients with cardiac hypertrophy

INTRODUÇÃO: O eletrocardiograma (ECG) é uma ferramenta diagnóstica primordial em cardiologia. Através de técnicas de pós-processamento do ECG digital é possível extrair dados do clássico vetorcardiograma (VCG) e ampliar as possibilidades do método. OBJETIVO: Comparar os dados matriciais do ECG entre os pacientes portadores de hipertrofia ventricular patológica (Cardiomiopatia Hipertrófica e Amiloidose Cardíaca) e o grupo controle utilizando duas matrizes diferentes de transformação de ECG para VCG: a matriz inversa de Dower (iDower) e a matriz de Kors. MÉTODOS: Foram analisados VCGs sintetizados a partir das duas matrizes, iDower e Kors, e tais exames foram obtidos para o grupo hipertrófico, composto por 11 pacientes, e para o grupo controle, com cinco pacientes. O tempo de coleta do ECG foi de 10 minutos e, para cada paciente, foram analisados 10 VCGs, referentes a cada minuto da coleta. Foram analisados a amplitude cúbica (mV), por meio da análise da variação dos valores, e o ângulo do complexo QRS e da onda T, por meio da análise de dados circulares. RESULTADOS: Em relação aos ângulos, as médias dos ângulos de cada grupo tiveram maior proximidade no grupo controle e menor no grupo hipertrófico. A variação destes ângulos foi maior utilizando a matriz de Kors para o grupo controle (Figura.1); no entanto, no grupo hipertrófico a maior variação no observada na matriz iDower (Figura 2). Em relação à amplitude cúbica das ondas, a variação foi pequena para ambos os grupos, porém com proximidade maior na onda T, e não houve diferença notável em relação às matrizes. CONCLUSÃO: Analisando as variáveis angulares e de amplitude cúbica nas ondas QRS e T para o grupo controle e hipertrófico, não foi observada diferença significativa entre os grupos. Tal proximidade entre as variáveis também foi observada considerando as matrizes de transformação.

Phytochemical characterization and biological activities of Plectranthus barbatus Andrews.

Plectranthus barbatus Andrews (Lamiaceae) is widely distributed in the world and has a range of popular therapeutic indications. This work aimed to evaluate the phytochemical characterization of two leaf extracts of P. barbatus, and their antimicrobial, antineoplastic and immunomodulatory potential. After collection, herborization and obtainment of the P. barbatus aqueous extract (PBA) and acetone:water 7:3 P. barbatus organic extract (PBO), the phytochemical characterization was performed by high-performance liquid chromatography (HPLC). The antimicrobial activity was performed to determine the minimum inhibitory concentration (MIC) against eight bacterial strains using the microdilution test and the fungus Trichophyton rubrum by disc diffusion assay and microdilution test. Cytotoxicity was assessed by MTT and trypan blue methods in normal peripheral blood mononuclear cells (PBMCs) at concentrations ranged between 0.1 to 100 µg.mL-1 and in neoplastic cell lines Toledo, K562, DU-145 and PANC-1 at 1, 10 and 100 µg.mL-1 . Immunomodulatory activity, was evaluated by sandwich ELISA of proinflammatory cytokines at BALB/c mice splenocytes cultures supernatant. Both extracts presented flavonoids, cinnamic derivatives, steroids and ellagic acid. PBO showed bacteriostatic activity against Acinetobacter baumannii (MIC = 250 µg.mL-1) clinical isolate and PBA fungistatic activity against Trichophyton rubrum (MIC = 800 µg.mL-1). The extracts did not exhibit toxicity to PBMCs and neoplastic cells (IC50 > 100 µg.mL-1). Additionally, PBO at 100 µg.mL-1 significantly inhibited IFN-γ and IL-17A cytokines (p = 0.03). Plectranthus barbatus is a potential candidate for therapeutic use due to its low toxicity in healthy human cells and exhibits biological activities of medical interest as bacteriostatic, fungistatic and immunomodulatory.

Molecular characteristics of oocytes and somatic cells of follicles at different sizes that influence in vitro oocyte maturation and embryo production.

During the last 10 to 15 yr, in vitro research to predict antral follicle growth and oocyte maturation has delivered interesting advances in the knowledge of processes regulating follicle growth and developmental competence of oocytes. This review discusses the contribution of cumulus and mural granulosa cells in the process of oocyte maturation and cumulus expansion in cumulus-oocyte complexes (COCs) from follicles of different sizes and shows that differences in gene expression in oocytes, granulosa, and theca cells of small and large follicles impact the success of in vitro blastocyst development. In addition, the molecular mechanisms by which COC metabolism and antioxidant defense provide oocyte competence are highlighted. Furthermore, new insights and perspectives on molecular and cellular regulation of in vitro oocyte maturation are emphasized.

Association between visceral/subcutaneous adipose tissue ratio and plasma inflammatory markers and score for cardiovascular risk prediction in a Brazilian cohort: Pró-Saúde Study

Visceral adipose tissue (VAT) is associated with various metabolic disorders, and adipokines, secreted by adipose tissue, are involved in their pathogenesis. This study investigated associations between VAT/subcutaneous adipose tissue (SAT) ratio, inflammatory markers, and cardiovascular (CV) risk-score in adults. Plasma levels of adipokines, plasma lipid profile, blood pressure, and body composition (using dual-emission x-ray absorptiometry) were determined. CV risk-score based on the American College of Cardiology and the American Heart Association (ACC/AHA) score was calculated in a sample of 309 Brazilian civil servants aged <60 years. Participants' VAT/SAT ratio were categorized into quartiles. Among males, plasma leptin (2.8 ng/mL) and C reactive protein (CRP) (0.2 mg/dL) (P<0.05) levels were higher at P75 and P50 than P5, and the highest calculated CV risk-score was observed at P75 (7.1%). Among females, higher plasma adiponectin levels were observed at P25 (54.3 ng/mL) compared with P75 (36 ng/mL) (P<0.05). Higher plasma CRP levels were observed at P75 (0.4 mg/dL) compared with P5 (0.1 mg/dL) (P<0.05). Higher CV risk-score was observed at P75 (2.0%) compared with P5 (0.7%). In both sexes, VAT and VAT/SAT ratio were directly associated with plasma leptin, CRP, and CV risk-score, and inversely associated with adiponectin; SAT was directly associated with plasma leptin and CRP (P<0.01); interleukin (IL)-10 and CRP were directly associated with adiponectin and leptin, respectively (P<0.05). Among men only, IL-10 (inversely) and CRP (directly) were associated with CV risk-score (P=0.02). Our results strengthened the relevance of the VAT/SAT ratio in cardiovascular risk.

Association between socioeconomic markers and adult telomere length differs according to sex: Pro-Saúde study.

Understanding the social determinants of telomere length is critical to evaluate the risk of early biological aging. We investigated sex differences on the association between socioeconomic status (SES) and demographic markers and leukocyte telomere length (LTL) in Brazilian adults. This cross-sectional study was conducted in a subsample (women=228; men=200) nested within the Pro-Saúde study, a prospective cohort study of university civil servants in Rio de Janeiro, Brazil (2012-2013). Adjusted multivariate models were used to test the relationship between SES markers (marital status, educational attainment, father's educational attainment, race/skin color, household income, and childhood experience of food deprivation) and LTL. After adjusting for age and potential health-related confounders, lower educational attainment was associated with shorter LTL among men (ß=-0.05, 95% confidence interval (CI)=95%CI: -0.10, 0.00, P=0.03). In women, LTL was inversely associated with unmarried status (ß=-0.05, 95%CI: -0.09, 0.00, P=0.03), lower father's educational attainment (ß=-0.05, 95%CI: -0.13, 0.00, P=0.04), and childhood experience of food deprivation (ß=-0.07, 95%CI: -0.13, 0.00, P=0.04). Our findings suggested that the association between SES markers and LTL differs according to sex. SES markers able to induce lifelong stress, reflected in LTL, appeared to be more related to individual factors in men, whereas in women they were family-related.

Achados clínicos, histopatológicos e moleculares da mielopatia degenerativa canina: relato de caso / Clinical, histopathological and molecular findings of canine degenerative myelopathy: case report

Objetivou-se descrever os achados clínicos, histopatológicos e moleculares associados à MDC em um cão da raça Pastor-Suiço. O cão possuía uma paraparesia progressiva em membros pélvicos e foi submetido a avaliações clínicas, pelas quais se obteve, entre outros diferenciais, o diagnóstico presuntivo de MDC. Com a evolução dos sinais, o tutor optou pela eutanásia. Os achados histopatológicos da medula espinhal foram compatíveis com uma degeneração segmentar axonal e mielínica. O diagnóstico molecular foi realizado por meio da extração do DNA obtido por swab oral. Uma PCR foi otimizada utilizando-se primers descritos em literatura para amplificar a região do gene SOD1. A amostra foi, então, submetida a sequenciamento unidirecional, que revelou que o animal em questão era homozigoto para o alelo A para a mutação c.118G>A no éxon 2 do gene SOD1. O diagnóstico clínico presuntivo da MDC no presente caso foi esclarecido por meio dos achados histopatológicos, associados aos achados clínicos, e da sua caracterização molecular. Ressalta-se a contribuição deste relato, que traz aspectos clínicos, histopatológicos e moleculares associados à MDC na raça Pastor-Suíço, para a qual, até o presente momento, na literatura consultada, não há relato dessa enfermidade.(AU)

The objective of this study was to describe the clinical, histopathological and molecular findings associated with MDC in a Swiss Shepherd dog. The dog had a progressive paraparesis in pelvic limbs and was submitted to clinical evaluations where, among other differentials, the presumptive diagnosis of MDC was obtained. With the progression of the nervous deficits tutor opted for euthanasia. The histopathological findings of the spinal cord were compatible with axonal and myelinic segmental degeneration. Molecular diagnosis was performed by extracting the DNA obtained by oral swab. PCR was optimized using primers described in the literature to amplify the SOD1 gene region. The sample was then subjected to one-way sequencing which revealed that the animal in question was homozygous for the A allele for the c.118G>A mutation in exon 2 of the SOD1 gene. The presumptive diagnosis of MDC in the present case was clarified by histopathological findings, as well as by its molecular characterization. The contribution of this report brings clinical, histopathological and molecular aspects associated with canine degenerative myelopathy in the Swiss Shepherd breed, that until this moment, in the literature consulted, there is no report of this disease in the breed mentioned.(AU)

Neighborhood food environment and consumption of fruit and leafy vegetables: Pro-Saude Study, Brazil.

OBJECTIVE: To explore the relationship between availability of different types of retail food stores and consumption of fruit and leafy vegetables. STUDY DESIGN: This study is a cross-sectional study. METHODS: Data were derived from 2032 adults living in the city of Rio de Janeiro, Brazil, participating in the longitudinal Pro-Saude Study. Exposure to street markets, fruit and vegetable stores, groceries and markets, unhealthy food outlets, restaurants, and supermarkets within 1600 m buffers was obtained by georeferencing residential addresses. Consumption of fruit and leafy vegetables was assessed via two single questions, categorized as 'yes' (≥4 days/week) and 'no' (<3 days/week). Multiple logistic regression models were used to assess relationships of interest adjusted for surrounding average monthly income, sex, age, education, and family income per capita. RESULTS: Except for supermarkets, the presence of a greater number of retail food stores - irrespective of the type - was associated with higher odds of consumption of fruit and leafy vegetables than with areas with a lower number (e.g. odds ratio = 1.47; 95% confidence interval = 1.13-1.91). CONCLUSIONS: The greater availability of several types of retail food stores close to participants' residences was associated with higher consumption of fruit and leafy vegetables in Rio de Janeiro.

Assessment of the acid phosphatase CP01850 from Corynebacterium pseudotuberculosis in DNA and subunit vaccine formulations against caseous lymphadenitis / Avaliação da fosfatase ácida recombinante CP01850 de Corynebacterium pseudotuberculosis em formulações vacinais de subunidade e de DNA contra a linfadenite caseosa

The target cp1002_RS01850 from Corynebacterium pseudotuberculosis was used to construct a DNA and recombinant subunit vaccine against caseous lymphadenitis. Recombinant protein rCP01850 was expressed in Escherichia coli using pAE vector, and DNA vaccine was engineered with pTARGET vector. BALB/c mice were divided in five groups containing eight animals each, inoculated with: pTARGET/cp01850 as DNA vaccine (G1); rCP01850 plus Al (OH)3 as recombinant subunit vaccine (G2); pTARGET/cp01850 and a boost with rCP01850 plus Al (OH)3 (G3); pTARGET (G4); or Al (OH)3 (G5). Mice were inoculated and blood samples were collected on days 0, 21, and 42 for the analysis of total IgG, IgG1 and IgG2a by ELISA. In each group, five animals were challenged with Mic-6 C. pseudotuberculosis strain, and three were used for cytokine quantification by qPCR. Although no group has been protected by vaccines against lethal challenge, G2 showed an increase in the survival rate after challenge. Significantly higher levels of IL-4, IL-12, IFN-γ, total IgG, IgG1 and IgG2a were also detected for G2, evidencing a mixed Th1/Th2 immunological profile. In conclusion, despite no protection level provided by different vaccinal strategies using cp1002_RS01850 from C. pseudotuberculosis, G2 developed a Th1/Th2 immune response with an increase in survival rate.(AU)

O alvo cp1002_RS01850 de Corynebacterium pseudotuberculosis foi utilizado para construir uma vacina recombinante de subunidade e de DNA contra a linfadenite caseosa. A proteína recombinante rCP01850 foi expressa em Escherichia coli usando o vetor pAE, e a vacina de DNA foi construída com o vetor pTARGET. Camundongos BALB/c foram divididos em grupos de oito animais, inoculados com: pTARGET/cp01850 como vacina de DNA (G1); rCP01850 e Al (OH)3 como vacina recombinante de subunidade (G2); pTARGET/cp01850 e um boost com rCP01850 e Al (OH)3 (G3); pTARGET (G4); ou Al (OH)3 (G5). Os animais foram inoculados e amostras de sangue foram coletadas nos dias 0, 21, e 42 do experimento para a análise de IgG total, IgG1 e IgG2a por ELISA. De cada grupo, cinco animais foram desafiados com a cepa Mic-6 de C. pseudotuberculosis, e três foram usados para a quantificação de citocinas por qPCR. Apesar de nenhum grupo ter sido protegido pelas vacinas testadas contra o desafio letal, G2 apresentou taxa de sobrevida e níveis de IL-4, IL-12, IFN-γ, IgG total, IgG1 e IgG2a significativamente mais altos, evidenciando um perfil imunológico misto Th1/Th2. Conclui-se que apesar das diferentes estratégias vacinais utilizando cp1002_RS01850 de C. pseudotuberculosis não terem sido capazes de gerar proteção, G2 desenvolveu uma resposta Th1/Th2 e elevou a taxa de sobrevida.(AU)

Effects of epidermal growth factor and progesterone on development, ultrastructure and gene expression of bovine secondary follicles cultured in vitro.

The aims of this study were to investigate the effects of epidermal growth factor (EGF) and progesterone on the development, viability and the gene expression of bovine secondary follicle culture in vitro for 18 days. Secondary follicles (∼0.2 mm) were isolated from ovarian cortex and individually cultured at 38.5 °C, with 5% CO2 in air, for 18 days, in TCM-199+ (n = 63) alone (control medium) or supplemented with 10 ng/mL progesterone (n = 64), 10 ng/mL EGF (n = 61) or both EGF and progesterone (n = 66). The effects of these treatments on growth, antrum formation, viability, ultrastructure and mRNA levels for GDF-9, c-MOS, H1foo and cyclin B1 were evaluated, significantly different (p < 0.05). The results showed that there was a progressive increase in follicular diameter in all treatments, but only follicles cultured in medium supplemented with EGF had increased significantly in diameter when compared to follicles cultured in the control medium at the end of the culture period, significantly different (p < 0.05). A positive interaction between EGF and progesterone was not observed. In addition, the presence of EGF, progesterone or both in culture medium did not influence the rate of follicle survival and antrum formation. However, the presence of only progesterone in cultured medium increased the expression of mRNAs for GDF9 and cyclin B1 in oocytes. EGF also significantly increased the levels of mRNAs for cMOS and GDF9 when compared to follicles cultured in control medium. Ultrastructural analyzes showed that cultured follicles in all treatments maintained the integrity of granulosa cells. In conclusion, the EGF promotes the development of secondary follicles cultured in vitro for 18 days and increases the expression of cMOS and GDF9, while progesterone alone or in association with EGF have not a positive effect on follicular growth. However, progesterone increases the expression of GDF9 and cyclin B1 in oocytes.

Association between socioeconomic markers and adult telomere length differs according to sex: Pro-Saúde study

Understanding the social determinants of telomere length is critical to evaluate the risk of early biological aging. We investigated sex differences on the association between socioeconomic status (SES) and demographic markers and leukocyte telomere length (LTL) in Brazilian adults. This cross-sectional study was conducted in a subsample (women=228; men=200) nested within the Pro-Saúde study, a prospective cohort study of university civil servants in Rio de Janeiro, Brazil (2012-2013). Adjusted multivariate models were used to test the relationship between SES markers (marital status, educational attainment, father's educational attainment, race/skin color, household income, and childhood experience of food deprivation) and LTL. After adjusting for age and potential health-related confounders, lower educational attainment was associated with shorter LTL among men (β=-0.05, 95% confidence interval (CI)=95%CI: -0.10, 0.00, P=0.03). In women, LTL was inversely associated with unmarried status (β=-0.05, 95%CI: -0.09, 0.00, P=0.03), lower father's educational attainment (β=-0.05, 95%CI: -0.13, 0.00, P=0.04), and childhood experience of food deprivation (β=-0.07, 95%CI: -0.13, 0.00, P=0.04). Our findings suggested that the association between SES markers and LTL differs according to sex. SES markers able to induce lifelong stress, reflected in LTL, appeared to be more related to individual factors in men, whereas in women they were family-related.

Bloqueio de ganglio estrelado para tratamento de taquicardia ventricular refratária em paciente portador de cardiomiopatia chagásica / Starry ganglion block for treatment of refractory ventricular tachycardia in a patient with Chagas cardiomyopathy

INTRODUÇÃO: A maioria dos pacientes com doença de Chagas mostra perda do controle cardíaco parassimpático o que pode ser a causa de arritmias malignas e morte súbita. O conceito de tratar problemas cardíacos com bloqueio simpático existe desde que descobriu-se que a denervação simpática cardíaca esquerda reduziu a incidência de fibrilação ventricular (FV) em experimentos com cães. O bloqueio do gânglio estrelado evoluiu de denervação simpática e atualmente é aventado como uma importante terapia adjuvante no tratamento de tempestades elétricas. RELATO DE CASO: Paciente do sexo masculino, 50 anos, acompanhado em um serviço terciário de cardiologia devido a cardiomiopatia chagásica forma arritmogênica. Foi submetido a implante de cardiodesfibrilador implantável (CDI) em 2015 devido a episódio de taquicardia ventricular sustentada (TVS) induzida em estudo eletrofisiológico. Há 1 mês, paciente procura o serviço com queixa de dor torácica em queimação sendo admitido em TVS monomórfica com frequência de 130 bpm. Foi submetido a cardioverão química e elétrica com recorrência da taquicardia, a qual foi parcialmente controlada com terapias antitaquicardia do dispositivo. Optou-se pelo bloqueio do gânglio estrelado esquerdo com solução de bupivacaina e xylocaina, guiado por Ultrassom, e após 24 horas do procedimento evoluiu com apenas 1 episódio de TVS lenta revertida com ATP. Manteve-se assintomático sendo submetido a ablação epicárdica e endocárdica com mapeamento eletroanatômico 7 dias após. DISCUSSÃO: Tempestade elétrica pode ocorrer em até 20% de pacientes com CDI e está associada com morbidade e mortalidade significativas. Geralmente é acompanhada por um surto simpático frequentemente resistente a antiarrítmicos, terapia medicamentosa, necessitando de bloqueio simpático. Neste contexto, a denervação simpática cardíaca esquerda demonstrou reduzir a carga de choque de CDI em pequenas séries de pacientes com doença cardíaca estrutural e taquiarritmias recorrentes. O bloqueio do gânglio estrelado pode ser alcançado através da infusão de anestésico periganglionar e esta prática tende a estabilizar um paciente por tempo suficiente para realização de ablação por cateter. CONCLUSÃO: O bloqueio simpático do gânglio estrelado restaura o equilíbrio do sistema nervoso autonômico sendo uma alternativa de baixo risco para estabilização dos pacientes até a realização da terapia final. Infelizmente, ainda permanece subutilizado devido à falta de conhecimento e compreensão da eficácia e duração do efeito, além de ser necessário mais estudos em pacientes chagásicos. (AU)