Circuit training reduces cardiometabolic risk factors in women / El entrenamiento en circuito reduce los factores de riesgo cardiometabólico en mujeres / Treinamento em circuito reduz os fatores de risco cardiometabólicos em mulheres

ABSTRACT Introduction Combined training is more effective than an isolated modality in reducing cardiometabolic risk indicators. Objective To evaluate the effect of circuit training volume on anthropometric and biochemical risk indicators for cardiometabolic diseases in overweight women. Methods Thirty-two participants underwent 24 weeks of circuit training with free weights combined with aerobic exercise. The training volume during the 24 weeks was used to distribute the women into moderate-volume physical activity (MVA), low-volume physical activity (LVA) and control (CON) groups. Anthropometric indices (body mass, body mass index (BMI), waist circumference (WC), waist-hip ratio (WHR)), blood glucose, insulin, insulin resistance (HOMA-IR), total cholesterol (TC), triglycerides, HDL-c, and LDL-c were evaluated at the beginning of the program and after 12 and 24 weeks. Results There was no interaction between training volume and time for any of the variables studied, but the intervention time influenced body mass (p=0.013) and BMI (p=0.012), and there was a tendency for participation time to reduce body mass (p=0.063) and BMI (p=0.062) after six months of intervention. The volume of the physical activity affected HDL-c (p=0.037), being significant (p=0.030) in the comparison between the MVA and CON groups. Additionally, there was a downward trend in HDL-c after six months of intervention (p=0.073), with a smaller reduction observed in the MVA group, indicating a protective role of moderate physical activity in the reduction of this lipid fraction. The association between physical activity volume and participation time resulted in a clinical improvement in total cholesterol (χ2 = 5.453, p = 0.02), with a higher probability of reaching clinically adequate values in the MVA group (OR = 0.126; 95%CI 0.019 - 0.827). Conclusion Training volume improved cardiometabolic risk factors in overweight women. Level of evidence II; Therapeutic Studies - Investigating the Results of Treatment.

RESUMEN Introducción El entrenamiento combinado es más eficiente que la modalidad aislada en indicadores de riesgo cardiometabólico. Objetivo Evaluar el efecto del volumen de entrenamiento en circuito sobre indicadores antropométricos y bioquímicos con riesgo de enfermedades cardiometabólicas en mujeres con sobrepeso. Métodos Treinta y dos participantes se sometieron a 24 semanas de entrenamiento en circuito con pesos libres combinados con ejercicio aeróbico. El volumen de entrenamiento durante las 24 semanas se utilizó para distribuir a las mujeres en los grupos: actividad física de volumen moderado (AVM), actividad física de volumen bajo (AVB) y control (CON). Se evaluaron los índices antropométricos masa corporal, índice de masa corporal (IMC), circunferencia de la cintura (CC), relación cintura-cadera (RCC), glucemia, insulina, resistencia a la insulina (HOMA-IR), colesterol total (CT), triglicéridos, HDL-c y LDL-c al inicio del programa y después de las semanas 12 y 24. Resultados No hubo interacción entre el volumen y el tiempo de entrenamiento para ninguna de las variables estudiadas, pero el tiempo de intervención influyó en la masa corporal (p=0,013) y en el IMC (p=0,012), y el tiempo de participación tendió a reducir la masa corporal (p=0,063) y el IMC (p=0,062), después de seis meses de intervención. El volumen de actividad física afectó al HDL-c (p =0,037), siendo significativo (p=0,030) en la comparación entre AVM y CON. Además, hubo una tendencia a la reducción del HDL-c después de seis meses de intervención (p=0,073), observándose la menor reducción en AVM, lo que indica el papel protector de la actividad física de volumen moderado en la reducción de esta fracción lipídica. La actividad física y el tiempo de participación mostraron una mejora clínica en colesterol total (χ2 = 5,453, p = 0,02), con mayor probabilidad de alcanzar valores clínicamente adecuados de AVM (OR = 0,126; IC95% 0,019 - 0,827). Conclusión El volumen de entrenamiento atenuó los factores de riesgo cardiometabólico en mujeres con sobrepeso. Nivel de Evidencia II; Estudios terapéuticos: investigación de los resultados del tratamiento.

ABSTRACT Introdução O treinamento combinado é mais eficiente do que a modalidade isolada com relação aos indicadores de risco cardiometabólico. Objetivos Avaliar o efeito do volume de treinamento em circuito sobre indicadores antropométricos e bioquímicos com risco de doenças cardiometabólicas em mulheres com excesso de peso. Métodos Trinta e duas participantes foram submetidas a 24 semanas de treinamento em circuito, com pesos livres combinados com exercício aeróbico. O volume de treinamento durante as 24 semanas foi utilizado para distribuir as mulheres nos grupos: atividade física de volume moderado (AVM), atividade física de baixo volume (AVB) e controle (CON). Os índices antropométricos massa corporal, índice de massa corporal (IMC), circunferência de cintura (CC), relação cintura-quadril (RCQ), glicemia, insulina, resistência à insulina (HOMA-IR), colesterol total (CT), triglicerídeos, HDL-c e LDL-c, foram avaliados no início do programa e depois de 12 e 24 semanas. Resultados Não houve interação entre o volume de treinamento e o tempo para nenhuma das variáveis estudadas, mas o tempo de intervenção influenciou a massa corporal (p = 0,013) e o IMC (p = 0,012), e o tempo de participação tendeu a reduzir a massa corporal (p = 0,063) e o IMC (p = 0,062), depois de seis meses de intervenção. O volume de atividade física afetou o HDL-c (p = 0,037), sendo significativo (p = 0,030) na comparação entre AVM e CON. Adicionalmente, verificou-se tendência de redução HDL-c depois seis meses de intervenção (p = 0,073), sendo a menor redução observada no AVM, que indica o papel protetor de atividade física de volume moderado na redução dessa fração lipídica. A associação entre o volume de atividade física e o tempo de participação mostrou melhora clínica do colesterol total (χ2= 5,453, p = 0,02), com maior probabilidade de atingir valores clinicamente adequados de AVM (OR = 0,126; IC de 95% 0,019 - 0,827). Conclusão O volume de treinamento atenuou os fatores de risco cardiometabólico em mulheres com excesso de peso. Nível de evidência II; Estudos terapêuticos - Investigação dos resultados do tratamento.

Curcumin supplementation reduces blood glucose and serum lipids of Brazilian women with high waist circumference: a randomized clinical trial

ABSTRACT Objective: To evaluate the effect of curcumin supplementation on the body compositions and biochemical parameters of Brazilian women with high waist circumferences. Materials and methods: This is a blind, randomized, placebo-controlled clinical trial conducted in 2016 with 35 Brazilian women with high waist circumference (WC). In total, 80 participants were randomized [38 in the placebo group (PG) and 42 in the supplemented group (SG)], but at the end of the protocol, 20 individuals in the PG and 15 in the SG were evaluated. The sample consumed one capsule of curcumin (500 mg/day) (Curcumin C3 Complex®) or a placebo for 90 days. Body weight, height, body mass index, WC, body fat, fat free mass, fasting glucose (FG), lipid profile [triglycerides (TGs), total cholesterol (TC), HDL-c and LDL-c], physical activity level and food intake (energy, carbohydrate, total fat and protein) data were evaluated before and after the intervention. Results: Curcumin supplementation reduced body mass (p < 0.040) but did not alter other anthropometric parameters or body composition (p ≥ 0.050). In relation to the biochemical profile, the SG saw reductions in FG (p < 0.001), TGs (p < 0.001) and TC (p = 0.001) compared with the PG. At the baseline and during the intervention, the practice of physical activity and food intake did not differ between the SG and PG (p ≥ 0.050). Conclusion: Curcumin supplementation improved the blood glucose and lipid profile of Brazilian women with high WC, without altering body composition. New studies with larger sample sizes and longer durations are important for identifying more robust data regarding the proposal of this work.

Curcumin supplementation reduces blood glucose and serum lipids of Brazilian women with high waist circumference: a randomized clinical trial.

Objective: To evaluate the effect of curcumin supplementation on the body compositions and biochemical parameters of Brazilian women with high waist circumferences. Methods: This is a blind, randomized, placebo-controlled clinical trial conducted in 2016 with 35 Brazilian women with high waist circumference (WC). In total, 80 participants were randomized [38 in the placebo group (PG) and 42 in the supplemented group (SG)], but at the end of the protocol, 20 individuals in the PG and 15 in the SG were evaluated. The sample consumed one capsule of curcumin (500 mg/day) (Curcumin C3 Complex®) or a placebo for 90 days. Body weight, height, body mass index, WC, body fat, fat free mass, fasting glucose (FG), lipid profile [triglycerides (TGs), total cholesterol (TC), HDL-c and LDL-c], physical activity level and food intake (energy, carbohydrate, total fat and protein) data were evaluated before and after the intervention. Results: Curcumin supplementation reduced body mass (p < 0.040) but did not alter other anthropometric parameters or body composition (p ≥ 0.050). In relation to the biochemical profile, the SG saw reductions in FG (p < 0.001), TGs (p < 0.001) and TC (p = 0.001) compared with the PG. At the baseline and during the intervention, the practice of physical activity and food intake did not differ between the SG and PG (p ≥ 0.050). Conclusion: Curcumin supplementation improved the blood glucose and lipid profile of Brazilian women with high WC, without altering body composition. New studies with larger sample sizes and longer durations are important for identifying more robust data regarding the proposal of this work.

Expression of the cancer stem cell marker OCT4 is associated with worse prognosis and survival in cutaneous melanoma.

Cutaneous melanoma has an aggressive clinical presentation, showing rapid rate of growth and metastatic dissemination due to the permanence of cancer stem cells. The present study was to evaluate the expression of the self-renewal regulatory factor and the clinical significance of the transcription factor OCT4 in melanoma. Melanoma tissues were stained by immunohistochemistry and the correlation between the expression of this marker was determined through clinical-pathological variables and survival outcomes. Positive expression of nuclear and cytoplasmic OCT4 was observed in 49% and 41.2% of cases, respectively. The positive expression of nuclear OCT4 in melanoma was significantly associated with prognostic factors, such as Breslow depth, Clark's level, ulceration and metastasis. Survival of patients was 56% compared to positive nuclear OCT4 expression and 94.2% when compared to the low expression of the gene. Nuclear OCT4 positive genotype indicated aggressive tumor behavior with a worse clinical outcome, which indicates OCT4 as a useful biomarker in the prognosis of melanoma.

Endothelial nitric oxide concentrations in the saliva of jiu-jitsu athletes / Concentrações de óxido nítrico endotelial na saliva de atletas de jiu-jitsu / Concentraciones de óxido nítrico endotelial en la saliva de atletas de jiu-jitsu

ABSTRACT Introduction This article discusses the production of nitric oxide under the influence of sport-specific physical training, measured by the salivary nitrite of Jiu-Jitsu athletes. Objectives To verify the potential of the sport to produce optimal levels of nitric oxide stimulated by exertion, and to quantify training-related nitric oxide concentrations. Method The study participants were 14 volunteer athletes from the Tatame project (extension project), who were monitored for nine months in their training routine, providing samples of unstimulated saliva. Samples were collected each month, in three periods of the day: in the morning upon waking, immediately before training, and immediately after training. Salivary nitrite was quantified by the colorimetric Griess assay. Training heart rates were also monitored in order to establish training intensity. Results Mean monthly salivary nitrite levels showed a significant correlation with mean monthly heart rates, suggesting that salivary nitrite responds to training. However, salivary nitrite concentrations measured immediately after training were always lower than in the pre-training period. Conclusion The post-training reduction in concentrations was due to the nature of the sport studied, since because it involves a fight, the intense sympathetic stimulation inhibited salivary gland activity and irrigation, preventing salivary nitrite from producing an increase in circulating nitric oxide. Level of evidence IV; Case series.

RESUMO Introdução Este artigo discute a produção de óxido nítrico sob interferência do treinamento físico esportivo, medida pelo nitrito salivar em atletas de Jiu-Jitsu. Objetivos Verificar as potencialidades da modalidade na produção de níveis ideais de óxido nítrico estimulado pelo esforço e quantificar as concentrações de óxido nítrico relacionadas com o treinamento. Métodos O estudo teve a participação de 14 atletas voluntários do Projeto Tatame (projeto de extensão), que foram acompanhados por nove meses em sua rotina de treinamento e forneceram amostras de saliva não estimulada. Foram coletadas amostras a cada mês, em três períodos do dia: pela manhã ao acordar, imediatamente antes do treino e imediatamente após o treino. O nitrito salivar foi quantificado pelo método colorimétrico de Griess. Também foram monitoradas as frequências cardíacas de treinamento a fim de se estabelecer a intensidade do treinamento realizado. Resultados As médias mensais de nitrito salivar demonstraram correlação significativa com as médias mensais de frequência cardíaca, dando indícios de que o nitrito salivar responde ao treinamento. Contudo, as concentrações de nitrito salivar medidas imediatamente após o treinamento sempre eram menores com relação ao período anterior do treino. Conclusão A redução da concentração depois do treinamento deveu-se à natureza da modalidade estudada, visto que, por ser uma luta, a forte carga de estimulação simpática inibiu a atividade das glândulas salivares, bem como sua irrigação, impedindo que o nitrito salivar produzisse aumento do óxido nítrico circulante. Nível de evidência IV; Série de casos.

RESUMEN Introducción Este artículo analiza la producción de óxido nítrico bajo la influencia del entrenamiento físico deportivo, medida por el nitrito salival de atletas de Jiu-Jitsu. Objetivos Verificar el potencial de la modalidad en la producción de niveles ideales de óxido nítrico estimulado por el esfuerzo y cuantificar las concentraciones de óxido nítrico relacionadas con el entrenamiento. Métodos El estudio contó con la participación de 14 atletas voluntarios participantes del Proyecto Tatame (proyecto de extensión) a quienes se les dio seguimiento durante nueve meses en su rutina de entrenamiento y proporcionaron muestras de saliva no estimulada. Se recolectaron muestras cada mes, en tres períodos del día: en la mañana al despertar, inmediatamente antes del entrenamiento e inmediatamente después del entrenamiento. El nitrito salival se cuantific ó por el método colorimétrico de Griess. Las frecuencias cardíacas de entrenamiento se monitorearon para establecer la intensidad del entrenamiento realizado. Resultados Los promedios mensuales de nitrito salival mostraron una correlación significativa con los promedios mensuales de frecuencia cardíaca, lo que indica que el nitrito salival responde al entrenamiento. Sin embargo, las concentraciones de nitrito salival medidas inmediatamente después del entrenamiento siempre fueron más bajas en comparación con el período anterior al entrenamiento. Conclusión La reducción de la concentración después del entrenamiento se debió a la naturaleza de la modalidad estudiada, ya que, por ser una lucha, la fuerte carga de estimulación simpática inhibió la actividad de las glándulas salivales, así como su irrigación, evitando que el nitrito salival produjera aumento de óxido nítrico circulante. Nivel de evidencia IV; Serie de casos.

Waist Circumference Above 80 cm Predicts Increased Systolic Blood Pressure in Healthy Young Adult Women

Abstract Background: Among anthropometric measures for assessing adiposity-related risk, waist circumference (WC) is simple and fast to perform. Cut-off values for WC proposed by the International Diabetes Federation (IDF), and the Adult Treatment Panel III of the National Cholesterol Education Program (NCEP-ATP III) are categorized by gender and are not age-specific. Objective: To analyze the association between WC and cardiometabolic risk factors in adult women. Methods: A total of 164 healthy adult women were grouped by WC according to IDF and NCEP-ATP III cutoff values. Continuous variables were described as mean ± standard deviation or median (interquartile range). The Shapiro-Wilk test was used to assess the normality of data. Variables were analyzed by unpaired Student's t-test, Mann-Whitney U and Kruskal-Wallis tests. The correlation of WC categories with systolic (SBP) and diastolic (DBP) blood pressure, fasting blood glucose, high-density lipoprotein cholesterol (HDL-c), and triglycerides were examined by Spearman's rho correlation coefficient and linear regression analysis. A p value < 0.05 was considered statistically significant. Results: Increased WC showed a significant correlation with SBP, DBP, glucose, HDL-c, and triglycerides. In bivariate linear regression, approximately 63.0 % of the variability of SBP (≥ 130 mmHg) among the age group 20-40 years was predicted by increased WC according to both criteria. Conclusion: A WC above 80 cm in women aged 20-40 years strongly predicted variability in SBP, calling attention to the importance of measuring WC for the monitoring and prevention of cardiovascular and metabolic diseases in women in this age group.

Salivary cortisol and nitrite concentrations in school teachers: A longitudinal pilot study.

OBJECTIVE: Occupational/mental stressors, which may be evaluated with measurements of salivary cortisol concentrations, affect the endothelial function and has implications on cardiovascular health. Nitric oxide (NO) is known to have an important role in cardiac function and may also be assessed in the saliva, but its participation in physiological responses to hypothalamic-pituitary-adrenal (HPA) axis stimulation is still not completely understood. The aim of the present study was to test the hypothesis that salivary NO (as nitrite) and cortisol concentrations in school teachers vary throughout the academic year. METHODS: Saliva samples were collected from 13 teachers distributed across five secondary schools. The samples were collected at 6:30 a.m., 11:30 a.m., and 5:30 p.m. in the months of March, July, and December. Salivary cortisol levels were measured by enzyme immunoassay (EIA), and salivary NO concentration was determined by the quantification of nitrite. The area under the curve in relation to ground (AUC) was calculated to assess the cortisol and nitrite concentrations throughout the day. RESULTS: No significant changes were observed in cortisol or nitrite concentrations across the three periods of the school year, while cortisol and nitrite levels showed a low positive and significant correlation (r=0.3455, p=0.0336). CONCLUSION: The results suggest that changes in salivary cortisol levels are accompanied by changes in salivary nitrite levels. This is the first time that such association has been demonstrated. These results encourage further studies aimed to confirm the importance of salivary NO measurement in relation to occupational stress and cardiovascular health.

Nitric Oxide Concentration and Other Salivary Changes after Insertion of New Complete Dentures in Edentulous Subjects.

Objective. To assess changes in levels of salivary nitric oxide (NO) after insertion of new complete dentures and its association with clinical and salivary parameters. Methods. Nineteen fully edentulous subjects were included, mean age 64.4. Unstimulated whole saliva was collected before and after insertion of the dentures, at follow-up visits, and after 12 months. The concentration of the final stable NO product (nitrite) was measured by a colorimetric assay based on the Griess reaction. Clinical parameters were assessed during all clinical visits. Results. Functional adaptation to the dentures progressively improved, with no complaints at the long-term follow-up. NO concentration was not influenced by the level of functional adaptation, presence of injuries to the mucosa, salivary flow, and saliva viscosity. Pairwise comparison showed a reduction in NO concentration at the first follow-up compared to baseline values but differences were not statistically significant. Significant differences were observed in NO concentrations at the long-term follow-up when compared to the first (p = 0.024) and second (p = 0.027) visits. Conclusion. NO concentration reduced after denture insertion and returned to baseline levels in the long-term follow-up. This appears to be an autonomic response of the body and provides valuable complementary information for the management of the edentulous patient.

Estrogen signaling in the proliferative endometrium: implications in endometriosis.

Even though the physiological role of estrogen in the female reproductive cycle and endometrial proliferative phase is well established, the signaling pathways by which estrogen exerts its action in the endometrial tissue are still little known. In this regard, advancements in cell culture techniques and maintenance of endometrial cells in cultures enabled the discovery of new signaling mechanisms activated by estrogen in the normal endometrium and in endometriosis. This review aims to present the recent findings in the genomic and non-genomic estrogen signaling pathways in the proliferative human endometrium specifically associated with the pathogenesis and development of endometriosis.

Estrogen signaling in the proliferative endometrium: implications in endometriosis / Sinalização pelo estrogênio no endométrio proliferativo: implicações na endometriose

SUMMARY Even though the physiological role of estrogen in the female reproductive cycle and endometrial proliferative phase is well established, the signaling pathways by which estrogen exerts its action in the endometrial tissue are still little known. In this regard, advancements in cell culture techniques and maintenance of endometrial cells in cultures enabled the discovery of new signaling mechanisms activated by estrogen in the normal endometrium and in endometriosis. This review aims to present the recent findings in the genomic and non-genomic estrogen signaling pathways in the proliferative human endometrium specifically associated with the pathogenesis and development of endometriosis.

RESUMO Embora esteja bem estabelecido o papel fisiológico do estrogênio no ciclo reprodutivo feminino e na fase proliferativa do endométrio, as vias de sinalização por meio das quais a ação do estrogênio é exercida no tecido endometrial são ainda pouco conhecidas. Nesse sentido, o avanço nas técnicas de cultura celular e a manutenção de células endometriais em cultivo possibilitaram a descoberta de novos mecanismos sinalizadores ativados pelo estrogênio no endométrio normal e na endometriose. Esta revisão tem o objetivo de apresentar as descobertas recentes envolvendo as vias de sinalização genômica e não genômica do estrogênio no endométrio proliferativo humano, especificamente associadas à patogênese e ao desenvolvimento da endometriose.

Citopatogenicidade de Trichomonas vaginalis: uma via de sinalização carboidrato-dependente / Trichomonas vaginalis' pathogenic effect and cell signaling in a carbohydrate-dependent manner

Trichomonas vaginalis é um agente infectante da microbiota vaginal que vem sendo correlacionado ao câncer cervical. Um receptor denominado alectina-1 (Gal 1) pode ser expresso em células epiteliais cervicais humanas se ligando à glicofosfolipídica (LPG) de T. vaginalis. A interação de T. vaginalis com as células epiteliais é mediada por cadeias galactose e N-acetilglucosamina (LacNac). Gal 1 se liga aos sítios poly-LacNAC e está relacionada com a aderência de T. vaginalis à célula humana. A sinalização ocorre por intermédio de sítios da proteína Src (SH2) que se associam, ocorrendo sob os domínios de PI3K que fosforilam a membrana de lípides fosfatidilinositol (PIP e PIP2). Aderindo-se às membranas citoplasmáticas e secretando enzimas, T. vaginalis pode ocasionar a ruptura do envoltório celular podendo fagocitar células epiteliais em meio vaginal. O núcleo N-acetilactosamina de Gal 1 pode mediar a regulação do crescimento celular com a ajuda da proteína GRB2; entretanto, Gal 1 pode contribuir para a supressão da inflamação por meio da indução de apoptose pelas células T ativadas. (AU)

Trichomonas vaginalis is an infectious agent of the vaginal flora which has been associated with cervical cancer. Galectin-1 (Gal 1) is a cell receptor expressed in cervical epithelial cells binding T. vaginalis? lipophosphoglican (LPG). Interaction between T. vaginalis and the epithelial cell is mediated by poly-LacNac domains (galactoside and acetil-lactosamin) and is related to cell adherence as well. Cell signaling occurs by the time Src (SH2) domains are correlated with this interaction and PI3K phosphorilation brings up phosphatidil inositol lipid membranes (PIP and PIP2). T. vaginalis adheres to cytoplasm membrane and secrets specific enzymes that probably lead to membrane rupture. Moreover this parasite may phagocyte epithelial cells in vaginal discharge. Gal 1 nucleus called N-acetil-lactosamin can mediate growth development through GRB2 protein and may contribute to inflammation suppression owing to apoptosis induction of activated T cells.(AU)

Arginine homozygosity in codon 72 of p53 correlates with failure to imatinib response in chronic myeloid leukemia.

PURPOSE: In this study, the role of the polymorphism at codon 72 of tumor protein p53 gene (TP53) was investigated regarding the response to treatment with imatinib in chronic myeloid leukemia (CML). METHODS: A total of 85 patients with CML were treated according to the Brazilian National Cancer Institute (INCA) guidelines and at the end of the 18th month a blood sample were collected for genotyping. Genomic DNA was extracted and TP53 codon 72 genotyping was performed by allele-specific polymerase chain reaction (AS-PCR), which detects argine or proline alleles. RESULT: Of the 85 CML samples, 27 samples were homozygous for arginine (Arg/Arg), 12 homozygous for proline (Pro/Pro) and 46 samples heterozygous (Arg/Pro). TP53 codon 72 polymorphism was in Hardy-Weinberg equilibrium (χ(2)=1.17, P=0.37). We did not find significant association between codon 72 polymorphism and age at diagnosis and sex (P=0.76 and P=0.33, respectively). High Sokal score are significantly associated with Arg/Arg genotype carriers (Odds ratio, OR=4.09; 95% confidence interval, CI 1.01=15.89; P=0.036). The arginine allele in homozygosis also have an increased risk of failure response to imatinib when compared with both, the heterozygous (Arg/Pro) and proline homozygous patients (P=0.021; OR=2.99, 95% CI=1.16-7.67). Additionally, interaction analysis with age at diagnosis revealed that among patients over 40-yr old, Arg/Arg genotype was significantly associated with non-responder patients (P=0.007; OR=5.13, 95% CI=1.5-17.55). CONCLUSION: The findings suggest that in CML patients, TP53 codon 72 polymorphism may contribute to a high Sokal score and failure to imatinib treatment.

In vitro mutagenicity and blood compatibility of paclitaxel and curcumin in poly (DL-lactide-co-glicolide) films.

Curcumin is considered to be a potential component for drug-eluting stents due to its anti-inflammatory properties. In this study we compared the mutagenicity and blood compatibility of curcumin to first generation drug eluting stent components: paclitaxel and sirolimus. The Ames test was used to assess mutagenicity. Blood compatibility was tested by measuring platelet activation and fibrinogen adsorption on poly (DL-lactide-co-glycolide, PLGA) films. We discovered that there was no significant increase in the number of revertants/plate following treatment with curcumin (up to 0.5mg/plate) or sirolimus (up to 0.5 µg/plate). However, a significant induction in the frequency of bacterial his(+) revertant colonies by paclitaxel at concentrations of 0.02, 0.05, 0.1, 0.2 and 0.5 µg/plate was observed. We also discovered a significant reduction in platelet activation by PLGA films containing 30% and 50% by weight curcumin. A similar reduction in platelet activation was also observed for PLGA films containing 1% by weight paclitaxel. In addition, we observed an increase of fibrinogen adsorption to PLGA-films containing curcumin. This would compromise the potential use of curcumin as a component of drug-eluting stents. Moreover, our data challenges the current view that paclitaxel does not significantly induce mutagenesis.

Feeder-free culture of human embryonic stem cell line BG01V/hOG using magnetic field-magnetic nanoparticles system.

PURPOSE: Human embryonic stem (hES) cells are useful tools for regenerative medicine. Maintaining hES cells for research and clinical purposes remains a challenge. The hES cells have typically been grown on a mouse or human cell feeder layer, but these methods harbor potential health problems for the recipient. A culture system using magnetic field and iron oxide nanoparticles were previously demonstrated to sustain mouse embryonic stem cells in vitro. Now, by using the BG01v/hOG cell line, we could assess the effect of this culture system on the stemness of an embryonic stem cell of human origin. METHODS: Using a variant hES cell line, BG01V/hOG, expressing an emerald green fluorescent protein (EmGFP), we grown these cells in the presence of serum-free medium supplemented with magnetic nanoparticles functionalized with citrate. The cells were positioned over a circular magnet (4000 Gauss) and monitored daily by fluorescence microscopy. RESULTS: We discovered that hES cells can proliferate when labeled with magnetic nanoparticles and in the presence of a magnetic field without losing pluripotency. CONCLUSION: These results establish an alternative method for maintaining hES cells which would minimize health concerns as well as label cells for subsequent clinical tracking.

In vitro antiproliferative effects of the indole alkaloid vallesiachotamine on human melanoma cells.

In course of a screening for small molecules presenting potential anticancer properties, a known monoterpene indole alkaloid named vallesiachotamine was isolated from the leaves of Palicourea rigida (Rubiaceae) collected in the Brazilian Cerrado. The structure was determined by spectroscopic methods, mainly 1D- and 2D-NMR and its biological activities were investigated on cultured human (SK-MEL-37) melanoma cells. In vitro cytotoxicity was evaluated by the 3-(4, 5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The inhibitory concentration (IC(50)) was 14.7 ± 1.2 µM for 24 h of drug exposure. Flow cytometry analysis revealed that vallesiachotamine induced G0/G1 arrest and increased the proportion of sub-G1 hypodiploid cells (at 11 µM and 22 µM) and this effect was not dependent on time of incubation. At these concentrations, a typical ladder was observed by agarose gel electrophoresis of the extracted DNA. Treatment of cells with 50 µM vallesiachotamine for 24 h caused extensive cytotoxicity and necrosis. Our results demonstrated that the indole alkaloid vallesiachotamine exhibited important cytotoxicity toward human melanoma cells and that apoptosis and necrosis might be responsible for the observed events.

TP53 codon 72 polymorphism in pigmentary phenotypes.

The p53 protein exerts different cellular functions, and recent findings have demonstrated its influence on the cascade of skin pigmentation during UV exposure. Among TP53 gene polymorphisms, the most studied is the G to C transversion in exon 4 at codon 72, which results in three distinct genotypes, Arg/Arg, Pro/Pro and Arg/Pro, each one encoding different p53 isoforms. Therefore, this study aimed to determine the relationship between TP53 codon 72 polymorphism and skin protection against sunburn. Genomic DNA was extracted from peripheral blood samples and genotyping was performed by PCR and confirmed by restriction enzyme digestion. The genotype frequency was 50% for Arg/Arg and 14.6% for Pro/Pro genotype. The frequency of heterozygous subjects was 35.4%. In our population, p53 genotypes were in Hardy-Weinberg (HW) equilibrium (X2 HM less than 3.84), showing a predominance of arginine allele (total Arg allele frequency of 68%). No significant association between p53 genotype and skin colour, hair or eye colour and susceptibility to sun exposure was found. However, further analysis demonstrated a significant association between the genotype Pro/Pro and blue/green eyes among participants who presented redness (P=0.016). Our findings indicate susceptibility to sun exposure when this phenotype (eye colour) occurs simultaneously with Pro/Pro genotype.

Curcumin associated magnetite nanoparticles inhibit in vitro melanoma cell growth.

Curcumin is a natural product possessing therapeutic properties but the low water solubility of this compound limits its use. We have successfully incorporated curcumin into a bilayer of dodecanoic acid attached to magnetite nanoparticles in an effort to maximize solubility and delivery efficiency. Curcumin/magnetite nanoparticles were characterized using diffused reflectance infra-red fourier transform spectroscopy (DRIFTS) and X-ray powder diffraction (XRD). Moreover curcumin associated magnetite nanoparticles inhibited in vitro melanoma cell growth. An inhibitory concentration (IC50) of 66.0 +/- 3.0 microM (48 +/- 2.2 microg-iron/mL) was observed for the curcumin/magnetite nanoparticles. Fluorescent microscopy revealed that curcumin associated magnetite nanoparticles were internalized by the melanoma cells and remained in the cytoplasm. The curcumin/magnetic nanoparticles synthesized in this study possess magnetic and water solubility properties making this a novel curcumin formulation with therapeutic potential.

Embryonic stem-like cells derived from in vitro produced bovine blastocysts

The aim of this work was to study the derivation of bovine embryonic stem-like (ES-like) cells from the inner cell mass (ICM) of in vitro produced blastocysts. The ICMs were mechanically isolated and six out of seventeen (35 percent) ICMs could attach to a monolayer of murine embryonic fibroblasts (MEF). Ten days after, primary outgrowths were mechanically dissected into several small clumps and transferred to a new MEF layer. Cells were further propagated and passaged by physical dissociation over a 60 days period. The pluripotency of the bovine ES-like cells was confirmed by RT-PCR of Oct-4 and STAT-3 gene markers. The colonies were weakly stained for alkaline phosphatase and the mesoderm and endoderm differentiation gene markers such as GATA-4 and Flk-1, respectively, were not expressed. Embryoid bodies were spontaneously formed at the seventh passage. Results showed that bovine ES-like cells could be obtained and passaged by mechanical procedures from the fresh in vitro produced blastocysts.

Magnetic field-magnetic nanoparticle culture system used to grow in vitro murine embryonic stem cells.

The in vitro growth of embryonic stem cells (ESCs) is usually obtained in the presence of murine embryonic fibroblasts (MEF), but new methods for in vitro expansion of ESCs should be developed due to their potential clinical use. This study aims to establish a culture system to expand and maintain ESCs in the absence of MEF by using murine embryonic stem cells (mECS) as a model of embryonic stem cell. Magnetic nanoparticles (MNPs) were used for growing mESCs in the presence of an external magnetic field, creating the magnetic field-magnetic nanoparticle (MF-MNP) culture system. The growth characteristics were evaluated showing a doubling time slightly higher for mESCs cultivated in the presence of the system than in the presence of the MEF. The undifferentiated state was characterized by RT-PCR, immunofluorescence, alkaline phosphatase activity and electron microscopy. Murine embryonic stem cells cultivated in presence of the MF-MNP culture system exhibited Oct-4 and Nanog expression and high alkaline phosphatase activity. Ultrastructural morphology showed that the MF-MNP culture system did not interfere with processes that cause structural changes in the cytoplasm or nucleus. The MF-MNP culture system provides a tool for in vitro expansion of mESCs and could contribute to studies that aim the therapeutic use of embryonic stem cells.

Morphological alterations and G0/G1 cell cycle arrest induced by curcumin in human SK-MEL-37 melanoma cells

The aim of this work was to study the effect of curcumin on cell cycle in the human SK-MEL-37 melanoma cell line. In addition, morphological and structural analyses were also performed. Flow cytometric analysis showed a G0/G1 arrest at 5 µM after 24 h exposure and a concentration-dependent increase in the proportion of sub-G0 hypodiploid cells. Typical apoptotic events were also observed by the fluorescence microscopy, transmission and scanning electronic microscopy. Loss of mitochondrial membrane potential was not detected. Results suggested that curcumin could arrest human melanoma cells at G0/G1 phase and induce a mitochondrial-independent apoptotic pathway.

O melanoma é um tipo agressivo de câncer cujo tratamento culmina com o estabelecimento de resistência aos quimioterápicos empregados. Portanto, é importante o desenvolvimento de novos agentes farmacológicos que sejam menos tóxicos e que não provoquem quimiorresistência. As inúmeras propriedades terapêuticas da curcumina vêm sendo confirmadas através de estudos sobre o seu mecanismo de ação em células cultivadas. No presente estudo, empregamos células de melanoma humano da linhagem SK-MEL-37, que desenvolveram resistência in vitro à doxorubicina e cisplatina, drogas normalmente utilizadas na clínica. Investigamos o efeito da curcumina sobre o ciclo celular através de citometria de fluxo. Além disso, análises morfológicas e estruturais também foram realizadas. Os resultados demonstraram que o tratamento com uma concentração de 5 ?M de curcumina provocou uma parada na subfase G0/G1. Além disso, observou-se um aumento dose-dependente na proporção de células hipodiplóides em sub-G0. Eventos apoptóticos típicos foram observados por microscopia de fluorescência, microscopia eletrônica de transmissão e microscopia eletrônica de varredura. Não foi detectada alteração no potencial de membrana mitocondrial. Os resultados indicam que futuros estudos poderão tornar possível a utilização da curcumina como um modulador para agentes quimioterápicos empregados na clínica no tratamento do melanoma.