ANRIL, H19 and TUG1: a review about critical long non-coding RNAs in cardiovascular diseases

Cardiovascular diseases are the leading cause of death worldwide. They are non-transmissible diseases that affect the cardiovascular system and have different etiologies such as smoking, lipid disorders, diabetes, stress, sedentary lifestyle and genetic factors. To date, lncRNAs have been associated with increased susceptibility to the development of cardiovascular diseases such as hypertension, acute myocardial infarction, stroke, angina and heart failure. In this way, lncRNAs are becoming a very promising point for the prevention and diagnosis of cardiovascular diseases. Therefore, this review highlights the most important and recent discoveries about the mechanisms of action of the lncRNAs ANRIL, H19 and TUG1 and their clinical relevance in these pathologies. This may contribute to early detection of cardiovascular diseases in order to prevent the pathological phenotype from becoming established.

Variante da Subunidade Beta 3 da Proteína G ( GNB3 ) Está Associada a Alterações Bioquímicas em Pacientes Brasileiros com Hipertensão / G Protein Subunit Beta 3 (GNB3) Variant Is Associated with Biochemical Changes in Brazilian Patients with Hypertension

Resumo Fundamento Genes e suas variantes associadas a fatores ambientais contribuem para o desenvolvimento do fenótipo hipertenso. O gene da subunidade beta 3 da proteína G ( GNB3 ) está envolvido no processo de sinalização intracelular e suas variantes têm sido relacionadas à suscetibilidade à hipertensão arterial. Objetivo Determinar a associação da variante GNB3 (rs5443:C>T) com a hipertensão arterial, parâmetros bioquímicos, idade e obesidade em indivíduos hipertensos e normotensos de Ouro Preto, Minas Gerais. Método A identificação das variantes foi realizada por PCR em tempo real, utilizando o sistema TaqMan®, em amostras de 310 pacientes (155 hipertensos e 155 normotensos). Análises bioquímicas (função renal, perfil lipídico e glicemia) foram realizadas a partir do soro por meio de espectrofotometria UV/Vis e eletrodo íon-seletivo. Foi utilizado um modelo de regressão logística múltipla para identificar fatores associados à hipertensão arterial. A análise das variáveis contínuas com distribuição normal foi realizada usando o teste t de Student não pareado; dados não normais foram analisados usando o teste de Mann-Whitney. Valores de p < 0,05 foram considerados significativos. Resultados A variante rs5443:C>T não esteve associada à hipertensão arterial na população avaliada (p = 0,88). Em relação às medidas bioquímicas, o alelo T esteve associado a níveis elevados de triglicerídeos, glicose e ácido úrico em indivíduos hipertensos (p < 0,05). Conclusão Os presentes resultados mostram a importância do diagnóstico genético para prevenir as causas e consequências de doenças e sugerem que a variante GNB3 rs5443:C>T pode estar associada a alterações no perfil bioquímico em indivíduos hipertensos.

Abstract Background Genes and their variants associated with environmental factors contribute to the development of the hypertensive phenotype. The G protein beta 3 subunit gene (GNB3) is involved in the intracellular signaling process, and its variants have been related to susceptibility to arterial hypertension. Objective To determine the association of the GNB3 variant (rs5443:C>T) with arterial hypertension, biochemical parameters, age, and obesity in hypertensive and normotensive individuals from Ouro Preto, Minas Gerais, Brazil. Method The identification of variants was performed by real-time PCR, using the TaqMan® system, in 310 samples (155 hypertensive and 155 normotensive). Biochemical analyses (renal function, lipid profile and glycemia) were performed from the serum using UV/Vis spectrophotometry and ion-selective electrode. A multiple logistic regression model was used to identify factors associated with arterial hypertension. The analysis of continuous variables with normal distribution was performed using the unpaired Student's t test; non-normal data were analyzed using Mann-Whitney. P < 0.05 was considered significant. Results The rs5443:C>T variant was not associated with arterial hypertension in the evaluated population (p = 0.88). Regarding biochemical measures, the T allele was associated with high levels of triglycerides, glucose and uric acid in hypertensive individuals (p < 0.05). Conclusion These results show the importance of genetic diagnosis to prevent the causes and consequences of diseases and imply that the GNB3 rs5443:C>T variant may be associated with changes in the biochemical profile in hypertensive individuals.

G Protein Subunit Beta 3 (GNB3) Variant Is Associated with Biochemical Changes in Brazilian Patients with Hypertension. / Variante da Subunidade Beta 3 da Proteína G ( GNB3 ) Está Associada a Alterações Bioquímicas em Pacientes Brasileiros com Hipertensão.

BACKGROUND: Central Illustration : G Protein Subunit Beta 3 (GNB3) Variant Is Associated with Biochemical Changes in Brazilian Patients with Hypertension. BACKGROUND: Genes and their variants associated with environmental factors contribute to the development of the hypertensive phenotype. The G protein beta 3 subunit gene (GNB3) is involved in the intracellular signaling process, and its variants have been related to susceptibility to arterial hypertension. OBJECTIVE: To determine the association of the GNB3 variant (rs5443:C>T) with arterial hypertension, biochemical parameters, age, and obesity in hypertensive and normotensive individuals from Ouro Preto, Minas Gerais, Brazil. METHOD: The identification of variants was performed by real-time PCR, using the TaqMan® system, in 310 samples (155 hypertensive and 155 normotensive). Biochemical analyses (renal function, lipid profile and glycemia) were performed from the serum using UV/Vis spectrophotometry and ion-selective electrode. A multiple logistic regression model was used to identify factors associated with arterial hypertension. The analysis of continuous variables with normal distribution was performed using the unpaired Student's t test; non-normal data were analyzed using Mann-Whitney. P < 0.05 was considered significant. RESULTS: The rs5443:C>T variant was not associated with arterial hypertension in the evaluated population (p = 0.88). Regarding biochemical measures, the T allele was associated with high levels of triglycerides, glucose and uric acid in hypertensive individuals (p < 0.05). CONCLUSION: These results show the importance of genetic diagnosis to prevent the causes and consequences of diseases and imply that the GNB3 rs5443:C>T variant may be associated with changes in the biochemical profile in hypertensive individuals.

Design and synthesis of new 1,2,3-triazoles derived from eugenol and analogues with in vitro and in vivo activity against Trypanosoma cruzi.

Chagas disease (CD) is a neglected tropical disease endemic in 21 countries and affects about 8 million people around the world. The pharmacotherapy for this disease is limited to two drugs (Benznidazole and Nifurtimox) and both are associated with important limitations, as low cure rate in the chronic phase of the disease, high toxicity and increasing resistance by Trypanosoma cruzi. Recently, we reported a bioactive 1,2,3-triazole (compound 35) active in vitro (IC50 42.8 µM) and in vivo (100 mg/kg) against T. cruzi Y strains and preliminary in silico studies suggested the cysteine protease cruzain as a possible target. Considering these initial findings, we describe here the design and synthesis of new 1,2,3-triazoles derivatives of our hit compound (35). The triazoles were initially evaluated against healthy cells derived from neonatal rat cardiomyoblasts (H9c2 cells) to determine their cytotoxicity and against epimastigotes forms of T. cruzi Y strain. The most active triazoles were compounds 26 (IC50 19.7 µM) and 27 (IC50 7.3 µM), while benznidazole was active at 21.6 µM. Derivative 27 showed an interesting selectivity index considering healthy H9c2 cells (>77). Promising activities against trypomastigotes forms of the parasite were also observed for triazoles 26 (IC50 20.74 µM) and 27 (IC50 8.41 µM), mainly 27 which showed activity once again higher than that observed for benznidazole (IC50 12.72 µM). While docking results suggested cruzain as a potential target for these compounds, no significant enzyme inhibition was observed in vitro, indicating that their trypanocidal activity is related to another mode of action. Considering the promising in vitro results of triazoles 26 and 27, the in vivo toxicity was initially verified based on the evaluation of behavioral and physiological parameters, mortality, effect in body weight gain, and through the measurement of AST/ALT enzymes, which are markers of liver toxicity. All these evaluations pointed to a good tolerability of the animals, especially considering triazole 27. A reduction in parasitemia was observed among animals treated with triazole 27, but not among those treated with derivative 26. Regarding the dosage, derivative 27 (100 mg/kg) was the most active sample against T. cruzi infection, showing a 99.4% reduction in parasitemia peak. Triazole 27 at a dosage of 100 mg/kg influenced the humoral immune response and reduced myocarditis in the animals, bringing antibody levels closer to those observed among healthy mice. Altogether, our results indicate compound 27 as a new lead for the development of drug candidates to treat Chagas disease.

Synthesis, characterization and antiproliferative effects of naphtho [2,3-b] thiophen-4,9-quinone on bladder tumor cells.

Naphthoquinones are natural plants products or synthesized compounds. They have α, ß-cyclic aromatic dienones structure with a naphthalene skeleton. Little is known about naphthoquinone and nothing about naphtho [2,3-b] thiophen-4,9-quinone effects on bladder cancer. In this study, a naphthoquinone containing a hetero sulfur atom was synthesized using classical synthetic method. The molecular structure was elucidated by NMR techniques and the antitumor effects were evaluated on bladder tumor cell lines with different TP53 status using tripan blue and MTT cytotoxic method, quantification of reactive oxygen species (ROS), wound healing, cell morphology and cell cycle progression assays. The results showed selective cytotoxicity, colonies reduction, morphological change, inhibition of the cell migration process, induction of ROS production and cell cycle arrest. Naphtho [2,3-b] thiophen-4,9-quinone presents antiproliferative activity regardless TP53 status and may be a promising agent in the treatment of bladder cancer, as they have an oxidizing effect and interfere with cell cycle.

LncRNA JHDM1D-AS1 is a key biomarker for progression and modulation of gemcitabine sensitivity in bladder cancer cells

Long non-coding RNAs are frequently found to be dysregulated and are linked to carcinogenesis, aggressiveness, and chemoresistance in a variety of tumors. As expression levels of the JHDM1D gene and lncRNA JHDM1D-AS1 are altered in bladder tumors, we sought to use their combined expression to distinguish between low-and high-grade bladder tumors by RTq-PCR. In addition, we evaluated the functional role of JHDM1D-AS1 and its association with the modulation of gemcitabine sensitivity in high-grade bladder-tumor cells. J82 and UM-UC-3 cells were treated with siRNA-JHDM1D-AS1 and/or three concentrations of gemcitabine (0.39, 0.78, and 1.56 µM), and then submitted to cytotoxicity testing (XTT), clonogenic survival, cell cycle progression, cell morphology, and cell migration assays. When JHDM1D and JHDM1D-AS1 expression levels were used in combination, our findings indicated favorable prognostic value. Furthermore, the combined treatment resulted in greater cytotoxicity, a decrease in clone formation, G0/G1 cell cycle arrest, morphological alterations, and a reduction in cell migration capacity in both lineages compared to the treatments alone. Thus, silencing of JHDM1D-AS1 reduced the growth and proliferation of high-grade bladder-tumor cells and increased their sensitivity to gemcitabine treatment. In addition, the expression of JHDM1D/JHDM1D-AS1 indicated potential prognostic value in the progression of bladder tumors.

Naringin: antitumor potential in silico and in vitro on bladder cancer cells

Introduction: Urothelial carcinoma is a significant public health problem. Transitional cell carcinoma (TCC) is the most common subtype, accounting for approximately 90 % of all bladder cancers. Chemotherapeutic protocols have been studied, but some present high toxicity and low tolerability. Naringin is a polyphenolic compound found mainly in citrus fruits, which antitumor activity has been studied in several types of cancer. However, there is little information about naringin effects on bladder cancer. This study aimed to evaluate the antitumor potential of naringin in silico and in vitro using two bladder cancer cell lines. Method: In silico analysis was carried out by PASS Online software. In vitro , the effects of naringin treatment (12.5 - 400 µM) were evaluated regarding its cytotoxicity, clonogenic survival, morphological alterations, cell cycle progression, migration, and mutagenicity Results: In silico analyses predicted antitumor activity through several mechanisms of action. In vitro results showed naringin presented cytotoxic effects, reduced the number of colonies, inhibited cell migration, and changed the morphology and cell cycle progression of the two cell lines evaluated. However, naringin did not present mutagenic effects. Conclusions: Naringin has antiproliferative activity and is a promising candidate for bladder cancer treatment.(AU)

Introducción: El carcinoma urotelial es un problema de salud pública importante. El carcinoma de células de transición es el subtipo más común y representa aproximadamente el 90 % de todos los cánceres de vejiga. Se han estudiado protocolos quimioterapéuticos, pero algunos presentan alta toxicidad y baja tolerabilidad. La naringina es un compuesto polifenólico que se encuentra principalmente en los cítricos, cuya actividad antitumoral se ha estudiado en varios tipos de cáncer. Sin embargo, hay poca información sobre los efectos de la naringina en el cáncer de vejiga. Este estudio tuvo como objetivo evaluar el potencial antitumoral de la naringina in silico e in vitro utilizando dos líneas celulares de cáncer de vejiga. Método: El análisis in silico se llevó a cabo mediante el software PASS Online. In vitro, se evaluaron los efectos del tratamiento con naringina (12,5 - 400 µM) en cuanto a su citotoxicidad, supervivencia clonogénica, alteraciones morfológicas, progresión del ciclo celular, migración y mutagenicidad. Resultados: los análisis in silico predijeron la actividad antitumoral a través de varios mecanismos de acción. Los resultados in vitro mostraron que la naringina presentó efectos citotóxicos, redujo el número de colonias, inhibió la migración celular y cambió la morfología y la progresión del ciclo celular de las dos líneas celulares evaluadas. Sin embargo, la naringina no presentó efectos mutagénicos. Conclusiones: la naringina tiene actividad antiproliferativa y es un candidato prometedor para el tratamiento del cáncer de vejiga.(AU)

Effects and implications of the COVID-19 pandemic on medicine use by employees of a Brazilian public university: a cross-sectional study.

BACKGROUND: During the COVID-19 pandemic, universities have had to adopt remote education, a strategy that caused sudden changes of routine for everyone involved in academia. OBJECTIVE: To assess the profile of medicine use by the employees of a Brazilian public university during the COVID-19 pandemic. DESIGN AND SETTING: Cross-sectional study at a Brazilian public university. METHODS: Employees were invited to answer an online self-administered questionnaire, containing questions on sociodemographic features, medicine use, mental health and lifestyle habits during the COVID-19 pandemic. The outcome variable was the use of medicines stratified according to occupation. Descriptive, bivariate and multivariate (Poisson regression) statistical analyses were performed. RESULTS: A total of 372 employees participated in the study and use of medicine was reported by 53.2%. Among professors, suicide attempts (prevalence ratio [PR], 1.81; 95% confidence interval [CI], 1.20-2.74), physical activity (PR, 1.53; 95% CI, 1.11-2.11) and poor self-rated health (PR, 1.29; 95% CI, 1.01-1.66); and among technicians, decreased workload during the COVID-19 pandemic (PR, 1.41; 95% CI, 1.00-1.99), excess body weight (PR, 1.39; 95% CI, 1.02-1.88) and poor self-rated health (PR, 1.48; 95% CI, 1.14-1.92) were positively associated with use of medicines. In addition, among technicians, engaging in physical activity (PR, 0.60; 95% CI, 0.46-0.78) was a protective factor against medicine use. CONCLUSION: The profile of medicine use among these employees was similar to that of the Brazilian population. However, some associated factors may have been influenced by the COVID-19 pandemic, thus highlighting the need to examine this topic in a longitudinal study.

Effects and implications of the COVID-19 pandemic on medicine use by employees of a Brazilian public university: a cross-sectional study

Abstract BACKGROUND: During the COVID-19 pandemic, universities have had to adopt remote education, a strategy that caused sudden changes of routine for everyone involved in academia. OBJECTIVE: To assess the profile of medicine use by the employees of a Brazilian public university during the COVID-19 pandemic. DESIGN AND SETTING: Cross-sectional study at a Brazilian public university. METHODS: Employees were invited to answer an online self-administered questionnaire, containing questions on sociodemographic features, medicine use, mental health and lifestyle habits during the COVID-19 pandemic. The outcome variable was the use of medicines stratified according to occupation. Descriptive, bivariate and multivariate (Poisson regression) statistical analyses were performed. RESULTS: A total of 372 employees participated in the study and use of medicine was reported by 53.2%. Among professors, suicide attempts (prevalence ratio [PR], 1.81; 95% confidence interval [CI], 1.20-2.74), physical activity (PR, 1.53; 95% CI, 1.11-2.11) and poor self-rated health (PR, 1.29; 95% CI, 1.01-1.66); and among technicians, decreased workload during the COVID-19 pandemic (PR, 1.41; 95% CI, 1.00-1.99), excess body weight (PR, 1.39; 95% CI, 1.02-1.88) and poor self-rated health (PR, 1.48; 95% CI, 1.14-1.92) were positively associated with use of medicines. In addition, among technicians, engaging in physical activity (PR, 0.60; 95% CI, 0.46-0.78) was a protective factor against medicine use. CONCLUSION: The profile of medicine use among these employees was similar to that of the Brazilian population. However, some associated factors may have been influenced by the COVID-19 pandemic, thus highlighting the need to examine this topic in a longitudinal study.

Modulation of non-coding RNAs by natural compounds as a potential therapeutical approach in oral cancer: a comprehensive review

Oral cancer is a disease with high incidence and mortality worldwide, and its treatment still needs to be improved. The search for new therapies using natural products is strongly supported, given the wide chemical range of these compounds. In addition, phytochemicals can exert antitumor activities by several mechanisms of action, including the modulation of non-coding RNAs. Thus, in this review, we discussed the role of non-coding RNAs, including circular RNAs, microRNAs, and long non-coding RNAs, in oral cancer and presented their potential as treatment targets using natural products. Some natural products capable of being used to treat oral cancer have been suggested.

Resveratrol effects in bladder cancer: A mini review.

Bladder cancer has a high incidence worldwide and is the most common genitourinary cancer. The treatment of bladder cancer involves surgery and chemotherapy; however high failure rates and toxicity are observed. In this context, the search of new drugs aiming a more effective treatment is extremely necessary. Natural products are an important source of compounds with antiproliferative effects. Resveratrol is a naturally occurring plant polyphenol whose anticancer activity has been demonstrated in different types of cancer. This review summarizes the in vitro and in vivo studies using models of bladder cancer treated with resveratrol and discusses its different mechanisms of action.

Vancomycin-loaded N,N-dodecyl,methyl-polyethylenimine nanoparticles coated with hyaluronic acid to treat bacterial endophthalmitis: Development, characterization, and ocular biocompatibility.

Vancomycin-loaded N,N-dodecyl,methyl-polyethylenimine nanoparticles coated with hyaluronic acid (VCM-DMPEI nanoparticles/HA) were synthesized as an adjuvant for the treatment of bacterial endophthalmitis. The nanoparticles were formulated by experimental statistical design, thoroughly characterized, and evaluated in terms of bactericidal activity and both in vitro and in vivo ocular biocompatibility. The VCM-DMPEI nanoparticles/HA were 154 ± 3 nm in diameter with a 0.197 ± 0.020 polydispersity index; had a + 26.4 ± 3.3 mV zeta potential; exhibited a 93% VCM encapsulation efficiency; and released 58% of the encapsulated VCM over 96 h. VCM and DMPEI exhibited a synergistic bactericidal effect. The VCM-DMPEI nanoparticles/HA were neither toxic to ARPE-19 cells nor irritating to the chorioallantoic membrane. Moreover, the VCM-DMPEI nanoparticles/HA did not induce modifications in retinal functions, as determined by electroretinography, and in the morphology of the ocular tissues. In conclusion, the VCM-DMPEI nanoparticles/HA may be a useful therapeutic adjuvant to treat bacterial endophthalmitis.

Bacterial vaginosis: prevalence, risk profile and association with sexually transmitted infections / Vaginose bacteriana: prevalência, perfil de risco e associação com infecções sexualmente transmissíveis / Vaginosis bacteriana: prevalencia, perfil de riesgo y asociación con infecciones de transmisión sexual

Background and Objectives: Bacterial vaginosis (BV) is the most common cause of vaginal discharge in the world. The study aimed to estimate the prevalence and to identify risk factors associated with bacterial vaginosis. Methods: A cross-sectional study was conducted in Ouro Preto, Brazil, between February and December 2017. Three hundred and forty-one women aged 18 years or older, users of the Brazilian Unified Health System, participated in this study. Women who used oral or topical antibiotics in the four weeks prior to the sample collection and women who had undergone a total hysterectomy were excluded from the study. After signing the Informed Consent Form and filling out a questionnaire containing sociodemographic, behavioral and sexual data, the participants were directed to the collection room, where the nurse collected the samples for the preventive examination of the cervix and also two vaginal swabs. Vaginal swabs and cervical samples were analyzed for cytological abnormalities and BV using Gram staining and cytology. Pathogens causing sexually transmitted infections (STIs) were identified by Polymerase Chain Reaction (PCR). For the analysis of the data, statistical package STATA version 10.0 was used. This study was approved by the Research Ethics Committee of the Federal University of Ouro Preto (UFOP). Results: During the study, 341 women were evaluated. The prevalence of BV using Gram staining (32.5% [CI95% 27.7-37.7%]) and cytology (27.7% [CI95% 23.032.8%]) was similar, however, the sensitivity of cytology was lower (77.8%). Risk factors associated with BV were smoking (IRR 1.5 [CI95%: 1.1  2.1]), use of an intrauterine device (IRR 2.8 [CI95%: 1.2 - 6.5]), and past medical history of BV (IRR 1.5 [CI95%: 1.1 - 2.1]). Correlation between the presence of BV and Trichomonas vaginalis (TV) infection (r=0.24) was observed. Conclusion: The prevalence of BV was affected by life habits and was prevalent in women with TV. Thus, behavioral and social prevention approaches to women with diverse risk profiles may help mitigate TV/BV prevalence and recurrence of BV.(AU)

Contexte et objectifs: La vaginose bactérienne (VB) est la cause la plus fréquente de pertes vaginales dans le monde. Le but de cette étude était d'évaluer la prévalence et les facteurs associés à la vaginose bactérienne. Méthodes: Il s'agit d'une approche descriptive, transversale et quantitative réalisée à Ouro Preto, Minas Gerais, Brésil, entre février et décembre 2017. 341 femmes ont participé à cette étude, âgées de 18 ans ou plus, utilisatrices du Système de santé unifié. Les femmes ayant utilisé des antibiotiques oraux ou topiques dans les quatre semaines précédant le prélèvement et les femmes ayant subi une hystérectomie totale ont été exclues de l'étude. Après avoir signé le formulaire de consentement éclairé et rempli un questionnaire contenant des données sociodémographiques, comportementales et sexuelles, les participants ont été dirigés vers la salle de collecte, où l'infirmière a prélevé les échantillons pour l'examen préventif du col de l'utérus. et aussi deux écouvillons vaginaux. Les échantillons de frottis vaginaux et cervicaux ont été analysés pour les anomalies cytologiques et VB en utilisant la coloration de Gram et la cytologie. Les agents pathogènes causant des infections sexuellement transmissibles (IST) ont été identifiés par réaction en chaîne par polymérase. Pour l'analyse des données, le progiciel statistique STATA version 10.0 a été utilisé. Cette étude a été approuvée par le Comité d'éthique de la recherche de l'Université fédérale d'Ouro Preto (UFOP). Résultats: Au cours de l'étude, 341 femmes ont été évaluées. La prévalence de la VB avec coloration de Gram (32,5% [IC 95% 27,7 - 37,7%]) et de la cytologie (27,7% [IC 95% 23,0 - 32,8%]) était similaire, cependant la sensibilité cytologique était plus faible (77,8%). Les facteurs de risque associés à la VB étaient le tabagisme (IRR 1,5 [IC 95%: 1,1 - 2,1]), l'utilisation d'un dispositif intra-utérin (IRR 2,8 [IC 95%: 1,2 - 6,5] ) et antécédents médicaux de VB (IRR 1,5 [IC 95%: 1,1 - 2,1]). Il y avait une corrélation entre la présence d'une infection à VB et Trichomonas vaginalis (TV) (r = 0,24). Conclusion: La prévalence de la VB était affectée par le mode de vie et l'infection TV. Ainsi, les approches de prévention comportementale et sociale pour les femmes présentant des profils de risque différents peuvent aider à atténuer la prévalence de la TV / VB et la récurrence de la VB.(AU)

Justificativa e Objetivos: A vaginose bacteriana (VB) é a causa mais comum de corrimento vaginal no mundo. O objetivo desse estudo foi avaliar a prevalência e os fatores associados à vaginose bacteriana. Métodos: Trata-se de um descritivo, de forma transversal e abordagem quantitativa realizado em Ouro Preto, Minas Gerais, Brasil, entre fevereiro a dezembro de 2017. Participaram desse estudo 341 mulheres com idade igual ou superior a 18 anos, usuárias do Sistema Único de Saúde (SUS). Mulheres que usaram antibióticos orais ou tópicos nas quatro semanas anteriores à coleta e mulheres que haviam sido submetidas a uma histerectomia total foram excluídas do estudo. Após a assinatura do Termo de Consentimento Livre e Esclarecido e preenchimento de questionário contendo dados sócio-demográfico, comportamental e sexual, as participantes foram encaminhadas para a sala de coleta, onde a enfermeira realizou a coleta das amostras para o exame preventivo do colo do útero e também de dois swabs vaginais. As amostras de esfregaço vaginal e cervical foram analisadas quanto às anormalidades citológicas e VB usando coloração de Gram e citologia. Patógenos causadores de infecções sexualmente transmissíveis (ISTs) foram identificados por Reação em Cadeia da Polimerase (PCR). Para a análise dos dados foi utilizado o pacote estatístico STATA versão 10.0. O presente estudo foi aprovado pelo Comitê de Ética em Pesquisa da Universidade Federal de Ouro Preto (UFOP). Resultados: Durante o estudo, 341 mulheres foram avaliadas. A prevalência de VB com coloração de Gram (32,5% [IC95% 27,7 - 37,7%]) e citologia (27,7% [IC95% 23,0 - 32,8%]) foi semelhante, porém a sensibilidade da citologia foi menor (77,8%). Os fatores de risco associados ao VB foram tabagismo (IRR 1,5 [IC95%: 1,1 - 2,1]), uso de dispositivo intrauterino (IRR 2,8 [IC 95%: 1,2 - 6,5]) e história médica pregressa de VB (IRR 1,5 [IC95%: 1,1 - 2.1]). Observou-se correlação entre a presença de infecção por VB e Trichomonas vaginalis (TV) (r = 0,24). Conclusão: A prevalência de VB foi afetada por hábitos de vida e infecção por TV. Assim, abordagens de prevenção comportamental e social para mulheres com diversos perfis de risco podem ajudar a mitigar a prevalência de TV / VB e recorrência de VB.(AU)

Polymeric micelles containing resveratrol: development, characterization, cytotoxicity on tumor cells and antimicrobial activity

Antimicrobial and antitumor activities of resveratrol, a compound found mainly in grapes, have already been demonstrated. However, its low bioavailability is a limiting factor for therapeutic application. Polymeric micelles can be an approach to solve this problem since they can encapsulate hydrophobic substances. We developed and characterized micellar formulations containing resveratrol and evaluated their cytotoxic and antimicrobial effects. The formulations were prepared by the cold dispersion method with different concentrations of F127 (5 or 10% w/w) and resveratrol (500 or 5000 µM). The formulations were characterized according to size, polydispersity index, pH, encapsulation rate and in vitro release. Cytotoxic effect was evaluated on a bladder cancer cell line and antimicrobial effect was evaluated on E. coli, S. aureus and C. albicans. One of the formulations (10% w/w of F127 and 5000 µM of resveratrol) was a monodispersed solution with high encapsulation rate, thus it was chosen for the cytotoxicity and antimicrobial assays. MS- 10+RES-3 was able to preserve the antimicrobial and cytotoxic activity of resveratrol. This is the first study that evaluated antimicrobial potential and cytotoxicity of micelles containing resveratrol on bladder cancer cells and the results showed that micellar nanostructures could ensure the maintenance of the biological activity of resveratrol.

Antibacterial substances from leaves of Protium spruceanum (Burseraceae): in vitro and in silico evaluation

Due to the increase of bacterial resistance, the search for new antibiotics is necessary and the medicinal plants represent its most important source. The aim of this study was to evaluate the antibacterial property of extract and fractions from Protium spruceanum leaves, against pathogenic bacteria. By means of diffusion and microdilution assays, the crude extract was active against the nine bacteria tested being the hydromethanolic fraction the most active. During phytochemical procedures, procyanidin (1) and catechin (2) were identified as the main antibacterial constituents of this fraction. In silico results obtained using PASSonline tool indicated 1 and 2 as having good potential to interact with different targets of currently used antibiotics. These results no indicated potential to none DNA effect and indicated the cell wall as mainly target. Electrophoresis result supported that had no DNA damage. Cell wall damage was confirmed by propidium iodide test that showed increased membrane permeability and by cell surface deformations observed in scanning electronic microscopy. The in vitro assays together with the in silico prediction results establish the potential of P. spruceanum as source of antibacterial compounds that acts on important bacterial targets. These results contribute to the development of natural substances against pathogenic bacteria and to discovery of new antibiotics.

Increase of binucleated cells in the oral mucosa: a study on the use of psychotropics by students of a Brazilian institution.

This study aimed to assess the prevalence of medical and nonmedical use of psychiatric medication among undergraduate students of health sciences from a public university in Brasil. Another objective was to determine the frequency of nuclear morphological abnormalities in the buccal mucosa of students using psychiatric drugs. A cross-sectional study based on a Web survey was carried out with 375 health sciences undergraduate students from schools of Pharmacy, Physical Education, Nutrition, and Medicine. Additionally, spontaneous genetic damages in exfoliated cells of the buccal mucosa of 41 individuals by counting micronucleus (MN) and binucleated (BN) cells frequencies were evaluated. The results showed 76 (20.3%) of students reported the use of psychotropic drugs after enrolling in university. The majority of these students were from Pharmacy and Medicine programs, females, aged between 18-25 years old, nonsmokers, alcohol addicts, and with a family history of mental illness. In addition, Medical students, individuals with high-income, who live alone and are in the last period of the program are more likely to use psychotropic drugs. Moreover, exposure to psychiatric medication was able to increase the number of binucleated cells. These results provide evidence that the use of psychoactive drugs is increased in the academic context and may be related to the failure of the cell cycle.

Increase of binucleated cells in the oral mucosa: a study on the use of psychotropics by students of a Brazilian institution

SUMMARY This study aimed to assess the prevalence of medical and nonmedical use of psychiatric medication among undergraduate students of health sciences from a public university in Brasil. Another objective was to determine the frequency of nuclear morphological abnormalities in the buccal mucosa of students using psychiatric drugs. A cross-sectional study based on a Web survey was carried out with 375 health sciences undergraduate students from schools of Pharmacy, Physical Education, Nutrition, and Medicine. Additionally, spontaneous genetic damages in exfoliated cells of the buccal mucosa of 41 individuals by counting micronucleus (MN) and binucleated (BN) cells frequencies were evaluated. The results showed 76 (20.3%) of students reported the use of psychotropic drugs after enrolling in university. The majority of these students were from Pharmacy and Medicine programs, females, aged between 18-25 years old, nonsmokers, alcohol addicts, and with a family history of mental illness. In addition, Medical students, individuals with high-income, who live alone and are in the last period of the program are more likely to use psychotropic drugs. Moreover, exposure to psychiatric medication was able to increase the number of binucleated cells. These results provide evidence that the use of psychoactive drugs is increased in the academic context and may be related to the failure of the cell cycle.

RESUMO O objetivo deste estudo foi avaliar a prevalência de uso médico e não médico de medicação psiquiátrica entre estudantes de graduação em Ciências da Saúde de uma universidade pública do Brasil. Outro objetivo foi determinar a frequência de anormalidades nucleares morfológicas na mucosa bucal desses estudantes. Realizou-se um estudo transversal baseado na Web com 375 estudantes de graduação em Ciências da Saúde das escolas de Farmácia, Educação Física, Nutrição e Medicina. Adicionalmente, danos genéticos espontâneos em células esfoliadas da mucosa bucal de 41 indivíduos foram analisados para estudo da frequência de micronúcleos e células binucleadas. Os resultados mostraram que 76 (20,8%) dos estudantes relataram que usaram drogas psicotrópicas após se matricularem na universidade. O uso desses medicamentos foi maior entre estudantes de cursos de Farmácia e Medicina, sexo feminino, idade entre 18 e 25 anos, não tabagistas, alcoolistas e histórico familiar de doença mental. Além disso, estudantes de Medicina, indivíduos com alta renda, que moram sozinhos e estão no último período do curso são mais propensos a usar drogas psicotrópicas. Ademais, a exposição à medicação psiquiátrica foi capaz de aumentar o número de células binucleadas. Esses resultados fornecem evidências de que o uso de drogas psicotrópicas aumenta no contexto acadêmico e pode estar relacionado à falha do ciclo celular.

Anti-serum validation for use in immunohistochemistry for Trypanosoma cruzi detection.

INTRODUCTION: The detection of Trypanosoma cruzi in tissue samples is important in many situations, such as testing of the reactivation of the infection. The detection of T. cruzi nests in endomyocardial biopsies (EMB) may be useful to evaluate graft rejection. Given their scarcity, such nests are not routinely identified. To increase the diagnosis sensitivity, immunohistochemistry (IHC) may serve as a promising strategy. Here, we validate an antiserum for the detection of T. cruzi infection by IHC. METHODS: We used 1) positive controls (PCs) - 13 EMB, 12 skin biopsies, and 1 heart with T. cruzi nests as sections stained with hematoxylin and eosin (HE); 2) negative controls - a) 10 explant hearts and 10 EMB with no amastigote nests or clinical/laboratory signs of chagasic infection; and b) eight samples with leishmaniasis, toxoplasmosis, or histoplasmosis; and 3) Cases - 31 EMB of chagasic patients with no parasite nests in HE sections but detected positive for T. cruzi DNA by polymerase chain reaction. As a primary antibody, a hyperimmune serum from T. cruzi-infected rabbits was used. RESULTS: IHC results were positive for 21 of 26 PCs (80.8%) and one case of cutaneous leishmaniasis. In 4 of 31 cases, IHC revealed nests (12.9%), which were undetected by conventional histological examination. CONCLUSIONS: This study shows that IHC with the tested antiserum increases the sensitivity of the diagnosis and may be recommended for routine use in EMB analyses of cardiac transplant patients with Chagas disease.

Anti-serum validation for use in immunohistochemistry for Trypanosoma cruzi detection

Abstract INTRODUCTION: The detection of Trypanosoma cruzi in tissue samples is important in many situations, such as testing of the reactivation of the infection. The detection of T. cruzi nests in endomyocardial biopsies (EMB) may be useful to evaluate graft rejection. Given their scarcity, such nests are not routinely identified. To increase the diagnosis sensitivity, immunohistochemistry (IHC) may serve as a promising strategy. Here, we validate an antiserum for the detection of T. cruzi infection by IHC. METHODS: We used 1) positive controls (PCs) - 13 EMB, 12 skin biopsies, and 1 heart with T. cruzi nests as sections stained with hematoxylin and eosin (HE); 2) negative controls - a) 10 explant hearts and 10 EMB with no amastigote nests or clinical/laboratory signs of chagasic infection; and b) eight samples with leishmaniasis, toxoplasmosis, or histoplasmosis; and 3) Cases - 31 EMB of chagasic patients with no parasite nests in HE sections but detected positive for T. cruzi DNA by polymerase chain reaction. As a primary antibody, a hyperimmune serum from T. cruzi-infected rabbits was used. RESULTS: IHC results were positive for 21 of 26 PCs (80.8%) and one case of cutaneous leishmaniasis. In 4 of 31 cases, IHC revealed nests (12.9%), which were undetected by conventional histological examination. CONCLUSIONS: This study shows that IHC with the tested antiserum increases the sensitivity of the diagnosis and may be recommended for routine use in EMB analyses of cardiac transplant patients with Chagas disease.

Microdissecção e captura a laser na investigação do gene TP53 em tecidos incluídos em parafina / Laser-capture microdissection for TP53 gene analysis in paraffin-embedded tissues

INTRODUÇÃO: Microdissecção e captura a laser (MCL) é uma técnica de desenvolvimento recente que permite a coleta de células individuais ou pequeno conjunto de células para análise molecular. Atualmente, no Brasil, há raros microscópios para MCL, de modo que a divulgação dos procedimentos inerentes a essa técnica é oportuna para destacar seu amplo potencial para diagnóstico e investigação. OBJETIVO: Este trabalho descreve a padronização dos procedimentos de MCL e de extração de DNA de material fixado em formalina e incluído em parafina. MATERIAL E MÉTODOS: Foram estudados o éxon 8 do gene TP53 e o gene da ciclofilina em amostras de tecido normal e de neoplasias de fígado e rim provenientes de modelo de carcinogênese química induzida em rato. A extração do DNA foi comprovada por reação em cadeia da polimerase (nested-PCR). RESULTADOS: Foram padronizados os procedimentos de preparo dos cortes histológicos, de microdissecção e captura a laser e de obtenção de seqüências gênicas pela reação de nested-PCR para tecidos incluídos em parafina. Obtivemos amplificação de 48,3 por cento das amostras para o éxon 8 do gene TP53 e 51,7 por cento para o gene da ciclofilina. Considerando pelo menos um dos dois segmentos gênicos, foram amplificadas 79,3 por cento das amostras. DISCUSSÃO E CONCLUSÃO: A extração de DNA de tecidos fixados em formalina e incluídos em parafina e a técnica de nested-PCR foram adequadamente padronizadas para produtos gênicos de interesse, obtidos de material coletado por MCL. Esses procedimentos podem ser úteis para a obtenção de seqüências de DNA de arquivos para análise molecular.

BACKGORUND: Laser-capture micro-dissection (LCM) is a recently developed procedure that provides single cells or specific cell groups for molecular analysis. Currently, there are few LCM systems in Brazil, in such a way that it is necessary to disseminate the technical procedures inherent to the methodology, and also to characterize its enormous potential for diagnosis and research. OBJECTIVE: This study describes the standardization of LCM and DNA extraction from formalin fixed and paraffin-embedded tissues. MATERIAL AND METHOD: The gene TP53 exon 8 and the cyclophilin gene were studied in normal and neoplastic liver and kidney samples from a chemical carcinogenesis model in rat. DNA extraction was confirmed by nested-PCR. RESULTS: Histological sections preparation for LCM and the nested-PCR procedures were standardized; 48.3 percent amplifications of the gene TP53 exon 8 and 51.7 percent of the cyclophilin gene samples were obtained. When at least one of the gene segments was considered, 79.3 percent samples presented amplification. DISCUSSION AND CONCLUSION: Procedures for DNA extraction from formalin fixed and paraffin-embedded tissues collected by LCM were standardized. They can be useful for DNA collection for molecular studies.