Cbfa1 expression in vascular smooth muscle cells may be elevated by increased nitric oxide/iNOS / A expressão de Cbfa1 nas células do músculo liso vascular pode ser elevada pelo aumento do óxido nítrico/iNOS

ABSTRACT Introduction: Vascular calcification is a common complication of chronic kidney disease. Osteoblast differentiation factor (Cbfa1) is present in histologic sections of arteries from patients with end-stage renal disease. Vascular smooth muscle cells (VSMC) can dedifferentiate to osteoblast-like cells, possibly by up-regulation of Cbfa1. There is evidence that the production of nitric oxide (NO) may have an important role in the regulation of osteoblast metabolism. The aim of this study is to evaluate whether increased NO/iNOS expression causes an increase in cbfa1 expression in VSMC. Methods: VSMC were obtained from renal artery of Wistar male rats, treated for 72 hours with lipopolysaccharide (LPS), ß-glycerophosphate (BGF), a donor of phosphate and aminoguanidine (AG), an inhibitor of iNOS, in the following groups: CTL (control), LPS, BGF, LPS + BGF, and LPS + AG. NO synthesis was determined by chemiluminescence. Cbfa1 and iNOS mRNA expressions were analyzed by RT-PCR, Cbfa1 protein expression by immunohistochemistry and cellular viability by acridine orange. Results: Cbfa1 and iNOS mRNA expressions were higher in LPS and LPS+ BGF vs CTL (p < 0.05), and they were lower in LPS+AG vs LPS (p < 0.05). The Cbfa1 in the groups LPS and LPS+BGF also resulted in a higher value compared to CTL (p < 0.05), and in LPS+AG it was lower compared to LPS (p < 0.05). NO was higher in LPS and LPS+BGF compared to CTL group (p < 0.05) and lower in LPS + AG compared to LPS group (p < 0.05). Cellular viability showed no statistical difference among groups. Conclusion: This study showed that increased NO/iNOS expression causes an increase in cbfa1 expression in VSMC.

RESUMO Introdução: A calcificação vascular é uma complicação comum da doença renal crônica. O fator de diferenciação osteoblástica (Cbfa1) está presente em cortes histológicos das artérias de pacientes com doença renal em estágio terminal. As células do músculo liso vascular (CMLV) podem desdiferenciar para células do tipo osteoblastos, possivelmente pela regulação positiva da Cbfa1. Há evidências de que a produção de óxido nítrico (NO) pode ter um papel importante na regulação do metabolismo dos osteoblastos. O objetivo deste estudo é avaliar se o aumento da expressão de NO/iNOS causa um aumento na expressão de cbfa1 nas CMLV. Métodos: As CMLV foram obtidas da artéria renal de ratos machos Wistar, tratados por 72 horas com lipopolissacarídeo (LPS), ß-glicerofosfato (BGF), um doador de fosfato e aminoguanidina (AG), um inibidor da iNOS, nos seguintes grupos: CTL (controle), LPS, BGF, LPS + BGF e LPS + AG. A síntese de NO foi determinada por quimioluminescência. As expressões de mRNA de Cbfa1 e iNOS foram analisadas por RT-PCR, a expressão da proteína Cbfa1 por imunohistoquímica e viabilidade celular por laranja de acridina. Resultados: As expressões de mRNA de Cbfa1 e iNOS foram maiores em LPS e LPS + BGF v.s. CTL (p < 0,05) e menores em LPS + AG v.s. LPS (p <0,05). O Cbfa1 nos grupos LPS e LPS + BGF também resultou em um valor maior em comparação ao CTL (p < 0,05), e no LPS + AG foi menor em comparação ao LPS (p < 0,05). NO foi maior no LPS e LPS + BGF em comparação ao grupo CTL (p < 0,05) e menor no LPS + AG em comparação ao grupo LPS (p < 0,05). A viabilidade celular não mostrou diferença estatística entre os grupos. Conclusão: Este estudo mostrou que o aumento da expressão de NO/iNOS causa um aumento na expressão de cbfa1 nas CMLV.

Oxidative stress and immune system analysis after cycle ergometer use in critical patients

OBJECTIVE: The passive cycle ergometer aims to prevent hypotrophy and improve muscle strength, with a consequent reduction in hospitalization time in the intensive care unit and functional improvement. However, its effects on oxidative stress and immune system parameters remain unknown. The aim of this study is to analyze the effects of a passive cycle ergometer on the immune system and oxidative stress in critical patients. METHODS: This paper describes a randomized controlled trial in a sample of 19 patients of both genders who were on mechanical ventilation and hospitalized in the intensive care unit of the Hospital Agamenom Magalhães. The patients were divided into two groups: one group underwent cycle ergometer passive exercise for 30 cycles/min on the lower limbs for 20 minutes; the other group did not undergo any therapeutic intervention during the study and served as the control group. A total of 20 ml of blood was analysed, in which nitric oxide levels and some specific inflammatory cytokines (tumour necrosis factor alpha (TNF-α), interferon gamma (IFN-γ) and interleukins 6 (IL-6) and 10 (IL-10)) were evaluated before and after the study protocol. RESULTS: Regarding the demographic and clinical variables, the groups were homogeneous in the early phases of the study. The nitric oxide analysis revealed a reduction in nitric oxide variation in stimulated cells (p=0.0021) and those stimulated (p=0.0076) after passive cycle ergometer use compared to the control group. No differences in the evaluated inflammatory cytokines were observed between the two groups. CONCLUSION: We can conclude that the passive cycle ergometer promoted reduced levels of nitric oxide, showing beneficial effects on oxidative stress reduction. As assessed by inflammatory cytokines, the treatment was not associated with changes in the immune system. However, further research in a larger population is necessary for more conclusive results.

Estimativa de pessoas com diagnóstico de câncer no Brasil: dados da Pesquisa Nacional de Saúde, 2013 / Estimated number of people diagnosed with cancer in Brazil: data from the National Health Survey, 2013

RESUMO: Objetivo: Descrever o perfil dos pacientes que referiram diagnóstico médico de câncer e descrever os tipos de câncer mais prevalentes, segundo variáveis selecionadas. Métodos: Estudo descritivo que utilizou dados da Pesquisa Nacional de Saúde (PSN) de 2013 para estimar prevalências e respectivos valores do intervalo de confiança (IC95%). Resultados: Menos de 2% dos adultos referiram diagnóstico médico de câncer, sendo mais relatado por mulheres, por pessoas com mais de 60 anos, entre brancos, em residentes da zona urbana e moradores da Região Sul. O câncer de próstata foi o mais referido entre os homens; entre as mulheres, o câncer de mama foi o mais prevalente. A menor idade média do primeiro diagnóstico foi identificada para câncer de colo de útero (35,4 anos; IC95% 30,3 - 40,6), e a maior, para câncer de próstata (65,7 anos; IC95% 64,2 - 67,0). Conclusão: Os achados deste estudo são importantes para o planejamento dos serviços de saúde e do seu acesso, pois revelam diferenças, principalmente regionais.

ABSTRACT: Objective: To describe the profile of patients who reported a medical diagnosis of cancer and describe the most prevalent types of cancer, according to selected variables. Methods: A descriptive study that used data from the National Survey of Health, 2013, to estimate prevalence and their values of confidence interval (95%CI). Results: Less than 2% of adults reported a medical diagnosis of cancer, with most reported by women, people over 60, among whites, residents in the village and residents of South Prostate cancer was the most reported among men and breast among women. The lowest average age of first diagnosis was identified for cervical cancer (35.4 years; 95%CI 30.3 - 40.6) and the highest for prostate (65.7 years; 95%CI 64.2 - 67.0). Conclusion: The findings of this study are important for the planning of health services and access, as they show differences mainly regional.

Monitoring the effectiveness of root canal procedures on endotoxin levels found in teeth with chronic apical periodontitis

Objective: The aim of this study was to monitor the effectiveness of root canal procedures by using different irrigants and intracanal medication on endotoxin levels found in root canals of teeth with chronic apical periodontitis. Material and Methods: Thirty root canals of teeth with pulpal necrosis associated with periapical lesions were selected and randomly divided into groups according to the irrigants used: GI - 2.5% NaOCl, GII - 2% chlorhexidine (CHX) gel, and GIII - saline solution (SS) (all, n=10). Samples were collected with sterile/apyrogenic paper points before (S1) and after root canal instrumentation (S2), after use of 17% ethylenediaminetetraacetic acid (EDTA) (S3), and after 30 days of intracanal medication (Ca(OH)2+SS) (S4). A turbidimetric kinetic Limulus Amebocyte Lysate assay was used for endotoxin measurement. Results: Endotoxins were detected in 100% of the root canals investigated (30/30), with a median value of 18.70 EU/mL. After S2, significant median percentage reduction was observed in all groups, irrespective of the irrigant tested: 2.5% NaOCl (99.65%) (GI), 2% CHX (94.27%) (GII), and SS (96.79%) (GIII) (all p<0.05). Root canal rinse with 17% EDTA (S3) for a 3-minute period failed to decrease endotoxin levels in GI and a slight decrease was observed in GII (59%) and GIII (61.1%) (all p>0.05). Intracanal medication for 30 days was able to significantly reduce residual endotoxins: 2.5% NaOCl (90%) (GI), 2% CHX (88.8%) (GII), and SS (85.7%) (GIII, p<0.05). No differences were found in the endotoxin reduction when comparing s2 and s4 treatment groups. Conclusion: Our findings demonstrated the effectiveness of the mechanical action of the instruments along with the flow and backflow of irrigant enduring root canal instrumentation for the endotoxin removal from root canals of teeth with chronic apical periodontitis. Moreover, the use of intracanal medication for 30 days contributed for an improvement ...

Antimicrobial peptides and nitric oxide production by neutrophils from periodontitis subjects

Neutrophils play an important role in periodontitis by producing nitric oxide (NO) and antimicrobial peptides, molecules with microbicidal activity via oxygen-dependent and -independent mechanisms, respectively. It is unknown whether variation in the production of antimicrobial peptides such as LL-37, human neutrophil peptides (HNP) 1-3, and NO by neutrophils influences the pathogenesis of periodontal diseases. We compared the production of these peptides and NO by lipopolysaccharide (LPS)-stimulated neutrophils isolated from healthy subjects and from patients with periodontitis. Peripheral blood neutrophils were cultured with or without Aggregatibacter actinomycetemcomitans-LPS (Aa-LPS), Porphyromonas gingivalis-LPS (Pg-LPS) and Escherichia coli-LPS (Ec-LPS). qRT-PCR was used to determine quantities of HNP 1-3 and LL-37 mRNA in neutrophils. Amounts of HNP 1-3 and LL-37 proteins in the cell culture supernatants were also determined by ELISA. In addition, NO levels in neutrophil culture supernatants were quantitated by the Griess reaction. Neutrophils from periodontitis patients cultured with Aa-LPS, Pg-LPS and Ec-LPS expressed higher HNP 1-3 mRNA than neutrophils from healthy subjects. LL-37 mRNA expression was higher in neutrophils from patients stimulated with Aa-LPS. Neutrophils from periodontitis patients produced significantly higher LL-37 protein levels than neutrophils from healthy subjects when stimulated with Pg-LPS and Ec-LPS, but no difference was observed in HNP 1-3 production. Neutrophils from periodontitis patients cultured or not with Pg-LPS and Ec-LPS produced significantly lower NO levels than neutrophils from healthy subjects. The significant differences in the production of LL-37 and NO between neutrophils from healthy and periodontitis subjects indicate that production of these molecules might influence individual susceptibility to important periodontal pathogens.

Translational research into gut microbiota: new horizons in obesity treatment / Pesquisa translacional em microbiota intestinal: novos horizontes no tratamento da obesidade

Obesity is a pandemic which has been rapidly developing for three decades. When a population is submitted to the same nutritional stress, some individuals are less susceptible to diet-induced weight gain and hyperglycemia. This observation suggests that other mechanisms are involved which are not directly related to the human genome. The human gut contains an immense number of microorganisms, collectively known as the microbiota. Evidence that gut microbiota composition can differ between obese and lean humans has led to the speculation that gut microbiota can participate in the pathophysiology of obesity. Different mechanisms have been proposed to explain the link between gut flora and obesity. The first mechanism consists in the role of the gut microbiota to increase energy extraction from indigestible dietary polysaccharides. The second, consists in the role of gut flora to modulate plasma lipopolysaccharide levels which triggers chronic low-grade inflammation leading to obesity and diabetes. A third mechanism proposes that gut microbiota may induce regulation of host genes that modulate how energy is expended and stored. However, further studies are needed to clarify a number of issues related to the relationship between the gut microbiota and obesity.

A obesidade é uma pandemia que afeta milhões de pessoas em todo o mundo. Quando uma população é submetida ao mesmo estresse nutricional, alguns indivíduos são menos suscetíveis ao ganho de peso induzido pela dieta e à hiperglicemia. Essa observação sugere que outros mecanismos não diretamente relacionados ao genoma humano estejam envolvidos. O intestino humano é colonizado por milhões de bactérias, que coletivamente constituem a flora comensal normal. A evidência de que a composição da flora intestinal pode ser diferente em humanos magros e obesos levou à especulação de que a flora intestinal pode participar na fisiopatologia da obesidade. Diferentes mecanismos foram propostos para tentar explicar a correlação entre flora intestinal e obesidade. O primeiro mecanismo consiste no papel da flora intestinal na extração de energia de polissacarídeos não digeríveis. O segundo mecanismo envolve a modulação dos níveis de lipopolissacarídeo pela flora intestinal, o que desencadeia uma inflamação crônica subclínica que acarreta obesidade e diabetes. Um terceiro mecanismo propõe que a flora intestinal pode induzir a regulação de genes do hospedeiro que modulam como a energia é gasta e armazenada. Entretanto, estudos adicionais são necessários para estabelecer o papel da flora intestinal no desenvolvimento da obesidade.

Efeito do processo inflamatório sobre as propriedades regenerativas das células-tronco do ligamento periodontal / Effect of inflammation on the regenerative properties of stem cells from periodontal ligament

Estudos recentes mostraram que o ligamento periodontal abriga populações celulares com características de células-tronco adultas, as quais são denominadas como células progenitoras ou indiferenciadas de origem mesenquimal. O sucesso da regeneração dos tecidos periodontais depende de uma série de eventos biológicos, os quais envolvem a migração e a proliferação das células indiferenciadas para o interior do defeito periodontal, e sua subsequente diferenciação em osteoblastos, cementoblastos e fibroblastos, acompanhada pela síntese de componentes das matrizes teciduais. Alguns estudos na área médica mostraram que as propriedades biológicas das células-tronco podem ser alteradas após a exposição tecidual a um processo inflamatório crônico, comprometendo o seu potencial regenerativo. Desta maneira, o presente estudo teve como objetivo realizar uma revisão da literatura das evidências disponíveis a respeito do impacto da doença periodontal inflamatória crônica sobre as propriedades regenerativas das células mesenquimais indiferenciadas, localizadas nas estruturas periodontais remanescentes.

Bacterial pathogenesis and mediators in apical periodontitis: [review]

Apical periodontitis is a group of inflammatory diseases caused by microorganisms (mainly bacteria) infecting the necrotic root canal system. The pathogenesis of different types of apical periodontitis and even the same type in different individuals is unlikely to follow a stereotyped fashion with regard to the involved bacterial mediators. Disease pathogenesis is rather complex and involves a multitude of bacteria- and host-related factors. This review article discusses the bacterial pathogenesis of acute and chronic apical periodontitis, with the main focus on the bacterial mediators conceivably involved in the different stages of the infectious process, including secreted products (enzymes, exotoxins, N-formyl-methionyl-leucyl-phenylalanine peptides, heat-shock proteins and metabolic end-products) and structural components (lipopolysaccharide, peptidoglycan, lipoteichoic acid, lipoproteins, fimbriae, flagella, outer membrane proteins and vesicles, DNA, and exopolysaccharides). Knowledge of the bacterial factors involved in the pathogenesis of apical periodontitis is important to the understanding of the disease process and to help establishing proper therapeutic measures to inactivate this bacterial "artillery".

Lesões perirradiculares compreendem um grupo de doenças inflamatórias causadas por microrganismos (principalmente bactérias) infectando o sistema de canais radiculares com polpa necrosada. É improvável que a patogênese dos diferentes tipos de lesão perirradicular, e até mesmo daquelas do mesmo tipo, mas em diferentes indivíduos, obedeça um padrão estereotipado com relação aos mediadores bacterianos envolvidos. A patogênese destas doenças é complexa e envolve inúmeros fatores relacionados às bactérias e ao hospedeiro. Este artigo de revisão discute a patogênese bacteriana das lesões perirradiculares agudas e crônicas, enfatizando os fatores bacterianos que estão possivelmente envolvidos nos diferentes estágios do processo infeccioso, incluindo produtos secretados (enzimas, exotoxinas, peptídeos N-formilados, proteínas de choque térmico e produtos terminais do metabolismo) e componentes estruturais (lipopolissacarídeo, peptidoglicano, ácido lipoteicóico, lipoproteínas, fímbrias, flagelos, proteínas e vesículas de membrana externa, DNA e exopolissacarídeos). O conhecimento dos fatores bacterianos envolvidos na patogênese das lesões perirradiculares é importante para o entendimento do processo patológico bem como para ajudar no estabelecimento de medidas terapêuticas adequadas para desativação desta "artilharia" bacteriana.

Las enfermedades gíngivo-periodontales y sus repercusiones sistémicas: planeamiento estratégico interdisciplinario / Gingivo-periodontal diseases and their systemic repercussions: an interdisciplinary strategic planning

La periodontitis es una enfermedad causada por un grupo de bacterias Gram-negativas presentes en las superficies de las raíces de los dientes en forma de biofilm. Los lipopolisacáridos (LPS) y otras sustancias acceden al tejido gingival, iniciando y perpetuando la inmunoinflamación, resultando en la producción de altos niveles de citoquinas proinflamatorias. Esto induce el desarrollo de la matriz de metaloproteínas las cuales destruyen el tejido conectivo de la encía y del ligamento periodontal y de prostaglandinas las que median en la reabsorción del hueso alveolar. La periodontitis puede aumentar la susceptibilidad a ciertas enfermedades sistémicas por diferentes vías. Los LPS y las bacterias Gram-negativas viables del biofilm y citoquinas proinflamatorias del tejido periodontal inflamado pueden ingresar en la circulación en número suficiente y de esta manera influir en la salud general y en la susceptibilidad a ciertas enfermedades. En este caso, en particular, se desarrollará la relación de la enfermedad periodontal con las patologías sistémicas y el cambio de paradigma para una atención interdisciplinaria de nuestros pacientes. Esta ampliación del criterio de enfermedad periodontal debe reflejarse en una trasdisciplinariedad donde los integrantes del equipo de salud trabajen en coordinación para prevenir o tratar las diferentes patologías asociads con las enfermedades gíngivo-periodontales

Periodontitis y enfermedad cardiovascular ateroesclerótica / Periodontitis and atherosclerotic cardivascular disease

Los factores de riesgo tradicionales como la hipertensión, dislipidemia, diabetes, tabaquismo, etc., no explican la etiología y patogénesis de las complicaciones de la aterosclerosis en todos los pacientes, motivo por el cual la etiología infecciosa de esta patología ha recibido gran atención últimamente. La periodontitis, infección crónica producida por bacterias Gram (-), se ha relacionado con las enfermedades cardiovasculares ateroscleróticas. Ambas patologías comparten factores de riesgo y mecanismos patogénicos que explicarían la posible asociación entre ellas. La presencia de un gran reservorio de bacterias y mediadores inflamatorios relacionados con la periodontitis, son los posibles mecanismos biológicos que relacionan ambas patologías, y la base para la hipótesis de que la periodontitis es un posible factor de riesgo para las enfermedades cardiovasculares ateroscleróticas