Suppression of nitrosative stress and inflammation of the knee joint synovium in collagen type ii-induced rheumatoid arthritis by the inhibition of glycogen synthase kinase-3ß / Supresión del estrés nitrosativo e inflamación de la sinovial de la articulación de la rodilla en la artritis reumatoide inducida por colágeno tipo ii mediante la inhibición de la glucógeno sintasa quinasa-3 ß

SUMMARY: Rheumatoid arthritis (RA), an inflammatory autoimmune disease that causes cartilage degradation and tissue destruction, can affect synovial joints such as the knee joint. The link between the nitrosative stress enzyme inducible nitric oxide synthase (iNOS) and the cytokine interleukin-1 (IL-1β) in RA-induced knee joint synovial membrane damage with and without the incorporation of the GSK3β inhibitor TDZD-8 has never been studied. As a result, we used active immunization method with collagen type II (COII) for twenty one days to induce RA in rats. TDZD-8 (1 mg/kg; i.p.) was given daily into matched immunized rats for three weeks after day 21 (COII+TDZD-8). Blood and tissue samples were taken 42 days after immunization. A dramatic increase in rheumatoid factor (RF) blood levels, as well as considerable synovial tissue damage and inflammatory cell infiltration of the synovial membrane, were used to validate the onset of RA following COII immunization. COII immunization increased tissue levels of iNOS protein and IL- 1β mRNA and protein expression, which TDZD-8 suppressed considerably (p<0.0001). Furthermore, there was a significantly (p<0.001) positive correlation between iNOS, inflammatory biomarkers, and RF. We concluded that TDZD-8 reduced RA-induced IL-1β -iNOS axis-mediated arthritis in the rat knee joint synovium.

RESUMEN: La artritis reumatoide (AR), es una enfermedad autoinmune inflamatoria que causa la degradación del cartílago y la destrucción del tejido, pudiendo afectar las articulaciones sinoviales, como la articulación de la rodilla. No se ha estudiado el vínculo entre la óxido nítrico sintasa inducible por la enzima del estrés nitrosativo (iNOS) y la citocina interleucina-1 (IL-1β) en el daño de la membrana sinovial de la articulación de la rodilla provocado por AR con y sin la incorporación del inhibidor de GSK3β TDZD-8. Utilizamos el método de inmunización activa con colágeno tipo II (COII) durante veintiún días para inducir AR en ratas. Se administró TDZD-8 (1 mg/kg; i.p.) diariamente a ratas inmunizadas emparejadas durante tres semanas después del día 21 (COII+TDZD- 8). Se tomaron muestras de sangre y tejido 42 días después de la inmunización. Se observó un gran aumento de los niveles sanguíneos del factor reumatoideo (FR), así como un daño considerable del tejido sinovial e infiltración de células inflamatorias en la membrana sinovial, para validar la aparición de la AR después de la inmunización con COII. La inmunización con COII aumentó los niveles tisulares de la proteína iNOS y la expresión de proteína y ARNm de IL-1β, que TDZD-8 suprimió considerablemente (p<0,0001). Además, hubo una correlación positiva significativa (p<0,001) entre iNOS, biomarcadores inflamatorios y FR. Concluimos que TDZD- 8 redujo la artritis mediada por el eje IL-1β-iNOS inducida por la AR en la sinovial de la articulación de la rodilla de rata.

Induction of brain injury and depression in rats by chronic unpredictable stress associated with the augmentation of nitrosative stress and apoptosis / Inducción de lesión cerebral y depresión en ratas por estrés crónico impredecible asociado con el aumento del estrés nitrosativo y la apoptosis

SUMMARY: Repeated stress is a risk factor for memory impairment and neurological abnormalities in both humans and animals. We sought to investigate the extent of (i) brain tissue injury; (ii) nitrosative and oxidative stress in brain tissue homogenates; (iii) apoptotic and survival biomarkers in brain tissue homogenates; and (iv) immobility and climbing abilities, induced over a period of three weeks by chronic unpredictable stress (CUS). Wistar rats were either left untreated (Control group) or exposed to a variety of unpredictable stressors daily before being sacrificed after 3 weeks (model group). Assessment of depression-like behavior was performed and animals were then culled and harvested brain tissues were stained with basic histological staining and examined under light microscopy. In addition, brain tissue homogenates were prepared and assayed for these parameters; inducible nitric oxide synthase (iNOS), malondialdehyde (MDA), superoxide dismutase (SOD), caspase-3, and B-cell lymphoma 2 (Bcl-2). Histology images showed CUS induced profound damage to the cerebral cortex as demonstrated by severe neuronal damage with shrunken cells, disrupted atrophic nuclei, perineuronal vacuolation and swollen glial cells. CUS also significantly (p<0.05) induced iNOS, MDA, and caspase-3, whereas SOD and Bcl-2 brain tissue levels were inhibited by CUS. In addition, data from the depression-like behavior, forced swimming test showed significant (p<0.05) increase in animal immobility and decrease in climbing ability in the model group of rats. Thus, here we demonstrated a reliable rat model of chronic stress-induced brain injury, which can further be used to investigate beneficial drugs or agents used for a period of three weeks to protect against CUS-induced brain damage.

RESUMEN: El estrés crónico es un factor de riesgo para el deterioro de la memoria y las anomalías neurológicas tanto en humanos como en animales. Intentamos investigar el alcance de lesión del tejido cerebral; (ii) estrés nitrosativo y oxidativo en homogeneizados de tejido cerebral; (iii) biomarcadores apoptóticos y de supervivencia en homogeneizados de tejido cerebral; y (iv) inmovilidad y habilidades de escalada, inducidas durante un período de tres semanas por estrés crónico impredecible (ECI). Se dejaron sin tratamiento (grupo control) ratas Wistar, o se expusieron a una variedad de factores estresantes impredecibles diariamente antes de ser sacrificadas después de 3 semanas (grupo modelo). Se realizó una evaluación del comportamiento similar a la depresión y luego se sacrificaron los animales y se tiñeron los tejidos cerebrales con tinción histológica básica y se examinaron con microscopía óptica. Además, se prepararon homogeneizados de tejido cerebral y se analizaron los siguientes parámetros; óxido nítrico sintasa inducible (iNOS), malondialdehído (MDA), superóxido dismutasa (SOD), caspasa- 3 y linfoma de células B 2 (Bcl-2). Las imágenes histológicas mostraron que el CUS indujo un daño profundo en la corteza cerebral como lo demuestra el daño neuronal severo con células encogidas, núcleos atróficos alterados, vacuolación perineuronal y células gliales inflamadas. ECI también indujo significativamente (p <0,05) iNOS, MDA y caspase-3, mientras que los niveles de tejido cerebral SOD y Bcl-2 fueron inhibidos por ECI. Además, los datos del comportamiento similar a la de- presión, la prueba de natación forzada mostró un aumento significativo (p <0,05) en la inmovilidad animal y una disminución en la capacidad de escalada en el grupo modelo de ratas. Por lo tanto, aquí demostramos un modelo confiable de daño cerebral crónico en rata inducido por el estrés, que se puede utilizar para investigar medicamentos o agentes beneficiosos usados durante un período de tres semanas para proteger el daño cerebral inducido por ECI.

Acute effect of passive cycle-ergometry and functional electrical stimulation on nitrosative stress and inflammatory cytokines in mechanically ventilated critically ill patients: a randomized controlled trial

Early mobilization is beneficial for critically ill patients because it reduces muscle weakness acquired in intensive care units. The objective of this study was to assess the effect of functional electrical stimulation (FES) and passive cycle ergometry (PCE) on the nitrous stress and inflammatory cytometry in critically ill patients. This was a controlled, randomized, open clinical trial carried out in a 16-bed intensive care unit. The patients were randomized into four groups: Control group (n=10), did not undergo any therapeutic intervention during the study; PCE group (n=9), lower-limb PCE for 30 cycles/min for 20 min; FES group (n=9), electrical stimulation of quadriceps muscle for 20 min; and FES with PCE group (n=7), patients underwent PCE and FES, with their order determined randomly. The serum levels of nitric oxide, tumor necrosis factor alpha, interferon gamma, and interleukins 6 and 10 were analyzed before and after the intervention. There were no differences in clinical or demographic characteristics between the groups. The results revealed reduced nitric oxide concentrations one hour after using PCE (P<0.001) and FES (P<0.05), thereby indicating that these therapies may reduce cellular nitrosative stress when applied separately. Tumor necrosis factor alpha levels were reduced after the PCE intervention (P=0.049). PCE and FES reduced nitric oxide levels, demonstrating beneficial effects on the reduction of nitrosative stress. PCE was the only treatment that reduced the tumor necrosis factor alpha concentration.

Radiological aspects in computed tomography as determinants in the diagnosis of pulmonary tuberculosis in immunocompetent infants / Aspectos radiológicos na tomografia computadorizada determinantes no diagnóstico da tuberculose pulmonar em lactentes imunocompetentes

Abstract Objective: To describe the chest computed tomography (CT) findings in immunocompetent children under 36 months of age with pulmonary tuberculosis. Materials and Methods: This was a descriptive case series conducted in the city of Rio de Janeiro, Brazil, between January 2004 and July 2013, involving 20 young children who underwent CT after undergoing chest X-rays that did not provide a definitive diagnosis. Results: All of the participants had lymph node enlargement and consolidations. In 15 cases (75%), the consolidations were accompanied by atelectasis. Pulmonary cavitation was seen in 10 cases (50%), and cavitation within consolidations was seen in 7 (35%). The areas of cavitation and parenchymal destruction were not seen on conventional chest X-rays. Conclusion: The radiological presentation of pulmonary tuberculosis in young children differs from that described in older children and adults. CT is an effective method for the early diagnosis of pulmonary tuberculosis in immunocompetent infants, allowing the rapid institution of specific treatment, which is crucial for halting disease progression, as well as for preventing local and systemic complications.

Resumo Objetivo: Descrever achados radiológicos na tomografia computadorizada (TC) de tórax de crianças imunocompetentes menores de 36 meses com tuberculose pulmonar. Materiais e Métodos: Esta série de casos foi desenvolvida na cidade do Rio de Janeiro, no período de janeiro de 2004 a julho de 2013, onde 20 pacientes foram submetidos a TC após a realização de radiografias de tórax que não definiram o diagnóstico. Resultados: Todos os participantes tiveram linfonodomegalias e consolidações. Em 15 casos (75%) as consolidações tiveram atelectasia associada. Escavações pulmonares ocorreram em 10 casos (50%), havendo consolidações em 7 (35%). Áreas de escavação e destruição parenquimatosa em fase inicial não foram observadas nas radiografias simples. Conclusão: A apresentação radiológica de tuberculose pulmonar em lactentes não foi a mesma descrita em crianças maiores e adultos. A TC é um método aplicável para o diagnóstico precoce de tuberculose pulmonar em lactentes imunocompetentes, permitindo a rápida instituição de tratamento específico, que é crucial para interromper a progressão da doença e prevenir suas complicações locais e sistêmicas.

Metabolic syndrome components are associated with oxidative stress in overweight and obese patients

ABSTRACT Objective: The aim of this study is to evaluate the influence of the body mass index (BMI) and the metabolic syndrome (MetS) parameters on oxidative and nitrosative stress in overweight and obese subjects. Subjects and methods: Individuals were divided into three groups: the control group (G1, n = 131) with a BMI between 20 and 24.9 kg/m2, the overweight group (G2, n = 120) with a BMI between 25 and 29.9 kg/m2 and the obese group (G3, n = 79) with a BMI ≥ 30 kg/m2. Results: G3 presented higher advanced oxidation protein products (AOPPs) in relation to G1 and G2 (p = 0.001 and p = 0.011, respectively) whereas G2 and G3 had lower levels of nitric oxide (NO) (p = 0.009 and p = 0.048, respectively) compared to G1. Adjusted for the presence of MetS to evaluate its influence, the levels of AOPPs did not differ between the groups, whereas NO remained significantly lower. Data adjusted by the BMI showed that subjects with higher triacylglycerol levels had higher AOPPs (p = 0.001) and decreased total radical-trapping antioxidant parameter/uric Acid (p = 0.036). Subjects with lower high-density lipoprotein (HDL) levels and patients with higher blood pressure showed increased AOPPs (p = 0.001 and p = 0.034, respectively) and lower NO levels (p = 0.017 and p = 0.043, respectively). Subjects who presented insulin resistance had higher AOPPs (p = 0.024). Conclusions: Nitrosative stress was related to BMI, and protein oxidation and nitrosative stress were related to metabolic changes and hypertension. MetS components were essential participants in oxidative and nitrosative stress in overweight and obese subjects.

Evaluación de estrés nitrosativo y oxidativo en mujeres con diferentes manifestaciones clínicas del síndrome antifosfolipídico / Evaluation of nitrosative stress and oxidative in women with different clinical manifestations of the antiphospholipid syndrome

Introducción:El síndrome antifosfolipídico (SAF) es una enfermedad autoinmune caracterizada por la presencia persistente de anticuerpos antifosfolípidos (aAFL) y manifestaciones clínicas de trombosis y/o morbilidad gestacional que se asocian con estrés nitrosativo/ oxidativo y disminución de la capacidad antioxidante, alterando el desarrollo gestacional. Objetivo: Evaluar algunos biomarcadores de estrés nitrosativo/oxidativo del suero de mujeres con diferentes manifestaciones clínicas del SAF y sus efectos en células endoteliales. Métodos: Se incluyeron sueros de 48 mujeres divididas en dos grupos con y sin aAFL. Se evaluó la concentración de nitritos, la capacidad antioxidante y la actividad de la enzima paraoxonasa

Biomarcadores emergentes para diferentes patologías humanas / Emerging biomarkers for different human pathologies / Biomarcadores emergentes para diferentes doenças humanas

El objetivo de esta actualización fue revisar los hallazgos concernientes a la posible utilidad clínica de nuevos biomarcadores de estrés oxidativo-nitrosativo, especialmente aquellos derivados del metabolismo lipídico, como herramientas a implementar en el diagnóstico diferencial, la prognosis, el riesgo o la evaluación de estrategias terapéuticas, en enfermedades crónicas de tipo degenerativo. Se analizó la información disponible sobre el uso de marcadores emergentes en casos de infertilidad masculina por varicocele, pacientes con neurodegeneración debida a mal de Alzheimer, de Parkinson, o demencias de origen vascular, enfermos con cáncer de pulmón, próstata o mama, diabéticos tipo 2 y personas expuestas a agroquímicos en forma profesional. Como conclusión surge la posibilidad de implementar muchos de estos biomarcadores en el laboratorio bioquímico-clínico con promisoria y confiable utilidad en varias de estas situaciones patológicas, considerando que la mayoría de ellos se pueden determinar con relativa baja dificultad y requieren muestras obtenibles por métodos no invasivos o mínimamente invasivos.

This review aimed to analyze the possible clinical utility of various oxidative-nitrosative stress biomarkers, mostly derived from lipid metabolism, as a tool to be implemented in the differential diagnosis, prognosis, risk assessment, or the evaluation of the therapeutical strategies for chronic illnesses of the degenerative type. The available information concerning the utility of the biomarkers on male infertility due to varicocele, neurodegenerative disease such as Alzheimer, Parkinson or vascular dementia, prostate, lung and breast cancer, professional sprayers exposed to agrochemicals or type 2 diabetes patients was discussed. The main conclusion is that many of these emerging biomarkers could be implemented in clinica /biochemical analyses of human samples due to the fact that they can be obtained from non-invasive or minimal-invasive procedures and are easily determined.

O objetivo desta atualização é rever os achados sobre a possível utilidade clínica de novos biomarcadores de estresse oxidativo-nitrosativo, especialmente aqueles derivados de metabolismo lipídico, como ferramentas para implementar no diagnóstico diferencial, prognose, risco, ou avaliação de estratégias terapêuticas, em doenças crônicas de tipo degenerativo. Foi analisada a informação disponível sobre o uso de marcadores emergentes em casos de infertilidade masculina por varicocele, pacientes com neurodegeneração devida à doença de Alzheimer, de Parkinson ou demência de origem vascular, pacientes com câncer do pulmão, próstata ou mama, diabéticos tipo 2 e pessoas expostas a produtos agroquímicos devido a sua profissão. Como conclusão surge a possibilidade de implementar muitos destes biomarcadores no laboratório bioquímico- clínico com promissora e confiável utilidade em várias dessas situações patológicas, considerando-se que a maior parte deles pode ser determinada com relativa baixa dificuldade e requerem amostras obtidas através de métodos não invasivos ou minimamente invasivos.