Resumo Este estudo foi realizado para avaliar o efeito da Glutamina, como um dipeptídeo ou forma de aminoácido livre, na progressão de queimaduras em ratos. Trinta ratos Wistar machos foram queimados com um pente de metal aquecido em água fervente (98 °C) por três minutos, criando quatro áreas retangulares queimadas separadas por três interesespaços não queimados (zona de estase) em ambos os lados do dorso. Os animais foram randomizados em três grupos (n = 10): solução salina (G1-Controle) e grupos tratados que receberam glutamina via oral como dipeptídeo (G2-Dip) ou aminoácido livre (G3-FreeAA). Dois e sete dias após a queimadura, as lesões foram fotografadas para avaliação da evolução da necrose entre os espaços não queimados. Sete dias após a lesão, foi dosada a glutationa sérica e realizada análise histopatológica. Pelas fotografias, houve uma redução significativa na progressão da necrose no G3-Free-AA entre os dias dois e sete. A análise histopatológica no dia 7 mostrou uma zona de estase significativamente maior sem necrose e número mais elevado de fibroblastos em G2-Dip e G3-FreeAA em comparação com G1-Controle. Os níveis plasmáticos de glutationa foram maiores no G2-Dip em relação ao G1-Controle, e houve tendência a níveis mais elevados no G3-FreeAA. A redução das lesões histológicas, maior produção de fibroblastos, maior quantidade de glutationa podem ter beneficiado a evolução da necrose da queimadura, que mostrou maior preservação dos interespaços.

Abstract This study was carried out to evaluate the effect of Glutamine, as a dipeptide or a free amino acid form, on the progression of burn injuries in rats. Thirty male Wistar rats were burned with a comb metal plate heated in boiling water (98 °C) for three minutes, creating four rectangular full-thickness burn areas separated by three unburned interspaces (zone of stasis) in both dorsum sides. The animals were randomized into three groups (n=10): saline solution (G1-Control) and treated groups that orally received Glutamine as dipeptide (G2-Dip) or free amino acid (G3-FreeAA). Two and seven days after burn injury, lesions were photographed for unburned interspaces necrosis evolution assessment. Seven days after injury, glutathione seric was measured and histopathological analysis was performed. By photographs, there was a significant reduction in necrosis progression in G3-Free-AA between days two and seven. Histopathological analysis at day 7 showed a significantly higher stasis zone without necrosis and a higher number of fibroblasts in G2-Dip and G3-FreeAA compared with G1-Control. Also, glutathione serum dosage was higher in G2-Dip. The plasmatic glutathione levels were higher in the G2-Dip than the G1-Control, and there was a trend to higher levels in G3-FreeAA. The reduction in histological lesions, greater production of fibroblasts, and greater amounts of glutathione may have benefited the evolution of burn necrosis, which showed greater preservation of interspaces.

NMDAR (N-methyl-D-aspartate receptor) is one subtype of ionotrophic glutamate receptor which is extensively distributed in the central nervous system (CNS). In the mammalian CNS, NMDAR serves prominent roles in the pathophysiologic process of cerebral ischemia. This study aimed to investigate the pattern of expression of protein and gene of the excitatory neurotransmitter NMDAR in experimental focal cerebral ischemia and the hole of neuroprotection with hypothermia and ketoprofen. 120 rats were randomly divided into 6 groups (20 animals each): control - no surgery; sham - simulation of surgery; ischemic - focal ischemia for 1 hour, without reperfusion; ischemic + intraischemic hypothermia; ischemic + previous intravenous ketoprofen, and ischemic + hypothermia and ketoprofen. Ten animals from each experimental group were used to establish the volume of infarct. Transient focal cerebral ischemia was obtained in rats by occlusion of the middle cerebral artery with an intraluminal suture. The infarct volume was measured using morphometric analysis of infarct areas defined by triphenyl tetrazolium chloride and the patterns of expression of the protein and gene NMDA were evaluated by immunohistochemistry and quantitative real-time PCR, respectively. Increases in the protein and gene NMDA receptor in the ischemics areas were observed and these increases were reduced by hypothermia and ketoprofen. The increase in the NMDA receptor protein and gene expression observed in the ischemic animals was reduced by neuroprotection (hypothermia and ketoprofen). The NMDA receptor increases in the ischemic area suggests that the NMDA mediated neuroexcitotoxicity plays an important role in cell death and that the neuroprotective effect of both, hypothermia and ketoprofen is directly involved with the NMDA.

NMDAR (N-metil-D-aspartato) es un tipo de receptor de glutamato ionotrópico y está ampliamente distribuido en el sistema nervioso central (SNC). En el SNC de mamíferos, NMDAR se destaca de manera importante en el proceso fisiopatológico de la isquemia cerebral. Este estudio tuvo como objetivo investigar el patrón de expresión de proteínas y genes para el NMDA neurotransmisor excitatorio experimental de la isquemia cerebral focal y el vacío en la neuroprotección con hipotermia y ketoprofeno. Se dividieron 120 ratas aleatoriamente en grupos de 6 animales cada uno (20): Control - sin cirugía; Sham - simulación de cirugía; isquémicas - isquemia focal durante 1 hora, sin reperfusión isquémica; hipotermia intra-isquémica; isquemia; previa aplicación de ketoprofeno intravenoso, e hipotermia isquémica y ketoprofeno. Diez animales de cada grupo experimental fueron utilizados para establecer el volumen de infarto.La isquemia cerebral focal transitoria fue obtenida en ratas mediante oclusión de la arteria cerebral media con una sutura intraluminal. El volumen de infarto fue medido mediante análisis morfométrico de las áreas de infarto definidas por cloruro de trifenil tetrazolio y patrones de expresión de la proteína y el gen de NMDA, fueron evaluados por inmunohistoquímica y PCR cuantitativa en tiempo real, respectivamente. Se observaron aumentos en la proteína y en el gen del receptor de NMDA en las áreas isquémicas y estos aumentos fueron reducidos por la hipotermia y ketoprofeno. El aumento de la proteína del receptor de NMDA y la expresión génica observada en los animales isquémicos fue reducido mediante hipotermia y ketoprofeno. Los aumentos del receptor de NMDA en el área isquémica sugiere que la neuro excitotoxicidad mediada por NMDA desempeña un papel importante en la muerte celular y que el efecto neuroprotector de ambos, hipotermia y ketoprofeno está directamente relacionado al NMDA.

The aim was to analyze the protein expression of apoptotic genes caspase-3, caspase-8 and bcl-2 with the immunohistochemistry technique, correlating with tumor grade (I, II and III) and with the patient survival in order to understand the basic mechanism of tumoral transformation. The immunohistochemistry reactions on 50 samples of squamous cell carcinoma were carried out with the avidin-biotin immunoperoxidase method and antigen recovery. The analyses were made using the graduation method "in crosses" (0 to 4 crosses - no stain to more than 75% of positives cells) and in categories (low, intermediate, high) of the cytoplasm immunoreactivity of the epidermoid penile carcinoma cells. It was observed a statistically significant difference when the expression of caspase-3 were compared with the grades I and II of the tumor (p=0.0010) and when comparing the patient survival with the grades I and II of the tumor (p=0.0212). The protein bcl-2 was more expressed than caspase-3 and caspase-8 proteins, suggesting that the apoptotic rate in this carcinoma is low. The higher expression of the anti-apoptotic protein bcl-2 suggests a higher preservation of the tumoral cells.

El objetivo fue analizar la expresión de las proteínas de genes de apoptosis caspasa-3, caspasa-8 y Bcl-2-con la técnica de inmunohistoquímica, en correlación con el grado tumoral (I, II y III) y la supervivencia del paciente con el fin de comprender el mecanismo básico de la transformación tumoral. Se analizaron las reacciones inmunohistoquímicas sobre 50 muestras de carcinoma de células escamosas mediante el método de la inmunoperoxidasa avidina-biotina y la recuperación de antígeno. Los análisis se realizaron utilizando el método de graduación "en cruces" (0 a 4 cruces - no tinción a más del 75% de las células positivas) y en categorías (baja, media, alta) de la inmunorreactividad citoplasmática de las células de carcinoma epidermoide de pene. Se observó una diferencia estadísticamente significativa cuando la expresión de la caspasa-3 se comparó con los grados I y II del tumor (p = 0,0010) y cuando se comparan la supervivencia de los pacientes con los grados I y II del tumor (p = 0,0212). La proteína bcl-2 se expresa más que la caspasa-3 y caspasa-8, lo que sugiere que la tasa de apoptosis en este carcinoma es baja. La mayor expresión de la proteína anti-apoptótica bcl-2 sugiere una mayor preservación de las células tumorales.

The objective of this study was to analyze the submandibular glands of rats (Rattus norvegicus) submitted to experimental chronic alcoholism at electronic transmission microscopy. The experiment was programmed for a 15-month period, and in every five months, three animals from each group were sacrificed. Twenty-seven male rats, with average weight of 300g, were used and divided into three experimental groups with the following characteristics: 1) alcoholic: received ethanol, daily, at 6% for 15 months; 2) isocaloric: received diluted sucrose at a concentration of 87.14 g/L, corresponding energetically to the caloric content of the alcoholic group; and 3) control: received tap water "ad libitum" as liquid diet. The animals were fed with the same amount of Purina ration as a conventional weekly solid diet. It was observed several ultrastructural alterations in the alcoholic animals mainly in seromucous cells: dilation and enlargement of Golgi complex cisterns; the presence of digestive vacuoles, a large number of electronlucent vesicles and an increase in intercellular space, an increase between basolateral interdigitation cell and an enlargement of connective tissue. It was observed at some cells of the intercalary ducts at a lower part of lateral membranes, only after 15-month treatment, the presence of wide and clear vesicles with electronlucent material located among cell secretion granules of granular ducts. No alterations were observed in the animals of the control and isocaloric groups as well as the other parts of glandular parenchyma of animals of the alcoholic group

El objetivo de este estudio fue analizar las glándulas submandibulares de ratones (Rattus norvegicus) sometidos al alcoholismo crónico experimental, utilizando el microscopio electrónico de transmisión. El experimento fue programado para un período de 15 meses. Cada 5 meses, 3 animales de cada grupo fueron sacrificados. Utilizamos 27 ratones (machos) adultos, con peso medio de 400g los cuales fueron divididos en tres grupos experimentales: 1) Alcohólico, el cual recibió etanol diluido al 6% por 15 meses; 2) Isocalórico que recibió sacarosa diluida en concentración de 87,14 g/L, correspondiendo energéticamente al contenido calórico del grupo alcohólico, y 3) Control, el cual recibió agua de grifo "ad libitum"como dieta líquida. Los animales de todos los grupos fueron alimentados con una dieta sólida establecida semanalmente, que consistió en idéntica ración de Purina. Observamos varias alteraciones ultraestructurales, principalmente, en las células seromucosas: dilatación y ensanchamiento de las cisternas del complejo de Golgi, presencia de vacuolas digestivas, un gran número de vesículas electrodensas, aumento en el espacio intercelular, aumento entre las interdigitalizaciones basolaterales de las células y ensanchamiento del tejido conjuntivo. Además, observamos en algunas células de los ductos interpuestos en la posición más baja de sus membranas laterales, tras 15 meses de tratamiento, la presencia de vesículas claras y llenas de material electrodenso localizados entre los gránulos de secreción de las células de los ductos granulares. No observamos ninguna alteración en los animales los grupos control e isocalóricos, así como en otras partes del parénquima glandular de los animales del grupo alcoholizado

It has been described that chronic ingestion of ethanol leads to morphological changes in different tissues. In the present work we carried out a morphometric analysis of seromucous acini and granular ducts of submandibular glands from rats submitted to experimental chronic alcoholism. Male Wistar rats received an alcoholic diet containing ethanol 6% (v/v). At 5, 10 and 15 months after the beginning of the treatment, samples of submandibular glands were collected and the area of seromucous acini and granular ducts were analysed. The area of seromucous acini cells and granular ducts cells were also measured. The results indicated that chronic ethanol consumption significantly reduces the area of seromucous acini cells and granular ducts cells. On the other hand, there was an enlargement in the area of the seromucous acini and granular ducts after chronic ethanol consumption. We concluded that the effects of alcohol accentuated as the period of treatment with alcohol increased

Ha sido reconocido que la ingesta crónica de etanol causa alteraciones morfológicas en varios tejidos. En el presente trabajo fue realizado un análisis morfométrico de los acinos seromucosos y de los conductos granulosos de las glándulas submandibulares de ratones sometidos a alcoholismo crónico experimental. Ratones Wistar machos fueron sometidos a dieta alcohólica con etanol al 6% (v/v). A los 5, 10 y 15 meses tras el inicio del tratamiento, fueron recolectadas muestras de las glándulas submandibulares para analizar el área de los ácinos seromucosos y conductos granulosos. Los resultados indicaron que el consumo crónico de etanol reduce significativamente el área de las células de los acinos seromucosos y de las células de los conductos granulosos. Además, hubo un ensanchamiento en el área de los acinos seromucosos y de los conductos granulosos tras el consumo crónico de etanol. Concluimos que los efectos del alcohol fueron más graves mientras mayor fue el periodo de tratamiento