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The effects of voluntary running on the skeletal muscle of rats with pulmonary arterial hypertension (PAH) were tested in the present study. PAH was induced in rats by a single injection of monocrotaline (MCT, 60 mg/kg). Rats in the sedentary hypertension (HS) group had their tolerance to physical exertion reduced throughout the experiment, while those in the sedentary control (SC), exercise control (EC), exercise hypertension (EH) and median exercise (EM) groups maintained or increased. Despite that, the muscular citrate synthase activity was not different between groups. The survival time was higher in the EH (32 days) than in the SH (28 days) (p = 0.0032). SH and EH groups showed a lower percentage of muscle fiber and a higher percentage of extracellular matrix compared to control groups (p < 0.0001). However, the EM and EH groups presented higher percentage of muscle fiber and lower percentage of extracellular matrix than SH group (p < 0.0001). Regarding muscular gene expression, the SH and EM groups showed a lower expression of PGC1-α (p = 0.0024) and a higher expression of VEGF (p = 0.0033) compared to SC, while PGC1-α was elevated in the EH. No difference between groups was found for the carbonylated protein levels (p > 0.05), while the TNF-α/IL-10 ratio was augmented in the EH (p = 0.0277). In conclusion, voluntary running augments the proportion of fiber and affects the gene expression of inflammatory and mitochondrial biogenesis' markers in the skeletal muscle of rats with MCT-induced PAH, which benefits their survival and tolerance to physical effort.

RESUMO

Pulmonary arterial hypertension is associated with skeletal muscle myopathy and atrophy and impaired exercise tolerance. Aerobic exercise training has been recommended as a non-pharmacological therapy for deleterious effects imposed by pulmonary arterial hypertension. Aerobic physical training induces skeletal muscle adaptations via reduced inflammation, improved anabolic processes, decreased hypoxia and regulation of mitochondrial function. These benefits improve physical exertion tolerance and quality of life in patients with pulmonary arterial hypertension. However, the mechanisms underlying the therapeutic potential of aerobic exercise to skeletal muscle disfunctions in patients with pulmonary arterial hypertension are not well understood yet. This minireview highlights the pathways involved in skeletal muscle adaptations to aerobic exercise training in patients with pulmonary arterial hypertension.

RESUMO

Epithelial cells connect with each other by tight junctions (TJs) in several tissues. In epididymides, TJs proteins form the blood-epididymis barrier (BEB), which is crucial for male fertility. However, little is known about BEB morphological and physiological aspects in wild animals. This study examines the region-specific distribution pattern of TJs proteins in D. rotundus' epididymis, assessing their regulation in rainy and dry season. The expression of zonula occludens-1 (ZO-1), and claudins (Cldn)-1, -3, and -4 were evaluated by confocal immunofluorescence and ELISA analysis. Herein, ZO-1 was strictly expressed in TJs, whereas Cldns were expressed in TJs and basolateral membranes of epithelial cells. Their co-localization and intensity of expression varied in the epididymal regions examined. The effect of season on protein expression was detected mainly in TJ proteins located in the proximal regions. As such, in the initial segment (IS), Cldn-3 and -4 were detected at low levels in basolateral membranes in the rainy season compared to the dry season. Furthermore, in the distal IS, Cldn-1 expression was lower in TJs of epithelial cells during the rainy season than the dry season. ZO-1 expression was higher in the cauda region than the corpus region by ELISA analysis. Additionally, in the corpus region, ZO-1 expression was higher in TJs during dry season compared to the rainy season. Our study sheds light on the understanding of BEB in D. rotundus, improving the knowledge of their reproductive biology.

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Traumatic spinal cord injury results in severe neurological deficits, mostly irreversible. The cell therapy represents a strategy for treatment particularly with the use of stem cells with satisfactory results in several experimental models. The aim of the study was to compare the treatment of spinal cord injury (SCI) with and without mesenchymal stem cells (MSC), to investigate whether MSCs migrate and/or remain at the site of injury, and to analyze the effects of MSCs on inflammation, astrocytic reactivity and activation of endogenous stem cells. Three hours after SCI, animals received bone marrow-derived MSCs (1×107 in 1mL PBS, IV). Animals were euthanized 24 hours, 7 and 21 days post-injury. The MSC were not present in the site of the lesion and the immunofluorescent evaluation showed significant attenuation of inflammatory response with reduction in macrophages labeled with anti-CD68 antibody (ED1), decreased immunoreactivity of astrocytes (GFAP+) and greater activation of endogenous stem cells (nestin+) in the treated groups. Therefore, cell transplantation have a positive effect on recovery from traumatic spinal cord injury possibly due to the potential of MSCs to attenuate the immune response.

A lesão medular resulta em déficits neurológicos graves, a maioria irreversíveis. A terapia celular representa uma estratégia para o tratamento, especialmente com a utilização de células-tronco, com resultados satisfatórios em vários modelos experimentais. O objetivo do estudo foi comparar o tratamento de lesões da medula espinal (SCI), com e sem o uso de células-tronco mesenquimais (MSC), para investigar se as MSCs migram e/ou permanecem no local de lesão, e para analisar os efeitos de MSCs sobre a inflamação, reatividade astrocitária e ativação das células-tronco endógenas. Três horas depois da SCI, os animais receberam as MSC derivadas da medula óssea (1 × 107 em 1 mL de PBS, IV). Os animais foram sacrificados 24 horas, 7 e 21 dias pós-lesão. As MSC não estavam presentes no local da lesão e a avaliação por imunofluorescência demonstrou atenuação significativa da resposta inflamatória com redução em macrófagos marcados com anticorpo anti CD68 (ED1), diminuição da imunorreatividade de astrócitos (GFAP +) e maior ativação das células-tronco endógenas (nestin+) nos grupos tratados. Assim, o transplante de células teve efeito positivo sobre a recuperação de lesão traumática da medula espinal, possivelmente devido ao potencial das MSCs para atenuar a resposta imunológica.

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RESUMO

Traumatic spinal cord injury results in severe neurological deficits, mostly irreversible. The cell therapy represents a strategy for treatment particularly with the use of stem cells with satisfactory results in several experimental models. The aim of the study was to compare the treatment of spinal cord injury (SCI) with and without mesenchymal stem cells (MSC), to investigate whether MSCs migrate and/or remain at the site of injury, and to analyze the effects of MSCs on inflammation, astrocytic reactivity and activation of endogenous stem cells. Three hours after SCI, animals received bone marrow-derived MSCs (1×107 in 1mL PBS, IV). Animals were euthanized 24 hours, 7 and 21 days post-injury. The MSC were not present in the site of the lesion and the immunofluorescent evaluation showed significant attenuation of inflammatory response with reduction in macrophages labeled with anti-CD68 antibody (ED1), decreased immunoreactivity of astrocytes (GFAP+) and greater activation of endogenous stem cells (nestin+) in the treated groups. Therefore, cell transplantation have a positive effect on recovery from traumatic spinal cord injury possibly due to the potential of MSCs to attenuate the immune response.(AU)

A lesão medular resulta em déficits neurológicos graves, a maioria irreversíveis. A terapia celular representa uma estratégia para o tratamento, especialmente com a utilização de células-tronco, com resultados satisfatórios em vários modelos experimentais. O objetivo do estudo foi comparar o tratamento de lesões da medula espinal (SCI), com e sem o uso de células-tronco mesenquimais (MSC), para investigar se as MSCs migram e/ou permanecem no local de lesão, e para analisar os efeitos de MSCs sobre a inflamação, reatividade astrocitária e ativação das células-tronco endógenas. Três horas depois da SCI, os animais receberam as MSC derivadas da medula óssea (1 × 107 em 1 mL de PBS, IV). Os animais foram sacrificados 24 horas, 7 e 21 dias pós-lesão. As MSC não estavam presentes no local da lesão e a avaliação por imunofluorescência demonstrou atenuação significativa da resposta inflamatória com redução em macrófagos marcados com anticorpo anti CD68 (ED1), diminuição da imunorreatividade de astrócitos (GFAP +) e maior ativação das células-tronco endógenas (nestin+) nos grupos tratados. Assim, o transplante de células teve efeito positivo sobre a recuperação de lesão traumática da medula espinal, possivelmente devido ao potencial das MSCs para atenuar a resposta imunológica.(AU)

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A fatty diet is regarded as one of the most important risk factors related to the etiology of colorectal cancer, and this effect is linked to the quantity and principal types of fatty acids consumed. In this study, the chemopreventive effects of different oils on rats were investigated. Forty Wistar rats received 1,2-dimetilhidrazine (DMH) and were divided into 4 groups fed normal lipid diets to which 4% olive, fish, flaxseed, or soybean oils (control) were added. The group fed with fish oil presented higher levels of eicosapentaenoic acid (EPA) and docosahexaenoic acid in hepatic tissue and greater levels of linolenic acid and EPA in adipose tissue compared to the other treatments. In the proximal portion of the colon, lower levels of aberrant crypt foci were found in the fish and flaxseed oil groups; however, this behavior was not observed in the middle and distal regions. Via a benchmarking method, the fish oil group showed a greater transforming growth factor ß expression and lower interleukin-8 expression in relation to the other treatments. Fish oil in a normal lipid diet demonstrated a limited protective effect on the colonic precancerous mucosa in carcinogen-treated rodents, whereas it had a beneficial effect on inflammatory modulation.

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FUNDAMENTO: Métodos convencionais de dissector atualmente requerem consideráveis custos financeiros, técnicos e operacionais para estimar o número de células, incluindo cardiomiócitos, em uma área de 3D. OBJETIVO: Usar a microscopia de fluorescência em um método de dissector modificado para determinar o número de miócitos no tecido cardíaco em condições normais e patológicas. MÉTODOS: O estudo empregou camundongos Wistar machos com quatro meses de idade e peso de 366,25 ± 88,21 g randomizados em grupos controles (GC, n = 8) e infectados (GI, n = 8). Os animais do GI foram inoculados com cepa Y de T. cruzi (300.000 tripomastigotas/50 g). Após oito semanas, os animais foram pesados e sacrificados. Os Ventrículos Esquerdos (VE) foram removidos para análise estereológica da densidade numérica de cardiomiócitos (Nv [c]) e o número total dessas células no VE (N [c]). Esses parâmetros foram estimados usando um dissector fluorescente (DF) e comparados com os métodos convencionais de dissector óptico (DO) e dissector físico (DFi). RESULTADOS: Em ambos os métodos de dissector, os animais do GI apresentaram queda significativa de Nv[c] e N[c] em comparação com os animais do GC (P > 0,05). Uma correlação forte, igual ou superior a 96%, foi obtida entre DF, DO e DFi. CONCLUSÃO: O método DF parece ser igualmente confiável para determinar Nv[c] e N[c] em condições normais e patológicas, apresentando algumas vantagens em relação aos métodos convencionais de dissector: redução de cortes histológicos e imagens na análise estereológica, redução do tempo de análise das imagens, a construção de DF em microscópios simples, utilizando o modo de epifluorescência, distinção de planos de dissector em ampliações inferiores.

BACKGROUND: Conventional disector methods currently require considerable financial, technical and operational costs to estimate the number of cells, including cardyomyocytes, in a 3D area. OBJECTIVE: To use fluorescence microscopy in a modified disector method to determine the number of myocytes in cardiac tissue in normal and pathological conditions. METHODS: The study employed four-month-old male Wistar rats with weight of 366.25 ± 88.21g randomized in control (CG, n=8) and infected (IG, n=8) groups. IG animals were inoculated with T. cruzi Y strain (300,000 trypomastigotes/50g wt). After eight weeks, the animals were weighted and euthanized. The left ventricles (LV) were removed for stereological analysis of numerical density of cardiomyocytes (Nv[c]) and total number of these cells in the LV (N[c]). These parameters were estimated using a fluorescent disector (FD) and compared with the conventional optical (OD) and physical (PD) disector methods. RESULTS: In both disector methods, IG animals presented significant decrease of Nv[c] and N[c] compared to CG animals (P< 0.05). There was no significant difference in these variables despite the disector method applied in CG and IG animals (P> 0.05). A strong correlation, equal or above 96%, was obtained between FD, OD and PD. CONCLUSION: The FD method seems to be equally reliable to determine Nv[c] and N[c] in normal and pathological conditions and presents some advantages compared to conventional disector methods: reduction of histological slices and images in the stereological analysis, reduction of time to analyze the images, construction of FD in simple microscopes using the epifluorescence mode, distinction of disector planes in lower magnifications.

FUNDAMENTO: Métodos convencionales de disector actualmente requieren considerables costos financieros, técnicos y operativos para estimar el número de células, incluyendo cardiomiocitos, en un área de 3D. OBJETIVO: Usar la microscopia de fluorescencia en un método de disector modificado para determinar el número de miocitos en el tejido cardíaco en condiciones normales y patológicas. MÉTODOS: El estudio empleó ratones Wistar machos de cuatro meses de edad y peso de 366,25 ± 88,21 g randomizados en grupos controles (GC, n = 8) e infectados (GI, n = 8). Los animales del GI fueron inoculados con cepa Y de T. cruzi (300.000 tripomastigotas/50 g). Después de ocho semanas, los animales fueron pesados y sacrificados. Los Ventrículos Izquierdos (VI) fueron removidos para análisis estereológico de la densidad numérica de cardiomiocitos (Nv [c]) y el número total de esas células en el VI (N [c]). Esos parámetros fueron estimados usando un disector fluorescente (FD) y comparados con los métodos convencionales de disector óptico (OD) y disector físico (PD). RESULTADOS: En ambos métodos de disector, los animales del GI presentaron caída significativa de Nv[c] y N[c] en comparación con los animales del GC (P > 0,05). Una correlación fuerte, igual o superior a 96%, fue obtenida entre FD, OD y PD. CONCLUSIÓN: El método FD parece ser igualmente confiable para determinar Nv[c] y N[c] en condiciones normales y patológicas, presentando algunas ventajas en relación a los métodos convencionales de disector: reducción de cortes histológicos e imágenes en el análisis estereológico, reducción del tiempo de análisis de las imágenes, la construcción de FD en microscopios simples, utilizando el modo de epifluorescencia, distinción de planos de disector en ampliaciones inferiores.

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BACKGROUND: Conventional disector methods currently require considerable financial, technical and operational costs to estimate the number of cells, including cardyomyocytes, in a 3D area. OBJECTIVE: To use fluorescence microscopy in a modified disector method to determine the number of myocytes in cardiac tissue in normal and pathological conditions. METHODS: The study employed four-month-old male Wistar rats with weight of 366.25 ± 88.21g randomized in control (CG, n=8) and infected (IG, n=8) groups. IG animals were inoculated with T. cruzi Y strain (300,000 trypomastigotes/50g wt). After eight weeks, the animals were weighted and euthanized. The left ventricles (LV) were removed for stereological analysis of numerical density of cardiomyocytes (Nv[c]) and total number of these cells in the LV (N[c]). These parameters were estimated using a fluorescent disector (FD) and compared with the conventional optical (OD) and physical (PD) disector methods. RESULTS: In both disector methods, IG animals presented significant decrease of Nv[c] and N[c] compared to CG animals (P< 0.05). There was no significant difference in these variables despite the disector method applied in CG and IG animals (P> 0.05). A strong correlation, equal or above 96%, was obtained between FD, OD and PD. CONCLUSION: The FD method seems to be equally reliable to determine Nv[c] and N[c] in normal and pathological conditions and presents some advantages compared to conventional disector methods: reduction of histological slices and images in the stereological analysis, reduction of time to analyze the images, construction of FD in simple microscopes using the epifluorescence mode, distinction of disector planes in lower magnifications.

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The objective of this study was to quantify argyrophillic, argentaffin and insulin immunoreactive endocrine cells in the different segments of the small intestine of Didelphis aurita and measure probable differences in the number of these cells between adult and post-pubertal animals. Biological material consisted of ten male and female opossums specimen, divided in two groups according to weigh. The utilized staining techniques were Grimelius, modified Masson-Fontana and direct immunoperoxidase. Results indicated a predominance of argyrophillic cells in the small intestine of opossums from class 1 and 2, with an average of 52.58 and 56.15 cells mm-2, respectively; of which, the average number of total endocrine cells, argyrophillic and argentaffin cells decreased distally in the intestinal segments of opossums from classes 1 and 2. No significant difference was observed for the insulin immunoreactive cells between the intestinal segments of animals from class 2. A greater number of insulin immunoreactive cells was encountered in the jejunum and ileum of animals from class 2 when compared to the same segment in animals from class 1.

Os objetivos deste trabalho foram quantificar as células endócrinas argirófilas, argentafins e imunorreativas à insulina nos diferentes segmentos do intestino delgado de gambás Didelphis aurita e mensurar prováveis diferenças no número destas células entre animais adultos e pós-púberes. Dez exemplares de gambás D. aurita machos e fêmeas foram divididos em dois grupos de acordo com o peso. As técnicas de coloração utilizadas foram Grimelius, Masson-Fontana modificado e Imunoperoxidase direta. Os resultados indicaram um predomínio das células argirófilas no intestino delgado de gambás da classe 1 e 2, com uma média de 52,58 e 56,15 células mm-2, respectivamente. O número médio de células endócrinas totais, células argirófilas e argentafins decresceu distalmente nos segmentos intestinais dos gambás das classes 1 e 2. Nenhuma diferença significativa foi observada para as células imunorreativas à insulina entre os segmentos intestinais dos animais da classe 2. Foi encontrado maior número de células imunorreativas à insulina no jejuno e íleo de animais da classe 2 quando comparado ao mesmo segmento em animais da classe 1.

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Lizards of the family Gekkonidae display a variety of reproductive patterns, as evidenced by the presence of viviparous and oviparous species. The species Hemidactylus mabouia is oviparous. We examined, in vitellogenic females, oviductal structure by light microscopy after routine histological and histochemical techniques, as well as by scanning and transmission electron microscopy. The oviduct is composed of four different regions: the infundibulum, which opens into the coelomic cavity and receives the oocyte released at the time of ovulation; the uterine tube, where sperm storage takes place; the uterus, which is responsible for the eggshell production; and the vagina, the final portion of the oviduct that leads to the cloaca. The oviductal structure of H. mabouia is similar to that of other oviparous lizard species and can be useful for morphological comparative analysis among reptile species.

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