Doxycycline hyclate may damage testicular germinal epithelium in Wistar rats

Broad-spectrum antimicrobial doxycycline acts as an inhibitor of protein synthesis and it is widely used in the clinical treatment of various infections by microorganisms that are sensitive to the drug, as well as in animal feed. Its liposolubility guarantees its high tissue bioavailability, being associated with several biochemical changes in the organism and potentially adverse effects on reproduction. This study aims to evaluate the effects of the action of doxycycline on spermatogenesis to provide a complete analysis of the tubular and interstitial compartments and to identify possible changes in the testicular parenchyma. Adult male Wistar rats were divided into three groups: one control (water), and two treated with doxycycline at the doses of 10mg/kg and 30mg/kg, for 30 days. After euthanasia and sample processing, the following parameters were evaluated: a) tubular diameter and height of the seminiferous epithelium; b) volumetric proportions (%) and volumes (mL) of the components of the testicular parenchyma; c) counting testicular germ cell populations; d) evaluation of cell viability. The results of the comparative evaluation between the experimental groups demonstrated a significant increase in the diameter and area of the tubular lumen and a reduction in the count of spermatogonia in the experimental group that received doxycycline hyclate at a dose of 30mg/kg. In the same experimental group, an increase in the overall yield of spermatogenesis was found as a consequence of the increase in the mitotic index.

Comparison of the spermatogenic process in three different mice strains: Swiss, Balb/C and C57BL/6

Many studies have been trying to establish standard protocols for animal experimentation, especially for animal species or strains, to master research variables with high precision. The main mouse strains used in the field of the biology of reproduction are Swiss, Balb/c, and C57BL/6. Since some of the strains show reproduction limitations, such as the size of the litter, the present study aimed to compare their spermatogenic processes to verify differences regarding the testicular parenchyma and germ cell populations, which could explain low offspring production. In addition, the present study provides additional information concerning the testicular parenchyma of such strains, which consequently would help researchers to choose the most suitable strain for reproductive studies. Six adult male mice were used for each of the strains. After euthanasia, the testes were weighed, fixated with Karnovsky fixative, embedded in methacrylate, sectioned, and stained with toluidine blue/sodium borate 1%. Morphometrical analyses from the testicular parenchyma (seminiferous tubules and interstitium) were made using the software ImageJ. Germ and Sertoli cells populations were counted in seminiferous tubules cross-sections at stage I of the seminiferous epithelium cycle. The lowest body and testicular weights were observed in C57BL/6 animals, followed by Balb/c and Swiss, however, the relative testes, parenchyma, and albuginea weights were significantly lower only in C57BL/6. Despite the seminiferous tubules and seminiferous epithelium proportions were lower in Swiss animals, their relative amount related to the body weight was the same among strains. The total number of germ cells was higher in Swiss animals, reflecting higher spermatogenic yield and daily sperm production. Due to the lower relative number of Sertoli cells, the Swiss animals showed the highest Sertoli cell index and support capacity. On the other hand, the lowest pathological indexes regarding the germ cells were observed in Balb/c animals, followed by Swiss and C57BL/6. In the interstitium, the proportion of blood vessels was lower in Swiss mice, while the lymphatic cell proportion was lower in C57BL/6 animals. Moreover, the highest proportions of Leydig cells and macrophages were noticed in Swiss mice, which may indicate increased testosterone levels. Altogether, such observations must be taken into account when choosing any of the studied strains for reproduction studies.

Biometric, histomorphometric, and biochemical profile in atorvastatin calcium treatment of female rats with dexamethasone-induced osteoporosis / Perfil biométrico, histomorfométrico e bioquímico no tratamento com atorvastatina cálcica de ratas com osteoporose induzida com dexametasona

ABSTRACT Objective: To assess the effects of atorvastatin calcium in the treatment of dexamethasone-induced osteoporosis. Methods: Osteoporosis induction consisted of the administration of an intramuscular dose of 7.5 mg/kg of body weight of dexamethasone, once a week for four weeks, except for the control animals (G1). The animals were divided into the following groups: G1 (control group without osteoporosis), G2 (control group with untreated osteoporosis), G3 (control group with osteoporosis treated with sodium alendronate 0.2 mg/kg) and G4 (group with osteoporosis treated with atorvastatin calcium 1.2 mg/kg). Serum alkaline phosphatase, bone alkaline phosphatase, and biometric and bone histomorphometric assessments were performed after 30 and 60 days of treatment onset. Results: In relation to the biometric and histomorphometric analyses, at 60 days of treatment, G4 presented bone density (Seedor index), bone trabecular density, and cortical thickness of 0.222 ± 0.004 g/cm, 59.167 ± 2.401%, and 387,501 ± 8573 µm, respectively, with a positive and statistically significant difference (p < 0.05), in relation to G2. At 30 and 60 days of treatment, G4 presented statistically significant serum levels of alkaline phosphatase alkaline phosphatase (p < 0.05) that were higher than all groups (7.451 ± 0.173 µg/L and 7.473 ± 0.529 µg/L, respectively). Conclusion: Treatment with atorvastatin calcium demonstrated the ability of this drug to increase osteoblastic activity and bone tissue repair activity, acting differently from alendronate sodium, which demonstrated predominantly antirebsorptive activity.

RESUMO Objetivo: Avaliar os efeitos da atorvastatina cálcica no tratamento da osteoporose induzida com dexametasona. Métodos: A indução da osteoporose consistiu na administração de dexametasona na dose de 7,5 mg/kg de peso corporal, por via intramuscular, uma vez por semana durante quatro semanas, à exceção dos animais do grupo controle (G1). Os animais foram distribuídos nos seguintes grupos: G1 (grupo controle sem osteoporose), G2 (grupo controle com osteoporose sem tratamento), G3 (grupo controle com osteoporose tratado com alendronato de sódio 0,2 mg/kg) e G4 (grupo com osteoporose tratado com atorvastatina cálcica 1,2 mg/kg). Após 30 e 60 dias do início do tratamento dos animais, foram feitas as dosagens dos níveis séricos de fosfatase alcalina, fosfatase alcalina óssea, avaliação biométrica e histomorfométrica óssea. Resultados: Em relação às análises biométricas e histomorfométricas, aos 60 dias de tratamento o G4 apresentou densidade óssea (índice Seedor), densidade trabecular óssea e espessura da cortical de 0,222 ± 0,004 g/cm, 59,167 ± 2,401% e 387,501 ± 8,573 µm, respectivamente, com diferença positiva, estatisticamente significativa (p < 0,05), em relação ao grupo G2. Aos 30 e 60 dias de tratamento, o G4 apresentou níveis séricos de fosfatase alcalina óssea estatisticamente significativos (p < 0,05) e superiores a todos os grupos (7,451 ± 0,173 µg/L e 7,473 ± 0,529 µg/L, respectivamente). Conclusão: O tratamento com atorvastatina cálcica demonstrou a capacidade desse fármaco de aumentar a atividade osteoblástica e a atividade reparadora tecidual óssea, atuar de forma diferente do alendronato de sódio, que demonstrou atividade preponderantemente antirreabsortiva.

Avaliação dos níveis séricos de glicose e insulina no uso de alendronato de sódio e atorvastatina cálcica na osteoporose induzida com dexametasona / Evaluation of serum levels of glucose and insulin in the use of sodium alendronate and atorvastatin calcium in dexamethasone-induced osteoporosis

Introdução: Além da indução da osteoporose, os glicocorticoides ocasionam aumento da resistência à insulina e gliconeogênese hepática, tendo como consequência a hiperglicemia. Objetivo: Avaliar comparativamente os efeitos do alendronato de sódio e da atorvastatina cálcica nos níveis séricos de glicose e insulina na osteoporose induzida com dexametasona. Métodos: A indução da osteoporose consistiu na administração de dexametasona na dose de 7,5 mg/kg de peso corporal, uma vez por semana durante 4 semanas, à exceção dos animais do grupo controle (G1). Os animais foram distribuídos nos seguintes grupos: G1 (grupo controle sem osteoporose), G2 (controle com osteoporose sem tratamento), G3 (com osteoporose tratado com alendronato de sódio 0,2 mg/kg) e G4 (com osteoporose tratado com atorvastatina cálcica 1,2 mg/kg). No período de 30 e 60 após o início do tratamento, foram coletadas amostras de sangue para as dosagens dos níveis séricos de glicose e insulina. Resultados: Os grupos G2 e G3, quando comparados com o grupo normal G1, apresentaram aumento da glicemia e insulinemia durante todo o período experimental. O grupo G4 apresentou, com 30 dias, aumento da glicemia e insulinemia e, com 60 dias, aumento da glicemia e queda da insulinemia. Conclusão: Os resultados demonstraram o quadro de hiperglicemia consequente do aumento da resistência à insulina, presentes na indução da osteoporose pela dexametasona. O alendronato de sódio não ocasionou nenhuma melhora da glicemia e insulinemia. A atorvastatina cálcica ocasionou agravamento da hiperglicemia e hiperinsulinemia, potencializando o quadro de resistência à insulina e levando a uma insuficiência relativa de insulina característica do diabetes mellitus tipo 2.

Introduction: In addition to the induction of osteoporosis, glucocorticoids cause increased insulin resistance and hepatic gluconeogenesis resulting in hyperglycemia. Objective: Evaluate the effects of sodium alendronate and atorvastatin calcium on serum glucose and insulin levels in osteoporosis induced by dexamethasone. Methods: The induction of osteoporosis consisted of the administration of dexamethasone at a dose of 7.5 mg / kg body weight, once a week for 4 weeks, except for the control animals (G1). The animals were divided into the following groups: G1 (control group without osteoporosis), G2 (control with untreated osteoporosis), G3 (with osteoporosis treated with sodium alendronate 0.2 mg / kg) and G4 (with osteoporosis treated with atorvastatin calcium 1,2 mg / kg). In the 30 and 60 period after the start of the treatment blood samples were collected for dosages of serum glucose and insulin levels. Results: The G2 and G3 groups, when compared with the normal group G1, presented increased glycemia and insulinemia throughout the experimental period. The G4 group presented a 30-day increase in glycemia and insulinemia and at 60 days increased glycemia and decreased insulinemia. Conclusion: The results demonstrated the hyperglycaemia associated with the increase in insulin resistance present in the induction of osteoporosis by dexamethasone. Sodium alendronate did not cause any improvement in glycemia and insulinemia. Atorvastatin calcium caused worsening of hyperglycemia and hyperinsulinemia enhancing insulin resistance, leading to a relative insufficiency of insulin characteristic of type 2 diabetes mellitus.

Extract of the Bark of Bathysa cuspidataAttenuates the Development of Chemically-Induced Preneoplastic Colorectal Lesions in Rats

ABSTRACTThe aim of this study was to investigate the effect of the bark extractBathysa cuspidata on chemically induced preneoplastic colorectal lesions in Wistar rats. Forty male rats were randomly divided into four groups (n = 10 each): saline (control group, oral administration of saline solution 0.9%); dimethylsulfoxide (DMSO, vehicle control), B200 (treated with 200 mg/kg bark extract ofB. cuspidata), and B400 (treated with 400 mg/kg bark extract ofB. cuspidata). Administration of treatments was carried out by the gavage. The animals received four subcutaneous injections of 1,2-dimethylhydrazine (DMH, 40 mg/kg) in the initial two weeks of the experiment to induce preneoplastic colorectal lesions. After 15 weeks, the animals were euthanized and the presence of aberrant crypt foci (ACF), body weight, biochemical analyses, and oxidative stress markers were measured. The extract ofB. cuspidata decreased the levels of superoxide dismutase (SOD), but did not influence the levels of catalase (CAT), malondialdehyde (MDA), nitric oxide or protein carbonyl, compared with the saline group. The animals supplemented with a more concentratedB. cuspidata extract (B400) showed a significant reduction in the number of ACF in all the portions of the intestinal mucosa. The study demonstrated that the bark extract ofB. cuspidata at 400 mg/kg reduced the preneoplastic colorectal lesions in an animal model of colon cancer and that the effect could be dose-dependent.

Bark Extract of Bathysa cuspidata in the Treatment of Liver Injury Induced by Carbon Tetrachloride in Rats

The aim of this study was to investigate the effect of bark extract Bathysa cuspidata (BCE) in the reconstitution of hepatic parenchyma and stroma after dysfunction induced by the CCl4. Liver lesions were induced by intraperitoneal administration of CCl4 (1 mL/kg) every 48 h for 12 days. The animals were treated with B. cuspidata extract (BCE) administered by gavage for another 12 days. Forty-nine rats were randomized into seven treatment groups with seven animals receiving CCl4, BCE (200, or 400 mg/kg) and the vehicle DMSO alone, or in different combinations. The extract alone showed no evidence of hepatic toxicity. In general, rats acutely exposed to CCl4 without the treatment with BCE presented high ALT and AST serum levels and high tissue content of lipid hydroperoxides, malondialdehyde, and lipid droplets. BCE administration, especially at 400 mg/kg attenuated significantly all these parameters. Furthermore, the antioxidant activities of the enzymes superoxide dismutase and catalase were significantly increased in the groups receiving this extract. These results showed that the extract of B. cuspidata stem bark stimulated the antioxidant defense system and reduced the morphological and functional liver damage in Wistar rats previously exposed to CCl4.

Efeito da dieta hiperlipídica e do treinamento aeróbico na aterosclerose em camundongos apoE-/- / Effect of hyperlipidic diet and aerobic training on atherosclerosis in apoE-/- mice / Efecto de la dieta hiperlipídica y del entrenamiento aeróbico en la aterosclerosis en ratones apoE-/-

INTRODUÇÃO: A população brasileira tem seguido a tendência dos países desenvolvidos, cuja característica é o aumento do consumo de gorduras e sedentarismo. OBJETIVO: Investigar os efeitos da dieta hiperlipídica e do treinamento sobre o peso corporal, o consumo alimentar, a massa corporal magra, os lipídios plasmáticos, o peso do fígado e a aterosclerose em camundongos com deficiência de apolipoproteína E (apoE-/-). MÉTODOS: vinte e seis camundongos foram divididos em grupos/dieta: NS (normolipídica e sedentário), HS (hiperlipídica e sedentário), NT (normolipídica e treinamento) e HT (hiperlipídica e treinamento). Aferiu-se peso corporal (PC), consumo alimentar (CA) e peso relativo do fígado (PRF). Calculou-se a água da carcaça (A%) pela diferença do peso pré e pós-secagem, gordura (G%) por Soxhlet e proteína (P%) por Kjeldahl. Determinaram-se colesterol total (CT), triglicerídeos (TG) e HDL por método enzimático-colorimétrico e LDL e VLDL pela formula de Friedewald. Analisou-se a aterosclerose por fotomicrografia. Utilizou-se ANOVA e o método de Duncan, com P < 0,05. RESULTADOS: No PC inicial e final, não houve diferença de G% e P% entre os grupos (P > 0,05). A dieta hiperlipídica aumentou o CA (365,3 ± 5,09 vs. 340,16 ± 6,32) e LDL (452,20 ± 114,63 vs. 329,60 ± 77,54) nos animais sedentários e o CT (858,75 ± 140,84 vs. 320,87 ± 157,81) e o LDL (839,36±139,94 vs. 282,66±166,92) nos treinados (P < 0,05). Já o treinamento aumentou o CA (350,4 ± 7,81 vs. 340,16 ± 6,32) e A% (64,23 ± 2,46 vs. 62,62 ± 1,47) nos camundongos normolipídicos e diminuiu o CA (336,68 ± 6,23 vs. 365,3 ± 5,09), TG (22,75 ± 12,36 vs. 66,00 ± 21,88), HDL (14,83 ± 3,95 vs. 37,60 ± 13,12), VLDL (4,55 ± 2,47 vs. 13,20 ± 4,37) e aterosclerose (0,35 ± 0,13 vs. 0,56 ± 0,12) nos hiperlipídicos (P < 0,05). CONCLUSÃO: A dieta hiperlipídica pode desencadear distúrbios no metabolismo lipídico e, com isso, desencadear doenças cardiovasculares; porém, associada ao treinamento, pode diminuir a aterosclerose.

INTRODUCTION: The Brazilian population has followed the trend of developed countries, whose characteristic is the increased consumption of fats and sedentary lifestyle. OBJECTIVE: To investigate the effects of high fat diet and training on body weight, food intake, lean body mass, plasma lipids, liver weight and atherosclerosis in mice with apolipoprotein E (apoE-/-) deficiency. METHODS: 26 rats were divided into groups/diet: NS (normolipidic and sedentary), HS (high fat and sedentary), NT (normolipidic and training) and HT (high fat and training). Body weight (BW), food intake (FI) and relative liver weight (RLW) were measured in a Gehaka BG2000 scales. The water from the carcass (A%) was calculated from the weight difference between the pre-and post-drying ,the fat (% F) was calculated by Soxhlet, and the protein (P%) by Kjeldahl . We determined total cholesterol (TC), triglycerides (TG) and HDL by colorimetric enzymatic method and VLDL and LDL by the Friedewald formula. Atherosclerosis was analyzed by photomicrography. We used ANOVA as well as the Duncan's method, P<0.05. RESULTS: In the initial and final BW, there was no difference in F% and P% between groups (P>0.05). The high fat diet increased the FI (365,3±5,09 vs. 340,16±6,32) and LDL (452,20±114,63 vs. 329,60±77,54) in the sedentary animals and TC (858,75±140,84 vs. 320,87±157,81) and LDL (839,36±139,94 vs. 282,66±166,92) in trained ones (P<0.05). On the other hand, training increased the CA (350,4±7,81 vs. 340,16±6,32) and A% (64,23±2,46 vs. 62,62±1,47) in normolipidic mice and decreased FI (336,68±6,23 vs 365,3 ± 5,09), TG (22,75±12,36 vs. 66,00±21,88), HDL (14,83±3,95 vs. 37,60±13,12), VLDL (4,55±2,47 vs. 13,20±4,37) and atherosclerosis (0,35±0,13 vs. 0,56±0,12) in hyperlipidic mice (P<0.05). CONCLUSION: A high fat diet can trigger disturbances in the lipid metabolism and thereby trigger cardiovascular disease, but when associated with activity may decrease atherosclerosis.

INTRODUCCIÓN: La población brasileña ha seguido la tendencia de los países desarrollados, cuya característica es el aumento del consumo de grasas y el sedentarismo. OBJETIVO: Investigar los efectos de la dieta hiperlipídica y del entrenamiento sobre el peso corporal, el consumo de alimentos, la masa corporal delgada, los lípidos plasmáticos, el peso del hígado y la aterosclerosis en ratones con deficiencia de apolipoproteína E (apoE-/-). MÉTODOS: 26 ratones fueron divididos en grupos/dieta: NS (normolipídica y sedentario), HS (hiperlipídica y sedentario), NT (normolipídica y entrenamiento) y HT (hiperlipídica y entrenamiento). Se controlaron peso corporal (PC), consumo de alimentos (CA) y peso relativo del hígado (PRH). Se calculó el agua de la carcasa (A%) por la diferencia del peso antes y después del secado, grasa (G%) por Soxhlet y proteína (P%) por Kjeldahl. Se determinaron colesterol total (CT), triglicéridos (TG) y HDL por el método enzimático-colorimétrico, y LDL y VLDL por la fórmula de Friedewald. Por fotomicrografía se analizó la aterosclerosis. Se utilizó ANOVA y el método de Duncan, con P < 0,05. RESULTADOS: En el PC inicial y final, no hubo diferencia de G% y P% entre los grupos (P > 0,05). La dieta hiperlipídica aumentó el CA (365,3 ± 5,09 vs. 340,16 ± 6,32) y el LDL (452,20 ± 114,63 vs. 329,60 ± 77,54) en los animales sedentarios, y el CT (858,75 ± 140,84 vs. 320,87 ± 157,81) y el LDL (839,36±139,94 vs. 282,66±166,92) en los entrenados (P < 0,05). Desde otro punto de vista, el entrenamiento aumentó el CA (350,4 ± 7,81 vs. 340,16 ± 6,32) y A% (64,23 ± 2,46 vs. 62,62 ± 1,47) en los ratones normolipídicos y disminuyó el CA (336,68 ± 6,23 vs. 365,3 ± 5,09), TG (22,75 ± 12,36 vs. 66,00 ± 21,88), HDL (14,83 ± 3,95 vs. 37,60 ± 13,12), VLDL (4,55 ± 2,47 vs. 13,20 ± 4,37) y la aterosclerosis (0,35 ± 0,13 vs. 0,56 ± 0,12) en los hiperlipídicos (P < 0,05). CONCLUSIÓN: La dieta hiperlipídica puede causar disturbios en el metabolismo lipídico y, con eso, producir enfermedades cardiovasculares; no obstante, si está asociada con el entrenamiento, puede reducir la aterosclerosis.

Colon cancer and swimming exercise: effect on wistar rat testes

This study was undertaken to determine whether colon cancer (CC) and chronic swimming exercise alter rat testis. Eleven weeks old rats were distributed into control group (n=6) and the groups that were induced to develop CC by dimethylhydrazine injections (nEG, EG0, EG2 and EG4; n=10 each group). In the group nEG, the rats did not swim, whereas groups EG0, EG2 and EG4, underwent a swimming program with distinct loads (0, 2 and 4% of body mass, respectively) for 35 weeks. The morphometry, stereology and cell counts showed damage caused by the CC on the germ epithelium. These results were noteworthy since this was the first report to associate the CC with testicular damage. Swimming exercise had no significant role in reducing, or increasing the CC effects on the testis, despite having slightly improved the testis structure of the exercised rats without load. In conclusion, CC caused testis impairment, which could not be avoided by the swimming exercise.

Stereological analyses of the annual variation of captive bullfrogadult testes (Lithobates catesbeianus, Shaw 1802)

The occurrence of seasonality in bullfrog (Lithobates catesbeianus) spermatogenesis was analyzed by quantifying germ cell populations throughout the year, which were divided into bimesters, from November 2001 to October 2002. Body weight, liver weight, fat body weight and gonadal weight were taken and bimonthly grouped. Using these data, the gonadosomatic, hepatosomatic, and liposomatic indexes were calculated. All animals were kept in the same cement cages in a frog farm, and sampled every month. The cages were underthe influence of climate variables changes, such as photoperiod, temperature, rainfall and relative humidity.Body weight was higher during Spring and Summer, and a significant reduction has occurred between Julyand August. The highest gonadosomatic index values were registered from May to October, coinciding withthe period of gonadal preparation for reproduction. Also, the reproduction of bullfrog is directly related toenvironmental factors, mainly due to Summer and Winter humidity variations.

Efeito dos resíduos de café seco e fermentado por Monascus ruber no metabolismo de camundongos Apo E / Effect of dry coffee residues fermented with Monascus ruber on the metabolism of Apo E mice

FUNDAMENTO: A aterosclerose é uma doença inflamatória crônica de origem multifatorial que ocorre em resposta à agressão endotelial. O fungo Monascus ruber apresenta atividade hipocolesterolêmica e polifenóis presentes no resíduo de café apresentam atividade antioxidante, podendo auxiliar na prevenção de doenças cardiovasculares. O resíduo de café possui quantidade significativa de açúcares fermentescíveis, constituindo-se em substrato apropriado para o cultivo de fungos. OBJETIVO: O objetivo deste estudo foi avaliar o efeito dos resíduos de café seco e fermentado por Monascus ruber no metabolismo lipídico de camundongos knockout Apo E. MÉTODOS: O ensaio biológico foi realizado com 30 camundongos knockout para o gene Apo E, divididos em cinco grupos e submetidos a diferentes tratamentos. Foi realizada a prospecção fitoquímica e quantificação de compostos fenólicos dos resíduos fermentado e sem fermentar. O soro dos animais foi analisado utilizando kits enzimáticos e o tecido aórtico incluso em parafina e corado com H/E para realização da análise histopatológica. RESULTADOS: O resíduo de café sem fermentar 2%, em relação ao grupo controle, diminuiu em 42% o nível sérico de triacilgliceróis e em aproximadamente 41% a fração VLDL-c. Os grupos dos animais alimentados com 10% de resíduo não fermentado e 2% de resíduo fermentado diminuíram a área de lesão 10,5% e 15,4%, respectivamente, quando comparados ao controle. O resíduo fermentado apresentou um teor de compostos fenólicos superior ao resíduo não fermentado. CONCLUSÃO: O presente estudo mostra que a fermentação do resíduo de café apresenta potencial efeito benéfico sobre as doenças cardiovasculares, especialmente a aterosclerose.

BACKGROUND: Atherosclerosis is a chronic inflammatory disease of multifactorial origin, which occurs in response to endothelial injury. The fungus Monascus ruber has hypocholesterolemic activity, and the polyphenols present in coffee residue have an antioxidant activity and can help prevent cardiovascular diseases. Coffee residue has a significant amount of fermentable sugars, being an adequate substrate for growing fungi. OBJECTIVE: The objective of this study was to assess the effect of dry coffee residue fermented with Monascus ruber on the lipid metabolism of ApoE knockout mice. METHODS: The biological assay was performed with 30 ApoE knockout mice, divided into five groups and undergoing different treatments. The phytochemical prospection and quantification of phenolic compounds of the fermented and non-fermented coffee residues were performed. The sera of the animals were analyzed by using enzyme kits, and the aortic tissue was embedded in paraffin and stained with hematoxylin and eosin to undergo histopathological analysis. RESULTS: Comparing with the control group, the group receiving 2% non-fermented coffee residue showed a reduction of 42% in the serum levels of triacylglycerols and of approximately 41% in VLDL-c. The groups receiving 10% non-fermented coffee residue and 2% fermented coffee residue showed reductions in the lesion areas of 10.5% and 15.4%, respectively, as compared with the control group. The fermented coffee residue showed a higher content of phenolic compounds as compared with the non-fermented coffee residue. CONCLUSION: The present study showed that coffee residue fermentation has a potentially beneficial effect on cardiovascular diseases, especially atherosclerosis.

Testicular morphology of adult wistar rats treated with Rudgea viburnoides (Cham.) Benth. leaf infusion

The present study was carried out to investigate the effects of Rudgea viburnoides infusion on the body biometry and testicular morphometry and stereology of adult Wistar rats. Two groups received the infusion daily at the concentration of 3 or 6 mg/mL for 40 days. The control group received only water. Neither the biometrical parameters nor the tubular diameter and the height of the seminiferous epithelium showed any significant alterations in the treated animals. Leydig cells stereology did not show any significant alterations in the treated animals. These results indicated that R. viburnoides did not cause alterations in body biometry and testicular morphometry.

Effects of catuaba cristal® on the testis of wistar rats

The aim of this study was to evaluate the possible beneficial effects of Catuaba Cristal® (CC), an alcoholic drink made from wine and Erythroxylum catuaba Ar. Cam on testis. Wistar rats either received CC solution (n=8) or water (n=9). Results showed significant body weight reduction within the CC group, although, no weight changes were observed for liver, kidney, testis, epididymis, seminal vesicle and prostate. The volumetric proportion and volume of interstitial tissue and lymphatic space were reduced in the treated group. In the CC group, although the nuclear volume of Leydig cells (LC) decreased, the number of LC per testis increased. These results suggested that CC had no beneficial effect on spermatogenesis of Wistar rats.

Quantification of argyrophillic, argentaffin and insulin immunoreactive cells in the small intestine in the opossum Didelphis aurita (Wied-Neuwied, 1826) / Quantificação das células argirófilas, argentafins e imunorreativas à insulina nos diferentes segmentos do intestino delgado do gambá Didelphis aurita (Wied-Neuwied, 1826)

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.

Monitoramento por radiotelemetria da área de uso de onça parda reintroduzida no entorno do Parque Estadual da Serra do Brigadeiro - MG, Brasil / Radiotelemetry monitoring of the home range of cougar reintroduced at the surroundings of the State Park of Serra do Brigadeiro - MG, Brazil

Um exemplar macho, subadulto de onça parda reintroduzido foi monitorado por técnica de radiotelemetria. O monitoramento foi de 110 dias, realizado de forma aleatória, com intervalos descontínuos. Os dados do acompanhamento sugerem que a área utilizada por este espécime seja de, aproximadamente, 26km². Esta configura o menor índice já descrito para a espécie, porém, deve-se ressaltar a importância deste monitoramento básico, visto a incipiência do comportamento de um felino de grande porte reintroduzido em habitat natural após período em cativeiro.

One subadult male specimen of a reintroduced mountain lion was monitored using the radio telemetry technique. The monitoring lasted 110 days, being randomly distributed. Data of the monitoring through radiotelemetry suggested that the home range of this specimen was of approximately 26km². This sets as the smallest home range described for this species. However, the importance of this basic monitoring should be emphasized, since information of the behavior of a large feline reintroduced in its natural habitat after a period in captivity is incipient.

Ácido linoleico conjugado e exercício físico: efeitos na aterosclerose de camundongos apoe (-/-) / Conjugated linoleic acid and exercise: effects on atherosclerosis of apo e(-/-) Mice

Fundamento: Nenhum estudo foi encontrado em relação aos efeitos da combinação de ácido linoleico conjugado (CLA) e exercício físico na progressão de aterosclerose de camundongos APO E (-/-). Objetivo: Avaliar os efeitos da combinação de exercício físico e ácido linoleico conjugado (CLA) na progressão de aterosclerose de camundongos knockout para o gene da Apo E alimentados com dietas normo e hiperlipídica. Métodos: Camundongos knockout para Apo E foram alocados em quatro grupos/dieta: NS - dieta normolipídica e sedentário (n=5), HS - dieta hiperlipídica e sedentário (n=5), NECLA - dieta normolipídica com CLA e exercitado (n=8) e HECLA - dieta hiperlipídica com CLA e exercitado (n=8). O colesterol total e o HDL-C foram determinados através do método enzimático-colorimétrico. O LDL-C foi calculado pela fórmula de Friedewald. O fígado foi pesado e as lesões ateroscleróticas foram analisadas por fotomicrografia representativa da aorta. Utilizou-se ANOVA e Tukey ao nível de significância de 5%. Resultados: O grupo HECLA apresentou maiores valores de colesterol total e LDL-c que os grupos NECLA e NS (p<0,05). Em relação ao HDL-c, o grupo HS apresentou maior concentração que o grupo HECLA (p=0,019). O peso do fígado foi maior no grupo HECLA comparado com o NECLA (p=0,003). Em relação à progressão da aterosclerose, não foi encontrado diferenças significativas entre os grupos (p>0,05). Conclusões: A combinação exercício físico e CLA, independente do tipo de dieta, não foi eficiente na redução da progressão de aterosclerose de camundongos Knockout para o gene que expressa a apolipoproteina E.

Background: No studies were found regarding the effects of the combination of conjugated linoleic acid (CLA) and physical exercise in the progression of atherosclerosis in mice APO E (-/-). Objective: To evaluate the effects of the combination of exercise and conjugated linoleic acid (CLA) on the progression of atherosclerosis in mice knockout for the gene Apo E fed diets with normal and hyperlipidemic. Methods: Apo E knockout mice were divided into four groups / diet: NS - normolipídica diet and sedentary (n = 5), HS - high fat diet and sedentary (n = 5), NECLA - normolipídica diet with CLA and exercised (n = 8) and HECLA - High fat diet with CLA and exercised (n = 8). Total cholesterol and HDL-C were determined by enzymatic-colorimetric method. LDL-C was calculated using the Friedewald formula. The liver was weighed and atherosclerotic lesions were analyzed by representative photomicrographs of aorta. We used ANOVA and Tukey tests at a significance level of 5%. Results: The HECLA group had higher total cholesterol and LDL-C than groups NECLA and NS (p <0,05). In relation to HDL-C, the HS group had a higher concentration than the group HECLA (p = 0,019). Liver weight was higher in the group HECLA compared with NECLA (p = 0,003). Regarding the progression of atherosclerosis, was not found significant differences between groups (p> 0.05). Conclusions: The combination of exercise and CLA, regardless of the diet was not effective in reducing the progression of atherosclerosis Knockout mice for the gene that expresses the apolipoprotein E.

Treinamento em natação com baixa intensidade não protege músculo esquelético contra lesões induzidas por exercício exaustivo em ratos / Low-intensity swimming training does not protect the skeletal muscle

Enquanto o exercício aeróbico regular promove adaptações benéficas ao músculo esquelético, o exercício físico exaustivo induz lesões musculares. O objetivo deste estudo foi verificar se um programa de natação com baixa intensidade é capaz de proteger músculos esqueléticos contra lesões induzidas por exercício exaustivo. Ratos Wistar (peso: 376,50 ± 4,36g; idade: 90 dias) foram divididos aleatoriamente em quatro grupos: controle sedentário (CS); sedentário submetido a teste de exaustão (SE); treinado em natação (TN); treinado em natação submetido a teste de exaustão (TNE). Animais dos grupos TN e TNE foram submetidos a um programa de natação sem sobrecarga por 90 minutos/dia, cinco dias/semana, durante 17 semanas. Após este período, os grupos SE e TNE foram submetidos a um teste de exaustão em natação. Após eutanásia, fragmentos dos músculos sóleo e reto femoral foram coletados e submetidos à análise histológica e de proteínas de choque térmico (HSP70). Os resultados mostraram que o tempo até a exaustão foi maior no grupo TNE que no SE (125,0 ± 6,0 vs. 90,0 ± 8,5min, respectivamente, P < 0,05). Os níveis de lactato sanguíneo durante o teste e exaustão foram menores no grupo TNE que no SE (5,31 ± 0,22 vs. 8,76 ± 0,59mmol/L, respectivamente, P < 0,05). A frequência de fibras lesadas nos músculos foi maior nos grupos SE (sóleo: 34,86 ± 0,04; reto femoral: 37,57 ± 0,07) e TNE (sóleo: 41,57 ± 0,08; reto femoral: 39,57 ± 0,05), comparada aos grupos CS (sóleo: 13,88 ± 0,81; reto femoral: 16,75 ± 0,79) e TN (sóleo: 24,14 ± 0,06; reto femoral: 24,0 ± 0,05), respectivamente (P < 0,05). Não houve diferença significativa nos níveis de HSP70 dos músculos analisados entre os quatro grupos. Concluimos que apesar do treinamento em natação melhorar o desempenho dos animais no teste de exaustão, não promoveu proteção aos seus músculos esqueléticos contra as lesões induzidas pelo exercício exaustivo.

While regular aerobic exercise promotes beneficial adaptations to the skeletal muscle, acute exhaustive exercise causes structural damage to the skeletal muscle cells. The aim of this study was to verify whether a low-intensity swimming program protects the skeletal muscles against damage induced by exhaustive exercise. Male Wistar rats (weight: 376.50 4.36g; age: 90 days) were randomly divided into four groups: sedentary control (SC, N=8); sedentary submitted to exhaustive test (SE, N=7); swimming trained (TN, N=7); swimming trained submitted to exhaustive test (TNE, N=7). Animals of TN and TNE groups were submitted to a swimming regimen without overload for 90 min/day, 5 days/wk, during 17 weeks. Forty-eight hours after the last training session, animals from SE and TNE groups were submitted to an exhaustive exercise protocol. At sacrifice, fragments of soleus and rectus femoris muscles were collected and submitted to histological analysis and heat shock protein (HSP70) expression measurement. The results showed that the time until exhaustion was greater in the STE than in SE group (125.0 6.00 vs. 90.0 8.48 min, respectively, P<0.05). The levels of blood lactate during exhaustive exercise were lower in animals from TNE than SE (5.31 ± 0.22 vs. 876 ± 0.59 mmol/L, respectively, P<0.05)The frequency of damaged fibers in the muscles was greater in SE (soleus: 34.86±0.04; rectus femoris: 37.57 ± 0.07) and STE (soleus: 41.57±0.08; rectus femoris: 39.57 ± 0.05), compared to groups SC (soleus: 13.88±0.81; rectus femoris: 16.75 ± 0.79) and ST (soleus: 24.14±0.06; rectus femoris: 24.0 ± 0.05), respectively (P<0.05). There was no significant difference at the HSP70 levels of the analyzed muscles among the four groups (P>0.05). In conclusion, although a low-intensity swimming training increased the animals' performance in the exhaustive exercise test, it did not protect their skeletal muscles against damage induced by exhaustive exercise.

Microscopic morphology and testis morphometry of captivity-bred Adult bullfrogs (Lithobates catesbeianus Shaw, 1802)

The aim of this work was to study the testicular morphometry of captivity-bred adult bullfrogs. Fifteen young adult male were studied, in the rainy season and a lengthy photoperiod. The GSI was established at 0.15 percent. The nuclear diameter of germinative and Leydig cells, the nucleolus diameter of Sertoli cells and the area of cysts and tubules were determined and the mean number of ISPC, IISPC and SPT per cyst and the mean number of cysts per tubule was estimated. The nucleoplasmatic proportion of the nucleus of the Leydig cell was 76.22 percent, indicating less cytoplasmic activity. Eight generations of spermatogonia were found. The spermatogenesis efficiency in meiosis and in mitosis was 63 and 49 percent, respectively. The spermatogenesis of bullfrog fited in the pattern of other captivity Anurans, with differences as the morphology of Sertoli and Leydig cells nuclei.

A morfometria é uma importante ferramenta para a biologia estrutural, permitindo estudos estereológicos e análises quantitativas. Existem muitos pontos a serem esclarecidos sobre a morfometria testicular desta espécie, que objetivamos desvendar neste trabalho. Quinze machos adultos foram estudados, em período chuvoso e de fotoperíodo longo (dezembro, 2000). O IGS encontrado foi de 0.15 por cento. O diâmetro nuclear das células germinativas e da célula de Leydig, o diâmetro nucleolar das células de Sertoli e a área dos cistos e túbulos foram determinados. O número médio de ISPC, IISPC e SPT por cisto e o número médio de cisto por túbulo foi estimado. A proporção nucleoplasmática do núcleo da célula de Leydig foi de 76.22 por cento, indicando pouca atividade citoplasmática. Oito gerações de espermatogônia foram estimadas. A eficiência da espermatogênese na meiose e mitose foi de 63 por cento e49 por cento, respectivamente. A espermatogênese de rãtouro segue os padrões dos demais Anuros de cativeiro, apresentando diferenças nos núcleos das células de Sertoli e Leydig.

Investigation of liver tissue and biochemical parameters of adult wistar rats treated with Arctium lappa L

This study was carried out to evaluate the effects of Arctium lappa L. (burdock) on the liver of adult male Wistar rats as measured by light microscopy and biochemical parameters. The rats received the extract in water bottles at doses of 10 or 20 g/L daily for 40 days. There were no significant changes in the plasma levels of albumin, aspartate transaminase (AST), alanine transaminase (ALT), gamma glutamyl transferase (GGT), total protein, total cholesterol, urea, uric acid, triacylglycerol, calcium, phosphorus, chlorine and direct bilirubin. The morphological analysis did not reveal histopathological alterations in liver tissue. Both biochemical and morphological data did not indicate A. lappa toxicity.

A bardana (Arctium lappa L) é uma planta trazida do Japão e aclimatada no Brasil, e é extensamente utilizada na medicina popular em todo mundo. Este estudo foi realizado para avaliar os possíveis efeitos da A. lappa no fígado e nos parâmetros bioquímicos plasmáticos em ratos Wistar adultos. Estes receberam a infusão de bardana nas doses de 10 ou 20 g de folhas secas /L de água, por 40 dias. Não houve alteração significativa nos níveis plasmáticos de albumina, aspartato transaminase (AST), alanina transaminase (ALT), gamma glutamil transferase (GGT), proteínas totais, colesterol total, uréia, ácido úrico, triglicérides, cálcio, fósforo, bilirrubina direta e cloro. A análise morfológica não revelou alterações histopatológicas no fígado. Os dados bioquímicos e morfológicos não indicaram a toxicidade da bardana.

Liver morphology and morphometry and plasma biochemical parameters of Wistar rats that received leaf infusion of Rudgea viburnoides Benth. (Rubiaceae)

Rudgea viburnoides leaves are widely used in popular Brazilian medicine as a diuretic, antirheumatic, hypotensive and blood depurative tea. The present study was carried out to investigate the effects of this infusion on the liver and on the plasma biochemical parameters of Wistar rats. Two groups received the R. viburnoides leaf infusion at a daily dose of 10 or 20g dry-leaves/L water, during 40 days. The histopathological analysis did not show degenerated areas or infiltration of leucocytes. Hepatic morphometry showed accumulation of fat in the hepatocytes of the treated groups. There was no significant change in the plasma levels of urea, creatinin, uric acid, direct bilirubin, cholesterol, total proteins, albumin, gamma glutamyl tranferase (gamma-GT), alanine transaminase (ALT), aspartate transaminase (AST), chlorine, phosphate and calcium. A significant reduction in the plasma levels of triacylglycerol (TAG) occurred in the group that received the higher dose.

As folhas de Rudgea viburnoides Benth. são utilizadas na medicina popular como diuréticas, hipotensoras, anti-reumáticas, depurativas do sangue e em regimes de emagrecimento. O presente estudo foi delineado para avaliar o efeito da infusão das folhas de R. viburnoides nos parâmetros bioquímicos plasmáticos e na morfologia e morfometria hepática de ratos Wistar adultos. Dois grupos receberam a infusão das folhas, diariamente, nas dosagens de 10 e 20 g de folhas secas/L de água, durante 40 dias. O grupo controle recebeu a mesma quantidade de água. As análises histopatológicas não mostraram áreas degeneradas e infiltrados inflamatórios. A morfometria hepática mostrou acúmulo significativo de gordura nos hepatócitos dos animais tratados, principalmente no grupo que recebeu a maior dose da infusão (8,75 por cento de gotículas lipídicas), comparado com 0,25 por cento delas encontradas nos animais controles. Não foram observadas alterações nos níveis plasmáticos de uréia, creatinina, ácido úrico, bilirrubina direta, colesterol, proteínas totais, albumina, gama-glutamil transferase (GGT), alanina transaminase (ALT), aspartato transaminase (AST), cloro, fósforo e cálcio. Foi observada redução significativa de 20,6 por cento nos níveis plasmáticos de triacilglicerol no grupo que recebeu a infusão mais concentrada, comparada aos animais controle.