Caracterização de um modelo experimental de neuropatia em ratos diabéticos induzidos pela aloxana

Cem ratos norvégicus, machos, com aproximadamente 3 meses de idade foram distribuídos por sorteio em 2 grupos experimentais: Grupo Controle (GC): com 50 ratos sadios, não diabéticos e Grupo Diabético (GD): com 50 ratos diabéticos, induzidos pela aloxana, sem qualquer tratamento. Cada grupo foi dividido em 5 subgrupos com 10 ratos cada e sacrificados com 1, 3, 6, 9 e 12 meses de seguimento, respectivamente. Parâmetros clínicos (peso, ingestão hídrica e alimentar, e diurese) e laboratoriais (glicemia, glicose urinária e insulina) foram documentados em todos os momentos de avaliação. Um segmento do nervo ciático foi obtido de cada animal, em ambos os grupos, para estudo à MO. e ME. Alterações clínicas e laboratoriais significativas (P<0,01), compatíveis com diabetes grave, foram observadas em todos os animais do GD a partir do 4o dia após a indução. Ratos de ambos os grupos apresentaram alterações no número de fibras mielínicas e nos depósitos intraaxonais de glicogênio que não diferiram, estatisticamente, aos 1, 3 e 6 meses de seguimento. Entretanto, aos 9 e 12 meses, ratos do GD apresentaram diminuição significativa no número de fibras mielínicas, com aumento do número de fibras mielínicas de menor calibre, quando comparados com ratos do GC (P<0,05). Grânulos de glicogênio intraaxonais também foram mais acentuados em ratos do GD no 9o e 12o mês de seguimento. Não foram observadas diferenças na densidade de fibras amielínicas ou alterações ultraestruturais significativas entre os dois grupos, em relação aos espaços intraaxonais e endoneurais, bainhas de mielina e células de Schwann durante todo o estudo.

Experimental alcoholism and pathogenesis of prostatic diseases in UChB rats

Previous studies have shown that long-term alcohol treatment has negative effects on prostatic stromal-epithelial interaction. Thus, the aim of the present study was to analyze the histochemical, immunohistochemical and ultrastructural alterations that occur in the prostatic stroma and epithelium of rats submitted to chronic alcohol ingestion and alcohol abstinence, as well as to establish the relationship between these changes and prostatic diseases. Thirty male rats (10 Wistar and 20 UChB rats) were divided into three experimental groups: the control group received tap water, the alcoholic group received ethanol diluted to 10 degrees G.L. for 150 days, and the abstinent group received the same liquid diet as the alcoholic group up to 120 days of treatment and only tap water for 30 days thereafter. At the end of treatment, all animals were sacrificed and the ventral lobe of the prostate was removed and processed for histochemical, immunohistochemical and ultrastructural analyses. In addition, plasma testosterone levels were measured. The results showed, prostatic intraepithelial neoplasia, infolding of the epithelium towards the stroma, stromal hypertrophy and the presence of inflammatory cells in alcoholic animals. In the abstinent group, alterations were noted mainly in the stromal area. In conclusion, ethanol triggers alterations in prostatic epithelial and stromal compartments, affecting the stromal microenvironment and predisposing the organ to pathological processes. (C) 2006 International Federation for Cell Biology. Published by Elsevier Ltd. All rights reserved.

Bull semen varibles after experimental exposure with Bovine Herpesvirus type 5

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

O RATO COMO MODELO EXPERIMENTAL DO DIABETES - ESTUDO CLINICO-LABORATORIAL

In this study, 90 Wistar rats were used. They were equally divided into three experimental groups - control group (CG), diabetic group (DG) and treated diabetic group (TG). The analyzed parameters were clinical (behavior, activity, general aspect, weight, water ingestion and diuresis) and biochemical (fasting glycemia and urinary glycosis). The diabetes was induced by alloxan and, then, treated with insulin associated to oral hypoglycemic (acarbosis). Observations were made at 5 experimental moments, as it follows: 1, 3, 6, 9, and 12 months after the diabetes induction. The results were submitted to variance analysis, with 5% of significance level. The DG presented lower weight and higher diuresis level than the CG and TG. The water ingestion of the CG was similar to TG. The glycemia levels were higher in DG than in CG, at every experimental moment. The TG, however, presented glycemia similar to the CG, except for the dosages at 3, and 9 months. They urinary glycosis of the DG and TG were similar between themselves, but higher than the one of the CG.

Apical leakage after root canal filling with an experimental calcium hydroxide gutta-percha point

This study was an in vitro analysis of the apical seal of root canals filled with an experimental calcium hydroxide gutta-percha point. One hundred and ten recently extracted human permanent teeth with a single root were used. The root canals were biomechanically prepared, and some received a dressing of calcium hydroxide for 7 days. Root canals not dressed were filled by the lateral condensation technique with either regular gutta-percha or calcium hydroxide gutta-percha points. The dressed root canals were filled with the same sealers and regular gutta-percha points. The specimens were placed into a 2% methylene blue dye solution under vacuum, and the apical leakage was linearly evaluated. The experimental groups with calcium hydroxide dressing and the one with calcium hydroxide gutta-percha points exhibited significantly (p < 0.05) less leakage than the groups filled with regular gutta-percha points. The results obtained indicated that calcium hydroxide guttapercha points produced an improvement in the apical sealing quality of the root canal filling. Copyright © 1996 by The American Association of Endodontists.

Effects of chronic experimental alcoholism on the epithelium of the uterine horn of rats (Rattus norvegicus albinus)

Sixty adult tats (Rattus norvegicus albinus) of the same age (3 months) and with a mean body weight of 228 g were divided into two experimental groups. The control group received solid diet (Purina rat chow) and tap water ad libitum. The other (alcoholic group), received the same solid diet and was allowed to drink only sugar cane brandy dissolved in 30° Gay Lussac (v/v). At the end of periods of 90, 180 and 270 days of treatment, the animals were anaesthetized with ethyl ether during estrus, weighed and sacrificed. The final mean body weights were similar in the control and alcoholic groups. The results showed intense atrophy on the lining epithelium of the endometrium of uterine horns in the alcoholic group. Important ultrastructural epithelial alterations were also observed in the female alcoholic group, such as: intense lipid droplet accumulation, increased rough endoplasmic reticulum cisternae and mitochondrial size and presence of intraepithelial neutrophils. The secretory activity of these rats was reduced. Therefore, we concluded that alcohol acts as a toxin on the epithelial layer of the rat endometrium.

Avaliação experimental dos efeitos do anestésico local ropivacaína em nervos ciáticos

This study was undertaken to investigate the effects of ropivacaine after intrafascicular injection into the sciatic nerves of albino rabbits. Twenty adult albine rabbits were used, following sedation with intramuscular ketamine (50 mg/kg) for nerve exposure by lateral incision. We considered three experimental groups: Group I:sciatic nerve control; Group II: intrafascicular injection with 0.2 mL of physiologic saline solution in the left nerves and intrafascicular injection with 0.2 mL of local anesthetic ropivacaine into the rigth nerves. The specimens were colected at 48 h after drugs administration; Group III. intrafascicular injection with 0.2 mL of physiologic saline solution in the left nerves and intrafascicular injection with 0.2 mL of local anesthetic ropivacaine in the rigth nerves. The specimens were colected at 7 days after drugs administration. The sciatic nerves were removed from these animals and fixed in Karnowisky solution for 24 hours. After partial dehydration up to 95% ethanol, they were embedded in historesin (Leica). The tissue was then sectioned at 1-2μm. Sections were stained with haematoxylin-eosin (HE); toluidine blue (TB) or picrosirius-haematoxylin (PSH). Comparing with control group the histological evidence of inflammatory reaction (migration of macrophagic cells and eosinophils-appeared soon after injection, with intense proliferation of perineurial cells. The results show that after 7 days of intrafascicular injection there was a severe fibrosis and an increase on perineurial vascularization. In group 2 the inflammatory reaction was noted near the local of the injection. Furthermore in this experiment we observed an increase on the number of epineurial lipoblasts and adipocytes. This study demonstrated that the toxic effects of ropivacaine are transient. In many cases there was an initial fascicular recover and axonal regeneration after 7 days of the injection.

Methods of experimental induction of periapical inflammation. Microbiological and radiographic evaluation

Aim: To evaluate the influence of coronal filling and apical perforation on the induction of periapical inflammation. Methodology: Fifty-eight root canals in the teeth of dogs were divided into four groups. Groups I and II: root canals were exposed for 180 days; groups III and IV: root canals were exposed for 7 days and then the access cavity filled for 53 days. The root apices of groups I and III were perforated after the coronal opening, whilst those of groups II and IV remained intact. Standard radiographs were taken before and after the experimental periods. Digital images of the radiographs were created and then analysed by three examiners. After induction of periapical inflammation, the root canal contents were collected using paper points. Microbiologic evaluation of the type of microorganism was carried out by culture in different growth media. The radiographic and microbiologic data were statistically analysed using ANOVA at a 5% significance level. Results: There were a greater total number of microorganisms in groups I and II (P < 0.05). The number of anaerobes was greater than the number of aerobes (P < 0.05). The size of the periapical radiolucencies were not significantly different between the experimental groups. Conclusions: The different methods analysed induced similar areas of periapical radiolucency in dogs with predominantly anaerobic bacteria. However, the time required for induction was less when the method with coronal filling was used. © 2005 International Endodontic Journal.

The number of podocyte and slit diaphragm is decreased in experimental diabetic nephropathy

Purpose: To determine the number of podocyte, slit diaphragms, slit diaphragm extensions and GBM thickness in diabetic nephropathy. Methods: Sixty Rattus Wistarof both sexes weighing 200-300g were divided in two experimental groups: normal group 10 animals, and alloxan diabetic rats - 50 animals. Alloxan was administered in a single IV dose of 42mg/kg body weight. Body weight, water and food intake, diuresis, and blood and urine glucose were determined in both groups before alloxan injection and two weeks, six and twelve months after alloxan injection. Proteinuria was measured at 12 months in both groups. After 12 months animals were sacrificed, and the right kidney processed for electron microscopy. Results: Clear clinical and laboratory signs of severe diabetes were seen, in all alloxan-diabetic rats at all follow-up times. Glomerular basement membrane (GBM) thickening, podocyte number, and slit diaphragm number and extension were determined. GBM of all diabetic rats was significantly thicker (median=0.29μm; semi-interquartile range=0.065μm) than in the normal rats (0.23μm; 0.035μm). Diabetic rat podocyte number (8; 1), slit diaphragm number (4; 1), and slit diaphragm extension (0.021μm; 0.00435μm) were significantly lower than in normal rats (11; 1) and (7; 1.5), and (0.031μm; 0.0058μm). Diabetic rat proteinuria (0.060mg/24h; 0.037mg/24h) was higher than in normal rats (0.00185mg/24h; 0.00055mg/24h). Conclusion: Experimental diabetes is associated with significant (p<0.05) changes in podocyte foot process, slit number, slit diaphragm extension, and GBM thickness.

Efeito da mitomicina c no epitélio ciliar estudo experimental em coelhos com microscopia eletrônica de varredura e de transmissão

Purpose: To evaluate and correlate in the rabbit the possible changes caused by mitomycin C under the scleral flap in the ciliary epithelium with transmission electron microscopy (TEM) and scanning electron microscopy (SEM). Methods: The eyes of 32 albino rabbits were studied and divided in 4 experimental groups. The right eye (RE) was intended for the experimental groups and the left eye (LE) for the controls. Group I (G1) was formed by 8 eyes that received 0,5 mg/ml of mitomycin C under the scleral flap and were examined after 15 days. Group II (G2) differed from G1 only in the time of the exam, after 30 days. Group III (G3) was formed by 8 eyes that received 0,2 mg/ml of mitomycin C under the scleral flap and were examined after 15 days. Group IV (G4) differed from group 3 just in the time of the exam, after 30 days. In each eye the internal ciliary epithelium were examined with TEM. Results: The following changes in the internal ciliary epithelium were observed in groups G2, G3, and G4 with TEM: discontinuous and irregular basement membrane, more irregular and electron-dense nucleous, enlargement among interdigitation, edematous mitochondria and myelin figures. These alterations were not found in all the animals of the groups. Group G 1 did not present alterations. Roughness in groups G 1, G2, G3 and G4 were observed with SEM. In groups G 1 and G2 continuity solutions were also observed. Conclusion: Mitomycin C in 0,2 mg/ml and 0,5 mg/ml concentrations caused changes in the internal ciliary epithelium 15 and 30 days after, with TEM and SEM. There was no correlation between dosage, time and with TEM and SEM.

Experimental model of pulmonary carcinogenesis in Wistar rats

Purpose: To elaborate an experimental model of pulmonary carcinogenesis in Wistar rats. Methods: Male Rattus norvegicus albinus, Wistar lineage was carried through an intra-pulmonary instillation of the Benzo[a]pyrene (B[a]P) dilution in alcohol 70%, a polycyclic aromatic hydrocarbon widely known by its power of tumoral induction. Three experimental groups had been formed with 08 animals each: Control Group (Alcohol 70%); B[a]P Group 10 mg/kg; e B[a]P Group 20mg/ kg, submitted to euthanasia 08, 10, 12 and 14 weeks after the experimental procedure. The pulmonary sections had been colored by hematoxilin-eosin (HE) and submitted to the morphometrical analysis to describe the tissue alterations. Results: The presence of diffuse inflammatory alterations was observed in all groups, however, at the analysis of the pulmonary tissue of the experimental groups, it had been observed hyperplasic alterations (BALT hyperplasia), and in one of the animals of the experimental group 20mg/kg (12 weeks), it was noticed the presence of cellular epithelial tracheal pleomorphism, suggesting the adenocarcinoma formation in situ. Conclusion: The main secondary alterations to the intra-pulmonary instillation of B[a]P in Wistar rats were: cellular proliferation, inflammatory alterations of several degrees and nodular lymphoid hyperplasias. The association of an activator agent of the pulmonary metabolic reply is necessary to establish the ideal reply-dose to the development of the lung cancer.

Use of fibrin glue derived from snake venom in the repair of deep corneal ulcers - Experimental study in dogs (Canis familiaris, Linnaeus, 1758)

Fibrin glue has been researched as an alternative method for tissue synthesis and is known for its capability to promote hemostasis at the application site, good approximation of wound edges and fast healing. The current study consisted in the application of fibrin glue derived from snake venom as treatment for experimental corneal ulcers. Twenty-one dogs had their corneas experimentally prepared through lamellar keratectomy (of standardized diameter and depth). Animals were divided into seven groups of three animals each. Six experimental groups were periodically evaluated and collection was carried out on the 1st, 3rd, 7th, 15 th, 30th and 60th post-operative days, whereas one control group was evaluated throughout the experiment. Analyses consisted in the clinical evolution and in the histopathological study of all operated on eyes. Results indicated that fibrin glue was efficient in repairing keratectomy wounds in dogs and contributed to an earlier healing phenomenon, avoiding edema formation and keeping corneal clearness. The use of fibrin glue derived from snake venom showed to be easy to apply, feasible with animal models and of low cost, avoiding the lesion progress and allowing fast and appropriate corneal healing.

Experimental vaccination against Newcastle disease in Japanese quails (Coturnix coturnix japonica): Clinical and immunological parameters

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Response to experimental vaccination against newcastle disease in domestic ducks (Anas platyrhynchos): Clinical and immunological parameters

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Use of rubber tree (Hevea brasiliensis) latex biomembrane in diaphragmatic injuries in rabbits - an experimental study

The aim of this study was to evaluate the surgical use of the natural latex biomembrane in diaphragmatic injuries produced experimentally in rabbits. Fifteen healthy adult male and female New Zealand rabbits were employed. The rabbits were assigned to the experimental groups I, II, III, IV and V and analyzed on the 15th, 30th, 45th, 60th and 90th days post surgery, respectively. The surgical procedure consisted in the access to the diaphragm at the eighth right intercostal space, removal of a circle portion of approximately 1.5 cm in diameter following surgical repair with a latex membrane. Macroscopically, it was observed an excellent healing process during the experimental period. The clinical observations, complemented by the histological analysis, indicate that the latex membrane is useful for repair of traumatic inuries of the diaphragm of rabbits.