Experimental pancreas transplantation: the consequences of the portocaval shunt on the blood glucose, plasma insulin, and glucagon

A proposição foi realizada, no período pós-operatório imediato, em 40 ratos Wistar, distribuídos por sorteio em doi grupos: grupo NC, vinte ratos correspondentes ao grupo controle, não diabético, submetidos a operação simulada e o grupo PT, 20 ratos correspondentes ao grupo diabético que recebeu transplante de pâncreas heterotópico de ratos Wistar normais. Durante sete dias, antes do transplante, e 1, 3, 6, 12, 24, 48, 72, 96 horas após, determinava-se a glicose sanguínea, a insulina plasnática e o glucagon. Estes parâmetros eram obtidos também do grupo NC. Diabetes mellitus experimental era induzida pela administração intravenosa de aloxana. O grupo PT era imunosuprimido com ciclosporina A. O grupo NC apresentou níveis normais de glicose sanguínea, de insulina plásmica e de glucagon, durante todo o experimento. Foi encontrada nítida hiperinsulinemia no sangue venoso periférico do grupo PT. A insulina plasmática era significantemente maior no grupo PT comparada ao grupo NC começando 72 horas após o transplante. O glucagon plasmático, elevado no período pré-transplante, não se alterou após o transplante. Apesar de hiperinsulinemia e hiperglucagonemia, os níveis de glicose sanguínea eram elevados 6 horas após o transplante e mantiveram-se normais após este período. Considerando-se os níveis de glicose sanguínea 12 horas pós-transplante, não houve diferença estatisticamente significante entre os grupos PT e NC, até o sacrifício.

Vegetal polymer in repair of defects of the orbital floor: an experimental study in rabbits

Background: An experimental study was done to assess the ability of the vegetal polymer miniplates and screws to repair defects of the orbital floor.Methods: An artificial standard-sized defect was created in the bony floor of right orbit of 45 albino rabbits. The animals were divided into three experimental groups: control group (G1) involving animals with orbital floor defect and no treatment; titanium group (G2) containing animals with orbital floor defect repaired by titanium miniplates and screws; vegetal polymer group (G3) composed of animals with similar orbital floor defects repaired by vegetal polymer miniplates and screws. Throughout the course of the experiment, the animals were clinically evaluated. At 15, 30 and 60 days after surgery, the animals were killed. They were X-rayed immediately after the floor defect and at the moment of sacrifice. Histological and morphometric evaluation of inflammatory reaction and bone healing was done. Data were statistically evaluated.Results: No implants were extruded. Bone consolidation was similar in G2 and G3 and better than in G1 group animals. Inflammatory reaction was most pronounced in animals of G3 15 days after surgery, and it subsided over time.Conclusion: Vegetal polymer miniplates and screws induces small inflammatory reaction and had the ability to stimulate bone growth with good integration in the orbital floor defect allowing to consider the vegetal polymer adequate option to treat orbital floor defects. Future studies involving long-term follow-up and biomechanical tests to evaluate material resistance to traction are needed.

Agaricus blazei (Himematsutake) does not alter the development of rat diethylnitrosamine-initiated hepatic preneoplastic foci

The modifying potential of crude extracts of the mushroom Agaricus blazei Murrill (Himematsutake) on the development and growth of glutathione S-transferase placental form (GST-P)-positive liver foci (liver preneoplastic lesion) was investigated in adult male Wistar rats. Six groups of animals were used. Groups 2 to 5 were given a single i.p. injection of 200 mg/kg b.w. of diethylnitrosamine (DEN) and groups 1 and 6 were treated with saline at the beginning of the experiment. After 2 weeks, animals of groups 3 to 6 were orally treated with three dose levels of aqueous extracts of the mushroom A. blazei (1.2, 5.6, 11.5, and 11.5 mg/ml of dry weight of solids) for 6 weeks. All animals were subjected to two-thirds partial hepatectomy at week 3 and sacrificed at week 8. Two hours before sacrifice, ten animals of each group were administered a single i.p injection of 100 mg/kg of bromodeoxyuridine (BrdU). Apoptotic bodies and BrdU-positive hepatocyte nuclei were quantified in liver sections stained for hematoxylin and eosin (H&E) (eosinophilic foci) and simultaneously stained for GST-P expression (GST-P-positive foci), respectively. The 6-week treatment with A. blazei did not alter the development (number and size) of GST-P-positive foci and did not affect the growth kinetics of liver normal parenchyma or foci in DEN-initiated animals. Our results indicate that the treatment with aqueous extracts of the mushroom A. blazei during the post-initiation stage of rat liver carcinogenesis does not exert any protective effect against the development of GST-P-positive foci induced by DEN. (Cancer Sci 2003; 94: 188-192).

Thymic lymphomas in Wistar rats exposed to N-methyl-N-nitrosourea (MNU)

The Brazilian Agency for the Environment (IBAMA) recently adopted an alternative medium-term multiple-organ assay system with the Wistar rat strain for detection of the carcinogenic potential of pesticides. Originally, this initiation-promotion protocol was established in Japan with the isogenic Fischer 344 male rat. Among the initiating agents used in that assay, N-methyl-N-nitrosourea (MNU) rapidly induces malignant lymphoma and leukemia and early mortality of rats from different strains. This study was developed to evaluate whether the outbred Wistar rats are also similarly susceptible to MNU. Particularly, it aimed to evaluate the dose-response relationship and to register the MNUinduced pre-neoplasia and neoplasia that may develop in the lympho-hematopoietic system (LHS) of the Wistar rat within a medium-term period. Four groups of male Wistar rats were treated during 2 weeks with vehicle or with MNU (80, 160 or 240 mg/kg body weight, i.p.). After sacrifice at the 12th and 20th weeks, the thymus, spleen, bone marrow, cervical and mesenteric lymph nodes and liver were collected for analysis. At the 20th week, LHS malignant tumors and benign vascular tumors occurred only in the high- and intermediate-dose MNU-treated animals. Four animals treated with 240 mg/kg developed diffuse thymic lymphomas; two others, treated respectively with 240 mg/kg and 160 mg/kg, developed spleen hemangiomas. The present observations indicate that the Wistar strain is as susceptible as other strains to the early development of MNU-induced LHS (pre)neoplasia. Therefore, this strain seems suitable to be used as test system in bioassay protocols that adopt MNU as an initiating agent for carcinogenesis.

DNA damage and aberrant crypt foci as putative biomarkers to evaluate the chemopreventive effect of annatto (Bixa orellana L.) in rat colon carcinogenesis

Chemoprevention opens new perspectives in the prevention of cancer and other degenerative diseases. Use of target-organ biological models at the histological and genetic levels can markedly facilitate the identification of such potential chemopreventive agents. Colon cancer is one of the highest incidence rates throughout the world and some evidences have indicated carotenoids as possible agents that decrease the risk of colorectal cancer. In the present study, we evaluate the activity of annatto (Bixa orellaria L.), a natural food colorant rich in carotenoid, on the formation of aberrant crypt foci (ACF) induced by dimethy1hydrazine (DMH) in rat colon. Further, we investigate, the effect of annatto on DMH-induced DNA damage, by the comet assay. Male Wistar rats were given s.c. injections of DMH (40 mg/kg body wt.) twice a week for 2 weeks to induce ACE They also received experimental diets containing annatto at 20, 200 or 1000 ppm for five 5 weeks before (pre-treatment), or 10 weeks after (post-treatment) DMH treatment. In both protocols the rats were sacrificed on week 15th. For the comet assay, the animals were fed with the same experimental diets for 2 weeks. Four hours before the sacrifice, the animals received an s.c. injection of DMH (40 mg/kg body wt.). Under such conditions, dietary administration of 1000 ppm annatto neither induce DNA damage in blood and colon cells nor aberrant crypt foci in rat distal colon. Conversely, annatto was successful in inhibiting the number of crypts/colon (animal), but not in the incidence of DMH-induced ACF, mainly when administered after DMH. However, no antigenotoxic effect was observed in colon cells. These findings suggest possible chemopreventive effects of annatto through their modulation of the cryptal cell proliferation but not at the initiation stage of colon carcinogenesis. (c) 2005 Elsevier B.V. All rights reserved.

Effects of Diuron [3-(3,4-dichlorophenyl)-1,1-dimethylurea] on the urinary bladder of male Wistar rats

Diuron (3-(3,4-dichlorophenyl)-1,1-dimethylurea) is a substituted urea herbicide widely used on agricultural crops such as soy, cotton and sugar cane. In a previous long-term study this herbicide exerted carcinogenic activity on the urinary bladder mucosa of male Wistar rats. In general, the genotoxic and mutagenic potentials of Diuron are considered to be negative. The present study aimed to evaluate the mode of action of Diuron on the urinary bladder mucosa of male Wistar rats. Six-week old male Wistar rats were fed pelleted Nuvilab diet mixed with Diuron at 125, 500 and 2500 ppm. As a positive control, 8.3% sodium saccharin (NaS) was fed in the diet. Preceding the sacrifice of the animals at the 20th week, urinary pH was measured and the genotoxic potential of Diuron was evaluated by the comet assay. Histological urothelial lesions in the urinary bladder and in the renal pelvis mucosa, cell proliferation/apoptosis evaluations, and scanning electron microscopy (SEM) of the urinary bladder mucosa were also performed. No DNA changes were found in urothelial or peripheral blood cells, and urinary pH was comparable to controls in all Diuron groups. In the urinary bladder urothelium, the incidence of simple hyperplasia (SH) by light microscopy was significantly increased (7/10; p < 0.005) in the 2500 ppm Diuron group but not at the lower doses. By SEM, three of five animals treated with 2500 ppm Diuron showed urothelial cell necrosis and hyperplasia. In the renal pelvis, the incidence of SH was significantly increased in the Diuron 500 and 2500 ppm and in the NaS 8.3% groups. Cell proliferation was significantly increased in the Diuron 2500 ppm, (p < 0.05) and NaS 8.3% (p < 0.05) groups. The results indicate that a high dietary concentration of Diuron is associated with urothelial necrosis and continuous regenerative cell proliferation that leads to urothelial hyperplasia. (c) 2006 Elsevier B.V.. All rights reserved.

In vivo genotoxicity evaluation of a treated urban sewage sludge sample

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

Experimental paracoccidioidomycosis in the mouse. III. Histopathological and immunological findings after intravenous infection in the presence or absence of previous immunization

Cinqüenta camundongos suíços, brancos, com quatro semanas de idade, foram inoculados com 5x10(5) formas leveduriformes, viáveis de Paracoccidiodes brasiliensis (cepa 18). Dez destes animais tinham sido previamente imunizados com antígeno particulado de P. brasiliensis, durante quatro semanas, por injeção intradérmica. Os controles consistiram de 10 animais que foram somente imunizados e 10 inoculados com solução salina estéril. Os animais foram sacrificados após 2, 4, 7, 11 e 16 semanas. Estudamos: 1) resposta de hipersensibilidade retardada medida pelo teste do coxim plantar, 24 horas antes do sacrifício; 2) anticorpo- gênese específica avaliada pelo teste de imunodifusão dupla em gel de ágar; 3) histopatologia dos pulmões, fígado, baço, supra-renal e rins. Observamos: 1) os animais imunizados desenvolveram resposta imunecelular mais intensa que os infectados; 2) a infecção deprimiu a resposta imunecelular dos animais imunizados; 3) a histopatologia da infecção endovenosa revelou inflamação granulomatosa sistêmica e progressiva. Os animais infectados após imunização prévia apresentaram inflamação pulmonar menos extensa, com granulomas menores e com reduzido número de fungos. O presente modelo murino de paracoccidioidomicose mimetiza alguns achados da forma humana subaguda da micose (doença sistêmica com depressão da imunidade celular). O esquema de imunização extrapulmonar utilizado foi capaz de induzir certo grau de proteção do pulmão contra um desafio infeccioso pelo P. brasiliensis.

An acqueous extract of Bidens pilosa L. protects liver from cholestatic disease: experimental study in young rats

OBJETIVO: Testar o efeito hepatoprotetor do extrato aquoso de Bidens pilosa L. (EBP) na doença hepática induzida pela ligadura e ressecção do ducto biliar comum (LRDBC) em ratos jovens. MÉTODOS: Estudamos ratos Wistar com 21º. dia de vida (P21) divididos em quatro grupos de 10 animais, Grupo SA: operação simulada e água; Grupo SD: operação simulada e EBP (160mg de folhas frescas/100g de peso corporal/dia); Grupo LA: LRDBC e água e Grupo LD: LRDBC e EBP diariamente. O tempo de sono por pentobarbital (TSP), aspartato (AST) e alanina (ALT) aminotransferase foram determinadas após o sacrifício (P70). O Score de Ruwart (SR) para fibrose hepática foi atribuído para cada animal. Foi realizada análise de variância com dois fatores e pelo teste de Tukey para comparações múltiplas ou por teste não paramétrico. RESULTADOS: Houve diferença estatisticamente significativa entre LA e LD nas variáveis: TSP (médias LA=390; LD=173), AST (médias LA=8, LD=5), ALT (medianas LA=2; LD=1) e SR (medianas LA=2; LD=1). CONCLUSÃO: EBP poderá ser empregado na fitoterapia da doença hepática induzida pela colestase obstrutiva crônica, pois protege a função hepática, diminui a velocidade de necrose e a intensidade da fibrose hepática na obstrução biliar extra-hepática.

Comparative Study Between Porcine Small Intestinal Submucosa and Buccal Mucosa in a Partial Urethra Substitution in Rabbits

Purpose: Several urethral conditions may require tissue substitution. One collagen-base biomaterial that recently emerged as an option is small intestinal submucosa (SIS). The aim of this study was to compare the results of SIS and buccal mucosa for urethral substitution in rabbits.Materials and Methods: Thirty-six North Folk male rabbits were randomized into three groups. In all animals, a 10 x 5 mm urethral segment was excised, and the urethral defect was repaired using a one-layer SIS patch (group I [GI]); four-layer SIS (group II [GII]); or buccal mucosa (group III [GIII]). Urethrography was performed preoperatively and after 12 weeks. After sacrifice, graft retraction was objectively measured using Scion Image (R) computer analysis and by calculation of ellipse area. The grade of fibrosis, inflammatory reaction, vascular/epithelial regeneration, and collagen III/I ratio were analyzed by hematoxylin/eosin and Picrosirius red staining.Results: Urethrography confirmed a wide urethral caliber without any signs of strictures after surgery. Urethral fistulae was diagnosed in 8.3% of cases (1 animal each group). Average graft shrinkage was 55.2% in GI; 44.2% in GII; and 57.2% in GIII (p < 0.05). The intensity of chronic inflammation, fibrosis, epithelium regeneration, and neovascularization was similar in all groups (p > 0.05). Collagen III/I ratio was higher in GII (GI: 119.6; GII: 257.2 and GIII: 115.0); p < 0.01.Conclusions: The four-layer SIS is more advantageous than the one-layer SIS and buccal mucosa for urethral substitution in rabbits.

Análise da influência do ultrassom de baixa intensidade na região de reparo ósseo em ratos sob ausência de carga

Há evidências de que o ultrassom (US) de baixa intensidade pode acelerar a regeneração óssea. Este trabalho objetivou verificar a ação do US no defeito ósseo, criado experimentalmente em tíbias de ratos sob ausência de carga. Vinte Rattus novergicus albinus, Wistar adultos, divididos em: G1 (n=10), grupo experimental de 15 dias sem suspensão, e G2 (n=10), grupo experimental de 15 dias suspenso pela cauda, foram submetidos à osteotomia em ambas as tíbias e à aplicação do US, frequência de 1,5 MHz, ciclo de trabalho 1:4, 30 mW/cm², nas tíbias direitas por 12 sessões de 20 minutos. Após o sacrifício, as tíbias foram submetidas à análise da Densidade Mineral Óssea (DMO). Os resultados demonstraram DMO de 0,139±0,018 g/cm² para tíbia tratada; 0,131±0,009 g/cm² para tíbia controle no G1; e no G2 registrou-se 0,120±0,009 g/cm² para tíbia tratada e 0,106±0,017 g/cm² para tíbia controle. Houve diferença significante entre os grupos nos quais o G2 apresentou menor DMO, o que demonstra que a suspensão prejudica a manutenção das propriedades ósseas, e entre as tíbias tratadas e controles do G2, demonstrando que o US acelerou o processo de reparo, concluindo que a impossibilidade do estímulo mecânico causada pela não deambulação em um processo de reparo ósseo pode ser minimizada pela ação do US. No G1, a aplicação do US não teve influência significante no aumento da DMO, talvez pelo fato dos animais já terem estímulo mecânico suficiente à formação óssea.

Influence of cyclosporin A on quality of bone around integrated dental implants: a radiographic study in rabbits

To evaluate the influence of cyclosporin A (CsA) administration on bone around integrated dental implants assessed by a bone quality index and by quantitative subtraction radiography.A total of 36 machine surface commercial implants were placed in 18 adult rabbits. After a 3-month healing period without any disturbance, the animals were randomly divided into three groups of six animals each. Group A was sacrificed at this time. CsA was injected subcutaneously in an immunosuppressive dose of 10 mg/kg/day in a test group (Group T), and a Group B served as a control, receiving only vehicle. After 3 months of cyclosporin administration, the animals of both Groups B and T were sacrificed. Radiographs were obtained at implant surgery and at the day of sacrifice with a CMOS sensor. Bone quality around the implants was compared between the groups using a bone quality index and quantitative subtraction radiography.The bone analysis showed that in Group T, the bone quality changed dramatically from a dense cortical to a loose trabecular bone structure (P < 0.0001, chi(2) test) while in Groups A and B there were no significant differences. Quantitative digital subtraction radiography showed significantly (P < 0.05) lower gray shade values (radiographic density) in a region of bone formation around the implants in Group T (118 +/- 12) than in Groups A (161 +/- 6) and B (186 +/- 10).Within the limits of this study, CsA administration has a negative effect on the quality of bone around integrated dental implant.

Influence of homeopathic treatment with comfrey on bone density around titanium implants. A digital subtraction radiography study in rats

Objective: The objective of this study was to evaluate the influence of homeopathic treatment with comfrey (Shymphytum officinalis 6CH) on radiographic bone density and area around titanium implants.Material and methods: Forty-eight rats were divided into two groups of 24 animals each: a control group (C) and a test group (SO). Each animal received one titanium micro-implant placed in the tibia. The animals in Group SO were subjected to 10 drops of comfrey 6CH per day mixed into their drinking water until the day of sacrifice. Eight animals of each group were sacrificed at 7, 14 and 28 days post-surgery, respectively.Standardized digital radiographs were obtained on the day of implant installation (baseline images) and on the day of sacrifice (final images). Digital subtraction of the two corresponding images was performed to evaluate changes in bone density and the area related to change around the implant between baseline and final images.Results: Subtraction images demonstrated that a significant difference existed in mean shade of gray at 14 days post-surgery between Group SO (mean 175.3 +/- 14.4) and Group C (mean 146.2 +/- 5.2). Regarding the area in pixels corresponding to the bone gain in Group SO, the differences observed between the sacrifice periods and groups were only significant at 7 days sacrifice between Group SO (mean 171.2 +/- 21.9) and Group C (mean 64.5 +/- 60.4).Conclusion: Within the limits of this study, comfrey administration promotes an increase in radiographic bone density around titanium implants in the initial period of bone healing.

Effect of diabetes mellitus and insulin therapy on bone density around osseointegrated dental implants: a digital subtraction radiography study in rats

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

The effect of oestrogen and alendronate therapies on postmenopausal bone loss around osseointegrated titanium implants

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