Ruy Garcia Marques

Brazilian law for scientific use of animals

UNLABELLED The Brazilian scientific community claimed for a definitive systematization and for comprehensive and realistic national rules, to provide guidance and regulation, instead of sanctions, so that the question of scientific research involving animals could be better contemplated. This is beginning to occur now with Law no. 11.794, sanctioned by the President of the Republic on November 8, 2008. PURPOSE To describe the evolution of Brazilian regimentation for scientific use of animals and to analyze Law no. 11.794. METHODS The legislation about the use of animals in teaching and in scientific research in Brazil and in Rio de Janeiro State was identified and discussed. RESULTS Until now, there was no updated general and systematizing rule regarding animal vivisection and experimentation for didactic or scientific purposes. The only specific law dates back to 1979 and was not regimented. More recent laws equated the practice of scientific experiments to acts of abuse and mistreatment of animals, when alternative technology was available. Municipal laws that restricted the scientific practice of vivisection and experimentation with animals were created in the cities of Rio de Janeiro and Florianopolis. CONCLUSION With the claim and collaboration of the scientific community, the sanction of Law no. 11.794 regarding the scientific use of animals represented an invaluable advance in spite of the presence of some points that eventually may require another type of treatment. The new Law states that it will be regimented within 180 (one-hundred-and-eighty) days, when some of these points could be better elucidated.

Regeneration of splenic autotransplants.

Abstract. Splenic autotransplantation seems to be the only alternative for preservation of splenic tissue after total splenectomy. This work was carried out to analyze the morphologic regeneration of autotransplanted splenic tissue in Wistar rats and to determine the bacterial phagocytic function of their macrophages. We utilized an experimental model including young and adult rats, of both sexes, submitted to total splenectomy combined with autotransplantation in the greater omentum of slices of the whole mass of spleen. Sixteen weeks later animals were intravenously inoculated with a suspension of Escherichia coli AB1157. There was regeneration of autotransplanted splenic tissue in all animals. A similar morphological aspect among all animals was observed, with splenic tissue showing red and white pulps with a moderate architectural disarrangement. Macrophages containing bacterial aggregates were observed, as well as macrophages with hemosiderin pigments inside the cytoplasm. Blood vessels showed preserved walls, with no signs of vasculitis or thrombosis. The present results suggest that splenic autotransplants in the greater omentum of the rat acquire the macro- and microscopic architecture of a normal spleen, with reduced dimensions, and preserve bacterial phagocyte function.

Lipidic profile among rats submitted to total splenectomy isolated or combined with splenic autotransplant

PURPOSE To evaluate the alterations on plasmatic lipids levels among rats submitted to total splenectomy isolated or combined with splenic autotransplant receiving standard chow during the postoperative period. METHODS Thirty Wistar rats were divided into three groups: control (C)-- sham-operated, total splenectomy--isolated (TS) or combined with splenic autotransplantation (SA). Since the postoperative period, all animals received standard rat chow manipulated in accordance to the American Institute of Nutrition Rodents Diets (1993). The plasmatic levels of total cholesterol (TC), triglycerides (TG), high-density lipoprotein (HDL), low-density lipoprotein (LDL), very-low-density lipoprotein (VLDL), and glucose (GLUC) were analyzed before the surgical procedure and after 6 and 12 weeks. RESULTS All the animals presented significant increase of TG and VLDL levels. In relation to the other parameters there was no difference among the weeks 0 and 12 in the animals of group C. In TS group significant increase was observed in TC and GLUC levels during the experiment. In SA group TC, HDL, and GLUC levels remained unaffected while HDL levels increased. CONCLUSION Our findings suggest that isolated total splenectomy alters lipids metabolism in rats fed with standard chow and splenic autotransplantation is effective in restoring its control.

Bacterial phagocytosis by macrophage of autogenous splenic implant

Autogenous splenic implant seems to be the only alternative for preservation of splenic tissue after total splenectomy. This work was carried out to analyze the morphologic regeneration of autotransplanted splenic tissue in Wistar rats and to determine the bacterial phagocytic function of their macrophages. We utilized an experimental model with thirty-two rats, of both sexes, submitted to total splenectomy combined with autotransplantation in greater omentum of slices of the whole spleen mass. The animals were divided into two groups: I--young rats weighing 100 to 150 g; and II--adult rats weighing 250 to 300 g. Sixteen weeks later animals were intravenously inoculated with a suspension of Escherichia coli AB1157. Twenty minutes after inoculation, the animals were sacrificed and the splenic autotransplants were removed for morphological study. There was regeneration of autotransplanted splenic tissue in all animals. A similar morphological aspect among all animals was observed, with splenic tissue showing red and white pulps, lymphoid follicles, and marginal zone, with a moderate architectural disarrangement. Macrophages containing gram-negative bacterial aggregates as well as macrophages with hemosiderin pigments within the cytoplasm were observed. Blood vessels showed preserved walls, with no signs of vasculitis or thrombosis. The present results suggest that autogenous splenic implants in the greater omentum of the rat acquire the macro- and microscopic architecture of a normal spleen, with reduced dimensions, and preserve bacterial phagocyte function.

Bacterial clearance after total splenectomy and splenic autotransplantation in rats

Wistar rats submitted to isolated total splenectomy or total splenectomy combined with splenic autotransplantation were inoculated with 99mtechnetium-labeled Escherichia coli. Measurement of isotope uptake in the organs of the mononuclear phagocytic system showed a greater bacterial bloodstream clearance in rats with splenic autotransplantation. Although uptake of bacteria in the spleen was higher in the control group, the number of bacteria remaining in the bloodstream did not differ between groups. These results indicate that splenic autotransplantation preserves the phagocytic function of the spleen.

Morfologia e função fagocitária de implante esplênico autógeno regenerado em ratos

Purpose: To analyze the morphologic regeneration of autotransplanted splenic tissue in Wistar rats and to determine the bacterial phagocytic function of their macrophages. Methods: We utilized an experimental model including young and adult rats, of both sexes, submitted to total splenectomy combined with autotransplantation in the greater omentum of slices of the whole mass of spleen. Sixteen weeks later animals were intravenously inoculated with a suspension of Escherichia coli AB1157, and twenty minutes later killed with lethal dose of halothane and submitted to laparotomy for splenic autotransplants retrieval. Data were analyzed statistically by de Student-t test, with emphasis on the comparison of the extent level of autotransplanted splenic mass regeneration between young and adult animals of both sexes. Results: There was regeneration of autotransplanted splenic tissue in all animals. Young males and adult females presented greater regeneration. A similar morphological aspect among all animals was observed, with splenic tissue showing red and white pulps with a moderate architectural disarrangement, as well as lymphoid follicles. Blood vessels showed preserved walls, with no signs of vasculitis or thrombosis. Macrophages containing bacterial aggregates were observed, as well as macrophages with hemosiderin pigments inside the cytoplasm. Conclusion: The present results suggest that splenic autotransplant in the greater omentum of the rat acquires the macro- and microscopic architecture of a normal spleen, with reduced dimensions, and preserves bacterial phagocyte function.

Avaliação de métodos radiológicos na detecção de corpo estranho de madeira em modelo animal

ABSTRACT - Purpose: To determine the usefulness of different radiological methods in the diagnoses of wooden foreign bodies(FB). Methods: Eleven adult chickens were used. Each thigh received a puncture wound and in one of them a wooden splinter wasintroduced and left in place while in the contralateral it was introduced and removed (control group). After 7 days the animals wherekilled and the legs removed to be analyzed with conventional radiography (CR), ultrasonography (US), magnetic resonance andcomputed tomography. The results were viewed by 2 independent senior radiologists. Results: Sensitivity was: CR – 13.6%; US –63.6%; MR – 59.1%; and CT – 72.7%, with specificity of 100%, 100%, 95.5%, and 95.5%, respectively. The positive predictive valu efor CR and US was 100%, 95% for CT 95% and 93.8% for MR. CT had a negative predictive value of 78.3%, while US, MR, and CRhad 73.7%, 70.1%, and 53.7%, respectively. The accuracy for CT was 84.1%, followed by US – 81.8%, RM – 77.3%, and CR – 56.8%.Inflammatory reaction was histologically demonstrated in all thighs containing FB.

Infecção fulminante pós-esplenectomia

BACKGROUND Splenectomy performed at any age and for any reason increases the risk for death due to overwhelming infection. AIM To evaluate definition, etiology, incidence, risk factors and prophylaxis of overwhelming postsplenectomy infection, as well as the methods related to splenic tissue preservation when total splenectomy is necessary. METHOD Bibliographic review. RESULTS The etiological agents more frequently found are Streptococcus pneumoniae, Haemophilus influenza and type B, and Neisseria meningitidis. Other bacteria like Escherichia coli, Streptococcus b-hemolytic, Staphylococcus aureus and Pseudomonas sp represent a significant risk as well. In addition, a great variety of agents including other enteric Gram-negative microorganisms and non-bacteria pathogens can also be sporadically identified. The prophylaxis is based on three main aspects: patients education, immune prophylaxis and chemical prophylaxis. However these are not enough to prevent the higher risk of developing sepsis. When total splenectomy is unavoidable, heterotopic splenic autotransplantation seems to be the only alternative for splenic tissue preservation. According to clinical and experimental studies, the splenic autotransplanted tissue present a similar structure to a normal spleen and preserve the splenic immune function. CONCLUSIONS The high risk of overwhelming postsplenectomy infection, reduced the indication for total splenectomy, in trauma and several diseases. Prophylactic methods have been developed to minimize the effects of the sepsis. Several researches have been done to determine the immunocompetence of autogenous splenic grafts in response to bacteria invasion.ABSTRACT – Background – Splenectomy performed at any age and for any reason increases the risk for death due to overwhelming infection. Aim – To evaluate definition, etiology, incidence, risk factors and prophylaxis of overwhelming postsplenectomy infection, as well as the methodsrelated to splenic tissue preservation when total splenectomy is necessary. Method – Bibliographic review. Results – The etiological agents morefrequently found are Streptococcus pneumoniae , Haemophilus influenza and type B, and Neisseria meningitidis. Other bacteria like Escherichiacoli , Streptococcus β-hemolytic, Staphylococcus aureus and Pseudomonas sp represent a significant risk as well. In addition, a great variety ofagents including other enteric Gram-negative microorganisms and non-bacteria pathogens can also be sporadically identified. The prophylaxisis based on three main aspects: patient’s education, immune prophylaxis and chemical prophylaxis. However these are not enough to preventthe higher risk of developing sepsis. When total splenectomy is unavoidable, heterotopic splenic autotransplantation seems to be the onlyalternative for splenic tissue preservation. According to clinical and experimental studies, the splenic autotransplanted tissue present a similarstructure to a normal spleen and preserve the splenic immune function.

Intramuscular technique for gluteal augmentation: determination and quantification of muscle atrophy and implant position by computed tomographic scan.

Background: New surgical techniques for gluteal augmentation have improved final results. It is estimated that more than 35,000 patients have undergone augmentation gluteoplasty using implants. The authors sought to determine and quantify the presence of muscle atrophy, and to evaluate implant positioning using the intramuscular technique. Methods: Twenty-three female patients were selected prospectively for this study and underwent intramuscular gluteal augmentation using gluteal implants of a round or oval base. Computed tomographic scanning and three-dimensional volumetric reconstruction were used to investigate muscle atrophy and implant position, with comparison of the results between the preoperative scan and scans obtained 3, 6, and 12 months after surgery. Results: Three-dimensional reconstruction and volumetric analysis showed muscular atrophy. After 12 months of follow-up, 34 gluteal muscles (17 patients) were analyzed, with 4.3 percent atrophy remaining on the right side and 2.6 percent on the left side. Twenty-three patients were studied regarding position (46 gluteal implants). All oval base implants introduced in a vertical direction (seven patients) turned to an oblique direction, following the direction of muscle fibers by 3 months after surgery. Two patients showed rotation of the implant. Conclusions: The presence of a gluteal implant caused muscle atrophy. However, it did not lead to clinical or physical limitations. It is not important whether the implants are positioned vertically or obliquely, provided that they are symmetric. The technique proved to be safe in maintaining the intramuscular position of the implant, with good satisfaction for the patients. CLINICAL QUESTION/LEVEL OF EVIDENCE: Therapeutic, IV.

Importância da preservação de tecido esplênico para a fagocitose bacteriana

OBJECTIVE: Being the major lymphoid organ of human body, spleen performs critical immunological functions such as bacteria depuration from bloodstream and precocious antibody production against various antigenic particles. Spleen, liver and lung accounts for more than 95% of phagocytic activity in humans. METHODS: We used intravenous inoculation of Escherichia coli AB1157 in Wistar rats in order to evaluate this function. Young and adult rats of both sexes were submitted to total splenectomy and compared to animals not submitted to any surgical procedure. Sixteen weeks later, rats of both groups were challenged with a suspension of E. coli labeled with Tc-99m, and, after twenty minutes, they were killed. Liver, lung, spleen and a blood clot sample were removed for radioactivity determination. The statistical study was performed with Students t test. RESULTS: There were no significant differences in the radioactivity uptake obtained from young and adult splenectomized rats. However, these rats were associated with higher levels of blood counts than animals of the control group (p<0.0001) due to a larger bacteria remnant in the bloodstream. CONCLUSION: This finding suggests that some failure in mononuclear phagocytic system occurs in spleen absence adaptation, ratifying the necessity of development of alternative surgical techniques to total splenectomy in patients requiring spleen removal.