Pesquisas em animais: uma reflexão bioética

Animal models are an important tool in scientific research. It is known that this practice must be based on bioethical principles in order to impose limits about pain and suffering, or for inspecting proceedings and research site. This study has as aim to evaluate and present current legislation ruling animal research in Brazil, comparing it with other countries, as well as fostering bioethical reflection about this issue. For this, we present current Brazilian legislation and we have carried out international literature review and discussion. The articles were searched in PubMed data base, using the key words "bioethics" and "animal welfare". Nineteen articles from 1997 to 2013 were included. It was concluded the evident contribution of animal research for scientific research in several fields. It is morally relevant for humans, as rational beings, to secure humane treatment to animals contributing to research. The safest way to follow this path is establishing clear norms and studies well designed, involving society for assigning ethical norms. Thus, animal welfare would be guarantee.

Genotoxicity of anesthetics evaluated in vivo (animals)

The anesthesia has been improved all over the years. However, it can have impact on health, in both patients and animals anesthetized, as well as professionals exposed to inhaled anesthetics. There is continuing effort to understand the possible effects of anesthetics at molecular levels. Knowing the effects of anesthetic agents on genetic material could be a valuable basic support to better understand the possible mechanisms of these agents. Thus, the purpose of this review is to provide an overview on the genotoxic potential, evaluated in animal models, of many anesthetics that have already been used and those currently used in anesthesia.

Effect of IGF1, GH, and PIT1 markers on the genetic parameters of growth and reproduction traits in Canchim cattle

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

Diabetes reduces mesenchymal stem cells in fracture healing through a TNF alpha-mediated mechanism

Diabetes interferes with bone formation and impairs fracture healing, an important complication in humans and animal models. The aim of this study was to examine the impact of diabetes on mesenchymal stem cells (MSCs) during fracture repair.Fracture of the long bones was induced in a streptozotocin-induced type 1 diabetic mouse model with or without insulin or a specific TNF alpha inhibitor, pegsunercept. MSCs were detected with cluster designation-271 (also known as p75 neurotrophin receptor) or stem cell antigen-1 (Sca-1) antibodies in areas of new endochondral bone formation in the calluses. MSC apoptosis was measured by TUNEL assay and proliferation was measured by Ki67 antibody. In vitro apoptosis and proliferation were examined in C3H10T1/2 and human-bone-marrow-derived MSCs following transfection with FOXO1 small interfering (si)RNA.Diabetes significantly increased TNF alpha levels and reduced MSC numbers in new bone area. MSC numbers were restored to normal levels with insulin or pegsunercept treatment. Inhibition of TNF alpha significantly reduced MSC loss by increasing MSC proliferation and decreasing MSC apoptosis in diabetic animals, but had no effect on MSCs in normoglycaemic animals. In vitro experiments established that TNF alpha alone was sufficient to induce apoptosis and inhibit proliferation of MSCs. Furthermore, silencing forkhead box protein O1 (FOXO1) prevented TNF alpha-induced MSC apoptosis and reduced proliferation by regulating apoptotic and cell cycle genes.Diabetes-enhanced TNF alpha significantly reduced MSC numbers in new bone areas during fracture healing. Mechanistically, diabetes-enhanced TNF alpha reduced MSC proliferation and increased MSC apoptosis. Reducing the activity of TNF alpha in vivo may help to preserve endogenous MSCs and maximise regenerative potential in diabetic patients.

Pyridinic analog of the natural product (-)-spectaline as potential adjuvant for the treatment of central nervous system disorders

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

Analysis of generalized interictal discharges using quantitative EEG

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

Genetic parameter estimates and principal component analysis on performance and carcass traits of a terminal pig sire line

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

Competitive interactions between C. albicans, C. glabrata and C. krusei during biofilm formation and development of experimental Candidiasis

In this study, we evaluated the interactions between Candida albicans, Candida krusei and Candida glabrata in mixed infections. Initially, these interactions were studied in biofilms formed in vitro. CFU/mL values of C. albicans were lower in mixed biofilms when compared to the single biofilms, verifying 77% and 89% of C. albicans reduction when this species was associated with C. glabrata and C. krusei, respectively. After that, we expanded this study for in vivo host models of experimental candidiasis. G. mellonella larvae were inoculated with monotypic and heterotypic Candida suspensions for analysis of survival rate and quantification of fungal cells in the haemolymph. In the groups with single infections, 100% of the larvae died within 18 h after infection with C. albicans. However, interaction groups achieved 100% mortality after 72 h of infection by C. albicans-C. glabrata and 96 h of infection by C. albicans-C. krusei. C. albicans CFU/mL values from larvae hemolymph were lower in the interacting groups compared with the monoespecies group after 12 h of infection. In addition, immunosuppressed mice were also inoculated with monotypic and heterotypic microbial suspensions to induce oral candidiasis. C. albicans CFU/mL values recovered from oral cavity of mice were higher in the group with single infection by C. albicans than the groups with mixed infections by C. albicans-C. glabrata and C. albicans-C. krusei. Moreover, the group with single infection by C. albicans had a higher degree of hyphae and epithelial changes in the tongue dorsum than the groups with mixed infections. We concluded that single infections by C. albicans were more harmful for animal models than mixed infections with non-albicans species, suggesting that C. albicans establish competitive interactions with C. krusei and C. glabrata during biofilm formation and development of experimental candidiasis.

The 2',4'-dihydroxychalcone could be explored to develop new inhibitors against the glycerol-3-phosphate dehydrogenase from Leishmania species

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

Comparative analysis of the replication of bovine herpesvirus 1 (BHV1) and BHV5 in bovine derived-neuron-like cells

Members of the subfamily Alphaherpesvirinae use the epithelium of the upper respiratory and/or genital tract as preferential sites for primary replication. However, bovine herpesvirus 5 (BoHV5) is neurotropic and neuroinvasive and responsible for meningoencephalitis in cattle and in animal models. A related virus, BoHV1 has also been occasionally implicated in natural cases of neurological infection and disease in cattle. The aim of the present study was to assess the in vitro effects of BoHV1 and BoHV5 replication in neuron-like cells. Overall, cytopathic effects, consisting of floating rounded cells, giant cells and monolayer lysis, induced by both viruses at 48 h postinfection (p.i.) resulted in a loss of cell viability and high virus titres (r = 0.978). The BoHV1 Cooper strain produced the lowest titres in neuron-like cells, although viral DNA was detected in infected cells during all experiments. Virus replication in infected cells was demonstrated by immunocytochemistry, flow cytometry and qPCR assays. BoHV antigens were better visualized at 48 h p.i. and flow cytometry analysis showed that SV56/90 and Los Angeles antigens were present at higher levels. In spite of the fact that BoHV titres dropped at 48 h p.i, viral DNA remained detectable until 120 h p.i. Sensitive TUNEL (terminal deoxynucleotidyltransferase-mediated dUTP-biotin nick end labeling) and annexin V assays were used to identify apoptosis. BoHV5 induced death in approximately 50 % of cells within 24 h p.i., similar to what has been observed for BoHV1 Los Angeles. Infection with the BoHV1 Cooper strain resulted in 26.37 % of cells being in the early stages of apoptosis; 63.69 % of infected cells were considered viable. Modulation of mitochondrial function, as measured by mitochondrial membrane depolarization, was synchronous with the virus replication cycle, cell viability and virus titres at 48 h p.i. Our results indicate that apoptosis plays an important role in preventing neuronal death and provides a bovine-derived in vitro system to study herpesvirus-neuron interactions.

Carreadores lipídicos nanoestruturados para incorporação de complexos de cobre(ii) aplicáveis no estudo frente ao Mycobacterium tuberculosis

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Efeito de biomateriais no crescimento e funcionalidade de células progenitoras da polpa dentária

Pós-graduação em Odontologia Restauradora - ICT

Transplante multivisceral em suínos: modelo de pesquisa e treinamento

Objective: To present a model for research and training in multivisceral transplantation in pigs. Methods: Eight Large White pigs (four donors and four recipients) were operated. The multivisceral transplant with stomach, duodenum, pancreas, liver and intestine was performed similarly to transplantation in humans with a few differences, described below. Anastomoses were performed as follows: end-to-end from the supra-hepatic vena cava of the graft to the recipient juxta diaphragmatic vena cava; end-to-end from the infra-hepatic vena cava of the graft to the inferior (suprarenal) vena cava of the recipient; and endto-side patch of the aorta of the graft to the infrarenal aorta of the recipient plus digestive reconstruction. Results: The performance of the multivisceral transplantion was possible in all four animals. Reperfusions of the multivisceral graft led to a severe ischemia-reperfusion syndrome, despite flushing of the graft. The animals presented with hypotension and the need for high doses of vasoactive drugs, and all of them were sacrificed after discontinuing these drugs. Conclusion: Some alternatives to minimize the ischemia-reperfusion syndrome, such as the use of another vasoactive drug, use of a third pig merely for blood transfusion, presence of an anesthesia team in the operating room, and reduction of the graft, will be the next steps to enable experimental studies.

P-215-Quantitative variation of some gut bacteria in Crohn's disease can be related to the patient's gender

Background: Imbalance in bacterial species composition of the gut microbiota is one of the factors associated with the cause or complication of the symptoms of Crohn's disease (CD). This disequilibrium consists in the reduction of biodiversity, decrease of genus such as Bifidobacterium and elevation of species such as Escherichia coli. Human microbiota varies among subjects of a same population irrespective of their health condition and among individuals living in distinct geographic locations. In animal models, sex related differences could also be observed in gut bacterial species composition under some pathological conditions. Experiments conducted with mice have demonstrated that the manifestation of type 1 diabetes (T1D) could be under the influence of the animal sex and its serum level of testosterone, which in turn could be modulated by a particular gut microbiota. Considering the existence of similar features between T1D and CD, such as strong genetic component and malfunctioning of the immune system, we investigated whether differences could be observed in the gut microbiota dysbiosis of male and female CD patients. Methods: Fifty and 5 gut mucosal biopsies from 25 adult CD patients (11 males and 14 females) and 43 specimens of an equivalent clinical material from 22 control subjects (11 males and 11 females) were screened for bacterial biodiversity by analyzing sequences of 16SrDNA V6 region. A number of 2-3 samples each from distinct gut segments (from ileum to rectum) were taken from each subject. The 16SrDNA sequences were obtained by sequencing PCR amplicons of the corresponding gene in the Ion torrent PGM sequencer. Identification and classification of the bacterial groups followed the Ribosomal Database Project (RDP) website pipeline. The relationships of the bacterial taxa with each of the study parameters was performed by compiling the data in a MS Excel and the level of statistical significance determined by the Chi-square test. Results: A total of 3203 16SrDNA sequences were detected in the 98 biopsies samples, the majority of which matching Proteobacteria, Firmicutes, Bacterioidetes, and Actinobacteria. The percentage of DNA sequences for each of these phyla found in Male control subjects/Male CD patients was 40.5/33, 32.7/32.4, 20.8/24.5, and 4.4/4,4 for Proteobacteria, Firmicutes, Bacterioidetes, and Actinobacteria, respectively. In Female comparisons, these values were 35.6/42, 39.2/26.3, 19.8/23.3, 5.2/7. Both Male and Female CD patients presented higher numbers of sequences of Actinobacteria and Bacterioidetes than those of control subjects of the same gender. Case-control differences for Firmicutes could be observed only in female comparisons and, for Proteobacteria, although case-control differences were observed in both genders, the nature of difference was distinct, since while in CD female patients a higher number of sequences matching this phylum was detected, in males a reduced number was observed, in comparison with controls. The species responsible for the Proteobacteria variation in both gender was Escherichia coli. Conclusions: The data presented above suggest that any analysis of dysbiosis in CD must take in account the patient's gender, an observation particularly relevant for Escherichia coli, whose association with CD has been most intensively investigated and for which the present study shows a reverse quantitative variation regarding the patients' gender.

Análise quantitativa das descargas epileptiformes generalizadas e da neuroimagem de pacientes com epilepsia generalizada idiopática

Pós-graduação em Fisiopatologia em Clínica Médica - FMB