Avaliação do potencial efeito cicatrizante gástrico e antiartrítico da mangiferina, uma xantona glicosilada da Mangifera indica

Rheumatoid arthritis (RA) is a chronic illness with great potential to cause comorbidities resulting from cumulative inflammatory activities, and it contributes with the increase of disabilities and death of patients. It affects 1 to 2% of world population and usually occurs between 30 and 50 years of age. Among existing therapeutic options for the disease non-steroidal anti-inflammatory drugs (NSAIDs) still play an important part in the treatment, being widely used by patients to relieve pain and stiffness. However, this class of drugs causes many adverse gastrointestinal effects, such as dyspepsia, heartburn, nausea and vomit, and its use is one of the most common causes of peptic ulcers. Mangiferin (a glicosilated xanthone extracted mainly from the bark of Mangifera indica L.) is the main compound of an aqueous extract made from the bark stem of the mango tree. Previous studies conducted by our research group prove the anti-inflammatory action of mangiferin on an animal model of periodontitis, and its gastroprotective action has been described before. Considering these informations this study evaluated mangiferin’s potential on the treatment of RA and on gastric ulcer healing in animal models, and analyzed toxicity parameters to assure efficacy and safety of the compound as potential new drug for the treatment of the disease. RA was induced in rats by subcutaneous injection of bovine collagen and Freund’s complete adjuvant. This method presented low incidence of RA in rats, but we were able to induce the disease in 60 to 70% of the animals. Due to the wide use of NSAIDs and its potential to cause peptic ulcers, we induced gastric ulcer on arthritic rats to analyze mangiferin’s gastric healing effect. After 14 days of treatment we noticed small increase of the lesion area of animals treated with mangiferin or ibuprofen, when compared to the animals... (Complete abstract click electronic access below)

Analysis of generalized interictal discharges using quantitative EEG

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

A standartized research protocol for platelet-rich plasma (PRP) preparation in rats

Introduction: The urgent need for studies using standardized protocols to evaluate the real biological effects of PRP has been emphasized by several authors. Objective: The purpose of this study was to standardize a methodology for autologous Platelet-Rich Plasma (PRP) preparation in rats. Material and methods: Twentyfour, 5 to 6-month-old, male rats, weighing 450 to 500 g were used. After general anesthesia, 3.15 ml of blood was collected from each animal, via cannulation of the jugular vein. A standardized technique of double centrifugation was used to prepare PRP. PRP samples and peripheral blood platelets were then manually counted using a Neubauer chamber. Student’s t-test was used to compare the differences between the number of platelets in peripheral blood and PRP samples (p < 0.05). In addition, PRP and peripheral blood smears were stained to see platelets’ morphology. Results: All surgical procedures were well tolerated by the animals and they were healthy during the entire experimental period. PRP samples showed higher significantly platelet concentrations than peripheral blood samples (2,677,583 and 683,680 respectively). Conclusion: Within the limits of this study, it can be concluded that the method used produced autologous PRP with appropriated platelet quantity and quality, in rats.

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.

Experimental model of intestinal endometriosis: pilot study

Aims: The objective of this study is to create an experimental model of intestinal endometriosis in pigs, which might allow better understanding of deep infiltrating endometriosis and development of new treatment techniques. As secondary objective, we intend to create endometrial implants accessible by transrectal ultrasonography (TRUS). Study Design: Surgical experimental study in swine. Place and Duration of Study: This study was performed at the Instituto de Ensino e Pesquisa do Hospital Sírio-Libanês, São Paulo, Brazil, between January 2012 and December 2012. Methodology: Two sexually mature female minipigBR pigs underwent two laparotomies (each animal). The first laparotomy was performed to implant two fragments of autologous endometrium in the rectal wall. The second one was performed thirty days later to visualize, measure and obtain tissue of the site of the implants for histopathology study. A TRUS study was performed prior to the second surgery. The Institution’s Animal Utilization Study Committee approved the study. Results: In the first laparotomy a 5-cm segment of right uterine horn was resected. The endometrium was separated from the myometrium through sub-endometrial saline injection. Two endometrial fragments (1.0 x 2.0 cm) were dissected and sutured in the intra peritoneal anterior rectal wall of the animals. Thirty days later, all implants were identified during preoperative TRUS. “En-bloc” resection of the intestinal segment with the implants was performed during the second surgery. The autologous implants of endometrium invaded the muscular layer in one of the two animals. Conclusion: We demonstrated that the creation of an animal model of deep infiltrating endometriosis with intestinal involvement is feasible through a simple surgical technique. We believe that this model can be applied in experimental and clinical studies but further studies are necessary to refine the technique.

Efeitos do treinamento aeróbio e de força sobre a área de secção transversal (AST) das fibras do músculo plantar de ratos

Recent research seeking to elucidate the possible effects of different types of physical training on the morphological adaptations of skeletal muscle. Although it is relatively easy to study the effects of exercise training in humans, such research becomes limited due to the invasive nature of the biopsies and the risk inherent in the use of human subjects. Thus, the application of animal models of training has been considered an appropriate strategy for the study of muscular adaptations in response to exercise. Objective: This study used a rodent model to determine the possible effects of aerobic and strength training on the CSA of fibers of the plantaris muscle. Methods: 24 male Wistar rats (80 to 120 days, 250 to 400 g) were randomly divided into 3 groups: aerobic training (TA, n = 8), strength training (ST, n = 8) and control (CO, n = 8). The animals in groups TA and TF were subjected to 8 weeks of training, while the animals of group C remained without any stimulus from start to finish the training period. At the end of the experiment, the animals were sacrificed and right plantar muscles dissected and removed. For morphological and morphometric analysis of muscle fibers was performed staining was performed H.E. Results: There was no significant difference in initial body weight between experimental groups. After 8 weeks of training, the TA group showed a significant reduction in final body weight, compared to CO and TF groups. With respect to the CSA of fibers of the plantaris muscle, no significant difference between the groups CO and TA. On the other hand, the strength training promoted a significant increase in AST of the group TF in compared with the groups CO and TA. Conclusion: Strength training used in this study promoted an increase in CSA of fibers of the plantaris muscle. On the other hand, animals submitted to aerobic training showed no changes in the CSA of the fibers, however, there was reduction in PC animals. The data strongly suggest the use of animal model of strength training used in this study as an appropriate strategy for studying the hypertrophic response of skeletal muscle.

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

Pós-graduação em Fisiopatologia em Clínica Médica - FMB

Aerosol Exposure of White-Tailed Deer (Odocoileus virginianus) to Mycobacterium Bovis

Tuberculosis due to Mycobacterium bovis affects both captive and free-ranging Cervidae in the United States. Various animal models have been developed to study tuberculosis of both humans and animals. Generally, tuberculosis is transmitted by aerosol and oral routes. Models of aerosol exposure of large animals to M. bovis are uncommon. In order to develop a reliable method of aerosol exposure of white-tailed deer (Odocoileus virginianus) to M. bovis, 12 healthy white-tailed deer, aged 8–10 mo, were infected by aerosol exposure to 2x105 to 1x106 colony forming units (CFU) (high dose, n=4) of M. bovis or 6x102 to 1.6 x 103 CFU (low dose, n=8) of M. bovis. Tuberculous lesions were more widely disseminated in deer receiving the high dose, while lesions in deer receiving the low dose were more focused on the lungs and associated lymph nodes (tracheobronchial and mediastinal). Aerosol delivery of M. bovis to white-tailed deer results in a reliable manner of experimental infection that may be useful for studies of disease pathogenesis, immune response, mycobacterial shedding, and vaccine efficacy.

Alterations in pulmonary structure by elastase administration in a model of emphysema in mice is associated with functional disturbances

Several experimental studies of pulmonary emphysema using animal models have been described in the literature. However, only a few of these studies have focused on the assessment of ergometric function as a non-invasive technique to validate the methodology used for induction of experimental emphysema. Additionally, functional assessments of emphysema are rarely correlated with morphological pulmonary abnormalities caused by induced emphysema. The present study aimed to evaluate the effects of elastase administered by tracheal puncture on pulmonary parenchyma and their corresponding functional impairment. This was evaluated by measuring exercise capacity in C57Bl/6 mice in order to establish a reproducible and safe methodology of inducing experimental emphysema. Thirty six mice underwent ergometric tests before and 28 days after elastase administration. Pancreatic porcine elastase solution was administered by tracheal puncture, which resulted in a significantly decreased exercise capacity, shown by a shorter distance run (-30.5%) and a lower mean velocity (-15%), as well as in failure to increase the elimination of carbon dioxide. The mean linear intercept increased significantly by 50% in tracheal elastase administration. In conclusion, application of elastase by tracheal function in C57Bl/6 induces emphysema, as validated by morphometric analyses, and resulted in a significantly lower exercise capacity, while resulting in a low mortality rate. (C) 2011 Sociedade Portuguesa de Pneumologia. Published by Elsevier Espana, S.L. All rights reserved.

Effects of diazepam on BOLD activation during the processing of aversive faces

This study aimed to measure, using fMRI, the effect of diazepam on the haemodynamic response to emotional faces. Twelve healthy male volunteers (mean age = 24.83 +/- 3.16 years), were evaluated in a randomized, balanced-order, double-blind, placebo-controlled crossover design. Diazepam (10 mg) or placebo was given 1 h before the neuroimaging acquisition. In a blocked design covert face emotional task, subjects were presented with neutral (A) and aversive (B) (angry or fearful) faces. Participants were also submitted to an explicit emotional face recognition task, and subjective anxiety was evaluated throughout the procedures. Diazepam attenuated the activation of right amygdala and right orbitofrontal cortex and enhanced the activation of right anterior cingulate cortex (ACC) to fearful faces. In contrast, diazepam enhanced the activation of posterior left insula and attenuated the activation of bilateral ACC to angry faces. In the behavioural task, diazepam impaired the recognition of fear in female faces. Under the action of diazepam, volunteers were less anxious at the end of the experimental session. These results suggest that benzodiazepines can differentially modulate brain activation to aversive stimuli, depending on the stimulus features and indicate a role of amygdala and insula in the anxiolytic action of benzodiazepines.

Metacontingencies, Experimentation and Nonhumans: Searching for Conceptual and Methodological Advances

Metacontingency has been described as the functional relation between interlocking behavioral contingencies, plus their direct and immediate effect, called aggregated product, and a selecting event dependent of such effect, called cultural consequence. The metacontingencies analysis enables the discussion of human behavior complexity in social systems. In the present study, we aimed to review and discuss: (a) the importance of basic behavioral processes analysis for the comprehension of social human phenomena; (b) the necessity of constructing and improving metacontingencies experimental models; (c) the current state of metacontingencies experimental investigations in humans; (d) the use of animal models as a way to control the effects of verbal behavior, among other variables, over cultural selection; (e) a concrete and illustrative proposal of an animal model of metacontingencies.

Cerebral amyloid angiopathy impact on endothelium

Cerebral amyloid angiopathy (CAA) is an age-associated disease characterized by amyloid deposition in cerebral and meningeal vessel walls. CAA is detected in the majority of the individuals with dementia and also in a large number of non-demented elderly individuals. In addition, CAA is strongly associated with Alzheimer's disease (AD) pathology. Mechanical consequences including intra-cerebral or subarachnoid hemorrhage remains CAA most feared complication, but only a small fraction of CAA results in severe bleeding. On the hand the non-mechanical consequences in cerebrovascular regulation are prevalent and may be even more deleterious. Studies of animal models have provided strong evidence linking the vasoactive A beta 1-40, the main species found in CAA, to disturbances in endothelial-dependent factors, disrupting cerebrovascular regulation Here, we aimed to review experimental findings regarding the non-mechanical consequences of CAA for cerebrovascular regulation and discuss the implications of these results to clinical practice. (C) 2012 Elsevier Inc. All rights reserved.

Human Adipose-Derived Mesenchymal Stromal Cells Injected Systemically Into GRMD Dogs Without Immunosuppression Are Able to Reach the Host Muscle and Express Human Dystrophin

Duchenne muscular dystrophy (DMD), a lethal X-linked disorder, is the most common and severe form of muscular dystrophies, affecting I in 3,500 male births. Mutations in the DMD gene lead to the absence of muscle dystrophin and a progressive degeneration of skeletal muscle. The possibility to treat DMD through cell therapy has been widely investigated. We have previously shown that human adipose-derived stromal cells (hASCs) injected systemically in SJL mice are able to reach and engraft in the host muscle, express human muscle proteins, and ameliorate the functional performance of injected animals without any immunosuppression. However, before starting clinical trials in humans many questions still need to be addressed in preclinical studies, in particular in larger animal models, when available. The best animal model to address these questions is the golden retriever muscular dystrophy (GRMD) dog that reproduces the full spectrum of human DMD. Affected animals carry a mutation that predicts a premature termination codon in exon 8 and a peptide that is 5% the size of normal dystrophin. These dogs present clinical signs within the first weeks and most of them do not survive beyond age two. Here we show the results of local and intravenous injections of hASCs into GRMD dogs, without immunosuppression. We observed that hASCs injected systemically into the dog cephalic vein are able to reach, engraft, and express human dystrophin in the host GRMD dystrophic muscle up to 6 months after transplantation. Most importantly, we demonstrated that injecting a huge quantity of human mesenchymal cells in a large-animal model, without immunosuppression, is a safe procedure, which may have important applications for future therapy in patients with different forms of muscular dystrophies.

Lipopolysaccharide-Induced Sickness Behaviour Evaluated in Different Models of Anxiety and Innate Fear in Rats

The fact that there is a complex and bidirectional communication between the immune and nervous systems has been well demonstrated. Lipopolysaccharide (LPS), a component of gram-negative bacteria, is widely used to systematically stimulate the immune system and generate profound physiological and behavioural changes, also known as sickness behaviour (e.g. anhedonia, lethargy, loss of appetite, anxiety, sleepiness). Different ethological tools have been used to analyse the behavioural modifications induced by LPS; however, many researchers analysed only individual tests, a single LPS dose or a unique ethological parameter, thus leading to disagreements regarding the data. In the present study, we investigated the effects of different doses of LPS (10, 50, 200 and 500 mu g/kg, i.p.) in young male Wistar rats (weighing 180200 g; 89 weeks old) on the ethological and spatiotemporal parameters of the elevated plus maze, light-dark box, elevated T maze, open-field tests and emission of ultrasound vocalizations. There was a dose-dependent increase in anxiety-like behaviours caused by LPS, forming an inverted U curve peaked at LPS 200 mu g/kg dose. However, these anxiety-like behaviours were detected only by complementary ethological analysis (stretching, grooming, immobility responses and alarm calls), and these reactions seem to be a very sensitive tool in assessing the first signs of sickness behaviour. In summary, the present work clearly showed that there are resting and alertness reactions induced by opposite neuroimmune mechanisms (neuroimmune bias) that could lead to anxiety behaviours, suggesting that misunderstanding data could occur when only few ethological variables or single doses of LPS are analysed. Finally, it is hypothesized that this bias is an evolutionary tool that increases animals security while the body recovers from a systemic infection.

Combined effect of AMPK/PPAR agonists and exercise training in mdx mice functional performance

The present investigation was undertaken to test whether exercise training (ET) associated with AMPK/PPAR agonists (EM) would improve skeletal muscle function in mdx mice. These drugs have the potential to improve oxidative metabolism. This is of particular interest because oxidative muscle fibers are less affected in the course of the disease than glycolitic counterparts. Therefore, a cohort of 34 male congenic C57Bl/10J mdx mice included in this study was randomly assigned into four groups: vehicle solution (V), EM [AICAR (AMPK agonist, 50 mg/Kg-1.day-1, ip) and GW 1516 (PPAR delta agonist, 2.5 mg/Kg-1.day-1, gavage)], ET (voluntary running on activity wheel) and EM+ET. Functional performance (grip meter and rotarod), aerobic capacity (running test), muscle histopathology, serum creatine kinase (CK), levels of ubiquitined proteins, oxidative metabolism protein expression (AMPK, PPAR, myoglobin and SCD) and intracellular calcium handling (DHPR, SERCA and NCX) protein expression were analyzed. Treatments started when the animals were two months old and were maintained for one month. A significant functional improvement (p<0.05) was observed in animals submitted to the combination of ET and EM. CK levels were decreased and the expression of proteins related to oxidative metabolism was increased in this group. There were no differences among the groups in the intracellular calcium handling protein expression. To our knowledge, this is the first study that tested the association of ET with EM in an experimental model of muscular dystrophy. Our results suggest that the association of ET and EM should be further tested as a potential therapeutic approach in muscular dystrophies.