Effect of melatonin intake on oxidative stress biomarkers in male reproductive organs of rats under experimental diabetes

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Peritoneal reactivity evaluation in horses subjected to experimental small colon enterotomy and treated with subcutaneous heparin

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

Anti-inflammatory effects of Brazilian ginseng (Pfaffia paniculata) on TNBS-induced intestinal inflammation: experimental evidence

Inflammatory bowel disease (IBD) is a chronic, relapsing, idiopathic inflammation of the gastrointestinal tract. Clinical studies suggest that the initiation of IBD is multifactorial, involving genetics, the immune system and environmental factors, such as diet, drugs and stress. Pfaffia paniculata is an adaptogenic medicinal plant used in Brazilian folk medicine as an anti-stress agent. Thus, we hypothesised that the P. paniculata enhances the response of animals subjected to colonic inflammation. Our aim was to investigate the intestinal anti-inflammatory activity of P. paniculata in rats before or after induction of intestinal inflammation using trinitrobenzenesulfonic acid (TNBS). The animals were divided into groups that received the vehicle, prednisolone or P. paniculata extract daily starting 14days before or 7days after TNBS induction. At the end of the procedure, the animals were killed and their colons were assessed for the macroscopic damage score (MDS), extent of the lesion (EL) and weight/length ratio, myeloperoxidase (MPO) activity and glutathione (GSH), cytokines and C-reactive protein (CRP) levels. Histological evaluation and ultrastructural analysis of the colonic samples were performed. Treatment with the 200mg/kg dose on the curative schedule was able to reduce the MDS and the EL. In addition, MPO activity was reduced, GSH levels were maintained, and the levels of pro-inflammatory cytokines and CRP were decreased. In conclusion, the protective effect of P. paniculata was related to reduced oxidative stress and CRP colonic levels, and due to immunomodulatory activity as evidenced by reduced levels of IL-1β, INF-γ, TNF-α and IL-6.

Does spinal block through tattooed skin cause histological changes in nervous tissue and meninges?: an experimental model in rabbits

Although there is no documented evidence that tattoo pigments can cause neurological complications, the implications of performing neuraxial anesthesia through tattooed skin are unknown. In this study, we aimed to assess whether spinal puncture performed through tattooed skin of rabbits determines changes over the spinal cord and meninges. In addition, we sought to evaluate the presence of ink fragments entrapped in spinal needles. Thirty-six young male adult rabbits, each weighing between 3400 and 3900 g and having a spine length between 38.5 and 39 cm, were divided by lot into 3 groups as follows: GI, spinal puncture through tattooed skin; GII, spinal puncture through tattooed skin and saline injection; and GIII, spinal puncture through skin free of tattoo and saline injection. After intravenous anesthesia with ketamine and xylazine, the subarachnoid space was punctured at S1-S2 under ultrasound guidance with a 22-gauge 2½ Quincke needle. Animals in GII and GIII received 5 μL/cm of spinal length (0.2 mL) of saline intrathecally. In GI, the needle tip was placed into the yellow ligament, and no solution was injected into the intrathecal space; after tattooed skin puncture, 1 mL of saline was injected through the needle over a histological slide to prepare a smear that was dyed by the Giemsa method to enable tissue identification if present. All animals remained in captivity for 21 days under medical observation and were killed by decapitation. The lumbosacral spinal cord portion was removed for histological analysis using hematoxylin-eosin stain. None of the animals had impaired motor function or decreased nociception during the period of clinical observation. None of the animals from the control group (GIII) showed signs of injuries to meninges. In GII, however, 4 animals presented with signs of meningeal injury. The main histological changes observed were focal areas of perivascular lymphoplasmacyte infiltration in the pia mater and arachnoid. There was no signal of injury in neural tissue in any animal of both groups. Tissue coring containing ink pigments was noted in all GI smears from the spinal needles used to puncture the tattooed skin. On the basis of the present results, intrathecal injection of saline through a needle inserted through tattooed skin is capable of producing histological changes over the meninges of rabbits. Ink fragments were entrapped inside the spinal needles, despite the presence of a stylet.

A new heterologous fibrin sealant as a scaffold to cartilage repair - experimental study and preliminary results

Autologous fibrin gel is commonly used as a scaffold for filling defects in articular cartilage. This biomaterial can also be used as a sealant to control small hemorrhages and is especially helpful in situations where tissue reparation capacity is limited. In particular, fibrin can act as a scaffold for various cell types because it can accommodate cell migration, differentiation, and proliferation. Despite knowledge of the advantages of this biomaterial and mastery of the techniques required for its application, the durability of several types of sealant at the site of injury remains questionable. Due to the importance of such data for evaluating the quality and efficiency of fibrin gel formulations on its use as a scaffold, this study sought to analyze the heterologous fibrin sealant developed from the venom of Crotalus durissus terrificus using studies in ovine experimental models. The fibrin gel developed from the venom of this snake was shown to act as a safe, stable, and durable scaffold for up to seven days, without causing adverse side effects. Fibrin gel produced from the venom of the Crotalus durissus terrificus snake possesses many clinical and surgical uses. It presents the potential to be used as a biomaterial to help repair skin lesions or control bleeding, and it may also be used as a scaffold when applied together with various cell types. The intralesional use of the fibrin gel from the venom of this snake may improve surgical and clinical treatments in addition to being inexpensive and adequately consistent, durable, and stable. The new heterologous fibrin sealant is a scaffold candidate to cartilage repair in this study.

Humoral Immune Responses of White-Tailed Deer (Odocoileus virginianus) to Mycobacterium bovis BCG Vaccination and Experimental Challenge with M. bovis

Monitoring of the kinetics of production of serum antibodies to multiple mycobacterial antigens can be useful as a diagnostic tool for the detection of Mycobacterium bovis infection as well as for the characterization of disease progression and the efficacy of intervention strategies in several species. The humoral immune responses to multiple M. bovis antigens by white-tailed deer vaccinated with BCG orally via a lipid-formulated bait (n = 5), orally in liquid form (n = 5), and subcutaneously (n = 6) were evaluated over time after vaccination and after experimental challenge with virulent M. bovis and were compared to the responses by unvaccinated deer (n = 6). Antibody responses were evaluated by using a rapid test (RT), a multiantigen print immunoassay (MAPIA), a lipoarabinomannan enzyme-linked immunosorbent assay (LAM-ELISA), and immunoblotting to whole-cell sonicate and recombinant antigen MPB83. MAPIA and RT detected minimal to no antibody responses over those at the baseline to multiple M. bovis antigens in vaccinated white-tailed deer after challenge. This was in contrast to the presence of more readily detectable antibody responses in nonvaccinated deer with more advanced disease. The LAM-ELISA results indicated an overall decrease in the level of production of detectable antibodies against lipoarabinomannan-enriched mycobacterial antigen in vaccinated animals compared to that in nonvaccinated animals after challenge. Immunoblot data were inconsistent but did suggest the occurrence of unique antibody responses by certain vaccinated groups to Ag85 and HSP70. These findings support further research toward the improvement and potential use of antibody-based assays, such as MAPIA, RT, and LAM-ELISA, as tools for the antemortem assessment of disease progression in white-tailed deer in both experimental and field vaccine trials.

Experimental Deer-To-Deer Transmission of Mycobacterium bovis

Objective—To determine whether Mycobacterium bovis can be transmitted from experimentally infected deer to uninfected in-contact deer. Animals—Twenty-three 6-month-old white-tailed deer. Procedure—On day 0, M bovis (2 X 108 colony-forming units) was administered by intratonsillar instillation to 8 deer; 3 control deer received saline (0.9% NaCl) solution. Eight in-contact deer were comingled with inoculated deer from day 21. On day 120, inoculated deer were euthanatized and necropsied. On day 180, 4 in-contact deer were euthanatized, and 4 new incontact deer were introduced. On day 360, all in-contact deer were euthanatized. Rectal, oral, and nasal swab specimens and samples of hay, pelleted feed, water, and feces were collected for bacteriologic culture. Tissue specimens were also collected at necropsy for bacteriologic culture and histologic analysis. Results—On day 90, inoculated and in-contact deer developed delayed-type hypersensitivity (DTH) reactions to purified protein derivative of M bovis. Similarly, new in-contact deer developed DTH reactions by 100 days of contact with original in-contact deer. Tuberculous lesions in in-contact deer were most commonly detected in lungs and tracheobronchial and medial retropharyngeal lymph nodes. Mycobacterium bovis was isolated from nasal secretions and saliva from inoculated and in-contact deer, urine and feces from in-contact deer, and hay and pelleted feed. Conclusions and Clinical Relevance—Mycobacterium bovis is efficiently transmitted from experimentally infected deer to uninfected in-contact deer through nasal secretions, saliva, or contaminated feed. Wildlife management practices that result in unnatural gatherings of deer may enhance both direct and indirect transmission of M bovis.

Evaluation of an Experimental Gel Containing Euclea natalensis: An In Vitro Study

Objective. To evaluate the effect of an experimental gel containing Euclea natalensis extract on dentin permeability. Methods. Thirty-six dentin discs, 1-mm-thick. The discs were prepared from the coronal dentin of extracted human third molars that were divided into 3 groups (n = 10). The dentin discs in each group were treated with the groups following experimental materials: (FG): 1.23% fluoride gel, pH 4.1; (EG): Euclea natalensis extract gel, pH 4.1; (CG): control gel, pH 4.1. The gels were applied to the occlusal slide of the dentin under the following conditions: after 37% phosphoric acid and before 6% citric acid. The hydraulic conductance (HC) of each condition was determined four times using a fluid flow apparatus (Flodec). The data were analyzed using Two-way ANOVA and Tukey's test (P < 0.05). Results. The greatest mean reduction in HC was produced in group EG dentin discs (61.2%; P < 0.05). Even after acid challenge with 6% citric acid the great reduction occurred in group EG (66.0%; P < 0.05) than other groups (CG-77.1%, FG-90.8%). Conclusion. E. natalensis gel not only reduced dentin permeability, but also resisted posttreatment citric acid challenge without changing its permeability. Further research has to confirm this promising result in the clinical situation.

0.1T magnetic resonance image in the study of experimental hydrocephalus in rats. Accuracy of the method in the measurements of the ventricular size

PURPOSE: To investigate the accuracy of 1.0T Magnetic Resonance Imaging (MRI) to measure the ventricular size in experimental hydrocephalus in pup rats. METHODS: Wistar rats were subjected to hydrocephalus by intracisternal injection of 20% kaolin (n=13). Ten rats remained uninjected to be used as controls. At the endpoint of experiment animals were submitted to MRI of brain and killed. The ventricular size was assessed using three measures: ventricular ratio (VR), the cortical thickness (Cx) and the ventricles area (VA), performed on photographs of anatomical sections and MRI. RESULTS: The images obtained through MR present enough quality to show the lateral ventricular cavities but not to demonstrate the difference between the cortex and the white matter, as well as the details of the deep structures of the brain. There were no statistically differences between the measures on anatomical sections and MRI of VR and Cx (p=0.9946 and p=0.5992, respectively). There was difference between VA measured on anatomical sections and MRI (p<0.0001). CONCLUSION: The parameters obtained through 1.0T MRI were sufficient in quality to individualize the ventricular cavities and the cerebral cortex, and to calculate the ventricular ratio in hydrocephalus rats when compared to their respective anatomic slice.

An experimental model for resistance exercise in rodents

This study aimed to develop an equipment and system of resistance exercise (RE), based on squat-type exercise for rodents, with control of training variables. We developed an operant conditioning system composed of sound, light and feeding devices that allowed optimized RE performance by the animal. With this system, it is not necessary to impose fasting or electric shock for the animal to perform the task proposed (muscle contraction). Furthermore, it is possible to perform muscle function tests in vivo within the context of the exercise proposed and control variables such as intensity, volume (sets and repetitions), and exercise session length, rest interval between sets and repetitions, and concentric strength. Based on the experiments conducted, we demonstrated that the model proposed is able to perform more specific control of other RE variables, especially rest interval between sets and repetitions, and encourages the animal to exercise through short-term energy restriction and "disturbing" stimulus that do not promote alterations in body weight. Therefore, despite experimental limitations, we believe that this RE apparatus is closer to the physiological context observed in humans.

Short implants (6 mm) installed immediately into extraction sockets: An experimental study in dogs

Aim: To evaluate the effect of implant length (6 mm vs. 11 mm) on osseointegration (bone-toimplant contact) of implants installed into sockets immediately after tooth extraction. Material and methods: In six Labrador dogs, the pulp tissue of the mesial roots of P-3(3) was removed and the root canals were filled. Flaps were elevated bilaterally, the premolars hemisectioned and the distal roots removed. Recipient sites were prepared in the distal alveolus and a 6 mm or an 11 mm long implant was installed at the test and control sites, respectively. Non-submerged healing was allowed. After 4 months of healing, block sections of the implant sites were obtained for histological processing and peri-implant tissue assessment. Results: No statistically significant differences were found between test and control sites both for hard and soft tissue parameters. The bone-to-implant contact evaluated at the apical region of the implants was similar as well. Although not statistically significant, the location of the top of the bony crest at the buccal aspect was more apical in relation to the implant shoulder at the test compared with the control sites (2.0 +/- 1.4 and 1.2 +/- 1.1 mm, respectively). Conclusions: Shorter implants (6 mm) present with equal osseointegration than do longer implants (11 mm).

Role of teeth adjacent to implants installed immediately into extraction sockets: an experimental study in the dog

Aim: To evaluate the influence of the presence of both adjacent teeth on the level of alveolar bony crest at sites where implants were installed into the socket immediately after tooth extraction. Material and methods: Six Labrador dogs were used. Extractions of all teeth from the second premolar to the first molar were performed in the right side of the mandible, after full-thickness flap elevation. In the left side of the mandible, an endodontic treatment of the mesial root of the third and fourth premolars was performed. Full-thickness flaps were elevated, the teeth hemisected, and the distal roots removed. Immediately after, implants were bilaterally installed with the margin flush to the buccal bony crest. The implants were placed in the center of the alveolus at the third premolars and toward the lingual bony plate of the alveolus at the fourth premolars. After 3 months of healing, the animals were euthanized. Results: All implants were integrated in mature bone. More bone resorption was observed at the test compared to the control sites. At the buccal aspect, a resorption of 2.8 +/- 0.5 and 1.6 +/- 0.4 mm at the third premolars and of 2.4 +/- 0.6 and 0.8 +/- 0.7 mm at the fourth premolars were found, at the test and control sites, respectively. At the lingual aspect, the bony crest was apically located in relation to the implant shoulder 1.5 +/- 0.3 and 0.5 +/- 0.5 mm at the third premolars and 1.6 +/- 0.6 and 0.3 +/- 1.1 mm at the fourth premolars, at the test and control sites, respectively. A lower buccal bone resorption was found at the control implants placed lingually. Conclusion: Multiple extractions of teeth adjacent to a socket into which implants were installed immediately after, tooth extraction induced more alveolar bone recession compared to sites where the adjacent teeth were preserved. Moreover, an implant placed more lingually yielded less recession of the buccal aspect of the implant.

Experimental arthritis exacerbates Aggregatibacter actinomycetemcomitans-induced periodontitis in mice

Aim This study aimed to investigate whether chronic antigen-induced arthritis (AIA) influences infection-induced periodontitis (PD) in mice and whether PD modifies the clinical course of AIA. The contribution of anti-TNF-a therapy was also evaluated. Materials and methods The PD was induced in C57BL/6 mice by oral infection with Aggregatibacter actinomycetemcomitans. AIA was induced after infection. Anti-TNF-a and chlorhexidine therapies were used to investigate the role of TNF-a and oral infection on PD and AIA interaction. Maxillae, knee joints, lymph nodes and serum samples were used for histomorphometric, immunoenzymatic and/or real time-PCR analyses. Results Antigen-induced arthritis exacerbated alveolar bone loss triggered by PD infection. In contrast, PD did not influence AIA in the evaluated time-points. PD exacerbation was associated with enhanced production of IFN-? in maxillae and expression of the Th1 transcription factor tBET in submandibular lymph nodes. Increased serum levels of IL-6 and C-reactive protein were also detected. Anti-TNF-a and antiseptic therapies prevented the development and exacerbation of infectious-PD. Anti-TNF-a therapy also resulted in reduced expression of IFN-?, TNF-a and IL-17 in maxillae. Conclusions Altogether, the current results indicate that the exacerbation of infection-induced PD by arthritis is associated with an alteration in lymphocyte polarization pattern and increased systemic immunoreactivity. This process was ameliorated by anti-TNF-a and antiseptic therapies.

Experimental Basis and New Insights for Cell Therapy in Chronic Obstructive Pulmonary Disease

Chronic Obstructive Pulmonary Disease (COPD) can be briefly described as air flow limitation and chronic dyspnea associated to an inflammatory response of the respiratory tract to noxious particles and gases. Its main feature is the obstruction of airflow and consequent chronic dyspnea. Despite recent advances, and the development of new therapeutic, medical and clinical approaches, a curative therapy is yet to be achieved. Therapies involving the use of tissue-specific or donor derived cells present a promising alternative in the treatment of degenerative diseases and injuries. Recent studies demonstrate that mesenchymal stem cells have the capacity to modulate immune responses in acute lung injury and pulmonary fibrosis in animal models, as well as in human patients. Due to these aspects, different groups raised the possibility that the stem cells from different sources, such as those found in bone marrow or adipose tissue, could act preventing the emphysematous lesion progression. In this paper, it is proposed a review of the current state of the art and future perspectives on the use of cell therapy in obstructive lung diseases.

Characterization of an extrapolation chamber for low-energy X-rays: Experimental and Monte Carlo preliminary results

The extrapolation chamber is a parallel-plate ionization chamber that allows variation of its air-cavity volume. In this work, an experimental study and MCNP-4C Monte Carlo code simulations of an ionization chamber designed and constructed at the Calibration Laboratory at IFEN to be used as a secondary dosimetry standard for low-energy X-rays are reported. The results obtained were within the international recommendations, and the simulations showed that the components of the extrapolation chamber may influence its response up to 11.0%. (C) 2011 Elsevier Ltd. All rights reserved.