Anatomical features of the urethra and urinary bladder catheterization in female mice and rats. An essential translational tool

PURPOSE: To present fundamental anatomical aspects and technical skills necessary to urethra and urinary bladder catheterization in female mice and rats. METHODS: Urethral and bladder catheterization has been widely utilized for carcinogenesis and cancer research and still remains very useful in several applications: from toxicological purposes as well as inflammatory and infectious conditions to functional aspects as bladder dynamics and vesicoureteral reflux, among many others. RESULTS: Animal models are in the center of translational research and those involving rodents are the most important nowadays due to several advantages including human reproducibility, easy handling and low cost. CONCLUSIONS: Although technical and anatomical pearls for rodent urethral and bladder access are presented as tackles to the advancement of lower urinary tract preclinical investigation in a broaden sight, restriction to female animals hampers the male microenvironment, demanding future advances.

Biomechanical effect of one session of low-level laser on the bone-titanium implant interface

Low-level laser (LLL) has been used on peri-implant tissues for accelerating bone formation. However, the effect of one session of LLL in the strength of bone-implant interface during early healing process remains unclear. The present study aims to evaluate the removal torque of titanium implants irradiated with LLL during surgical preparation of implant bed, in comparison to non-irradiation. Sixty-four Wistar rats were used. Half of the animals were included in LLL group, while the other half remained as control. All animals had the tibia prepared with a 2 mm drill, and a titanium implant (2.2 × 4 mm) was inserted. Animals from LLL group were irradiated with laser (gallium aluminum arsenide), with a wavelength of 808 nm, a measured power output of 50 mW, to emit radiation in collimated beams (0.4 cm2), for 1 min and 23 s, and an energy density of 11 J/cm2. Two applications (22 J/cm 2) were performed immediately after bed preparation for implant installation. Flaps were sutured, and animals from both groups were sacrificed 7, 15, 30, and 45 days after implant installation, when load necessary for removing implant from bone was evaluated by using a torquimeter. In both groups, torque values tended to increase overtime; and at 30 and 45 days periods, values were statistically higher for LLL group in comparison to control (ANOVA test, p < 0.0001). Thus, it could be suggested that a single session of irradiation with LLL was beneficial to improve bone-implant interface strength, contributing to the osseointegration process. © 2012 Springer-Verlag London Ltd.

HDAC inhibition decreases XIST expression on female IVP bovine blastocysts

During initial development, both X chromosomes are active in females, and one of them must be silenced at the appropriate time in order to dosage compensate their gene expression levels to male counterparts. Silencing involves epigenetic mechanisms, including histone deacetylation. Major X chromosome inactivation (XCI) in bovine occurs between hatching and implantation, although in vitro culture conditions might disrupt the silencing process, increasing or decreasing X-linked gene expression. In this study, we aimed to address the roles of histone deacetylase inhibition by trichostatin A (TSA) on female preimplantation development.We tested the hypothesis that by enhancing histone acetylation, TSA would increase the percentage of embryos achieving 16-cell stage, reducing percentage of embryos blocked at 8-cell stage, and interfere with XCI in IVF embryos. We noticed that after TSA treatment, acetylation levels in individual blastomeres of 8-16 cell embryos were increased twofold on treated embryos, and the samewas detected for blastocysts. Changes among blastomere levels within the same embryo were diminished on TSA group, as low-acetylated blastomeres were no longer detected. The percentage of embryos that reached the 5th cleavage cycle 118 h after IVF, analyzed by Hoechst staining, remained unaltered after TSA treatment. Then, we assessed XIST and G6PD expression in individual female bovine blastocysts by quantitative real-time PCR. Even though G6PD expression remained unaltered after TSA exposure, XIST expression was eightfold decreased, and we also detected a major decrease in the percentage of blastocysts expressing detectable XIST levels after TSA treatment. Based on these results, we conclude that HDAC is involved on XCI process in bovine embryos, and its inhibition might delay X chromosome silencing and attenuate aberrant XIST expression described for IVF embryos. © 2013 Society for Reproduction and Fertility.

Qualitative analysis of preputial and vaginal bacterial microbiota in owl monkeys (Aotus azarai infulatus) raised in captivity

Background: The aim of this study was to identify the aerobic bacteria of the preputial and vaginal microbiota in owl monkeys that have been raised in captivity and to evaluate the antimicrobial susceptibility of these bacteria by gender and social organization. Methods: Thirty clinically healthy Aotus azarai infulatus were used. A total of 134 samples were collected, 60 from the preputial mucosa and 74 from the vaginal mucosa. An automated system of bacterial identification was used. Results and Conclusions: Staphylococcus intermedius and Proteus mirabilis were the microorganisms that were most frequently identified according to gender and social organization. The antimicrobial susceptibility of the isolated gram-positive bacteria was similar in both sexes. However, the gram-negative strains had some differences. The aerobic bacterial population of the vaginal and preputial microbiota is similar in owl monkeys, and there are no differences in the number and bacterial species according to sex and social organization. © 2012 John Wiley & Sons A/S. Published by Blackwell Publishing Ltd.

Associação genética entre produção de mozzarela e produção de leite e teores de gordura e proteína em bubalinos

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Estudo da participação de reguladores negativos endógenos da atividade de STAT1 e STAT3 (SOCS1 e SOCS3) na doença periodontal experimental

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Expressão de MMP-13 e ativação de vias de sinalização intracelular em dois modelos de doença periodontal induzida experimentalmente em ratos

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Cicatrização de defeitos de deiscência periodontal associados ao aplainamento radicular tratados com enxerto de tecido conj untivo subepitelial ou membrana bioabsorvível: análise histológica e histométrica em cães

Pós-graduação em Odontologia - FOA

Platelet-Rich Plasma, Low-Level Laser Therapy, or Their Combination Promotes Periodontal Regeneration in Fenestration Defects: A Preliminary In Vivo Study

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

Metabolomics method to comprehensively analyze amino acids in different domains

Amino acids play essential roles in both metabolism and the proteome. Many studies have profiled free amino acids (FAAs) or proteins; however, few have connected the measurement of FAA with individual amino acids in the proteome. In this study, we developed a metabolomics method to comprehensively analyze amino acids in different domains, using two examples of different sample types and disease models. We first examined the responses of FAAs and insoluble-proteome amino acids (IPAAs) to the Myc oncogene in Tet21N human neuroblastoma cells. The metabolic and proteomic amino acid profiles were quite different, even under the same Myc condition, and their combination provided a better understanding of the biological status. In addition, amino acids were measured in 3 domains (FAAs, free and soluble-proteome amino acids (FSPAAs), and IPAAs) to study changes in serum amino acid profiles related to colon cancer. A penalized logistic regression model based on the amino acids from the three domains had better sensitivity and specificity than that from each individual domain. To the best of our knowledge, this is the first study to perform a combined analysis of amino acids in different domains, and indicates the useful biological information available from a metabolomics analysis of the protein pellet. This study lays the foundation for further quantitative tracking of the distribution of amino acids in different domains, with opportunities for better diagnosis and mechanistic studies of various diseases.

Morphology of ileal and jejunal pouches used as rectal substitutes

PURPOSE: Our aim was to determine whether the mucosa of a canine jejunal pouch used as a rectal substitute after proctocolectomy retains its morphologic features better than the mucosa of a canine ileal pouch so used. METHODS: Among ten dogs that underwent proctocolectomy, five had a jejunal pouch-distal rectal anastomosis and five an ileal pouch-distal rectal anastomosis. After six months, the animals were killed, the intestinal pouches and portions of unaltered distal ileum were removed, and a blinded, mucosal morphometric analysis was performed. RESULTS: The mucosa of the jejunal pouches had an overall thickness (mean ± standard deviation, anterior and posterior walls, 1,300 ± 140 μm), villous height (286 ± 46 μm), and crypt depth (790 ± 77 μm) greater than that of the ileal pouches (920 ± 170, 208 ± 47, and 530 ± 130 μm, respectively; P < 0.05). Moreover, the mucosal thickness of the jejunal pouches was similar to that of the distal ileum proximal to the pouch (1,200 ± 200 μm; P > 0.05), whereas the mucosal thickness of the ileal pouch was thinner (P < 0.05). CONCLUSIONS: The jejunal mucosa retains its major morphometric features when the jejunum is used as a rectal substitute after proctocolectomy. In contrast, the ileal mucosa atrophies when the ileum is so used.

A New Approach for Managing Bovine Tuberculosis: Veterinary Services’ Proposed Action Plan

Bovine tuberculosis (TB) is a serious disease with animal health, public health, and international trade consequences. The cooperative Federal-State-industry effort to eradicate bovine TB from cattle in the United States has made significant progress since the program’s inception in 1917. However, the goal of eradication remains elusive. This proposed action plan presents Veterinary Services’ (VS’) current thinking about changes we are considering for the TB program to address our current challenges. This action plan will: 1. Reduce the introduction of TB into the U.S. national herd from imported animals and wildlife by: o Applying additional requirements to cattle imports from Mexico o Enhancing efforts to mitigate risks from wildlife 2. Enhance TB surveillance by: o Crafting a comprehensive national surveillance plan o Accelerating diagnostic test development to support surveillance 3. Increase options for managing TB-affected herds by: o Conducting epidemiological investigations and assessing individual herd risk o Applying whole-herd depopulation judiciously and developing alternative control strategies o Applying animal identification (ID) standards to meet animal ID needs 4. Modernize the regulatory framework to allow VS to focus resources where the disease exists 5. Transition the TB program from a State classification system to a science-based zoning approach to address disease risk To succeed, this new approach will require VS’ continued partnership with State animal health and wildlife officials, other Federal agencies, industry, international partners, academia, and other stakeholders. Successful partnerships will allow us to use available resources efficiently to achieve program objectives and protect our nation’s herd. Implementation of the VS proposed action plan will benefit Federal and State animal health officials, the regulated industries, and producers by allowing a more rapid response that employs up-to-date science and can adapt rapidly to changing situations.

Surgical Performance During Laparoscopic Radical Nephrectomy Is Improved With Training in a Porcine Model

Background and Purpose: Becoming proficient in laparoscopic surgery is dependent on the acquisition of specialized skills that can only be obtained from specific training. This training could be achieved in various ways using inanimate models, animal models, or live patient surgery-each with its own pros and cons. Currently, there are substantial data that support the benefits of animal model training in the initial learning of laparoscopy. Nevertheless, whether these benefits extent themselves to moderately experienced surgeons is uncertain. The purpose of this study was to determine if training using a porcine model results in a quantifiable gain in laparoscopic skills for moderately experienced laparoscopic surgeons. Materials and Methods: Six urologists with some laparoscopic experience were asked to perform a radical nephrectomy weekly for 10 weeks in a porcine model. The procedures were recorded, and surgical performance was assessed by two experienced laparoscopic surgeons using a previously published surgical performance assessment tool. The obtained data were then submitted to statistical analysis. Results: With training, blood loss was reduced approximately 45% when comparing the averages of the first and last surgical procedures (P = 0.006). Depth perception showed an improvement close to 35% (P = 0.041), and dexterity showed an improvement close to 25% (P = 0.011). Total operative time showed trends of improvement, although it was not significant (P = 0.158). Autonomy, efficiency, and tissue handling were the only aspects that did not show any noteworthy change (P = 0.202, P = 0.677, and P = 0.456, respectively). Conclusions: These findings suggest that there are quantifiable gains in laparoscopic skills obtained from training in an animal model. Our results suggest that these benefits also extend to more advanced stages of the learning curve, but it is unclear how far along the learning curve training with animal models provides a clear benefit for the performance of laparoscopic procedures. Future studies are necessary to confirm these findings and better understand the impact of this learning tool on surgical practice.

Effects of selective bile duct ligation on liver parenchyma in young animals: histologic and molecular evaluations

Background/Purpose: The mechanisms of increased collagen production and liver parenchyma fibrosis are poorly understood. These phenomena are observed mainly in children with biliary obstruction (BO), and in a great number of patients, the evolution to biliary cirrhosis and hepatic failure leads to the need for liver transplantation before adolescence. However, pediatric liver transplantation presents with biliary complications in 20% to 30% of cases in the postoperative period. Intra-or extrahepatic stenosis of bile ducts is frequent and may lead to secondary biliary cirrhosis and the need for retransplantation. It is unknown whether biliary stenosis involving isolated segments or lobes may affect the adjacent nonobstructed lobes by paracrine or endocrine means, leading to fibrosis in this parenchyma. Therefore, the present study aimed to create an experimental model of selective biliary duct ligation in young animals with a subsequent evaluation of the histologic and molecular alterations in liver parenchyma of the obstructed and nonobstructed lobes. Methods: After a pilot study to standardize the surgical procedures, weaning rats underwent ligation of the bile ducts of the median, left lateral, and caudate liver lobes. The bile duct of the right lateral lobe was kept intact. To avoid intrahepatic biliary duct collaterals neoformation, the parenchymal connection between the right lateral and median lobes was clamped. The animals were divided into groups according to the time of death: 1, 2, 3, 4, and 8 weeks after surgical procedure. After death, the median and left lateral lobes (with BO) and the right lateral lobe (without BO [NBO]) were harvested separately. A group of 8 healthy nonoperated on animals served as controls. Liver tissues were subjected to histologic evaluation and quantification of the ductular proliferation and of the portal fibrosis. The expressions of smooth muscle alpha-actin (alpha-SMA), desmin, and transforming growth factor beta 1 genes were studied by molecular analyses (semiquantitative reverse transcriptase-polymerase chain reaction and real-time polymerase chain reaction, a quantitative method). Results: Histologic analyses revealed the occurrence of ductular proliferation and collagen formation in the portal spaces of both BO and NBO lobes. These phenomena were observed later in NBO than BO. Bile duct density significantly increased 1 week after duct ligation; it decreased after 2 and 3 weeks and then increased again after 4 and 8 weeks in both BO and NBO lobes. The portal space collagen area increased after 2 weeks in both BO and NBO lobes. After 3 weeks, collagen deposition in BO was even higher, and in NBO, the collagen area started decreasing after 2 weeks. Molecular analyses revealed increased expression of the alpha-SMA gene in both BO and NBO lobes. The semiquantitative and quantitative methods showed concordant results. Conclusions: The ligation of a duct responsible for biliary drainage of the liver lobe promoted alterations in the parenchyma and in the adjacent nonobstructed parenchyma by paracrine and/or endocrine means. This was supported by histologic findings and increased expression of alpha-SMA, a protein related to hepatic fibrogenesis. (C) 2012 Elsevier Inc. All rights reserved.

Ética da pesquisa em modelos animais

A utilização de animais em experimentos científicos é descrita desde o século V a.C. Avanços científicos na área da saúde são atribuídos a modelos animais. O status moral dos animais sempre foi debatido. OBJETIVOS: Este artigo visa à revisão histórica e resumo da legislação atual, para orientar pesquisadores ao utilizar modelos animais na pesquisa em otorrinolaringologia. MATERIAL E MÉTODOS: Pesquisa na base de dados Medline. RESULTADOS: no Brasil, por muitos anos não havia regulamentação para o uso de animais em experimentação. Eram seguidas normas de organizações nacionais e internacionais. Recentemente, foi sancionada a lei nº 11.794/08, que estabelece procedimentos para o uso científico de animais. Na otorrinolaringologia, os estudos com laringe utilizaram coelho, porco, cachorro, cobaias (Cavia porcellus) e camundongo; estudos para face coelho, rato e cachorro; rinoplastia com coelho; e orelha interna com ratos e cobaias (albinas). CONCLUSÕES: Os pesquisadores envolvidos em trabalhos científicos com animais devem conhecer os princípios da lei nº 11.794/08 e pesquisar quais animais são apropriados para cada subárea estudada seus modelos com maior aplicabilidade. Os otorrinolaringologistas, especialmente aqueles que se dedicam à pesquisa, necessitam estar sempre atentos para o respeito às regras éticas de utilização de animais em seus estudos.