Search

Autophagy is a pathway through which cells execute a plethora of functions, such as macromolecules and organelles quality control, recycling of building blocks and apoptosis. Numerous studies have shown in the past that autophagy is an important mechanism associated with the pathology of various neurodegenerative diseases, whose impairment may lead to several disease-characteristic phenotypes (e.g. misfolded protein and defective organelles accumulation). With this in mind, we aimed to investigate whether alterations in expression of autophagy-related proteins would show before hyperphosphorylation of Tau, a hallmark of Alzheimer's disease (AD). After analyzing 7 different proteins, we observed that, while Pink1 and p62 show an age-related reduction in the Ts65Dn mice respectively in the locus coeruleus and hippocampus, Parkin shows an age-genotype interaction-associated reduction in both brain areas. This suggests potential outcomes in pathways associated with Parkin that could relate to later stages of the disease development.

The association between advanced paternal age and impaired reproductive outcomes is still controversial. Several studies relate decrease in semen quality, impaired embryo/fetal development and offspring health to increased paternal age. However, some retrospective studies observed no alterations on both seminal status and reproductive outcomes in older men. Such inconsistency may be due to the influence of intrinsic and external factors, such as genetics, race, diet, social class, lifestyle and obvious ethical issues that may bias the assessment of reproductive status in humans. The use of the murine model enables prospective study and owes the establishment of homogeneous and controlled groups. This study aimed to evaluate the effect of paternal age on in vitro embryo development at 4.5 day post conception and on in vivo fetal development at 16 days of gestation. Murine females (2–4 months of age) were mated with young (4–6 months of age) or senile (18–24 months of age) males. We observed decreased in vitro cleavage, blastocyst, and embryo development rates; lighter and shorter fetuses in the senile compared to the young group. This study indicated that advanced paternal age negatively impacts subsequent embryo and fetal development.

RESUMEN Introducción: El correcto funcionamiento del eje hipotálamo-hipófisis-tiroides es indispensable para el crecimiento y desarrollo embrionario-fetal, al intervenir en la diferenciación de los tejidos, el desarrollo cerebral y somático, la maduración ósea y la regulación del metabolismo. El paso de las hormonas tiroideas maternas al feto a través de la placenta depende de transportadores transmembrana, enzimas desyodinasas (DIO2 y DIO3) y proteínas transportadoras (TTR). Objetivo: Identificar las zonas de expresión de DIO3 y TTR en la placenta de ratón Mus musculus E10.5, E12.5, E14.5. Métodos: La estructura placentaria y expresión de DIO3 y TTR fueron evaluadas con técnicas histoquímicas e inmunofluorescencia. Resultados: Desde E10.5 se encontraron las tres zonas placentarias, laberinto, zona de unión y decidua. En E12.5 se observó la conformación placentaria definitiva. DIO3 y TTR fueron detectadas en los tres estadios, con predominio en la zona del laberinto. Conclusión: DIO3 y TTR se expresan a lo largo del establecimiento y maduración de la placenta de ratón. El biomodelo murino es una herramienta útil para el estudio del transporte placentario de hormonas tiroideas desde la circulación materna a la fetal. ABSTRACT Introduction: Correct functioning of hypothalamic-pituitary-thyroid axis is essential for embryonic-fetal growth and development, as it is involved in tissue differentiation, brain and somatic development, bone maturation and metabolic regulation. Maternal thyroid hormones passage to the fetus through the placenta depends on transmembrane transporters, deiodinase enzymes (DIO2 and DIO3) and carrier proteins (TTR). Objective: Identify DIO3 and TTR expression within placental layers of Mus musculus E10.5, E12.5 and E14.5. Methods: Placental structure, DIO3 and TTR expression were evaluated using histochemistry and immunofluorescence techniques. Results: We found that the three placental layers, labyrinth zone, junctional zone, and decidua were present since E10.5. At E12.5 placental final conformation was observed. DIO3 and TTR were detected in the three stages with a predominance in the labyrinth. Conclusion: DIO3 and TTR are expressed throughout the establishment and maturation of mouse placenta. Mice are a useful tool for studying how thyroid hormones are transported from maternal t° fetal circulation at the placenta. RESUMO Introdução: O correto funcionamento do eixo hipotálamo-hipófise-tireoide é essencial para o crescimento e desenvolvimento embrionário-fetal, pois intervém na diferenciação dos tecidos, desenvolvimento cerebral e somático, maturação óssea e regulaçãodo metabolismo. A passagem dos hormônios tireoidianos maternos para o feto através da placenta depende de transportadores transmembranas, enzimas deiodinase (DIO2 e DIO3) e proteínas transportadoras (TTR). Objetivo: Identificar as zonas de expressão de DIO3 e TTR na placenta de rato Mus musculus E10.5, E12.5, E14.5. Métodos: A estrutura placentária e a expressão de DIO3 e TTR foram avaliadas com técnicas histoquímicas e imunofluorescência. Resultados: De E10.5 as três zonas placentárias, labirinto, zona de união e decídua foram encontradas. Em E12.5 a conformação definitiva da placenta foi observada. O DIO3 e o TTR foram detectados nas três fases, com predomínio na área do labirinto. Conclusão: DIO3 e TTR são expressos ao longo do estabelecimento e maturação da placenta de rato O biomodelo murino é uma ferramenta útil para o estudo do transporte placentário dos hormônios tireoidianos da circulação materna para a fetal.

Resumo A utilização de animais não humanos como ferramenta de pesquisa biomédica e em testes da indústria para consumo humano foi incorporada às práticas científicas e assimilada como fundamental. A revisão sistemática dos resultados de protocolos de fases pré-clínicas não é prática corrente, mas metanálises recentes questionam a capacidade de projeção desses dados para a espécie humana. Atualmente, junto com o questionamento científico há abrangente discussão ética sobre os conflitos inerentes à instrumentalização da vida do animal não humano, cujo ápice é alcançado na criação de animais transgênicos. O objetivo deste artigo é discutir a aplicação do conceito de vulnerabilidade ao animal não humano no contexto da experimentação e pensar as relações de poder implícitas nessas práticas. Como aplicação prática da teoria exposta, propõe-se implantar e desenvolver técnicas substitutivas ao modelo animal, que conjuguem ética e inovação. Abstract The use of non-human animals has been incorporated into scientific practices as an essential biomedical research tool and in industry tests for human consumption. The systematic review of protocol results of preclinical phases is not a common practice, but recent meta-analyses question the projection accuracy of these data for humans. Currently, along with scientific questioning, there is a comprehensive ethical discussion about the conflicts in the instrumentalization of non-human life, which reached its peak with the creation of transgenic animals. This article discusses the application of the concept of vulnerability to non-human animals in experiments and reflects on the implicit power relations of these practices. We propose to implement and develop alternative techniques to the animal model, combining ethics and innovation. Resumen El uso de animales no humanos como herramienta para la investigación biomédica y en pruebas de la industria para el consumo humano se ha incorporado a las prácticas científicas y se ha asimilado como fundamental. La revisión sistemática de los resultados de protocolos de fases preclínicas no es una práctica corriente, pero metaanálisis recientes cuestionan la capacidad proyección de estos datos a la especie humana. Actualmente, junto con el cuestionamiento científico, hay una discusión ética sobre los conflictos inherentes a la instrumentalización de la vida del animal no humano, que alcanza su ápice en la creación de animales transgénicos. Este artículo tiene como objetivo discutir la aplicación del concepto de vulnerabilidad al animal no humano en el contexto de la experimentación y proponer una reflexión sobre las relaciones de poder implícitas en estas prácticas. Como una aplicación práctica de la teoría expuesta, se propone implantar y desarrollar técnicas alternativas al modelo animal, que conjuguen ética e innovación.

Resumo Fundamento: A taxa de falha de enxerto de veia safena um ano após a cirurgia de revascularização do miocárdio varia de 10% a 25%. O objetivo deste estudo foi de investigar se a atorvastatina pode reduzir o acúmulo de células musculares lisas vasculares para inibir a hiperplasia intimal por meio da inibição da via p38 MAPK. Métodos: Quarenta e cinco ratos Sprague-Dawley foram randomizados em três grupos. Trinta ratos foram submetidos à cirurgia de enxerto de veia e randomizados para tratamento com veículo ou atorvastatina; quinze ratos foram submetidos à cirurgia sham. Detectamos a hiperplasia intimal por meio de coloração com hematoxilina-eosina e a expressão de proteínas relacionadas por meio de análise imuno-histoquímica e Western blot. Foram realizadas as comparações por análise de variância de fator único e pelo teste da diferença mínima significativa de Fisher, com p < 0,05 considerado significativo. Resultados: A íntima analisada pela coloração com hematoxilina-eosina era dramaticamente mais espessa no grupo controle que no grupo atorvastatina e no grupo sham (p < 0,01). Os resultados da coloração imuno-histoquímica de α-SMA demonstraram que a porcentagem de células positivas para α-SMA no grupo controle era mais alta que no grupo atorvastatina (p < 0,01). Nós também avaliamos α-SMA, PCNA, p38 MAPK e fosforilação de p38 MAPK após o tratamento com estatina por meio de análise de Western blot e os resultados indicaram que a atorvastatina não levou à redução de p38 MAPK (p < 0,05); no entanto, resultou na inibição da fosforilação de p38 MAPK (p < 0,01) e reduziu significativamente os níveis de α-SMA e PCNA, em comparação com o grupo controle (p < 0,01). Conclusão: Nós demonstramos que a atorvastatina pode inibir o acúmulo de células musculares lisas vasculares por meio da inibição da via p38 MAPK e é capaz de inibir a hiperplasia intimal em modelos de enxerto de veia em ratos. Abstract Background: The rate of saphenous vein graft failure one year after coronary artery bypass grafting ranges from 10% to 25%. The aim of this study was to explore whether atorvastatin can reduce accumulation of vascular smooth muscle cells to inhibit intimal hyperplasia via p38 MAPK pathway inhibition. Methods: Forty-five Sprague-Dawley rats were randomized to three groups. Thirty rats received a vein graft operation, and they were randomized to be treated with vehicle or atorvastatin; fifteen rats received a sham operation. We detected intimal hyperplasia by hematoxylin-eosin staining and related protein expression by immunohistochemical and Western blot analysis. Comparisons were analyzed by single-factor analysis of variance and Fisher's least significant difference test, with p < 0.05 considered significant. Results: The intima analyzed by hematoxylin-eosin staining was dramatically thicker in the control group than in the atorvastatin group and sham group (p < 0.01). The outcomes of immunohistochemical staining of α-SMA demonstrated that the percentage of α-SMA-positive cells in the control group was higher than in the atorvastatin group (p < 0.01). We also evaluated α-SMA, PCNA, p38 MAPK, and phosphorylation of p38 MAPK after statin treatment by Western blot analysis, and the results indicated that atorvastatin did not lead to p38 MAPK reduction (p < 0.05); it did, however, result in inhibition of p38 MAPK phosphorylation (p < 0.01), and it significantly reduced α-SMA and PCNA levels, in comparison with the control group (p < 0.01). Conclusion: We have demonstrated that atorvastatin can inhibit accumulation of vascular smooth muscle cells by inhibiting the p38 MAPK pathway, and it is capable of inhibiting intimal hyperplasia in a rat vein graft model.

Submitted by DSpace Unilasalle (dspace@unilasalle.edu.br) on 2021-08-02T19:11:58Z No. of bitstreams: 1 bclopes.etal2.pdf: 928490 bytes, checksum: bf669722d86cf476c34fed69f52984f4 (MD5) Made available in DSpace on 2021-08-02T19:11:58Z (GMT). No. of bitstreams: 1 bclopes.etal2.pdf: 928490 bytes, checksum: bf669722d86cf476c34fed69f52984f4 (MD5) Previous issue date: 2020 Submitted by DSpace Unilasalle (dspace@unilasalle.edu.br) on 2021-08-02T19:12:18Z No. of bitstreams: 1 bclopes.etal2.pdf: 928490 bytes, checksum: bf669722d86cf476c34fed69f52984f4 (MD5) Made available in DSpace on 2021-08-02T19:12:18Z (GMT). No. of bitstreams: 1 bclopes.etal2.pdf: 928490 bytes, checksum: bf669722d86cf476c34fed69f52984f4 (MD5) Previous issue date: 2020 Background Behavioral alterations, like mechanical and thermal hyperalgesia, and modulation of biomarkers in the peripheral and central nervous systems (CNS) are markers of chronic pain. Transcranial direct current stimulation (tDCS) with exercise is a promising therapy for pain due to its neuromodulatory capacity. Objective To assess the individual effects of tDCS, exercise, and the two combined on the nociceptive response and BDNF, IL-1β, and IL-4 levels in the CNS structures of rats in a chronic pain model. Methods For 8 consecutive days after the establishment of chronic neuropathic pain by inducing a constriction injury to the sciatic nerve (CCI), the rats received tDCS, exercise, or both treatments combined (20 min/day). The hyperalgesic response was assessed by von Frey and hot plate tests at baseline, 7, and 14 days after CCI surgery and immediately, 24 h, and 7 days after the end of treatment. The BDNF, IL-1β, and IL-4 levels were assessed in the cerebral cortex, brainstem, and spinal cord by enzyme-linked immunosorbent assay at 48 h and 7 days after the end of treatment. Results The CCI model triggered marked mechanical and thermal hyperalgesia. However, bimodal tDCS, aerobic exercise, and the two combined relieved nociceptive behavior for up to 7 days following treatment completion. Conclusions Bimodal tDCS, aerobic exercise, or both treatments combined promoted analgesic effects for neuropathic pain. Such effects were reflected by cytokine modulation throughout the spinal cord-brainstem-cerebral cortex axis.

Introduction During open surgical dislocated hip reduction, several anatomical structures, such as the round ligament, are approached. However, there is controversy over both the possibility of preserving the ligament and its functional importance. Materials and Methods This experimental study used skeletally immature rabbits as a model for congenital hip dislocation. Thirty-six rabbits comprised the sample that was submitted to the round ligament analysis. The sample was stratified for analysis (biomechanics, zymography, histology, and immunohistochemistry). Statistical analysis compared the unstable side to the control side of each rabbit. Results Biomechanical assays showed that the mean maximal strength of the round ligament on the unstable side was similar to that of the control side (p = 0.594), which was also the case with maximum deformation (p = 0.328). Histologically, there was a statistically significant increase in cellularity on the unstable side (p

Denture stomatitis is the most prevalent form of oral candidiasis and the most frequent oral lesion in removable prosthesis wearers. It is characterized by an inflammatory response of the denture-bearing mucosa, especially the palatal mucosa, and its clinical signs include chronic edema and erythema, and papillary hyperplasia. Despite having a multifactorial etiology, its main etiological agent is the infection by Candida albicans. Given its high treatment failure rates, an in vivo model of denture stomatitis should be established to test alternative treatments. The aim of this study is to review the existing denture stomatitis models and to provide an overview of the main methodological differences between them. Over the last 40 years, different animal models were developed in order to study denture stomatitis etiopathogenesis and to assess novel therapies. Many approaches, including the use of antibiotics and immunosuppressors, have to be further investigated in order to establish which protocol is more appropriate and effective for the development of an animal model of denture stomatitis.

[Resumen]: La artrosis (osteoarthritis, OA) es la forma más común de artritis, consistiendo en el desgaste y pérdida progresiva del cartílago articular hialino, siendo la principal causa de dolor y discapacidad a nivel mundial. Aunque ha sido ampliamente estudiada en modelos animales mamíferos, en el presente proyecto se propone el pez cebra (Danio rerio) como alternativa para el estudio de la enfermedad, ya que este posee una articulación sinovial en la mandíbula, muy similar a la de los humanos. Se ha visto que la modulación de los microARNs en la articulación puede tener un importante efecto en la OA, en concreto el miR-140, el cual está reducido en el tejido con OA. De este modo, se inyectará el anti-miR-140 en los embriones wild type para silenciarlo. Posteriormente, mediante una serie de tinciones histológicas y técnicas inmunohistoquímicas, se valorará, según el score OARSI, como afecta este microARN al cartílago articular y los cambios osteoartríticos que se producen en la mandíbula del pez cebra. Creando, de este modo, una línea para futuras investigaciones en el tratamiento de la enfermedad. [Resumo]: A artrose (osteoarthritis, OA) é a forma máis común de artrites, consistindo no desgaste e perdida progresiva do cartilaxe articular hialino, sendo a principal causa de dor e discapacidade a nivel mundial. Aínda que foi amplamente estudada en modelos animais mamíferos, no presente proxecto proponse o peixe cebra (Danio rerio) como alternativa para o estudo da enfermidade, xa que este posúe unha articulación sinovial na mandíbula, moi similar á dos humanos. Veuse que a modulación dos microARNs na articulación pode ter un importante efecto na OA, en concreto, o miR-140, o cal aparece reducido no tecido con OA. Deste modo, inxectarase o anti-miR-140 nos embrións wild type para silencialo. Posteriormente, mediante unha serie de tincións histolóxicas e técnicas inmunohistoquímicas, valorarase, segundo o score OARSI, como afecta este microARN ao cartilaxe articular e os cambios osteoartríticos que se produce na mandíbula do peixe cebra. Creando, deste modo, una liña para futuras investigacións no tratamento da enfermidade. [Abstract]: Osteoarthritis (OA) is the most common form of arthritis, consisting of the progressive wear and tear and loss of hyaline joint cartilage, and is the main cause of pain and disability worldwide. Although it has been extensively studied in mammalian animal models, this project proposes the zebrafish (Danio rerio) as an alternative for the study of the disease, as it has a synovial joint in the jaw, very similar of humans. It has been shown that modulation of miRNAs in the joint can have an important effect on OA, in particular miR-140, which is reduced in OA tissue. Thus, anti-miR-140 will be injected into wild type embryos to silence it. Subsequently, a series of histological staining and immunohistochemical techniques will be used to assess, according to the OARSI score, how this miRNA affects articular cartilage and the osteoarthritic changes that occur in the zebrafish jaw. Creating a line for future research into the treatment of the disease. Traballo fin de grao (UDC.CIE). Bioloxía. Curso 2021/2022

Background and purpose In preclinical studies, cannabidiol (CBD) mitigates fear memories by facilitating their extinction or interfering with their generalization and reconsolidation. The brain regions and mechanisms underlying these effects, and their temporal window, are still poorly understood. Here, we have investigated related questions in the dorsal hippocampus (DH) during contextual fear consolidation. Experimental approach Adult male Wistar rats received CBD (10-30 pmol) intra-DH immediately, 1 or 3 hr after fear conditioning. Effects of CBD on consolidation were inferred behaviourally and by analysing expression of the activity-regulated, cytoskeleton-associated (Arc) protein. The contribution of anandamide, CB1 , CB2 , 5-HT1A , A2A , and PPARγ receptors was also assessed. Key results CBD impaired memory consolidation when given immediately or 1 hr after fear conditioning, but not after 3 hr. Expression of Arc protein in DH was reduced by systemic CBD treatment in both cases. Immediately after fear conditioning, CBD effects were abolished by CB1 or CB2 receptor blockade, partly reduced by 5-HT1A or A2A antagonism, and remained unchanged after antagonism of PPARγ receptors. One hour after fear conditioning, CBD effects were prevented only by PPARγ receptor antagonism. Also, inhibition of fatty acid amide hydrolase by URB597, impaired memory consolidation when infused immediately, but not 1 hr after fear conditioning. Conclusions and implications CBD disrupts memory consolidation up to 1 hr after fear conditioning, allowing an extended window of opportunity to mitigate aversive memories after their acquisition. Our results suggest time-dependent participation of anandamide, CB1 , CB2 and PPARγ receptors in the DH, during this process.

Introdução: A constituição da memória se dá através de fases diante de estímulos variados. O som constitui importante fonte de tal estimulação. A extinção, com a qual informações anteriormente estabelecidas são suprimidas, representa um novo processo de aprendizagem. Objetivo: Investigar a possível interferência da Sonata K448 de Mozart na extinção da memória de medo ao som. Métodos: Foram utilizados camundongos da linhagem C57BL/6J, divididos em: G1: Mozart (n=7); G2: Ambiente (n=6); e G3: Controle (n=6). Apenas o grupo G1 foi exposto à música a partir da vida intrauterina. Nos dias 50º ao 53º após o nascimento, os animais foram submetidos à Habituação. No 54º dia, realizou-se um Treino Aversivo com G1 e G2. No 55º dia, iniciou-se o Teste de Extinção por doze dias consecutivos com os três grupos. No 81º dia, todos os grupos passaram pelo Teste de Recordação. Os testes foram gravados em vídeo para análise. Os dados foram analisados pelo teste ANOVA de medidas repetidas, sendo considerados significativos p≤0,05. Resultados: Não foi observado efeito principal dos grupos (F(2;16)=2,70; p=0,098), assim como não foi verificada interação grupo x dia (F(2,16;17,30)=1,58; p=0,147). Conclusão: Embora não se tenha observado diferença estatisticamente significativa entre os grupos, futuros estudos com amostras maiores podem ser oportunos para elucidar melhor a influência da música na memória. Introduction: The process of memory occurs through phases due to various stimuli. Sound is an important source of such stimulation. Extinction, with which previously established information is suppressed, represents a new learning process. Objective: To investigate the interference of Mozart's Sonata K448 in the extinction of the memory of fear of the sound. Methods: C57BL/6J mice were used, divided into: G1: Mozart (n=7); G2: Environment (n=6); and G3: Control (n=6). Only the G1 group was exposed to music from intrauterine life onwards. On days 50 to 53 after birth, the animals were subjected to Habituation. On the 54th day, an Aversive Training was performed only with G1 and G2. On the 55th day, the Extinction Test started, lasting twelve consecutive days with the three groups. On the 81st day, all groups passed the Recall Test. The tests were videotaped for analysis. Data were analyzed using the repeated measures ANOVA test, with p≤0.05 being considered significant. Results: There was no groups main effect (F(2;16)=2,70; p=0,098), nor an interaction between groups x days (F(2,16;17,30)=1,58; p=0,147). Conclusion: Although no statistically significant difference was observed between the groups, future studies with larger samples may be opportune to better elucidate the influence of music on memory. Introducción: La constitución de la memoria se da por fases ante diversos estímulos. El sonido es una fuente importante de tal estimulación. La extinción, con la que se suprime la información previamente establecida, representa un nuevo proceso de aprendizaje. Objetivo: Investigar la posible interferencia de la Sonata K448 de Mozart en la extinción de la memoria del miedo al sonido. Métodos: se utilizaron ratones C57BL/6J divididos en: G1: Mozart (n=7); G2: Ambiente (n=6); y G3: Control (n=6). Solo el grupo G1 estuvo expuesto a la música de la vida intrauterina. En los días 50 a 53 después del nacimiento, los animales fueron sometidos a Habituación. El día 54 se realizó un Entrenamiento Aversivo con G1 y G2. El día 55 se inició el Test de Extinción durante doce días consecutivos con los tres grupos. El día 81, todos los grupos pasaron la Prueba de Recordación. Las pruebas fueron grabadas en video para su análisis. Los datos fueron analizados mediante la prueba ANOVA de medidas repetidas, considerándose significativo p≤0,05. Resultados: No hubo efecto principal de los grupos (F(2;16)=2.70; p=0.098), así como tampoco hubo interacción grupo x día (F(2.16;17.30)=1.58 ; p=0.147). Conclusión: Aunque no se observó una diferencia estadísticamente significativa entre los grupos, futuros estudios con muestras más grandes pueden ser oportunos para dilucidar mejor la influencia de la música en la memoria.

Congenital anomalies and its causes, particularly, by external factors are the aim of the field called teratology. The external factors studied by teratology are known as teratogens and can be biological or environmental factors for example, chemicals, medications, recreational drugs, environmental pollutants, physical agents (e.g., X-rays and maternal hyperthermia) and maternal metabolic conditions. Proving the teratogenicity of a factor is a difficult task requiring epidemiology studies as well as experimental teratology evidence from the use of animal models, one of which is the chicken embryo. This model in particular has the advantage of being able to follow development live andin vivo, with rapid development hatching around 21 days, is cheap and easy to manipulate and to observe development. All this allows the chicken embryo to be used in drug screening studies, teratogenic evaluation and studies of mechanisms of teratogenicity. The chicken embryo shares morphological, biochemical and genetic similarities with humans as well as mammalian species, making them ideal to ascertain the actions of teratogens, as well as screen drugs to test for their safety. Pre-clinical trials for new drugs are carried out in rodents and rabbits, however, chicken embryos have been used to screen new compounds or analogs of thalidomide as well as to investigate how some drugs can lead to congenital malformations. Indeed, the chicken embryo has proved valuable in understanding how many congenital anomalies, seen in humans, arise following teratogen exposure. The aim of this review is to highlight the role of the chicken embryo as an experimental model for studies in teratology, exploring its use in drug screening studies, phenotypic evaluation and studies of teratogenic mechanisms of action. Here, we discuss many known teratogens, that have been evaluated using the chicken embryo model including some medicines, such as, thalidomide, valproic acid; recreational drugs including alcohol; environmental influences, such as viruses, specifically ZIKV, which is a newly discovered human teratogen. In addition, we discuss how the chicken embryo has provided insight on the mechanisms of teratogenesis of many compounds and also how this impact on drug safety.

Universidade do Estado do Par?. Programa de P?s-Gradua??o em Biologia Parasit?ria na Amaz?nia. Bel?m, PA, Brasil. Universidade do Estado do Par?. Programa de P?s-Gradua??o em Biologia Parasit?ria na Amaz?nia. Bel?m, PA, Brasil. Universidade do Estado do Par?. Programa de P?s-Gradua??o em Biologia Parasit?ria na Amaz?nia. Bel?m, PA, Brasil. Universidade Federal Rural da Amaz?nia. Programa de P?s-Gradua??o em Sa?de e Produ??o Animal na Amaz?nia. Bel?m, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Centro Nacional de Primatas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. OBJETIVO: Estabelecer e avaliar um modelo experimental com primatas neotropicais n?o humanos Sapajus apella para verificar m?todos diagn?sticos mais eficazes na detec??o da esquistossomose, subsidiando novos estudos que auxiliam na cura e controle dessa doen?a. MATERIAIS E M?TODOS: Foram utilizados seis primatas adultos da esp?cie Sapajus apella, distribu?dos randomicamente em dois grupos experimentais de igual n?mero. Desses, tr?s animais foram infectados por via intrad?rmica e tr?s por via transcut?nea, com suspens?o de cerc?rias obtidas de caramujos Biomphalaria glabrata infectados naturalmente. Para acompanhamento da infec??o, foram coletadas amostras de urina via sonda ou pun??o direta, com o animal anestesiado, e fezes diretamente das gaiolas. As amostras de urina foram examinadas pelo teste r?pido de urina (point-of-care cathodic circulating antigen ? POC-CCA) e as amostras de fezes por exames de sedimenta??o espont?nea das fezes (HPJ) e Kato-Katz. RESULTADOS: O uso das vias transcut?nea e intrad?rmica resultou na infec??o de dois dos tr?s animais avaliados (66,6%) em cada grupo experimental. O exame POC-CCA foi o primeiro a detectar positivos (21? dia), seguido dos exames de HPJ (35? dia) e Kato-Katz (42? dia). CONCLUS?O: O modelo analisado apresenta-se vi?vel para reprodu??o em diferentes estudos sobre esquistossomose mans?nica, pois os animais responderam de maneira satisfat?ria ? infec??o em ambas as vias utilizadas, sendo poss?vel verificar ainda a praticidade da via intrad?rmica quando comparada ? transcut?nea.

Orientador: Fernanda Klein Marcondes Tese (doutorado) - Universidade Estadual de Campinas, Instituto de Biologia Resumo: Dados da Organização Mundial de Saúde mostram que atualmente a depressão atinge 121 milhões de pessoas e deverá ser a doença mais comum no mundo em 2020. Evidências científicas mostram sua associação com o estresse crônico e alterações neuronais relacionadas à depressão, ansiedade, prejuízo no aprendizado e memória. A qualidade do ambiente em que o indivíduo está inserido e o suporte social de que dispõe representam os principais fatores na regulação dessas alterações. Com relação aos mecanismos fisiopatológicos envolvidos nos efeitos do estresse crônico, há estudos que demonstram uma importante associação entre estresse e hiperatividade do sistema renina angiotensina (SRA). Porém, os mecanismos envolvidos na associação entre estresse e depressão e nos efeitos protetores de condições ambientais favoráveis ainda não estão esclarecidos. Em modelo animal, a intervenção ambiental por meio da estimulação tátil ou enriquecimento ambiental tem sido proposta como um fator que poderia diminuir o estresse e promover o bem-estar. No experimento 1, o objetivo deste estudo foi avaliar os efeitos da intervenção ambiental, por meio da estimulação tátil (manipulação) ou enriquecimento ambiental, sobre as respostas hormonais e comportamentais induzidas pelo estresse crônico moderado e imprevisível (ECMI) em ratos Sprague-Dawley jovens. Nesse estudo foi evidenciado que o ECMI aumentou as concentrações séricas de corticosterona e plasmáticas de noradrenalina e adrenalina; induziu comportamentos análogos à depressão; prejudicou o aprendizado e memória e aumentou a concentração tecidual de angiotensina I, II e IV no hipotálamo. Tanto a manipulação quanto o enriquecimento ambiental atenuaram a secreção hormonal, os comportamentos análogos à depressão e o aumento de angiotensina II no hipotálamo; e cancelaram o prejuízo cognitivo induzido pelo ECMI. Com o objetivo de estudar se os efeitos da angiotensina II, mediados pelo receptor tipo 1 de angiotensina II (AT1), estão envolvidos nas respostas hormonais e no prejuízo cognitivo induzido pelo ECMI, no experimento 2, ratos machos Sprague-Dawley controles e estressados, tratados ou não com losartan (antagonista de receptor AT1) foram avaliados. Nesse experimento, a administração do losartan cancelou o aumento na secreção dos hormônios do estresse, atenuou o desamparo aprendido e o prejuízo cognitivo em animais estressados. Os resultados obtidos mostraram que a manipulação ou o enriquecimento ambiental produzem efeitos hormonais e comportamentais positivos capazes de melhorar o bem-estar animal e podem diminuir os efeitos deletérios induzidos por estresse crônico em ratos jovens. Além disso, sugerem que a atividade do sistema renina-angiotensina pode estar envolvida nos efeitos negativos desencadeados pelo estresse crônico Abstract: Data from the World Health Organization show that depression currently affects 121 million people and will probably be the most common disease in the world in 2020. Scientific evidences show that it is associated with chronic stress, neuronal changes depression-related, anxiety and impairment in learning and memory. There are also a significant association between stress and hyperactivity of the renin angiotensin system on behavioral changes and cardiovascular effects. It is also noteworthy that the quality of the environment in which the individual belongs and the social support represent the main factors that may regulated these effects. In addition, the pathophysiological mechanisms involved in the association between stress and depression, and the protective effects of favorable environmental conditions remain unclear. In animal models, environmental intervention through tactile stimulation (handling) or environmental enrichment have been proposed as factors that may reduce stress and promote well-being. In the experiment 1, the objective of this study was to evaluate the effects of environmental intervention, through tactile stimulation (handling) or environmental enrichment on behavioral and hormonal responses induced by chronic mild unpredictable stress (CMS) in young Sprague-Dawley rats. The results of this study showed that the CMS increased serum corticosterone, plasma norepinephrine and epinephrine concentrations; induced depression-like behaviors; impaired learning and memory and increased hypothalamus angiotensin I, II and IV. Handling and environmental enrichment attenuated stress hormones secretion, depression-like behaviors and increased hypothalamic angiotensin II levels; and cancelled impairment of learning and memory induced by CMS. Considering that the aim of study 1 was evaluate if the effects of angiotensin II, mediated by angiotensin II type 1 receptor (AT1), are involved in hormone responses and cognitive impairment induced by CMS, the experiment 2 were performed. In the second study, male Sprague-Dawley rats controls and stressed, treated or not with losartan (AT1 receptor antagonist) were evaluated. Results from the second study showed that the losartan administration cancelled the increase in stress hormones secretion, attenuated depression-like behaviors and reduced the impairment of learning and memory in stressed animals. The data obtained indicated that the handling or environmental enrichment produced positive hormonal and behavioral effects capable of improving the animal¿s welfare, diminishing the deleterious effects induced by chronic stress in adult rats. The results also suggest that renin-angiotensin system over activity seems to be involved in negative effects of CMS Doutorado Fisiologia Doutora em Biologia Funcional e Molecular

Orientadores: José Antônio Rocha Gontijo, Patrícia Aline Boer Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Ciências Médicas Resumo: Introdução: A síndrome dos ovários policísticos (SOP) é uma manifestação endócrina proteiforme e de etiologia desconhecida. Acomete, aproximadamente, 8 % das mulheres em idade reprodutiva. Dentre as principais características desta síndrome destaca-se o hiperandrogenismo, anovulação crônica e infertilidade. A SOP também apresenta alta prevalência de distúrbios que levam à disfunção metabólica, dentre as quais resistência à insulina, hiperinsulinemia, diabetes mellitus tipo 2, obesidade, dislipidemia e, hipertensão arterial. Ações não farmacológicas, como modificações na dieta e exercício físico, têm sido consideradas estratégicas como abordagens de primeira linha na prevenção e tratamento desta disfunção. Objetivo: Diante da problemática apresentada, o objetivo deste estudo foi avaliar em ratas fêmeas androgenizadas em estro persistente, a resposta metabólica, de receptores angiotensinérgicos e a resposta pressórica bem como o efeito do exercício físico aeróbico. Metodologia: Os animais foram divididos em quatro grupos: sedentários SOP (Sedsop), exercício SOP (Exesop), sedentário controle (Sedcon) e exercício controle (Execon). Os animais permaneceram com livre acesso à água e a ração padrão para roedores. Os animais foram submetidos à natação (temperatura termoneutro), com sobrecarga progressiva, durante uma hora por dia cinco vezes por semana, ao longo de oito semanas sucessivas. Avaliações antropométricas, cardiovasculares, metabólicas, renais e de vias intracelulares avaliando parâmetros tais como, massa corporal total, pressão arterial, frequência cardíaca, perfil lipídico, perfil glicêmico, resposta insulinêmica, filtração glomerular, manipulação tubular renal de sódio e expressão de receptores angiotensinérgicos foram realizadas em tempos bem definidos antes e após a submissão dos animais à atividade física programada. A sinalização intracelular foi avaliada em músculo esquelético através do estudo da expressão de proteínas IR, IRS-1, Akt, GLUT-4 bem como a determinação dos níveis de receptores de angiotensina II (AT1R e AT2R) em rins. Resultados: Os dados mostraram que os animais Sedsop apresentaram aumento da massa corporal, intolerância à glicose, hiperinsulinemia, hipertensão arterial e aumento de frequência cardíaca, significativamente maior após 8 semanas de acompanhamento, quando comparados aos grupos treinados. Aumento da massa corporal, resistência à insulina e hipertensão arterial se associaram positivamente com elevados níveis séricos de colesterol, triglicérides e queda dos níveis de HDL-C. Em relação ao estudo funcional renal, os grupos treinados (Exesop e Execon) apresentaram uma menor excreção renal de sódio comparado aos grupos sedentários. Nestes animais (Exesop) esta antinatriurese ocorreu principalmente pela redução da excreção de sódio nos segmentos proximais do nefro, parcialmente compensada pelo aumento da excreção de sódio nos segmentos pós-proximais. A menor excreção proximal de sódio nestes animais treinados foi associada à redução da excreção renal de potássio. Sugere-se que a redução natriurética nos grupos treinados tenha ocorrido como consequência da ativação de mecanismos homeostáticos compensatório se contrarreguladores em decorrência da redução da pressão arterial e frequência cardíaca. Na análise de AT1R e AT2R, os animais Sedsop mostraram menor expressão desses receptores. Em contrapartida, o grupo Exesop apresentou aumento significativo na expressão de AT2R se comparados ao Sedsop, o que repercutiu na elevação da razão AT1R/AT2R. O estudo das vias intracelulares da insulina em músculo esquelético mostrou que os animais Sedsop apresentaram significativas modificações na transdução do sinal em resposta à insulina denotada pela redução da expressão de IRS-1 e GLUT-4 sugerindo uma resistência à ação da insulina nestes animais. Conclusão: Esses dados demonstraram que o exercício físico programado não está associado à atenuação dos aspectos ligados a androgenização neste modelo específico, contudo, o programa de treinamento físico proporcionou consistente melhora nos parâmetros metabólicos e hemodinâmicos possivelmente, por modificações na resposta de vias intracelulares relacionadas à resposta insulínica nestes animais em estro permanente quando comparados ao grupo sedentário Abstract: The polycystic ovary syndrome (PCOS) is an endocrine heterogeneous disease with unknown etiology. It affects about 8% of women in reproductive age. Among the main features of this disease is the hyperandrogenism, infertility and chronic anovulation. The PCOS also have high prevalence of disorders that lead to the metabolic syndrome, such as obesity, insulin resistance, hyperinsulinemia, hyperlipidemia, hypertension and diabetes mellitus type 2. The pharmacological measures, such as diet and physical exercise, have been considered as a first-line strategic on disease prevention and treatment of that. Regarding this presented situation, the aim of this study was to assessusing an animal model of androgenization, PCOS relationship between metabolic syndrome and aerobic exercise effects on the chronic treatment of these disorders. The animals groups were sedentary PCOS (PCOSsed), PCOS exercise (PCOSexe), sedentary control (SEDcon) and exercise control (EXEcon). The animals had free access to water and food. The training methodology was swimming with pleasant water temperature, during 1 hour per day, 5 times a week, over 8 weeks with progressive overload. The anthropometric, metabolic, cardiovascular, renal and intracellularly analysis were carried out considering measures such as body mass ratings, blood pressure, heart rate, lipid profile, glycemic profile, insulin analyses, glomerular filtration, renal sodium handling and intracellular signaling pathways. In the skeletal muscle was evaluated intracellular signaling through the proteins IR, IRS-1, Akt, GLUT-4 and Angiotensin II receptors (AT1R and AT2R) in kidneys. The PCOSsed animals showed the parameters obesity, glucose intolerance, hyperinsulinemia, hypertension, and increased heart rate significantly higher than trained groups, after 8 weeks of experiments. The obesity, insulin resistance and hypertension results were positively correlated with high cholesterol, triglycerides and HDL-C levels. For the kidney function¿s test, the trained groups (PCOSexe and EXEcon) showed lower renal sodium excretion than sedentary groups. However, only in PCOSexe animals the decrease occurred primarily by reducing excretion in the proximal portion, which was offset by an increase in sodium excretion in the distal portion. Another renal response in animals PCOSexe observed was a decrease in renal excretion of potassium. Natriuretic reduction in trained animals may have occurred due to the homeostatic mechanism for compensatory reduction in blood pressure and heart rate. The analysis of AT1R and AT2R, PCOSsed animals have showed less expression of these receptors. On the other hand, the PCOSexe group has presented a significant increase in the expression of AT2R when compared to PCOSsed, which have influenced in the elevation of the reason AT1R/AT2R. The intracellular analysis of insulin pathways in skeletal muscle, PCOSsed animals showed changes in signal transduction pathways of insulin through the significant decrease of the expression of IRS-1 and GLUT-4 of suggesting a response of insulin resistance in these animals. These results have demonstrated that scheduled exercise did not reverse the infertility in animal androgenization model. Nevertheless, the physical training program provided consistent improvements in metabolic parameters and intracellular animals with PCOS when it was compared to the sedentary group Doutorado Fisiopatologia Médica Doutora em Ciências

Orientadores: Ilka de Fátima Santana Ferreira Boin, Artur Udelsmann Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Ciências Médicas Resumo: A oclusão vascular temporária do fluxo hepático é um dos procedimentos essenciais nas cirurgias hepáticas. Muitas pesquisas relacionadas à isquemia e reperfusão (I/R) em ratos Wistar, seja através de interrupção por pouco tempo, ou período maior, têm demonstrado sérias complicações causadas por lesões de I/R. Pesquisas demonstram grandes alterações metabólicas e fisiológicas quando analisadas durante, ou nas primeiras horas após o experimento. O objetivo deste trabalho foi avaliar as alterações hepáticas morfológicas e bioquímicas tardias, ocorridas após pinçamento total ou parcial do hilo hepático em ratos Wistar. Um total de 26 ratos Wistar, machos, peso médio 315,5 ± 61,5g foram utilizados em dois estudos. No primeiro estudo, 12 ratos foram divididos em dois grupos: Grupo Pinçamento do Ducto Biliar Comum (PDBC; n=6) foram submetidos à anestesia com tiopental sódico iv, à incisão abdominal até 2cm, tendo o ducto biliar isolado, dissecado e pinçado por 10 minutos. Após este tempo, a pinça foi retirada e a incisão fechada. Grupo Operação Simulada I (OSI; n=6) em condições de normalidade, os animais foram submetidos unicamente à anestesia e laparotomia e, posteriormente, a exames de controle. Nos dois grupos após o 28ºdia foram realizadas biópsias hepáticas e exames bioquímicos seguidos de eutanásia dos animais. Observamos que 83% dos animas do grupo PDBC apresentaram dilatação do colédoco, com alterações histológicas hepáticas: proliferação ductular, formação de septos, focos de necrose do parênquima, com formação de micro abcessos e alterações dos exames bioquímicos, quando comparados aos animais do grupo OSI (p < 0,05). No segundo estudo, 14 animais foram anestesiados com ketamina 5% (30mg/Kg) e xylazina 2% (30mg/Kg) via intraperitoneal. No grupo clampeamento intermitente do pedículo hepático (CIPH; n=7) os animais foram submetidos à incisão em U no abdome; o pedículo hepático foi isolado, dissecado e submetido a pinçamento, com micro pinça, intermitente por 4 ciclos de 5 minutos de isquemia, seguidos de 5 minutos de reperfusão, e a incisão foi fechada. No Grupo Operação Simulada II (OSII; n=7) os animais foram submetidos à anestesia, laparotomia e manipulação do pedículo hepático e,posteriormente, ao controle dos exames. Em todos os animais no 35o dia, após jejum de 12 horas, foi realizada nova anestesia para coleta da biópsia hepática e sangue para dosagem de alanina amino transferase (ALT) e de aspartato amino transferase (AST). A análise estatística foi realizada pelo teste Mann-Whitney para comparação de médias com significância de 5%. Observamos que todos os animais do grupo CIPH apresentaram dilatação do colédoco e aumento significativo nas enzimas hepáticas (p

Purpose: To compare the blood flow in the internal thoracic artery when dissected endoscopically in a conventional manner, in addition to develop a reliable experimental training model for the surgical team. Methods: Paired experimental study. Ten pigs were operated and had both internal thoracic arteries dissected, the right with a conventional technique and the left by video endoscopy. The main outcomes to be studied were flow, length, and time of dissection of each vessel. Results: Blood flow measurements were performed with mean heart rate of 100 ± 16 bpm and mean arterial pressure of 89.7 ± 13 mm Hg. The mean blood flow of endoscopic dissection of the internal thoracic artery was 170.2 ± 66.3 mL/min and by direct view was 180.8 ± 70.5 (p = 0.26). Thus, there was no statistically significant difference between the flows, showing no inferiority between the methods. Conclusions: The minimally invasive dissection of the internal thoracic artery was shown to be not inferior to the dissection by open technique in relation to the blood flow in the present experimental model. In addition, the model that we replicated was shown to be adequate for the development of the learning curve and improvement of the endoscopic abilities.

Orientador: Ana Cláudia Rossi Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Odontologia de Piracicaba Resumo: O objetivo do presente estudo foi observar o efeito de uma membrana de nanofibras poliméricas a base de poli-epsilon-caprolactona (PCL), impregnadas ou não com hidroxiapatita (HA), na reparação óssea em defeitos críticos produzidos artificialmente na calvária de ratos. Foram utilizados 36 ratos, divididos aleatoriamente em seis grupos de seis animais. Para a indução do defeito ósseo padronizado foram realizadas duas perfurações, por meio de trefina de 3 mm de diâmetro, na calvária dos animais, uma em cada lado, sendo que o lado direito recebeu os tratamentos (HA ou PCL-HA) e os controles (coágulo ou PCL) foram feitos no lado esquerdo. Decorridos 30, 60 e 90 dias da cirurgia, os animais foram mortos por aprofundamento da anestesia. As cabeças foram submetidas à tomografia, considerando um voxel de 0.12 mm, com campo visual de 06 × 16 cm e tempo de aquisição de 40s. Foram utilizados 120 kV, 8 mA e 36.12 mA/s na aquisição. Os diâmetros dos orifícios foram medidos por meio do software Invesalius 3.0. As cabeças foram fixadas com solução de formol tamponado (pH 7,2) em tampão fosfato de sódio a 0,1 M, durante 72 horas e submetidas à descalcificação em solução de EDTA a 7% e formol a 5% até verificação de completa descalcificação (aproximadamente 30 dias). Cortes semi-seriados de 7 ?m foram corados por Hematoxilina-Eosina e foram observadas as características histológicas do processo de cicatrização óssea nas lâminas nos diferentes tempos do estudo. A comparação quantitativa das medidas dos orifícios obtidas com a tomografia foi feita pelo teste de Kruskal-Wallis (teste de Dunn como post hoc), considerando um nível de significância de 5%. As imagens tomográficas revelaram uma tendência de redução do volume do orifício ao longo do tempo, a qual foi maior após 90 dias para todos os tratamentos. O tratamento com PCL+HA mostrou menor volume de orifício do que os outros tratamentos independentemente do período. As medidas histológicas mostraram maior formação de osso induzida pela membrana de PCL com ou sem HA do que os outros tratamentos independentemente do período. A HA mostrou maior aumento da cicatrização óssea com ou sem o PCL. Concluímos que a HA adicionada às nanofibras de PCL melhorou significativamente a cicatrização óssea em defeitos provocados na calvária de ratos Abstract: The aim of the present study was to observe the effect of a polymeric-nanofiber membrane of poly-epsilon-caprolactone (PCL) with or without hydroxyapatite (HA) on bone healing of critical defects induced in rat calvaria. 36 animals were randomly divided into six groups. The standardized bone defects were obtained by two perforations with 3.0 mm diameter trephine directly into the animals¿ calvaria, one in each side. The right side received the treatments (HA or PCL-HA) and the left side the controls (blood clot or PCL). After 30, 60 and 90 days of the surgical procedure, all animals were killed; the head was sectioned and submitted to tomography (voxel of 0.12 mm, visual field of 6x16 cm, acquisition time of 40s, 120 kV, 8mA and 36.12 mA/s). The perforation diameters were measured using the software Invesalius 3.0. After tomography, all heads were fixed by buffered formol (pH=7.2) in 0.1M sodium-phosphate buffer during 72 hours and submitted to decalcification in 7% EDTA/5% formol solution until complete decalcification (approximately 30 days). Semi-serial 7 µm cuts were stained with Hematoxicilin-Eosin. Histological characteristics of bone healing were observed according to groups and periods. Quantitative comparisons of perforation measurements from both tomography and histological analysis were performed by Kruskal-Wallis (Dunn post hoc) test with a 5% significance level. Tomography images revealed a tendency to reduce the perforation volume along time, which was higher after 90 days for all treatments. The treatment with PCL+HA showed the lower volumes of perforations than the other treatments irrespectively of the period. Histological measurements showed more osseous formation induced by treatment with PCL with or without HA than the other treatments irrespectively of the period. HA appear to increase bone healing with or without PCL. We concluded that HA added to PCL nanofibers significantly improved the bone healing in bone defects of rat calvaria Mestrado Anatomia Mestra em Biologia Buco-Dental

Orientador: Ricardo Kalaf Mussi Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Ciências Médicas Resumo: A cirurgia experimental surge como excelente alternativa em busca de resposta a importantes dúvidas da atividade médica. Contudo, modelos experimentais em animais de pequeno porte estão padronizados para procedimentos de curta duração, sendo necessárias adaptações para tempos operatórios prolongados. Objetivo: Avaliar a resposta inflamatória pulmonar e sistêmica imediata após período operatório de longa duração. Métodos: Ratos Wistar do grupo experimental foram anestesiados e submetidos à traqueostomia, toracotomia e permaneceram em ventilação mecânica por 3 horas. O grupo controle não foi submetido ao protocolo operatório. Os seguintes parâmetros foram avaliados: atividade da mieloperoxidase pulmonar, níveis de extravasamento de proteínas séricas pulmonares, relação peso pulmonar úmido/seco e medidas dos níveis séricos de citocinas. Resultados: Os animais operados apresentaram menor extravasamento de proteínas séricas nos pulmões comparados aos animais controle. A relação peso pulmonar úmido/seco e a atividade de mieloperoxidase não diferiram entre os grupos. As citocinas séricas IL-1ß, TNF?- e IL-10 não foram quantificáveis nos grupos, enquanto que IL-6 só foi detectada no soro dos animais operados. Conclusão: Padronizamos um modelo experimental de ventilação mecânica em ratos com tempo cirúrgico prolongado que não apresentou alterações inflamatórias locais (pulmonar) e sistêmicas significativas, permitindo avaliar a resposta inflamatória em outros procedimentos da cirurgia torácicaß Abstract: Experimental surgery has emerged as an excellent alternative to answer important questions raised in the daily medical activity. However, experimental models in small animals are standardized for short surgical procedures, thus adjustments are necessary for prolonged operative times. Purpose: To evaluate the immediate pulmonary and systemic inflammatory response after a long-term operative period. Methods: Wistar rats in the experimental group were anaesthetized and submitted to tracheostomy, thoracotomy and remained on mechanical ventilation during 3 h. Control animals were not submitted to the operative protocol. The following parameters have been evaluated: pulmonary myeloperoxidase activity, pulmonary serum protein extravasation, lung wet/dry weight ratio and measurement of levels of cytokines in serum. Results: Operated animals exhibited significantly lower serum protein extravasation in lungs compared with control animals. The lung wet/dry weight ratio and myeloperoxidase activity did not differ between groups. Serum cytokines IL-1ß, TNF-?, and IL-10 levels were not detected in groups, whereas IL-6 was detected only in operated animals. Conclusion: We have standardized an experimental model of mechanical ventilation in anaesthetized rats with a prolonged surgical time that did not produce significant local (pulmonary) and systemic inflammatory changes and permit to evaluate others procedures in thoracic surgery Mestrado Fisiopatologia Cirúrgica Mestre em Ciências

Orientador: José Andrés Yunes Dissertação (mestrado) - Universidade Estadual de Campinas, Instituto de Biologia Resumo: A leucemia linfoide aguda (LLA) é o câncer mais comum na infância. O transplante de células primárias de LLA humana em camundongos imunosuprimidos tem sido de suma importância para o entendimento da fisiopatologia da doença e para o teste de novos fármacos. Ao contrário de modelos animais de tumores sólidos, cujo volume é facilmente medido na superfície dos animais, a LLA infiltra órgãos inacessíveis ao exterior, daí a necessidade de definir métodos adequados para o monitoramento da progressão da doença. Resultados aqui apresentados indicam que proteínas secretadas pela LLA podem servir como marcadores quantitativos da carga leucêmica, facilmente aferidos por ELISA de amostras de plasma sanguíneo. Dentre três proteínas testadas (B2M, IGFBP2 e Hsp90), o ELISA de Hsp90 apresentou sensibilidade superior à análise da porcentagem de células leucêmicas no sangue dos animais, por citometria de fluxo de células marcadas com anti-huCD45. Os níveis de Hsp90 humano no plasma sanguíneo mostraram-se positivamente correlacionados com o porcentual de células leucêmicas na medula óssea e fígado e em menor grau com os níveis do baço e sangue periférico (SP) ao longo do tempo, tanto nas LLA de linhagem B quanto nas LLA-T. O ELISA de Hsp90 permite detectar a instauração da leucemia nos animais transplantados, até duas semanas antes da detecção pelo método tradicional de análise de sangue periférico por citometria de fluxo. Ao contrário do observado para IGFBP2, o tratamento dos animais leucêmicos com Dexametasona ou um inibidor da PI3K não interferiu nos níveis de Hsp90, que se mantiveram proporcionais à porcentagem de células leucêmicas huCD45+ no sangue periférico. No conjunto, os resultados demonstram que a análise do plasma dos animais por ELISA de Hsp90 é um método melhor do que os atualmente utilizados, para diagnóstico precoce e acompanhamento de LLA humana quando em níveis de doença residual mínima, ou seja, quando a porcentagem de células de LLA é inferior a 5% do total de células da medula óssea Abstract: Acute lymphoblastic leukemia (ALL) is the most common type of childhood cancer . The transplantation of human primary ALL cells in immunodeficient mice has been of much importance for understanding the disease's pathophysiology and testing new drugs. Unlike animal models of solid tumors whose volume is easily measured on the animal surface, the ALL infiltrates organs that are inaccessible to external antigens, hence the need to define more suitable methods for monitoring the disease's progression. Results presented here indicate that proteins secreted by the ALL can serve as quantitative markers of leukemic burden and are easily measured by ELISA of blood plasma samples. Among three tested proteins (B2M, IGFBP2 and Hsp90), Hsp90 ELISA analysis showed higher sensitivity than the analysis of leukemic cells on animal blood by flow cytometry of anti- huCD45 labeled cells. The levels of Hsp90 in human blood plasma were shown to be positively correlated with the percentage of leukemic cells in the bone marrow and liver and to a lesser extent with the levels in the spleen and peripheral blood (PB) over time, both in B-lineage ALL as in ALL-T. The Hsp90 ELISA allows the leukemia's engraftment detection in transplanted animals up to two weeks prior to detection by the traditional method of peripheral blood analysis by flow cytometry. Unlike observed for IGFBP2, treatment of leukemic animals with Dexamethasone or PI3K inhibitors did not interfere in Hsp90 levels, which remained proportional to the percentage of huCD45+ leukemic cells in the peripheral blood. Taken together, the results demonstrate that the analysis of animal plasma by Hsp90 ELISA is a better method than those currently used for early diagnosis and monitoring of human ALL on minimal residual disease levels, when the percentage of ALL cells is less than 5 % of the total bone marrow cells Mestrado Genética Animal e Evolução Mestre em Genética e Biologia Molecular

Many experimental models exist to better understand the necrosis of the femoral head etiology, both in terms of the species variety in which necrosis is induced and in the operative techniques used for treatment. Objective: This study has two main objectives, the first is to review the literature concerning experimental models of avascular necrosis of the growing femoral head, the second, to demonstrate the experimental pig model’s reproducibility using a pilot study. Methods: This was a bibliographic review to describe the attempts over time to find the best species and technique for induction that would reproduce ischemic necrosis of the growing femoral head in humans. Simultaneously, a pilot study was performed to verify the replication of induction in pigs, the species that has more similarities with the human hip. The pilot’s methodological analysis consists of conventional radiology and verification of possible anatomical, pathological changes. Results: In imaging exams; lateral sub-dislocation of the femur head and triangular appearance of the head were observed, characterizing its flattening; in macroscopic examination, the femoral head flattening with femoral neck widening and shortening was identified; in histology, the proliferation of articular cartilage with the presence of vascular granulation regenerative tissue, with osteoclasts and fibrocartilaginous tissue in the metaphyseal femoral neck region was identified. Conclusion: The experimental pig model can be used as a valuable tool for the reproducibility of anatomical, pathological changes in ischemic necrosis of the growing femoral head. The model is reproducible and feasible and can be beneficial for future studies on the anatomical pathology of necrosis of the growing femoral head. Level of Evidence III, Literature Review . RESUMO Na tentativa de compreender melhor a etiologia da necrose da cabeça femoral, existe uma diversidade de modelos experimentais tanto no que diz respeito à variedade das espécies em que é induzida a necrose quanto nas técnicas operatórias utilizadas para o tratamento. Objetivo: Este trabalho tem fundamentalmente dois objetivos: a revisão da literatura concernente aos modelos experimentais da necrose avascular da cabeça do fêmur em crescimento e demonstrar a reprodutibilidade do modelo experimental do suíno por meio de um estudo piloto. Métodos: Foi realizada uma revisão bibliográfica descrevendo as tentativas ao longo do tempo em buscar qual seria a melhor espécie e técnica para indução que reproduzisse a necrose isquêmica da cabeça do fêmur em crescimento nos humanos. Simultaneamente foi feito um estudo piloto para verificar a replicação da indução na espécie suína, o espécime cujo quadril tem mais similaridades com o humano. A análise metodológica do piloto consiste na radiologia convencional e verificação das possíveis alterações anátomo patológicas. Resultados: Nos exames por imagem, foram observadas sub-luxação lateral da cabeça do fêmur e aparência triangular da cabeça, caracterizando o achatamento da mesma; no exame macroscópico, identificamos o achatamento da cabeça femoral com alargamento e encurtamento do colo; na histologia, identificamos a proliferação da cartilagem articular com presença de tecido regenerativo vascular de granulação, com osteoclastos e tecido fibrocartilaginoso na região metafisária do colo femoral. Conclusão: Podemos inferir que o modelo experimental suíno pode servir como ferramenta valiosa para a reprodutibilidade das alterações anátomo patológicas da necrose isquêmica da cabeça femoral em crescimento. O modelo é reprodutível e factível, servindo para estudos futuros sobre a anátomo patologia da necrose da cabeça do fêmur em crescimento. Nível de Evidência III, Revisão da Literatura .

Introduction: Oral mucositis (OM) is considered the most frequent acute side effect of the antineoplasic treatment, with ulcerative lesions resulting from a painful symptomatology, affecting the oral cavity in response to the Antineoplastic treatment. In order to study these side effects, experiments in animal models are necessary, using antineoplastic drugs for the induction of OM and anesthetics, mainly Ketamine and Xylazine, to perform scarification of the cheeks. Objective: The goal is to report an experimental model of induced OM, without the use of anesthetics for the scarification stage of the animal cheeks. Methods: Fourty five male Wistar rats, 7 weeks old and weighting 220g, were used, divided into 2 groups; with OM induced by 5-Fluorouracil intraperitoneal administration. Two days later, Group I was physically contained, in contrast, Group II were anesthetized with Ketamine and Xylazine, focusing on irritating the cheek mucosa using the tip of a sterile needle, in order to potentialize the development of OM. The animals were euthanized with an anesthetic overdose. Results: Concerning the experiment of 5-Fluorouracil chemo-induced of OM, where the irritation was performed by physical containment, without the use of anesthetics (Ketamine and Xylazine), the animals had a longer survivability and a rapid improvement of the side effects induced by chemotherapy. Conclusion: This new model is promising, considering that the use of anesthetics (Ketamine and Xylazine) showed a high mortality rate. In the absence of anesthesia, all the animals survived until the end of the experiment involving chemotherapy model with 5-Fluorouracil and physical restraint. RESUMO Introdução: A mucosite oral (MO) é considerada o efeito colateral agudo mais frequente do tratamento antineoplásico, com lesões ulcerativas decorrentes de sintomatologia dolorosa, acometendo a cavidade oral em resposta ao tratamento antineoplásico. Para estudar esses efeitos colaterais, são necessários experimentos em modelos animais, utilizando fármacos antineoplásicas para a indução de MO e anestésicos, principalmente Cetamina e Xilazina, para realizar a escarificação das bochechas Objetivos: Relatar um novo modelo experimental de MO induzida, sem o uso de anestésicos, para a etapa de escarificação das bochechas dos animais. Métodos: Foram utilizados 45 ratos Wistar machos, com 7 semanas de idade e peso de 220g, divididos em 2 grupos com MO induzida pela administração intraperitoneal de 5-fluorouracil. Dois dias depois, o Grupo I foi contido fisicamente, ao contrário, o Grupo II foi anestesiado com Cetamina e Xilazina, com foco em irritar a mucosa da bochecha com a ponta de uma agulha estéril, a fim de potencializar o desenvolvimento de MO. Os animais foram eutanasiados com sobredose de anestésica. Resultados: Em relação ao experimento de 5-Fluorouracil quimioinduzido para MO, onde a irritação foi realizada por contenção física, sem o uso de anestésicos (Cetamina e Xilazina), os animais tiveram maior sobrevida e rápida melhora dos efeitos colaterais induzidos por quimioterapia. Conclusão: Este novo modelo é promissor, visto que o uso de anestésicos (Cetamina e Xilazina) apresentou elevada mortalidade. Porém, na ausência de anestesia, todos os animais sobreviveram até o final do experimento envolvendo este modelo de quimioterapia induzida com 5-fluorouracil e contenção física.

Orientador: Denise Vaz de Macedo Dissertação (mestrado) - Universidade Estadual de Campinas, Instituto de Biologia Resumo: O treinamento intensificado (overtraining - OT), resultado de um aumento na frequência de treinos diários com a consequente diminuição no período de descanso entre as sessões pode produzir dois estados diferenciados em relação ao desempenho: um estado de manutenção ou mesmo ligeiro aumento (funcional overreaching - FOR) ou um estado de perda do condicionamento previamente adquirido (non funcional overreaching - NFOR). Estudos anteriores do nosso grupo de pesquisa sugerem que a dinâmica mitocondrial está na gênese de ambos os processos. O objetivo deste trabalho foi analisar a possível existência de um sinal pró-apoptótico no músculo gastrocnêmio vermelho de animais classificados como NFOR em comparação com o grupo classificado como FOR após um protocolo de OT em esteira. Nossa hipótese é que a sinalização pró apoptótica induzida por um aumento nos níveis de estresse oxidativo explicaria o fluxo autofágico basal reduzido observado anteriormente na musculatura dos animais NFOR. A presente dissertação está dividida em 2 capítulos. No capítulo 1 apresentamos os resultados referentes a proteínas relacionadas ao processo autofágico e apoptótico induzidas por Espécies Reativas de Oxigênio (LC3 I e LC3 II, JNK1/2, p53, caspase 3) nos animais FOR e NFOR. O protocolo de overtraining foi realizado em colaboração cientifica com o Laboratório de Fisiologia de Equinos e Farmacologia (Unesp-Jaboticabal). O protocolo originalmente proposto (Hohl et al.,2009) prediz um período de 60h de descanso após o último teste de exaustão para a análise das amostras de ambos os grupos de animais. No entanto, a despeito da evidente existência de 2 grupos de animais diferenciados em relação ao desempenho, o sacrifício dos animais e a concomitante retirada das amostras de músculo imediatamente após o teste de desempenho claramente contaminaram a análise da resposta crônica frente ao protocolo de OT com a resposta aguda ao teste de exaustão. Essa conclusão foi evidenciada pela ausência de diferenças na atividade da enzima citrato sintase e razão LC3II/I nos grupos FOR e NFOR. Observamos nos animais NFOR uma redução do conteúdo de pró-caspase-3 mitocondrial com concomitante aumento de seu fragmento no citosol associado a maiores níveis de pró-caspase-3 citosolica. Esta alteração pode representar a clivagem da caspase-3 nas mitocôndrias e sua translocação para o citosol, sendo este um mecanismo associado a sinalização apoptótica. Observamos também maiores níveis da proteína p53 na fração citosólica do grupo NFOR. A localização dessas proteínas no citosol pode interferir na síntese de autofagossomos, uma vez que a ação sinérgica entre p53 e caspase-3 pode inibir proteínas do maquinário autofágico, evidenciado pela menor razão LC3II/I no grupo NFOR. Tal interferência poderia explicar a queda no desempenho no grupo NFOR. Somente a repetição do protocolo com um outro grupo de animais e a observância do tempo correto de descanso após o teste de exaustão podem confirmar ou refutar nossa hipótese. No capítulo 2 apresentamos a padronização de um protocolo de fracionamento subcelular de pequenas quantidades de amostras congeladas de tecido muscular esquelético. O congelamento de tecidos causa alterações estruturais nas células, induzindo rompimento de membranas celulares e de organelas, sendo as mitocôndrias muito sensíveis a esta condição. As análises por western blot das proteínas marcadoras das frações citosólica (alfa-tubulina), mitocondrial (VDAC1) e nuclear (histona H3) associadas a maior atividade enzimática da citrato sintase na fração mitocondrial indicaram que as modificações introduzidas no protocolo associadas aos cuidados relatados durante a homogeneização das amostras congeladas possibilitaram que quantidades pequenas (50mg) de músculo esquelético produzissem frações subcelulares com razoável grau de pureza e com maior retenção do conteúdo solúvel da fração mitocondrial. A possibilidade de utilizar pequenas quantidades de tecidos viabilizou as análises apresentadas no capítulo 1, da distribuição das proteínas relacionadas a sinalização pró-apoptótica nas frações citosolica, mitocondrial e nuclear. Além disso, possibilita o fracionamento de amostras provenientes de biopsias, facilitando o planejamento de experimentos com humanos Abstract: The intensified training (Overtraining - OT), resulting from an increase in the frequency of daily workouts with the consequent decrease in the rest period between sessions can produce two differentiated states related to performance: a state of maintenance or even slight increase (functional overreaching - FOR) or a state of loss of previously acquired conditioning (non-functional overreaching - NFOR). Previous studies from our research group suggest that mitochondrial dynamics is in the genesis of both processes. The objective of this work was to analyze the possible existence of a pro-apoptotic signal in the red gastrocnemius muscle of animals classified as NFOR in comparison with the group classified as FOR after an OT protocol. Our hypothesis is that the pro-apoptotic signaling induced by an increased oxidative stress levels would explain the reduced basal autophagic flow observed previously in muscle of NFOR animals. This dissertation is divided in 2 chapters. In Chapter 1 we present the results referring to proteins related to the autophagic and apoptotic process (LC3 I and LC3 II, JNK, p53, caspase-3) induced by reactive oxygen species in FOR and NFOR animals. The OT protocol was conducted in scientific collaboration with the Laboratory of Physiology of Equines and Pharmacology (UNESP-Jaboticabal). The protocol originally proposed (Hohl et al.,2009) predicted a 60h rest after the exhaustion test for the analysis of samples from both groups of animals. However, in this work the muscle samples were collected immediately after the exhaustion test and frozen in liquid nitrogen. Thus, the results presented here do not represent only the chronic response to the OT protocol, being contaminated with the acute effects of the incremental exhaustion test. This conclusion can be evidenced by the absence of differences in the citrate synthase activities and LC3II/I ratio in the FOR and NFOR groups. We observed a pro-apoptotic signaling coming from the mitochondria in NFOR animals, which presented reduction of the mitochondrial pró-caspase-3 content with concomitant increase of its fragment in the cytosol associated with higher levels of cytosolic prócaspase-3. This alteration may represent the cleavage of caspase-3 in mitochondria and its translocation to the cytosol, a mechanism associated with mitochondrial apoptotic signaling. We also observed higher levels of p53 in the cytosolic fraction of the NFOR group. The cytosolic localization of these proteins may interfere with the synthesis of autophagosomes, since the synergistic action between p53 and caspase-3 can inhibit autophagic machinery proteins, evidenced by the lowest LC3II/I ratio in the NFOR. Such interference could reduce the capacity of mitochondria degradation, increasing the content of dysfunctional mitochondria and decrease in performance in the NFOR group. In Chapter 2 we present the standardization of a subcellular fractionation protocol of small amounts of frozen samples of skeletal muscle tissue. Tissue freezing causes structural alterations in cells, inducing disruption of cell membranes and organelles, being mitochondria very sensitive to this condition. Western blot analyses of cytosolic (alpha-tubulin), mitochondrial (VDAC1) and nuclear (histone H3) fractions in association with citrate synthase activity in mitochondrial fraction indicated that the modifications introduced in the protocol associated with the care reported during the homogenization of frozen samples allowed to produce subcellular fractions with a reasonable degree of purity of small amounts (50mg) of skeletal muscle and with greater retention of the soluble content of the mitochondrial fraction. The possibility of using small amounts of tissues enabled the analyses presented in Chapter 1, of the distribution of proteins related to proapoptotic signaling in the cytosolic, mitochondrial and nuclear fractions without important contaminations among these fractions. In addition, it allows the fractionation of samples from biopsies, facilitating the planning of experiments with human beings Mestrado Bioquímica Mestre em Biologia Funcional e Molecular CAPES

Resumo Fundamento: A taxa de falha de enxerto de veia safena um ano após a cirurgia de revascularização do miocárdio varia de 10% a 25%. O objetivo deste estudo foi de investigar se a atorvastatina pode reduzir o acúmulo de células musculares lisas vasculares para inibir a hiperplasia intimal por meio da inibição da via p38 MAPK. Métodos: Quarenta e cinco ratos Sprague-Dawley foram randomizados em três grupos. Trinta ratos foram submetidos à cirurgia de enxerto de veia e randomizados para tratamento com veículo ou atorvastatina; quinze ratos foram submetidos à cirurgia sham. Detectamos a hiperplasia intimal por meio de coloração com hematoxilina-eosina e a expressão de proteínas relacionadas por meio de análise imuno-histoquímica e Western blot. Foram realizadas as comparações por análise de variância de fator único e pelo teste da diferença mínima significativa de Fisher, com p < 0,05 considerado significativo. Resultados: A íntima analisada pela coloração com hematoxilina-eosina era dramaticamente mais espessa no grupo controle que no grupo atorvastatina e no grupo sham (p < 0,01). Os resultados da coloração imuno-histoquímica de α-SMA demonstraram que a porcentagem de células positivas para α-SMA no grupo controle era mais alta que no grupo atorvastatina (p < 0,01). Nós também avaliamos α-SMA, PCNA, p38 MAPK e fosforilação de p38 MAPK após o tratamento com estatina por meio de análise de Western blot e os resultados indicaram que a atorvastatina não levou à redução de p38 MAPK (p < 0,05); no entanto, resultou na inibição da fosforilação de p38 MAPK (p < 0,01) e reduziu significativamente os níveis de α-SMA e PCNA, em comparação com o grupo controle (p < 0,01). Conclusão: Nós demonstramos que a atorvastatina pode inibir o acúmulo de células musculares lisas vasculares por meio da inibição da via p38 MAPK e é capaz de inibir a hiperplasia intimal em modelos de enxerto de veia em ratos.

Objective: to evaluate the effects of increasing pressures on the cutaneous blood flow in the skin of pigs. Methods: we conducted an experimental study in pigs submitted to subcutaneous magnetic implants (n=30). After healing, were applied external magnets with varying magnetic forces to the skin, generating compression. We evaluated the cutaneous circulation of the skin under compression by the Laser Speckle Contrast Imaging (LSCI) technique. We measured the depth of the implants by ultrasonography, and applied computational simulations to the calculation of the different pressure values, considering the different distances between implants and external magnets. Results: nineteen implants presented complications. The remaining 11 were submitted to different magnetic compression forces and perfusion analysis. Two linear regression models showed an inverse correlation between exerted pressure and cutaneous perfusion, with significant variation, mainly in the initial pressure increases, of up to 20mmHg. Conclusion: The main reduction in cutaneous blood flow resulted from initial increases of up to 20 mmHg. The results suggest that tissue ischemia can occur even in low-pressure regimes, which could contribute to the appearance of skin lesions, particularly ulcers related to medical devices. RESUMO Objetivo: avaliar os efeitos de pressões crescentes exercidas sobre a pele de porcos no fluxo sanguíneo cutâneo. Métodos: estudo experimental em porcos submetidos a implantes magnéticos subcutâneos (n=30). Após a cicatrização, foram aplicados sobre a pele, ímãs externos com forças magnéticas variadas, gerando compressão. A circulação cutânea da pele submetida à compressão foi avaliada pela técnica Laser Speckle Contrast Imaging (LSCI). A profundidade dos implantes foi medida por ultrassonografia, e simulações computacionais foram aplicadas para o cálculo dos diferentes valores de pressão, considerando-se as variadas distâncias entre implantes e ímãs externos. Resultados: dezenove implantes apresentaram complicações. Os 11 restantes foram submetidos à diferentes compressões magnéticas e análise de perfusão. Dois modelos de regressão linear mostraram uma correlação inversa entre pressão exercida e perfusão cutânea com variação significativa principalmente nos acréscimos iniciais de pressão até 20mmHg. Conclusão: a principal redução do fluxo sanguíneo cutâneo resulta dos acréscimos iniciais de pressão de até 20mmHg. Os resultados sugerem que a isquemia tecidual pode ocorrer mesmo em regimes de baixa pressão, o que poderia contribuir para surgimento de lesões de pele, particularmente as úlceras relacionadas a dispositivos médicos.

Objective: to describe and evaluate the acceptance of a porcine experimental model in venous cutdown on a medical education project in Southwest of Brazil. Method: a porcine experimental model was developed for training in venous cutdown as a teaching project. Medical students and resident physicians received theoretical training in this surgical technique and then practiced it on the model. After performing the procedure, participants completed a questionnaire on the proposed model. This study presents the model and analyzes the questionnaire responses. Results: the study included 69 participants who used and evaluated the model. The overall quality of the porcine model was estimated at 9.16 while the anatomical correlation between this and human anatomy received a mean score of 8.07. The model was approved and considered useful in the teaching of venous cutdown. Conclusions: venous dissection training in porcine model showed good acceptance among medical students and residents of this institution. This simple and easy to assemble model has potential as an educational tool for its resemblance to the human anatomy and low cost. RESUMO Objetivo: descrever e avaliar a aceitação de um modelo experimental porcino no aprendizado de dissecção venosa em projeto de educação médica no sudoeste do Brasil. Método: um modelo experimental porcino foi desenvolvido para treinamento em dissecção venosa como projeto de ensino. Estudantes de medicina e médicos residentes receberam treinamento teórico sobre esta técnica cirúrgica e em seguida a praticaram no modelo. Após realizar o procedimento, os participantes preencheram um questionário sobre o modelo proposto. Este estudo apresenta o modelo e analisa as respostas ao questionário. Resultados: o estudo contou com 69 participantes que utilizaram e avaliaram o modelo. A qualidade geral do modelo porcino foi estimada em 9,16 enquanto a correlação anatômica entre este e a anatomia humana recebeu o escore médio de 8,07. O modelo foi aprovado e considerado útil no ensino da dissecção venosa. Conclusão: o treinamento de dissecção venosa em modelo porcino apresentou boa aceitação entre estudantes e residentes de medicina desta Instituição. Este modelo simples e de fácil confecção, tem potencial como instrumento de aprendizado por sua semelhança com a anatomia humana, e baixo custo.

Several studies indicated that morphine administration impairs cognitive brain functions. Therefore, in the current study, we investigated the effect of subchronic exposure to morphine and its withdrawal on effort- and/or delay-based forms of cost-benefit decision making and alterations in p-CREB/CREB ratio, p-GSK3β/GSK3β ratio, and BDNF level during decision making in the amygdala. Our data displayed an impairment of both forms of cost-benefit decision making following subchronic exposure to morphine. However, preference of high reward/high effort and/or high delay reward increased after naloxone injection. In molecular section, levels of BDNF and p-CREB/CREB ratio increased during cost-benefit decision making while p-GSK3β/GSK3β ratio decreased in both forms of decision making. In morphine-treated rats, level of BDNF and p-CREB/CREB ratio reduced during both forms of decision making while p-GSK3β/GSK3β ratio increased during delay-based and did not have a significant difference with the control group during effort-based decision making. On the withdrawal day, BDNF level raised while p-GSK3β/GSK3β ratio attenuated compared to morphine-treated group in both form of decision making. In addition, p-CREB/CREB ratio increased only during delay-based decision making on the withdrawal day. In conclusion, our data revealed that subchronic exposure to morphine interferes with the cost-benefit decision making may be via changes in level of BDNF, p-CREB/CREB and p-GSK3β/GSK3β ratio in the amygdala.

NAD(P)+ transhydrogenase (NNT) links redox states of the mitochondrial NAD(H) and NADP(H) via a reaction coupled to proton-motive force across the inner mitochondrial membrane. NNT is believed to be ubiquitously present in mammalian cells, but its expression may vary substantially in different tissues. The present study investigated the tissue distribution and possible roles of NNT in the mouse brain. The pons exhibited high NNT expression/activity, and immunohistochemistry revealed intense NNT labeling in neurons from brainstem nuclei. In some of these regions, neuronal NNT labeling was strongly colocalized with enzymes involved in the biosynthesis of 5-hydroxytryptamine (5-HT) and nitric oxide ( NO), which directly or indirectly require NADPH. Behavioral tests were performed in mice lacking NNT activity (Nnt−/−, mice carrying the mutated NntC57BL/6J allele from the C57BL/6J strain) and the Nnt+/+ controls. Our data demonstrated that aged Nnt−/− mice (18–20 months old), but not adult mice (3–4 months old), showed an increased immobility time in the tail suspension test that was reversed by fluoxetine treatment, providing evidence of depressive-like behavior in these mice. Aged Nnt−/− mice also exhibited behavioral changes and impaired locomotor activity in the open field and rotarod tests. Despite the colocalization between NNT and NO synthase, the S-nitrosation and cGMP levels were independent of the Nnt genotype. Taken together, our results indicated that NNT is unevenly distributed throughout the brain and associated with 5-THergic and NOergic neurons. The lack of NNT led to alterations in brain functions related to mood and motor behavior/performance in aged mice.

Emotional disturbances emerge following alcohol withdrawal. The anxiolytic effect of alcohol may be one important motivation for its consumption in conditions where alcohol intake is anxiety reducing. Besides, early alcohol experience will predict future alcohol-related problems at some point in their lives. Rats classified according to their anxiety-like behavior phenotype show a higher preference for alcohol. Yet, despite preclinical studies have identified the behavioral and physiological effects of alcohol withdrawal, little has been shown on relapse to alcohol after a period of abstinence following intermittent long-term alcohol consumption in low-(LA) and high-anxiety (HA) phenotype rats. This study attempts to assess the role of emotional reactivity of early-aged LA and HA rats on later alcohol preference, through an operant response procedure. For this, a sweetened liquid fading procedure associated with a long-term and intermittent voluntary alcohol drinking was used, with the animals being tested on different withdrawal periods. Alcohol withdrawal symptoms were recorded, and blood alcohol levels were assessed at two intervals to examine the influence of anxiety phenotype on blood alcohol clearance. Data from HA control and HA withdrawn rats show that emotionality selectively influences alcohol preference. Blood alcohol decay was faster in HA than in LA alcohol pretreated rats. HA rats showed an ascending curve of alcohol intake, exhibiting a high level of alcohol drinking on withdrawal and withdrawal symptoms. Moreover, HA alcohol experienced rats show a high frequency of lever-pressing response during extinction, associated with a facilitation of bar-pressing recovery trials, an indication of alcohol-seeking behavior.

BACKGROUND New bone formation and tissue remodeling are the major challenges in implantology today. Titanium meshes have demonstrated reconstructive potential for vertical bone gain. However, the soft tissue healing is technically sensitive to the surgical procedure. The combined usage of collagen membrane and specification of the meshes may ensure greater predictability. Therefore, this study aims to evaluate the influence of collagen membrane on the quality of the new bone formation in guided bone regeneration (GBR) procedures with different titanium meshes. METHODS Twenty-eight Wistar rats were randomly allocated into four main experimental groups, according to mesh pore size in μm: Group P300 (titanium meshes, with 0.3-mm thickness and 3-mm pore size; n = 7); Group P175 (titanium meshes, with 0.3-mm thickness and 1.75-mm pore size; n = 7); Group P85: (titanium meshes, with 0.04-mm thickness and 0.85-mm pore size; n = 7); Group P15: (titanium meshes. with 0.04-mm thickness and 0.15-mm pore size; n = 7). The femurs of each animal were subdivided into test and control groups: Test: bovine bone graft associated with porcine collagen and collagen membrane was used; control: bovine bone graft associated with porcine collagen was used without association with collagen membrane. Bone quality evaluation by in vivo microtomography and histologic analysis were performed. RESULTS Bone volume formation was similar between groups (P >0.05). However, the titanium meshes with pore size >1 mm demonstrated higher mineral bone density in comparison with meshes with pore size

Microbiota-derived acetate coordinates innate immune responses during intestinal Clostridium difficile infection through its cognate receptor FFAR2. Acetate accelerates early neutrophil recruitment and increases ILC3 expression of the IL-1 receptor, boosting ILC3 production of IL-22 in response to neutrophil-derived IL-1β., Antibiotic-induced dysbiosis is a key predisposing factor for Clostridium difficile infections (CDIs), which cause intestinal disease ranging from mild diarrhea to pseudomembranous colitis. Here, we examined the impact of a microbiota-derived metabolite, short-chain fatty acid acetate, on an acute mouse model of CDI. We found that administration of acetate is remarkably beneficial in ameliorating disease. Mechanistically, we show that acetate enhances innate immune responses by acting on both neutrophils and ILC3s through its cognate receptor free fatty acid receptor 2 (FFAR2). In neutrophils, acetate-FFAR2 signaling accelerates their recruitment to the inflammatory sites, facilitates inflammasome activation, and promotes the release of IL-1β; in ILC3s, acetate-FFAR2 augments expression of the IL-1 receptor, which boosts IL-22 secretion in response to IL-1β. We conclude that microbiota-derived acetate promotes host innate responses to C. difficile through coordinate action on neutrophils and ILC3s., Graphical Abstract

The aim of this study was to evaluate the in vitro osteogenic potential of osteoblasts from neural crest-derived frontal bone (OB-NC) and mesoderm-derived parietal bone (OB-MS) and the bone formation induced by them when injected into calvarial defects. Calvarial bones were collected from newborn Wistar rats (3-day old) and characterized as frontal and parietal prior to OB-NC and OB-MS harvesting. The cells were cultured, and several parameters of osteoblast differentiation were evaluated. These cells, or PBS without cells (control), were locally injected into 5-mm rat calvarial defects (5 × 106 cells/defect) and after 4 weeks bone formation was evaluated by morphometric and histological analyses. The characterization of frontal and parietal bones assured the different embryonic origin of both cell populations, OB-NC and OB-MS. The OB-NC presented higher proliferation while the OB-MS presented higher alkaline phosphatase (ALP) activity, extracellular matrix mineralization and gene expression of runt-related transcription factor 2, Alp, bone sialoprotein and osteocalcin revealing their high osteogenic potential. µCT analysis indicated that there was higher amount of bone formation in defects injected with both OB-NC and OB-MS compared to the control. Moreover, the bone tissue formed by both cells displayed the same histological characteristics. Despite the distinct in vitro osteogenic potential, OB-NC and OB-MS induced similar bone repair in a rat calvarial defect model. Thus, osteoblasts, irrespective of their in vitro osteogenic potential linked to embryonic origins, seem to be suitable for cell-based therapies aiming to repair bone defects.

Investigate the pathways of glycerol-3-P (G3P) generation for triacylglycerol (TAG) synthesis in retroperitoneal (RWAT) and epididymal (EWAT) white adipose tissues from high-fat diet (HFD)-fed mice. Mice were fed for 8 weeks a HFD and glycolysis, glyceroneogenesis and direct phosphorylation of glycerol were evaluated, respectively, by 2-deoxyglucose uptake, phosphoenolpyruvate carboxykinase (PEPCK-C) activity and pyruvate incorporation into TAG-glycerol, and glycerokinase activity and glycerol incorporation into TAG-glycerol in both tissues. HFD increased body and adipose tissue mass and serum levels of glucose and insulin, which were accompanied by glucose intolerance. RWAT and EWAT from HFD-fed mice had increased rates of de novo fatty acid (FA) synthesis (52% and 255%, respectively). HFD increased lipoprotein lipase (LPL) activity and content in EWAT (107%), but decreased in RWAT (79%). HFD decreased the lipolytic response to norepinephrine (57%, RWAT and 25%, EWAT), β3-adrenoceptor content (50%), which was accompanied by a decrease in phosphorylated-hormone-sensitive lipase (~80%) and phosphorylated-adipocyte triacylglycerol lipase (~60%) in both tissues. HFD decreased the in vitro rates of glucose uptake (3.5- and 6-fold), as well as in glyceride-glycerol synthesis from pyruvate (~3.5-fold) without changes in PEPCK-C activity and content in RWAT and EWAT, but increased glycerokinase activity(~3-fold) and content (90 and 40%) in both tissues. The data suggest that direct phosphorylation of glycerol by glycerokinase may be responsible for maintaining the supply of G3P for the existing rates of FA esterification and TAG synthesis in RWAT and EWAT from HFD-fed mice, contributing, along with a lower lipolytic response to norepinephrine, to higher adiposity.

Background Based on the crosstalk of inflammation with apoptosis, autophagy, and endoplasmic reticulum (ER) stress, the main objective of this study was to explore the role of interleukin-6 (IL-6) on genes and proteins related to these phenomena in the livers of mice submitted to acute exhaustive exercise. Methods Reverse transcription-quantitative polymerase chain reaction and immunoblotting technique were used to evaluate the livers of wild-type (WT) and IL-6 knockout (KO) mice at baseline (BL) and 3 h after the acute exhaustive physical exercise (EE). Results Compared to the WT at baseline, the IL-6 KO had lower exhaustion velocity, mRNA levels of Mtor, Ulk1, Map1lc3b, and Mapk14, and protein contents of ATG5 and p-p70S6K/p70S6K. For the WT group, the EE decreased glycemia, mRNA levels of Casp3, Mtor, Ulk1, Foxo1a, Mapk14, and Ppargc1a, and protein contents of ATG5 and p-p70S6K/p70S6K, but increased mRNA levels of Sqstm1. For the IL-6 KO group, the EE decreased glycemia, mRNA levels of Casp3 and Foxo1a, and protein contents of pAkt/Akt and Mature/Pro IL-1beta, but increased mRNA levels of Sqstm1, and protein contents of p-AMPK/AMPK. Conclusion The inhibition of the hepatic autophagy markers induced by the acute EE was attenuated in IL-6 KO mice, highlighting a new function of this cytokine.

The physical exercise is a potential strategy to control age-related metabolic disorders, such as insulin resistance, impaired glucose homeostasis, and type 2 diabetes. Rho-kinase (ROCK) increases skeletal muscle glucose uptake through Insulin Receptor Substrate 1 (IRS1) phosphorylation. Here, we investigated the role of physical exercise in ROCK pathway in the skeletal muscle of Fischer middle-aged rats. Firstly, we observed the ROCK distribution in different skeletal muscle fiber types. ROCK signaling pathway (ROCK1 and ROCK2) and activity (pMYPT1) were higher in the soleus, which was associated with increased insulin signaling pathway (pIR, pIRS1, pPDK, pGSK3β). Middle-aged rats submitted to physical exercise, showed the upregulation of ROCK2 content and normalized RhoA (ROCK activator enzyme) levels in soleus muscle compared with middle-aged sedentary rats. These molecular changes in middle-aged exercised rats were accompanied by higher insulin signaling (pIRS1, pGSK3β, pAS160, GLUT4) in the soleus muscle. Reinforcing these findings, when pharmacological inhibition of ROCK activity was performed (using Y-27632), the insulin signaling pathway and glucose metabolism-related genes (Tpi, Pgk1, Pgam2, Eno3) were decreased in the soleus muscle of exercised rats. In summary, ROCK signaling seems to contribute with whole-body glucose homeostasis (∼50 %) through its higher upregulation in the soleus muscle in middle-aged exercised rats.

Orthostatic hypotension is one of the most common symptoms observed in Parkinson's disease (PD), a neurodegenerative disease caused by death of dopaminergic neurons in the substantia nigra pars compacta (SNc), and it is associated with denervation of the heart and impairment of the baroreflex. Here, we aimed to investigate if the impaired baroreflex was associated with lower activation of cardiovascular brainstem areas in a 6-hydroxydopamine (6-OHDA) animal model of PD. The PD model was generated with male Wistar rats by injection of 6-OHDA or vehicle into the striatum. After 20 or 60 days, the femoral vein and artery were cannulated to assess cardiovascular parameters during injection of sodium nitroprusside (SNP) or phenylephrine (Phe). Brainstem slices were submitted to immunohistochemistry and immunofluorescence. After 6-OHDA injection, 75% of the dopaminergic neurons in the SNc were absent, confirming establishment of the PD model. Intravenous (iv) injection of SNP generated reduced hypotension and tachycardia response, and the noncatecholaminergic (non C1) neurons of the rostral ventrolateral medulla (RVLM) were less activated. Additionally, iv injection of Phe increased blood pressure and bradycardia to the same extent and activated equivalent numbers of neurons in the nucleus of the solitary tract and the caudal ventrolateral medulla as well as cholinergic neurons of the dorsal motor nucleus of the vagus and the nucleus ambiguus between control and PD animals. In summary, these data showed that in the PD model, impairment of cardiovascular autonomic control was observed only during deactivation of the baroreflex, which could be related to reduced activation of non-C1 neurons within the RVLM.

Simple Summary Through a systematic review of reports where swine were used as animal biomodels for testing or researching new surgical techniques, we sought to determine the quality of the report of the methodologies carried out on the basis of the ARRIVE guidelines (Animal Research: Reporting of In Vivo Experiments) in a total of 108 studies from 2013 to 2018. In a large percentage of the articles, the information presented in the methodology of the studies showed incomplete data according to the ARRIVE guidelines recommendations for reporting the use of animals. There was a strong focus on descriptions of surgical techniques; however, sample size calculation, description of maintenance conditions, animal handling, and anesthetic and pain management protocols used were not very detailed. This could lead to the inability of others to replicate the described experiments. For this reason, we encourage authors to implement the ARRIVE guidelines to improve the quality of scientific reports and ensure animal welfare. Abstract Over the last two decades, pigs have become animal biomodels widely used for the investigation and practice of surgical techniques because of their great physiological and anatomical similarities to humans. Even though many of these studies must be carried out later in humans, the description of basic information is limited, making exact repetitions of the reported experimental methods impossible. In this review, 108 studies from 2013 to 2018 were considered to determine the quality of adherence to the ARRIVE guidelines in the reports of the methodologies. The majority of the studies lacked the details recommended in the ARRIVE guidelines regarding data directly related to the welfare of animals undergoing surgery and those about anesthetic protocols and analgesics. Information related to sample size calculation and housing and husbandry conditions was also very limited. We believe that the ARRIVE guidelines are an excellent tool for good-quality reporting. We encourage scientists to consistently use them as a tool to improve the quality of their scientific reports and, consequently, ensure animal welfare.

Objective To examine the effectiveness of stratification to identify and target antioxidant therapy for animal models of lethal sepsis and in patients who develop sustained hypotension. Methods Rats were subjected to sepsis induced by cecal ligation and puncture. Animals were divided into two groups: those with high and low plasma levels of interleukin-6. Following stratification, N-acetylcysteine plus deferoxamine or saline was administered to animals starting 3 and 12 hours after surgery. N-Acetylcysteine plus deferoxamine or placebo was administered within 12 hours of meeting the inclusion criteria in hypotensive patients. Results N-Acetylcysteine plus deferoxamine increased survival in the cecal ligation and puncture model when administered 3 and 12 hours after sepsis induction. When dividing animals that received antioxidants using plasma interleukin-6 levels, the protective effect was observed only in those animals with high IL-6 levels. The antioxidant effect of N-acetylcysteine + deferoxamine was similar in the two groups, but a significant decrease in plasma interleukin-6 levels was observed in the high-interleukin-6-level group. Compared with patients treated with antioxidants in the low-interleukin-6 subgroup, those in the high-interleukin-6 subgroup had a lower incidence of acute kidney injury but were not different in terms of acute kidney injury severity or intensive care unit mortality. Conclusion Targeting antioxidant therapy to a high inflammatory phenotype would select a responsive population.

Preclinical and clinical data indicate that cannabidiol (CBD), a non-psychotomimetic compound from the Cannabis sativa plant, can induce antipsychotic-like effects. In an animal model of schizophrenia based on the antagonism of NMDA receptors, the behavioral and molecular changes induced by repeated treatment with the NMDA receptor antagonist MK-801 were prevented when CBD was co-administered with MK-801. It is unknown, however, if CBD would reverse these changes once they have been established. Thus, in the present study we used male C57BL/6J mice, 6 weeks old, to evaluate whether daily CBD injection for seven days, starting after the end of the repeated treatment with MK-801 for 14 days, would reverse MK-801-induced deficits in the social interaction (SI) and novel object recognition (NOR) tests, which have been used to investigate the negative and cognitive symptoms of schizophrenia, respectively. We also assessed whether CBD effects would be blocked by pretreatment with AM251, a CB1 receptor antagonist, AM630, a CB2 receptor antagonist, or WAY100635, a 5-HT1A receptor antagonist. CBD and the second-generation antipsychotic clozapine, used as a positive control, attenuated the impairments in the SI and NOR tests induced by repeated administered MK-801. CBD effects were blocked by WAY100635, but not by AM251 or AM630. These data suggest that CBD induces antipsychotic-like effects by activating 5-HT1A receptors and indicate that this compound could be an interesting alternative for the treatment of negative and cognitive symptoms of schizophrenia.

Purpose The pineal gland plays an important role in biological rhythms, circadian and circannual variations, which are key aspects in several headache disorders. Overview Melatonin, the main pineal secreting hormone, has been extensively studied in primary and secondary headache disorders. Altered melatonin secretion occurs in many headache syndromes. Experimental data show pineal gland and melatonin both interfere in headache animal models, decreasing trigeminal activation. Melatonin has been shown to regulate CGRP and control its release. Discussion Melatonin has been used successfully as a treatment for migraine, cluster headaches and other headaches. There is a rationale for including the pineal gland as a relevant brain structure in the mechanisms of headache pathophysiology, and melatonin as a treatment option in primary headache.

Changes to neonatal nutrition result in long-lasting impairments in energy balance, which may be described as metabolic programing. Astrocytes, which are interconnected by gap junctions, have emerged as important players in the hypothalamic control of food intake. In order to study the effects of nutritional programming on glial morphology and protein expression, cross-fostered male Wistar rats at postnatal day 3 were assigned to three groups based on litter size: small litter (3 pups per dam, SL), normal litter (10 pups per dam, NL), and large litter (16 pups per dam, LL). Rats from the SL group exhibited higher body weight throughout the study and hyperphagia after weaning. LL animals exhibited hyperphagia, high energy efficiency and catch-up of body weight after weaning. Both the SL and LL groups at postnatal day 60 (PN60) exhibited increased levels of plasma leptin, the Lee index (as an index of obesity), adiposity content, immunoreactivity toward T-cell protein tyrosine phosphatase (TCPTP), and glial fibrillary acidic protein (GFAP) in the arcuate nucleus (ARC) of the hypothalamus. Astrocyte morphology was altered in the ARC of SL and LL animals, and this effect occurred in parallel with a reduction in immunoreactivity toward connexin 30 (CX30). The data obtained demonstrate that both neonatal over- and underfeeding promote not only alterations in the metabolic status but also morphological changes in glial cells in parallel with increasing TCPTP and changes in connexin expression.

Introduction: Nuocytes play an important role in Type 2 immunity. However, the contribution of ILC2s to allergic rhinitis remains to be clearly elucidated. Objective: To evaluate the role of nuocytes from mesenteric lymph node on allergic responses in mice. Methods: After intraperitoneal administration of interleukin IL-25 and IL-33 to wild-type and Il17br-/-Il1rl1-/- double-deficient mice, nuocytes were purified from the the nasal-associated lymphoid tissue and mesenteric lymph nodes. Then, we assessed productions of IL-5 and IL-13 in nuocytes’ cultures. Finally, we adoptively transferred the mesenteric lymph node-derived nuocytes from wild-type and Il17br-/-Il1rl1-/- mice to the murine model of allergic rhinitis to evaluate their roles in nasal allergic responses. Results: We showed that nuocytes in the mesenteric lymph nodes of wild-type mice were upregulated after application of IL-25 and IL-33, and were induced to produce IL-5 and IL-13. Numbers of sneezing and nasal rubbing as well as eosinophils were all enhanced after the adoptive transfer of wild-type nuocytes. Concentrations of IL-5, IL-13, IL-25 and IL-33 in nasal lavage fluid of allergic mice were also increased. However, nuocytes fromIl17br-/-Il1rl1-/- mice did not increase sneezing and nasal rubbing and eosinophilia, and upregulate the above cytokines in the nasal lavage fluid. Conclusion: The findings demonstrate that nuocytes from the mesenteric lymph nodes of wildtype mice promote allergic responses in a mouse model. Resumo Introdução: Os nuócitos desempenham um papel importante na imunidade do tipo 2. No entanto, a contribuição das interleucinas ILC2s na rinite alérgica ainda precisa ser elucidada. Objetivo: Avaliar o papel dos nuócitos de linfonodos mesentéricos nas respostas alérgicas em camundongos. Método: Após a administração intraperitoneal de interleucina (IL)-25 e IL-33 em camundongos do tipo selvagem e camundongos Il17br-/-Il1rl1-/- com deficiência dupla, os nuócitos foram purificados do tecido linfoide associado a mucosa nasal e linfonodos mesentéricos. Em seguida, avaliamos as produções de IL-5 e IL-13 em culturas de nuócitos. Finalmente, transferimos adotivamente os nuócitos derivados de linfonodos mesentéricos de camundongos do tipo selvagem e camundongos Il17br-/-Il1rl1-/- para o modelo murino de rinite alérgica para avaliar seu papel nas respostas alérgicas nasais. Resultados: Mostramos que os nuócitos nos linfonodos mesentéricos de camundongos do tipo selvagem estavam up-regulados após a aplicação de IL-25 e IL-33 e foram induzidos a produzir IL-5 e IL-13. Os espirros e friçcão nasal, bem como os eosinófilos, aumentaram após a transferência adotiva de nuócitos do tipo selvagem. As concentrações de IL-5, IL-13, IL-25 e IL-33 no líquido da lavagem nasal de camundongos alérgicos também estavam aumentadas. Entretanto, os nuócitos de camundongos Il17br-/-Il1rl1-/- não aumentaram os espirros e a friçcão nasal ou eosinofilia e up-regularam as citocinas acima no líquido de lavagem nasal. Conclusão: Os achados demonstram que os nuócitos dos linfonodos mesentéricos de camundongos selvagens promovem respostas alérgicas em um modelo de camundongo.

The rate of saphenous vein graft failure one year after coronary artery bypass grafting ranges from 10% to 25%. The aim of this study was to explore whether atorvastatin can reduce accumulation of vascular smooth muscle cells to inhibit intimal hyperplasia via p38 MAPK pathway inhibition.Forty-five Sprague-Dawley rats were randomized to three groups. Thirty rats received a vein graft operation, and they were randomized to be treated with vehicle or atorvastatin; fifteen rats received a sham operation. We detected intimal hyperplasia by hematoxylin-eosin staining and related protein expression by immunohistochemical and Western blot analysis. Comparisons were analyzed by single-factor analysis of variance and Fisher's least significant difference test, with p0.05 considered significant.The intima analyzed by hematoxylin-eosin staining was dramatically thicker in the control group than in the atorvastatin group and sham group (p0.01). The outcomes of immunohistochemical staining of α-SMA demonstrated that the percentage of α-SMA-positive cells in the control group was higher than in the atorvastatin group (p0.01). We also evaluated α-SMA, PCNA, p38 MAPK, and phosphorylation of p38 MAPK after statin treatment by Western blot analysis, and the results indicated that atorvastatin did not lead to p38 MAPK reduction (p0.05); it did, however, result in inhibition of p38 MAPK phosphorylation (p0.01), and it significantly reduced α-SMA and PCNA levels, in comparison with the control group (p0.01).We have demonstrated that atorvastatin can inhibit accumulation of vascular smooth muscle cells by inhibiting the p38 MAPK pathway, and it is capable of inhibiting intimal hyperplasia in a rat vein graft model.A taxa de falha de enxerto de veia safena um ano após a cirurgia de revascularização do miocárdio varia de 10% a 25%. O objetivo deste estudo foi de investigar se a atorvastatina pode reduzir o acúmulo de células musculares lisas vasculares para inibir a hiperplasia intimal por meio da inibição da via p38 MAPK.Quarenta e cinco ratos Sprague-Dawley foram randomizados em três grupos. Trinta ratos foram submetidos à cirurgia de enxerto de veia e randomizados para tratamento com veículo ou atorvastatina; quinze ratos foram submetidos à cirurgia sham. Detectamos a hiperplasia intimal por meio de coloração com hematoxilina-eosina e a expressão de proteínas relacionadas por meio de análise imuno-histoquímica e Western blot. Foram realizadas as comparações por análise de variância de fator único e pelo teste da diferença mínima significativa de Fisher, com p0,05 considerado significativo.A íntima analisada pela coloração com hematoxilina-eosina era dramaticamente mais espessa no grupo controle que no grupo atorvastatina e no grupo sham (p0,01). Os resultados da coloração imuno-histoquímica de α-SMA demonstraram que a porcentagem de células positivas para α-SMA no grupo controle era mais alta que no grupo atorvastatina (p0,01). Nós também avaliamos α-SMA, PCNA, p38 MAPK e fosforilação de p38 MAPK após o tratamento com estatina por meio de análise de Western blot e os resultados indicaram que a atorvastatina não levou à redução de p38 MAPK (p0,05); no entanto, resultou na inibição da fosforilação de p38 MAPK (p0,01) e reduziu significativamente os níveis de α-SMA e PCNA, em comparação com o grupo controle (p0,01).Nós demonstramos que a atorvastatina pode inibir o acúmulo de células musculares lisas vasculares por meio da inibição da via p38 MAPK e é capaz de inibir a hiperplasia intimal em modelos de enxerto de veia em ratos.

Several axonal growth inhibitors have already been identified following spinal cord injury, the most known being myelin-derived proteins, such as Nogo-A. The present study aimed to correlate the formation of glial scar with the beginning of growth inhibitor, Nogo-A, release in rats previously submitted to compressive spinal cord injury. For this, 12 male and female Wistar rats (250 ± 50g) were divided into 3 groups of 4 animals each, according to the animals' euthanasia time after spinal cord injury (G3 - three days; G5 - five days; G7 - seven days). Spinal cord injuries were induced by means of dorsal laminectomy of the T10 vertebra and epidural compression. Histopathological evaluation and immunoreactivity of the Nogo-A axonal growth inhibitor were performed. It was observed that there was the release of the axonal inhibitor Nogo-A after 24h after the occurrence of spinal cord injury, and that the glial scar must be maintained, in this time interval, in order to guarantee the rebalancing of the post-trauma environment. Thus, it is suggested that the glial scar should be maintained in the acute phase of the lesion, guaranteeing its numerous benefits for the rebalancing of the post-injured environment and, after 24 hours, when the release of the studied axonal growth inhibitor begins, it should be removed. Ya se han identificado varios inhibidores del crecimiento axonal después de una lesión de la médula espinal, siendo los más conocidos proteínas derivadas de la mielina, como Nogo-A. El presente estudio tuvo como objetivo correlacionar la formación de cicatrices gliales con el comienzo de la liberación del inhibidor del crecimiento. Axonal Nogo-A en ratas previamente sometidas a lesión medular por compresión. Para ello, se dividieron 12 ratas Wistar macho y hembra (250 ± 50g) en 3 grupos de 4 animales cada uno, según el tiempo de eutanasia de los animales tras la lesión medular (G3 - tres días; G5 - cinco días; G7 - siete días). Las lesiones medulares se indujeron mediante laminectomía dorsal de la vértebra T10 y compresión epidural. Se realizó la evaluación histopatológica y la inmunorreactividad del inhibidor del crecimiento axonal Nogo-A. Se observó que hubo liberación del inhibidor axonal Nogo-A a las 24 h de ocurrida la lesión medular, y que la cicatriz glial debe mantenerse, en este intervalo de tiempo, para garantizar el reequilibrio del postraumatismo. ambiente. Así, se sugiere que la cicatriz glial se mantenga en la fase aguda de la lesión, garantizando sus numerosos beneficios para el reequilibrio del ambiente post-lesionado y, a las 24 horas, cuando comience la liberación del inhibidor del crecimiento axonal estudiado, debería ser eliminado. Diversos inibidores de crescimento axonal já foram identificados acompanhando a lesão medular, sendo as mais conhecidas as proteínas derivadas de mielina, como o Nogo-A. O presente trabalho teve como objetivo correlacionar a formação da cicatriz glial com o início da liberação do inibidor de crescimento axonal Nogo-A em ratos previamente submetidos à lesão medular compressiva. Para isso, 12 ratos Wistar, machos e fêmeas (250 ± 50g), foram divididos em 3 grupos de 4 animais cada, de acordo com o tempo de eutanásia dos animais após a lesão medular (G3 - três dias; G5 - cinco dias; G7 - sete dias). Foi realizada a indução das lesões medulares por meio de laminectomia dorsal da vértebra T10 e compressão epidural. A avaliação histopatológica e da imunorreatividade do inibidor de crescimento axonal Nogo-A foram realizadas. Foi observado que houve a liberação do inibidor axonal Nogo-A a partir de 24h após a ocorrência de lesão medular, e que cicatriz da glia deve ser mantida, nesse intervalo de tempo, para que garanta o reequilíbrio do meio pós-trauma. Dessa forma, sugere-se que a cicatriz glial deve ser mantida na fase aguda da lesão, garantindo seus inúmeros benefícios ao reequilíbrio do ambiente pós-lesionado e, após 24h, quando se inicia a liberação do inibidor de crescimento axonal estudado, esta deve ser removida.

Submitted by DSpace Unilasalle (dspace@unilasalle.edu.br) on 2021-09-14T15:00:14Z No. of bitstreams: 1 coliveira.etal.pdf: 2666754 bytes, checksum: 924064e82040b2fb190e26cd7e99fe90 (MD5) Made available in DSpace on 2021-09-14T15:00:14Z (GMT). No. of bitstreams: 1 coliveira.etal.pdf: 2666754 bytes, checksum: 924064e82040b2fb190e26cd7e99fe90 (MD5) Previous issue date: 2020 In the present study, we investigated the effect of repeated neonatal morphine exposure and/or maternal deprivation(MD) on the nociceptive response and central biomarkers’ BDNF, IL-1β, and IL-4 levels at postnatal days 16(PND16), 30(PND30), and 60(PND60). At birth, the litters were standardized to contain 8 pups/dam (n = 58). From PND1 to PND10, the pups of the deprived groups were separated daily from their mothers for 3 h and divided into 5 groups: control(C), saline(S), morphine(M), deprived-saline(DS), and deprived-morphine(DM). The pups received subcutaneous injections of saline/morphine (5 μg) in the mid-scapular area between PND8 and PND14. Nociceptive responses were assessed by hot plate(HP) and tail-flick(TFL) tests and biomarker levels by ELISA. Thermal hyperalgesia(HP) was found in all assessments for the M, DS, and DM groups, and a decrease in nociceptive threshold(TFL) was found in the DS group at PND16; M and DM groups at PND30; and M, DS, and DM groups at PND60. There were interactions between treatment/deprivation/timepoint in all central biomarkers’ levels. The current study indicates that neonatal exposure to morphine and MD, which occurs in the pediatric ICU, can alter the nociceptive and neuroinflammatory responses.