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Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Objetivo: Avaliar uma correlação entre comportamentos de consumo e comportamentos sociais nas linhagens Wistar e SHR e verificar se manipulações farmacológicas (tratamento crônico e preventivo com nitroprussiato de sódio - NPS) e não-farmacológicas (sessões de treino de interação social) são capazes de promover uma melhora no desempenho/hedonia social com consequente normalização da autoadministração de sacarina na linhagem SHR. Métodos: Experimento 1: Ratos Wistar e SHR receberam veículo ou NPS (2,5 mg/kg) do dia 30 ao 60 pós-natais. Após o término do tratamento, foram submetidos aos testes de investigação social, interação social e autoadministração operante de sacarina. Experimento 2: ratos Wistar e SHR entre os 30 e 90 dias de vida foram submetidos ao treino de interação social, 3 vezes por semana, ou expostos ao campo aberto sozinho. Após o período de treinos, os animais passaram pelos testes comportamentais do experimento 1. Resultados: No experimento 1, ratos Wistar investigam mais o estímulo social em relação ao neutro, diferença não observada na linhagem SHR. Animais SHR tiveram aumento de locomoção e a maior quantidade de levantamentos em relação aos ratos Wistar, bem como os parâmetros de consumo elevados no teste de autoadministração operante de sacarina. No tratamento crônico e preventivo com NPS foi observada uma diminuição na quantidade de pressões na barra ativa no protocolo de razão progressiva. No segundo experimento, ratos SHR tiveram menor porcentagem de investigação social, e aqueles que foram submetidos aos treinos com outro animal interagiram menos que os que passaram pelo campo aberto sozinhos. O treino não teve efeito sobre o défict de interação social ou sobre a quantidade de levantamentos, mas em relação a atividade locomotora, os animais que foram treinados com outro animal no campo aberto percorreram menos quadrantes durante o teste, independentemente da linhagem. Também foi observado um comportamento consumatório elevado da linhagem SHR nos protocolos de autoadministração operante de sacarina; no entanto, o treino de interação social não teve efeito sobre as medidas de consumo. Nos dois experimentos observamos correlações negativas entre comportamentos de consumo e comportamentos sociais, indicando que os animais que interagem menos socialmente são aqueles que possuem medidas de consumo mais altas. Conclusões: Observamos uma correlação negativa entre comportamentos sociais e de consumo nos ratos, o que fortalece a nossa hipótese da existência de um tônus hedônico. No entanto, o comportamento social da linhagem SHR não foi atenuado pelas manipulações propostas. Isso indica que a diminuição de comportamentos de consumo, como induzido pelo tratamento preventivo com o NPS, não decorreu de um “ajuste” do desequilíbrio hedônico visto na linhagem.

The incidence of Parkinson’s disease (PD) is increasing worldwide. Although the PD hallmark is the motor impairments, nonmotor dysfunctions are now becoming more recognized. Recently, studies have suggested that baroreflex dysfunction is one of the underlying mechanisms of cardiovascular dysregulation observed in patients with PD. However, the large body of literature on baroreflex function in PD is unclear. The baroreflex system plays a major role in the autonomic, and ultimately blood pressure and heart rate, adjustments that accompany acute cardiovascular stressors on a daily basis. Therefore, impaired baroreflex function (i.e., decreased sensitivity or gain) can lead to altered neural cardiovascular responses. Since PD affects parasympathetic and sympathetic branches of the autonomic nervous system and both are orchestrated by the baroreflex system, understanding of this crucial mechanism in PD is necessary. In the present review, we summarize the potential altered central and peripheral mechanisms affecting the feedback-controlled loops that comprise the reflex arc in patients with PD. Major factors including arterial stiffness, reduced number of C1 and activation of non-C1 neurons, presence of central α-synuclein aggregation, cardiac sympathetic denervation, attenuated muscle sympathetic nerve activity, and lower norepinephrine release could compromise baroreflex function in PD. Results from patients with PD and from animal models of PD provide the reader with a clearer picture of baroreflex function in this clinical condition. By doing so, our intent is to stimulate future studies to evaluate several unanswered questions in this research area.

Introdução: O modelo de artrite em ratos induzido pela administração de Adjuvante Completo de Freund (CFA) é bem caracterizado na literatura e é rotineiramente empregado para busca de novos fármacos para o tratamento da dor inflamatória, incluindo a osteoartrite. A administração de CFA na articulação temporomandibular (ATM) tem sido utilizada como um modelo de dor orofacial inflamatória crônica. A lesão do tecido periférico decorrente da indução do modelo geralmente produz aumento da sensibilidade térmica, bem como da sensibilidade mecânica. Objetivo: Dessa forma, considerando a importância do uso de modelos confiáveis para investigação de novas opções de tratamento em estudos pré-clínicos, este estudo teve como objetivo avaliar o curso temporal do desenvolvimento da dor orofacial inflamatória crônica em ratos, induzida por injeção unilateral de CFA na ATM, procurando determinar parâmetros objetivos que justifiquem seu uso. Como objetivo secundário foram investigados os efeitos da administração de melatonina sobre a hiperalgesia induzida pela morfina em ratos. Materiais e Métodos: No experimento 1, 22 ratos Wistar adultos foram divididos em dois grupos: sham (solução salina) e dor (injeção intra-articular de 50 μl de CFA na ATM direita). A sensibilidade mecânica da região da ATM foi verificada usando um analgesímetro eletrônico von Frey no baseline e nos dias 7, 9, 11, 13, 15, 17, 19, 21, 23 e 25 após a indução do modelo. Além disso, os testes de acetona e Operant Orofacial Pain Assessment Device (OPAD) foram realizados para avaliar a sensibilidade térmica. Os comportamentos locomotor e exploratório, e do tipoansioso foram avaliados pelos testes do Campo Aberto e Labirinto em Cruz Elevado, respectivamente. Os níveis de IL-1β, IL-10 e BDNF foram mensurados pela técnica de ELISA no gânglio trigeminal. A avaliação qualitativa do grau de inflamação na área retrodiscal foi feita por meio de histologia e microscopia óptica. A pontuação foi baseada em um sistema pelo qual a porcentagem de células mononucleares infiltrantes foi classificada como ausente, leve, moderada e grave. No experimento 2, 70 ratos Wistar machos adultos foram divididos em oito grupos: Sham+Veículo+Salina (SVS); Sham+Veículo+Morfina (SVMo); Sham+Melatonina+Salina (SMeS); Sham+Melatonina+Morfina (SMeMo); Dor+Veículo+Salina (DVS); Dor+Veículo+Morfina (DVMo); Dor+Melatonina+Salina (DMeS); Dor+Melatonina+Morfina (DMeMo). Os animais foram tratados por oito dias consecutivos com morfina (5 mg/Kg s.c.) e/ou melatonina (50 mg/Kg i.p.), ou seus respectivos veículos. A hiperalgesia mecânica da região da ATM foi verificada pelo von Frey eletrônico no baseline e nos dias 13, 14, 18, 21 e 22 após a indução do modelo. Além disso, foram realizados os testes de acetona, OPAD Campo Aberto e Labirinto em Cruz Elevado. Os dados foram avaliados utilizando teste “t” e “U” de Mann-Whitney para dados não paramétricos e expressos como média ± erro padrão da média (EPM) ou mediana e intervalo interquartil, considerando significativos P < 0,05. O SPSS 20.0 para macOS foi usado para análise estatística. Este projeto foi avaliado e aprovado pela CEUA-HCPA (#20200689). Resultados: Experimento 1: foi observado o estabelecimento do modelo de dor nos seguintes parâmetros: 1. diminuição da latência de retirada da face no teste de von Frey entre os dias sete e 23 após a injeção de CFA; 2. diminuição do número de lambidas no teste OPAD; 3. redução do índice de ansiedade no Labirinto em Cruz Elevado; 4. aumento do número de rearings no Open Field; 5. redução dos níveis de BDNF no gânglio trigeminal; 6. aumento do espessamento do disco articular e maior escore de inflamação na ATM. Experimento 2: houve replicação do modelo de dor do experimento 1, indexado pela redução do limiar nociceptivo no teste de von Frey. Observou-se efeito do modelo, da morfina e da melatonina no teste do Labirinto em Cruz Elevado, enquanto que nos parâmetros do teste do Campo Aberto e teste da acetona, não houve efeito. Apesar do reduzido número de animais, foi possível observar o efeito antinociceptivo da morfina que foi incrementado pela administração conjunta da melatonina. Nos períodos e doses avaliadas não foi possível determinar a hiperalgesia e tolerância induzidas pela morfina. Por outro lado, os resultados demonstram que a melatonina não apresentou efeito analgésico agudo na dose utilizada, necessitando de administração repetida para induzir analgesia neste modelo de dor. Além disso, a melatonina induziu uma potencialização em relação a analgesia da morfina, demonstrando um excelente efeito sinérgico. Estes efeitos da melatonina se mantêm por pelo menos 24h após o final do tratamento. Outra observação importante é que a ação dos fármacos é estado específico, uma vez que os resultados distintos em animais com e sem dor. Conclusão: Esta dissertação demonstra que a administração intra-articular de CFA na ATM em ratos pode ser utilizada para avaliar alterações agudas e crônicas que mimetizam a artrite inflamatória da ATM. No entanto, os estudos crônicos são limitados a 25 dias após a injeção de CFA, pois a partir desse período a dor não está mais presente. No segundo experimento, os resultados preliminares permitem concluir que a melatonina é capaz de potencializar a analgesia induzida pela morfina, que permanece por pelo menos 24 horas após o final do tratamento, assim como a analgesia induzida pela melatonina per se, que necessita de doses repetidas para induzir efeito analgésico. Mais estudos estão em andamento para melhor entender os efeitos observados. Introduction: The arthritis model in rats induced by the administration of Complete Freund's Adjuvant (CFA) is well characterized in the literature, being routinely used to screen for new drugs for the treatment of inflammatory pain, including osteoarthritis. The administration of CFA into the temporomandibular joint (TMJ) has been used as a model of chronic inflammatory orofacial pain. Peripheral tissue injury resulting from the model usually produces increased thermal as well as mechanical sensitivity. Aim: Thus, considering the importance of using reliable models to investigate new treatment options in preclinical studies, this study aimed to evaluate the temporal course of the development of chronic inflammatory orofacial pain in rats, induced by unilateral injection of CFA into the TMJ, seeking to determine objective parameters that justify its use. As a secondary objective, the effects of melatonin administration on morphine-induced hyperalgesia in rats were investigated. Materials and methods: In experiment 1, 22 adult male Wistar rats were divided into two groups: sham (saline solution) and pain (intra-articular injection of 50 μl of CFA into the right TMJ). Mechanical sensitivity of the TMJ region was verified using an electronic von Frey analgesimeter at baseline and on days 7, 9, 11, 13, 15, 17, 19, 21, 23, and 25 after model induction. In addition, the Acetone and the Operant Orofacial Pain Assessment Device (OPAD) tests were performed to assess thermal sensitivity. The locomotor and exploratory, and anxiety-like behaviors were evaluated by the Open Field and Elevated Plus Maze tests, respectively. The levels of IL-1β, IL-10 and BDNF were measured by ELISA in the trigeminal ganglion. Qualitative assessment of the degree of inflammation in the retrodiscal area was performed using histology and light microscopy. Scoring was based on a system whereby the percentage of infiltrating mononuclear cells was classified as absent, mild, moderate, and severe. In experiment 2, 70 adult male Wistar rats were divided into eight groups: Sham+Vehicle+Saline (SVS); Sham+Vehicle+Morphine (SVMo); Sham+Melatonin+Saline (SMeS); Sham+Melatonin+Morphine (SMeMo); Pain+Vehicle+Saline (DVS); Pain+Vehicle+Morphine (DVMo); Pain+Melatonin+Saline (DMeS); Pain+Melatonin+Morphine (DMeMo). The animals were treated during eight consecutive days with morphine (5 mg/kg s.c.) and/or melatonin (50 mg/kg i.p.), or their respective vehicles. The mechanical sensitivity of the TMJ region was verified using an electronic von Frey analgesimeter at baseline and on days 13, 14, 18, 21 and 22 after model induction. In addition, the acetone and the OPAD tests were performed to assess thermal sensitivity. The locomotor and exploratory, and anxiety-like behaviors were evaluated by the Open Field and Elevated Plus Maze tests, respectively. Ethical approval was obtained from CEUA-HCPA (#20200689). Results: Experiment 1: the establishment of the pain model was observed in the following parameters: 1. decrease in face withdrawal latency in the von Frey test between days seven and 23 after CFA injection; 2. decrease in the number of licks in the OPAD test; 3. reduction of the anxiety index in the Elevated Plus Maze; 4. increase in the number of rearings in the Open Field; 5. reduction of BDNF levels in the trigeminal ganglion; 6. increased joint disc thickening and higher TMJ inflammation score. Experiment 2: the pain model from experiment 1 was replicated, indexed by the reduction of the nociceptive threshold in the von Frey test. There was an effect of the model, morphine and melatonin in the Elevated Plus Maze test, while no effects were observed in the parameters of the Open Field and the Acetone tests. Despite the small number of animals, it was possible to observe the antinociceptive effect of morphine that was increased by the combined administration of melatonin. The hyperalgesia and tolerance induced by morphine could not be observed at the current time and dose. On the other hand, the results demonstrate that melatonin did not present an acute analgesic effect at the dose employed, requiring repeated administration to induce analgesia in this model. In addition, melatonin induced a potentiation of the morphine analgesia, demonstrating an excellent synergistic effect. These effects of melatonin are maintained for at least 24 hours after the end of treatment. Another important observation is the state-specific effect, since drug-induced effects have temporally different effects between the sham and pain groups. In conclusion, this dissertation demonstrates that intra-articular administration of CFA into the TMJ in rats can be used to assess acute and chronic changes that mimic TMJ inflammatory arthritis. However, chronic studies are limited to 25 days after CFA injection, since after this period, pain is no longer present. In the second experiment, the preliminary results enable us to conclude that melatonin is capable of potentiating the morphine-induced analgesia, which persists for at least 24 hours after the end of treatment, as well as melatonin-induced analgesia per se, which requires repeated doses to induce an analgesic effect. More studies are underway aiming to better understand the observed effects.

O acidente vascular encefálico (AVE) é uma enfermidade decorrente da redução ou interrupção do fluxo sanguíneo cerebral, resultando em morte celular e deficits cognitivos e sensoriomotores. É uma das principais causas de morte e incapacidade no mundo. Após um AVE, é formada uma zona central de dano tecidual irreversível (núcleo isquêmico) e uma região recuperável em volta do núcleo, demonimada zona de penumbra. A hipertensão arterial sistêmica (HAS), caracterizada pelo aumento persistente da pressão arterial, é o principal fator de risco para o AVE e afeta negativamente a progressão da doença e a resposta ao tratamento. A HAS provoca uma série de alterações neurovasculares, como enrijecimento da parede de grandes artérias cerebrais, disfunção da barreira hematoencefálica e alteração de mecanismos de controle do fluxo sanguíneo cerebral. É importante utilizar modelos animais com comorbidades na pesquisa pré-clínica em AVE, de forma a torná-la mais translacional. Sendo assim, o presente trabalho teve como objetivo avaliar os efeitos de um modelo de AVE isquêmico focal permanente em ratos hipertensos sobre a função sensoriomotora e volume da lesão cerebral isquêmica. A pesquisa foi previamente aprovada pela Comissão de Ética no Uso de Animais do Instituto de Cardiologia, registrada nos protocolos UP 5806/20 e UP 5850/20. Ratos espontaneamente hipertensos (SHR) e Wistar Kyoto (WKY - controle) machos adultos foram anestesiados e submetidos a isquemia cerebral focal permanente por termocoagulação dos vasos piais. Avaliações comportamentais foram realizadas ao longo de 42 dias para determinar a recuperação sensoriomotora e a memória de habituação. No teste do cilindro, os animais apresentaram assimetria dos membros anteriores e observou-se uma recuperação total da simetria dos ratos normotensos (WKY), mas não houve recuperação nos ratos hipertensos (SHR). No teste de remoção de adesivo, houve recuperação sensoriomotora total nos ratos WKY e parcial nos ratos SHR. Na tarefa do campo aberto, a isquemia cerebral afetou a memória de habituação a curto prazo nos ratos WKY. Não foi observada memória de habituação a curto prazo nos ratos SHR submetidos ou não a isquemia cerebral. Observou-se uma redução do volume cerebral isquêmico nos ratos WKY 7 dias pós-AVE. Com estes resultados, pode-se observar que a HAS prejudica a recuperação sensoriomotora de ratos submetidos à isquemia cerebral. A redução do volume de lesão isquêmica observada nos ratos WKY pode explicar a recuperação sensoriomotora total observada nesses animais. O trabalho reforça a necessidade de se utilizar modelos animais com comorbidades na pesquisa em AVE. Stroke is a disease characterized by reduction or interruption of cerebral blood flow, leading to cell death and cognitive and sensorimotor deficits. It is a leading cause of death and disability worldwide. After a stroke, a central zone of irreversibly damaged tissue is formed (ischaemic core) and a salvageable surrounding area named ischaemic penumbra. Hypertension is characterized by sustained elevation of blood pressure and is the most prevalent risk factor for stroke and worsens stroke outcome and response to treatment. Hypertensions causes neurovascular alterations like stiffening of large cerebral arteries, blood brain barrier dysfunction and alterations of cerebral blood flow regulation mechanisms. It is important to use animal models with comorbidities in stroke research, to make it more translational. Therefore, the study aims to evaluate the effects of a model of permanent ischaemic stroke in hypertensive rats on sensorimotor function and cerebral infarct size. The study was approved by IC/FUC’s Ethical Committee on Animal Use (UP 5806/20 and 5850/20). Adult male spontaneously hypertensive (SHR) and Wystar Kyoto (WKY) rats were anesthetized and submitted to focal ischaemia by thermocoagulation of pial vessels. Behavioral assessments were performed over 42 days to evaluate sensorimotor recovery and habituation memory. In the cylinder task, there was a total recovery of symmetry of WKY rats, but there was no recovery in SHR rats. In adhesive removal task, there was a total sensorimotor recovery for WKY rats and partial recovery for SHR rats. Stroke impaired short-term habituation memory in open field task in WKY rats. The short-term habituation memory wasn’t observed in SHR rats. There was a bigger cerebral infarct size in SHR rats compared to WKY at 7 days after stroke. There results shows how hypertension impairs sensorimotor recovery in rats submitted to cerebral ischaemia. Cerebral infarct size reduction observed in WKY might explain the total sensorimotor recovery of this strain. The study reinforce the need of using animal models with comorbidities in stroke research.

O Transtorno do Espectro Autista (TEA) é um transtorno do neurodesenvolvimento caracterizado por prejuízos na comunicação e interação social, comportamentos repetitivos e interesses restritos. Estima-se que, mundialmente, 1% da população é afetada pela condição. A etiologia do TEA é desconhecida, porém sabe-se que existe uma contribuição complexa de fatores ambientais e genéticos para o desenvolvimento do transtorno. Alterações transcritômicas são consistentemente observadas no sistema nervoso central de indivíduos com TEA e não estão restritas a expressão de genes codificantes. Uma classe de RNAs não-codificantes de interesse especialmente recente é a dos RNAs circulares (circRNAs). Os circRNAs são abundantes no encéfalo e podem desempenhar funções variadas como competir pela ligação de miRNAs, estimular a transcrição do gene parental e ser traduzido em proteínas funcionais. O envolvimento dos circRNAs no TEA ainda é pouco explorado. Recentemente foram feitas as primeiras observações de circRNAs diferencialmente expressos (CDE) no córtex cerebral de indivíduos com TEA. Adicionalmente, CDE também foram observados em dois modelos animais de TEA. Modelos animais são essenciais para o estudo de transtornos do neurodesenvolvimento e podem auxiliar os pesquisadores a investigar a etiologia e os mecanismos biológicos subjacentes. A detecção em massa e a descoberta de novos circRNAs depende quase exclusivamente de softwares de identificação de circRNA. Entretanto, possíveis problemas na usabilidade dessas ferramentas são relatados na literatura e podem prejudicar os usuários ao consumir tempo e dificultar a reprodutibilidade das análises. Considerando o potencial envolvimento de circRNAs no TEA e a dependência metodológica de softwares para identificação, a presente tese analisa por bioinformática a expressão gênica de circRNAs no encéfalo de diferentes modelos animais de TEA, assim como desenvolver um método de avaliação e melhoramento da usabilidade das ferramentas de bioinformática utilizadas para a identificação dos circRNAs. No Capítulo I, identificamos milhares de circRNAs em diferentes modelos animais de TEA em camundongos e demonstramos que as alterações na expressão estão relacionadas principalmente às funcionalidades sinápticas. No Capítulo II, identificamos parcialmente a sequência do circRNA ciRS-7 no modelo animal de ratos induzidos por ácido valproico. Demonstramos que a expressão de ciRS-7 está elevada, assim como encontrada nos modelos Ash1L, BTBR e Kmt5b no Capítulo I, e que funções sinápticas estão entre os possíveis grupos funcionais afetados. Por fim, no Capítulo III, identificamos os problemas de usabilidade das ferramentas de identificação de circRNAs utilizadas nos Capítulos I e II. Também criamos um conjunto de regras para auxiliar os desenvolvedores desse tipo de software para criarem ferramentas mais utilizáveis para os usuários finais. Em conjunto, os resultados apresentam um panorama do envolvimento de circRNAs no TEA a partir de modelos animais. Além disso, apresentamos um novo método de avaliação de usabilidade de software que deve possibilitar que futuras análises estejam alinhadas com as necessidades e expectativas de uso de ferramentas de identificação de circRNAs. Autism Spectrum Disorder (ASD) is a neurodevelopmental disorder characterized by impairments in communication and social interaction, repetitive behaviors, and restricted interests. It is estimated that the condition affects 1% of the population worldwide. The etiology of ASD is unknown, but it is known that there is a complex contribution of environmental and genetic factors to the development of the disorder. Transcriptomic alterations are consistently observed in the central nervous system of individuals with ASD and are not restricted to the expression of coding genes. Circular RNAs (circRNAs) are a class of noncoding RNAs of recent interest. CircRNAs are abundant in the brain and can perform varied functions, such as competing for binding miRNAs, stimulating parental gene transcription, and being translated into functional proteins. The involvement of circRNAs in ASD is still poorly explored. Recently, the first observations of differentially expressed circRNAs (DECs) were made in the cerebral cortex of individuals with ASD. Additionally, DECs were also observed in two animal models of ASD. Animal models are essential for studying neurodevelopmental disorders and can help researchers investigate the etiology and underlying biological mechanisms. The mass detection and discovery of novel circRNAs rely almost exclusively on circRNA identification software. However, potential problems in the usability of these tools are reported in the literature and may harm users by consuming time and hindering the reproducibility of analyses. Considering the potential involvement of circRNAs in ASD and the methodological dependence on software for identification, this thesis analyzes by bioinformatics the gene expression of circRNAs in the brain of different animal models of ASD, as well as develops a method to evaluate and improve the usability of bioinformatics tools used for circRNA identification. In Chapter I, we identify thousands of circRNAs in different animal models of ASD in mice and demonstrate that the changes in expression are mainly related to synaptic functionalities. In Chapter II, we partially identify the sequence of the circRNA ciRS-7 in the valproic acid-induced rat model. We demonstrate that ciRS- 7 expression is elevated, as found in the Ash1L, BTBR, and Kmt5b models in Chapter I, and that synaptic functions are among the possible functional groups affected. Finally, in Chapter III, we identify the usability problems of the circRNAs identification tools used in Chapters I and II. We also create a set of rules to assist developers of such software in creating more usable tools for end users. Taken together, the results present an overview of the involvement of circRNAs in ASD from the perspective of animal models. In addition, we present a new software usability evaluation method that should enable future analyses to be aligned with the needs and expectations of using circRNAs identification tools.

A hipercolesterolemia familiar (HF) é uma doença prevalente que nos últimos anos vem sendo fortemente associada a alterações cerebrais características de doenças neurodegenerativas. A HF é causada por mutações no gene que codifica o receptor da lipoproteína de baixa densidade (LDLr), a principal carreadora de colesterol periférico. Estudos com camundongos nocautes para o LDLr (LDLr-/-) apontaram que a neuroinflamação e disfunção da barreira hematoencefálica (BHE) são eventos importantes no distúrbio cerebral associado à HF. Nesse sentido, foi demonstrado em camundongos LDLr-/- que a inibição da mTOR pode atenuar a quebra da BHE. Dessa forma, nesta dissertação nós hipotetizamos que a exposição dos animais modelos experimentais de HF à metformina, um fármaco que potencialmente modula a sinalização da mTOR via AMPK, poderia atenuar as alterações cerebrais causadas pela HF. Camundongos LDLr-/- e selvagens (WT) machos adultos receberam metformina (200 mg/Kg) ou salina diariamente por 30 dias. Todos os animais foram pesados semanalmente. Na última semana de tratamento, foram realizados o teste de campo aberto para avaliação da atividade locomotora e exploratória, os testes de reconhecimento e realocação de objetos para avaliação da memória e o teste de suspensão pela cauda para avaliação do fenótipo depressivo. Uma coorte de animais foi eutanasiada por exsanguinação via punção cardíaca ao fim do tratamento. Na sequência, o plasma heparinizado foi usado para dosagem de colesterol total e triglicerídeos. Outra coorte de animais foi perfundida com paraformaldeído para a coleta de cérebros fixados. O colesterol total dos camundongos LDLr-/- se mostrou aumentado em relação aos grupos WT. O tratamento com metformina aumentou os triglicerídeos e diminuiu o peso corporal nos camundongos LDLr-/- em comparação com os animais administrados com salina. Os camundongos LDLr-/- mostraram atividade hiperlocomotora, danos cognitivos e um fenótipo depressivo, o que foi parcialmente atenuado pela metformina. Não se observou diferença significativa entre os grupos com relação à imunorreatividade microglial no córtex pré-frontal ou hipocampo, mas a metformina pareceu melhorar a astrogliose no hipocampo de camundongos LDLr-/-. No geral, o tratamento com metformina mostrou efeito promissor contra o dano de memória e a reatividade astroglial observados em camundongos LDLr-/-, independentemente da redução do colesterol plasmático. Familial hypercholesterolemia (FH) is a prevalent disease that has been strongly associated with brain alterations characteristic of neurodegenerative diseases in recent years. FH is caused by mutations in the gene that encodes the low-density lipoprotein receptor (LDLr), the main carrier of peripheral cholesterol. Studies with LDLr knockout mice (LDLr-/-) showed that neuroinflammation and blood-brain barrier (BBB) breakdown are important events in FH-related brain dysfunction. In this sense, it was demonstrated in LDLr-/- mice that mTOR inhibition can attenuate BBB disruption. Thus, in this dissertation, we hypothesized that exposure of experimental FH animal models to metformin, a drug that potentially modulates mTOR signaling via AMPK, could attenuate the brain alterations caused by FH. LDLr-/- and wild-type (WT) male adult mice received a daily oral administration of metformin (200 mg/Kg) or saline for 30 days. All animals were weighed weekly. In the last week of treatment, mice performed the open field test to evaluate locomotor and exploratory activity, the novel object recognition, the object reallocation task for evaluation of memory, and the tail suspension test to evaluate a depressed phenotype. A cohort of animals was euthanized by exsanguination via cardiac puncture at the end of the treatment protocol. Heparinized plasma was used to determine total cholesterol and triglyceride levels. Another cohort of animals was perfused with paraformaldehyde to collect fixed total brains. Total cholesterol levels of the LDLr-/- mice were significantly higher than the WT groups. Metformin treatment increased triglyceride levels and decreased total body weight in LDLr-/- mice in comparison to saline administration. LDLr-/- mice showed hyperlocomotive activity, cognitive damage, and a depressed phenotype, which was in parts attenuated by metformin treatment. We did not observe significant differences between groups regarding microglia immunoreactivity in the pre-frontal cortex or hippocampus of the animals, but metformin treatment seemed to ameliorate astrogliosis in the hippocampus of LDLr-/- mice. Overall, metformin treatment showed a promising effect on the memory impairment and astrocyte reactivity observed in LDLr-/- mice, which was not via a cholesterol-lowering outcome.

Autism spectrum disorder (ASD) is a prevalent neurodevelopmental disorder characterized by several alterations, including disorganized brain cytoarchitecture and excitatory/inhibitory (E/I) imbalance. We aimed to analyze aspects associated with the inhibitory components in ASD, using bioinformatics to develop notions about embryonic life and tissue analysis for postnatal life. We analyzed microarray and RNAseq datasets of embryos from different ASD models, demonstrating that regions involved in neuronal development are affected. We evaluated the effect of prenatal treatment with resveratrol (RSV) on the neuronal organization and quantity of parvalbumin-positive (PV+), somatostatin-positive (SOM+), and calbindin-positive (CB+) GABAergic interneurons, besides the levels of synaptic proteins and GABA receptors in the medial prefrontal cortex (mPFC) and hippocampus (HC) of the ASD model induced by valproic acid (VPA). VPA increased the total number of neurons in the mPFC, while it reduced the number of SOM+ neurons, as well as the proportion of SOM+, PV+, and CB+ neurons (subregion-specific manner), with preventive effects of RSV. In summary, metabolic alterations or gene expression impairments could be induced by VPA, leading to extensive damage in the late developmental stages. By contrast, due to its antioxidant, neuroprotective, and opposite action on histone properties, RSV may avoid damages induced by VPA.

Os avanços significativos conquistados nas últimas décadas no entendimento da neurobiologia de transtornos mentais, como a esquizofrenia, infelizmente não resultaram em tratamentos mais eficazes para os pacientes acometidos por tais condições. Antagonistas de receptores glutamatérgicos do tipo NMDA induzem um estado psicótico em indivíduos saudáveis semelhante à psicose observada na esquizofrenia, além de serem amplamente utilizados para induzir sintomas psicóticos em modelos pré-clínicos da doença. A taurina é um aminoácido com ação neuromoduladora inibitória do sistema nervoso central, neuroprotetora e antioxidante. Nesse trabalho investigamos o potencial da taurina em prevenir os déficits induzidos por administração aguda de MK-801 (dizocilpina), um antagonista NMDA, em ensaios comportamentais relacionados à esquizofrenia em camundongos C57BL/6 e peixes zebra. Nos experimentos em roedores, os animais foram injetados intraperitonealmente (i.p.) com solução salina ou taurina (50, 100 e 200 mg/kg), e 30 minutos depois receberam outra injeção i.p. de solução salina ou MK-801 (0,15 mg/kg). Os testes comportamentais de inibição por pré-pulso da resposta de sobressalto e interação social foram realizados 30 minutos após a última injeção, enquanto a atividade locomotora foi avaliada continuamente desde a primeira injeção, finalizando 60 minutos após a última. Já no caso de peixes-zebra, os animais foram colocados em um béquer contendo 200 mL de água ou taurina (42, 150 ou 400 mg/L) durante 20 minutos, sendo subsequentemente expostos a água ou MK-801 (5 μM) durante mais 20 minutos. O teste de interação social foi realizado imediatamente após o término da última exposição. Como esperado, a administração de MK-801 causou hiperlocomoção e déficit de inibição por pré-pulso em camundongos, enquanto em peixes-zebra induziu hiperlocomoção e déficit de interação social. Curiosamente, camundongos tratados com MK-801 passaram mais tempo interagindo com os animais estímulo; taurina na dose de 50 mg/kg teve o mesmo efeito, o que pode estar relacionado aos seus efeitos ansiolíticos já documentados. Em nenhuma das espécies foram observados efeitos preventivos da taurina sobre as alterações comportamentais induzidas agudamente por MK-801, contrariando evidências prévias da literatura. Evidências recentes têm fomentado a ideia de que o curso da esquizofrenia pode ser modificado por estratégias de intervenção iniciadas precocemente, antes do primeiro surto psicótico e do estabelecimento do transtorno em sua forma plena. Deste modo, é uma perspectiva desse trabalho avaliar o tratamento precoce e contínuo com taurina em um modelo desenvolvimental de esquizofrenia para melhor elucidar o potencial preventivo dessa molécula. The significant advances made in recent decades in understanding the neurobiology of mental disorders such as schizophrenia have not, unfortunately, resulted in more effective treatments for patients afflicted with such conditions. Glutamate NMDA receptor antagonists induce a psychotic state in healthy individuals similar to the psychosis observed in schizophrenia and are also widely used to induce psychotic symptoms in preclinical models of the disease. Taurine is an amino acid that acts as an inhibitory neuromodulator of the central nervous system with neuroprotective and antioxidant properties. In this work we investigated the potential of taurine to prevent deficits induced by acute administration of MK-801 (dizocilpine), an NMDA antagonist, in behavioral assays with relevance to schizophrenia in C57BL/6 mice and zebrafish. In rodent experiments, animals were injected intraperitoneally (i.p.) with either saline or taurine (50, 100 e 200 mg/kg), and 30 minutes later they received another i.p. injection of saline or MK-801 (0.15 mg/kg). The behavioral tests of prepulse inhibition of the startle response and social interaction were performed 30 minutes after the last injection, while locomotor activity was assessed continuously from the first injection, ending 60 minutes after the last one. For zebrafish experiments, the animals were placed in a beaker containing 200 mL of tank water or taurine (42, 150 or 400 mg/L) for 20 minutes, being subsequently exposed to tank water or MK-801 (5 μM) for another 20 minutes. The social interaction test was performed immediately after the end of the last exposure. As expected, MK-801 administration induced hyperlocomotion and prepulse inhibition deficits in rodents, whereas in zebrafish it induced hyperlocomotion and social interaction deficit. Interestingly, mice treated with MK-801 spent more time interacting with the stimulus animals; taurine at a dose of 50 mg/kg had the same effect, which may be related to its already documented anxiolytic effects. Preventive effects of taurine on behavioral changes acutely induced by MK- 801 were not observed in any of the species, contradicting previous evidence in the literature. Recent evidence has fostered the idea that the course of schizophrenia can be modified by intervention strategies initiated early, before the first psychotic break and the establishment of the full-blown disorder. Thus, it is a perspective of this work to evaluate early and continuous treatment with taurine in a developmental model of schizophrenia to better elucidate the preventive potential of this molecule.

6-Nitrodopamine (6-ND) is released by rat vas deferens and exerts a potent contractile response that is antagonized by tricyclic antidepressants and α

Cannabidiol (CBD) is a non-intoxicating compound extracted from Cannabis sativa, showing antidepressant-like effects in different rodent models. However, inconsistent results have been described depending on the species and the strain used to assess depressive-like behavior. Moreover, only a few studies investigated the effect of CBD in female rodents. Therefore, we aimed to (i) investigate the effects of CBD in two different strains of mice (Swiss and C57BL/6) and a rat model of depression based on selective breeding (Flinders Sensitive and Resistant Lines, FSL and FRL) subjected to tests predictive of antidepressant-like effects and (ii) investigate the influence of sex in the effects of CBD in both mice and rats. CBD induced an antidepressant-like effect in male Swiss but not in female Swiss or C57BL/6 mice in the tail suspension test (TST). In male FSL rats, CBD produced an antidepressant-like effect 1 h post injection. However, in female FSL, CBD induced a bimodal effect, increasing the immobility time at 1 h and decreasing it at 2 h. In conclusion, strain, sex, and administration time affect CBD’s behavioral response to rodents exposed to tests predictive of antidepressant effects.

Despite the significant increase in the generation of SARS-CoV-2 contaminated domestic and hospital wastewater, little is known about the ecotoxicological effects of the virus or its structural components in freshwater vertebrates. In this context, this study evaluated the deleterious effects caused by SARS-CoV-2 Spike protein on the health of Danio rerio, zebrafish. We demonstrated, for the first time, that zebrafish injected with fragment 16 to 165 (rSpike), which corresponds to the N-terminal portion of the protein, presented mortalities and adverse effects on liver, kidney, ovary and brain tissues. The conserved genetic homology between zebrafish and humans might be one of the reasons for the intense toxic effects followed inflammatory reaction from the immune system of zebrafish to rSpike which provoked damage to organs in a similar pattern as happen in severe cases of COVID-19 in humans, and, resulted in 78,6% of survival rate in female adults during the first seven days. The application of spike protein in zebrafish was highly toxic that is suitable for future studies to gather valuable information about ecotoxicological impacts, as well as vaccine responses and therapeutic approaches in human medicine. Therefore, besides representing an important tool to assess the harmful effects of SARS-CoV-2 in the aquatic environment, we present the zebrafish as an animal model for translational COVID-19 research., Graphical abstract Unlabelled Image

To analyze the consumption of oxygen and to quantify the mitochondrial respiratory chain proteins (OXPHOS) in the gastrocnemius muscle of rats exposed to cigarette smoke and/or RT practitioners.Wistar rats were divided into groups: Control (C), Smoker (S), Exercise (E) and Exercise Smoker (ES). Groups F and ES were exposed to the smoke of 4 cigarettes for 30 min, 2× a day, 5× a week, for 16 weeks. Groups E and ES performed four climbs with progressive load, 1× per day, 5× per week, for 16 weeks. The gastrocnemius muscle was collected for analysis of OXPHOS content and oxygen consumption. Groups S (vs. C) and ES (vs. C and E) showed lower body weight gain when observing the evolution curve.The S rats showed a reduction in the NDUFB8 proteins of complex 1, SDHB of complex 2, MTC01 of complex 4 and ATP5A of complex 5 (ATP Synthase) compared to Group C. Additionally, S rats also showed increased consumption of O2 in Basal, Leak, Complex I and I/II combined measures compared to the other groups, suggesting that the activity of the mitochondria of these animals increased in terms of coupling and uncoupling parameters.Our data suggest that exposure to cigarette smoke for 16 weeks is capable of causing impairment of mitochondrial function with reduced expression of respiratory chain proteins in skeletal muscle. However, the RT was effective in preventing impairment of mitochondrial function in the skeletal muscle of rats exposed to secondary cigarette smoke.

Propolis is a honeybee product with various biological activities, including antidiabetic effects. We previously reported that artepillin C, a prenylated cinnamic acid derivative isolated from Brazilian green propolis, acts as a peroxisome proliferator-activated receptor γ (PPARγ) ligand and promotes adipocyte differentiation. In this study, we examined the effect of baccharin, another major component of Brazilian green propolis, on adipocyte differentiation. The treatment of mouse 3T3-L1 preadipocytes with baccharin resulted in increased lipid accumulation, cellular triglyceride levels, glycerol-3-phosphate dehydrogenase activity, and glucose uptake. The mRNA expression levels of PPARγ and its target genes were also increased by baccharin treatment. Furthermore, baccharin enhanced PPARγ-dependent luciferase activity, suggesting that baccharin promotes adipocyte differentiation via PPARγ activation. In diabetic ob/ob mice, intraperitoneal administration of 50 mg/kg baccharin significantly improved blood glucose levels. Our results suggest that baccharin has a hypoglycemic effect on glucose metabolic disorders, such as type 2 diabetes mellitus.

Orientador: Marcus Alexandre Finzi Corat Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Ciências Médicas Resumo: A anemia falciforme é uma doença hereditária que acomete milhões de pessoas Promove destruição crônica das células vermelhas (eritrócitos) do sangue, causando anemia, vaso-oclusão, isquemia e consequentes sintomas deintensa dor, susceptibilidade à infecções e, em alguns casos, a morte precoce. Nas últimas décadas, estudos têm mostrado que o aumento dos níveis de hemoglobina fetal (HbF) inibe o processo de falcização das Hemácias melhorando os sintomas de pacientes com anemia falciforme. Neste estudo, produzimos um modelo animal humanizado com traço falciforme e expressão concomitante de HbF persistente, associando as características genéticas dos camundongos transgênicos B-HPFH195 e Berk-SCM . O primeiro é animal transgênico com persistência de hemoglobina fetal, onde aprodução de HbF continua após o nascimento. A segunda linhagem corresponde ao modelo para doença falciforme, Hbatm1Paz Hbbtm1Tow Tg (HBA - HBBS ) 41Paz / J ( Berk - SCM) e mimetiza a doença humana em sua forma mais grave. Nestes animais a reprodução está comprometida, as fêmeas não levam a gestação ao termino e a maioria dos neonatos morrem horas depois do nascimento. Isto ocorre principalmente devido ao fato de que a (HbF) presente no transgene humano destes animais ser trocada pela HbS enquanto o feto ainda esta em gestação. O novo modelo desenvolvido foi analisado clinicamente e histologicamente para a caracterização e apresentou em sua constituição sanguínea hemoglobinas totalmente humanizadas com cadeias ?, ?s e ? persistente porém, ainda mantendo alterações histopatológicas semelhantes aos modelos da doença falciforme. Este novo animal representa um modelo potencial para uso em testes de drogas no aumento de HbF onde os resultados podem ser relevantes para o estudo e tratamento de hemoglobinopatias, bem como para compreender o comportamento das mutações hereditárias para a expressão persistente da cadeia de globina Y Abstract: Sickle cell disease is a hereditary disease that affects millions of people promotes chronic destruction of red blood cells (erythrocytes), causing anemia, vascular occlusion, ischemia, pain, and susceptibility to infections and in some instances, death premature. In recent decades, studies have shown that increased HbF levels play an important role in improving the symptoms of patients with sickle cell anemia. In this study, we produced and characterized a humanized animal model with sickle cell trait and concomitant expression of persistent HbF, using the genetic characteristics of transgenic mice B - HPFH195 with Berk¿SCM. This model is a transgenic mice model with hereditary persistence of HbF. The second strain constitutes a mouse with sickle cell disease, Hbatm1Paz Hbbtm1Tow Tg ( HBA - HBBs ) 41Paz /J ( Berk - SCM ), an animal model used to study SCD with human hemoglobin S ( HbS ) in its blood. The Berkeley mouse mimics the human disease in severe form, which compromises in its, females do not lead to pregnancy termination and most newborns die hours after birth. This occurs mainly due to the fact that fetal hemoglobin (HbF), present in the human transgene of these animals,is exchanged to HbS while the fetus still in gestation. The new model produced was analyzed clinically and histologically for proper characterization and presents in its constitution humanized blood, hemoglobin chains with ?, ? ?s and persistent, but still maintains histopathological changes similar to models of sickle cell disease. This model may be a potential model for use in drug in testing increase HbF. And results may be relevant for the study and treatment of hemoglobinopathies as well as for understanding the behavior of hereditary mutations for the persistent expression of the Y globin chain Mestrado Clínica Médica Mestra em Clínica Médica

High plasma angiotensin II (Ang II) levels are related to many diseases, including hypertension, and chronic kidney diseases (CKDs). Here, we investigated the relationship among prolonged Ang II infusion/AT1 receptor (AT1R) activation, oxidative stress, and endoplasmic reticulum (ER) stress in kidney tissue. In addition, we explored the chronic effects of Ang II on tubular Na+transport mechanisms. Male Wistar rats were subjected to sham surgery as a control or prolonged Ang II treatment (200 ng⋅kg–1⋅min–1, 42 days) with or without losartan (10 mg⋅kg–1⋅day–1) for 14 days. Ang II/AT1R induced hypertension with a systolic blood pressure of 173.0 ± 20 mmHg (mmHg,n= 9) compared with 108.0 ± 7 mmHg (mmHg,n= 7) in sham animals. Under these conditions, gene and protein expression levels were evaluated. Prolonged Ang II administration/AT1R activation induced oxidative stress and ER stress with increased Nox2, Nox4,CybaandNcf1mRNA expression, phosphorylated PERK and eIF2α protein expression as well asAtf4mRNA expression. Ang II/AT1R also raisedIl1b, Nfkb1andActa2mRNA expression, suggesting proinflammatory, and profibrotic effects. Regarding Na+tubular handling, Ang II/AT1R enhanced cortical non-phosphorylated and phospho/S552/NHE3, NHE1, ENaC β, NKCC2, and NCC protein expression. Our results also highlight the therapeutic potential of losartan, which goes beyond the antihypertensive effect, playing an important role in kidney tissue. This treatment reduced oxidative stress and ER stress signals and recovered relevant parameters of the maintenance of renal function, preventing the progression of Ang II-induced CKD.

Objective: To identify and describe comparativelythe chemical models of the induction of inflammatory bowel diseases (IBD) in rodents most used and that best mimic the pathogenesis in humans.Methods: Based on an integrative review in the MEDLINE and LILACS databases, it was investigated which experimental induction models were most cited in articles published from 2004 to 2020, with the descriptors "Colitis/CI", "Colitis model ulcerative" and "Intestinal inflammation model."All empirical articles that addressed one or more inflammation models in rats or mice were included.Results: 239 articles were identified; of these, only ten empirical articles were selected.The most used models were colitis induced by TNBS acid, DSS, and colitis induced by acetic acid (AA).Conclusion: It was possible to identify the most used models to promote the induction of intestinal inflammation in rats, and both models proved to be effective according to the limitations observed in the models described, suggesting the need for new works that use more well-defined protocols and that more fully represent the pathophysiological complexity of the disease. Objetivo: Identificar e descrever de forma comparativa os modelos químicos de indução das doenças inflamatórias intestinais (DII) em roedores mais utilizados e que melhor mimetizam a patogênese em humanos. Métodos: a partir de uma revisão integrativa nas bases de dados MEDLINE e LILACS, investigou-se quais os modelos de indução experimental mais citados nos artigos publicados no período de 2004 a 2020, com os descritores “Colite/CI", “Modelo de colite ulcerativa” e “Modelo de inflamação intestinal”. Foram incluídos todos os artigos empíricos que abordassem um ou mais modelos de inflamação em ratos ou camundongos. Resultados: 239 artigos foram identificados; destes, somente dez artigos empíricos foram selecionados. Os modelos mais utilizados foram o de colite induzida por ácido TNBS, por DSS e colite induzida por ácido acético (AA). Conclusão: Foi possivel identificar os modelos mais utilizados para promover a indução da inflamação intestinal em ratos e ambos os modelos se mostraram eficazes de acordo com seu protocolo de indução. Ficaram claras as limitações observadas nos modelos já descritos, sugerindo a necessidade de novos trabalhos que utilizem protocolos mais bem definidos e que representem de forma mais integral a complexidade fisiopatológica da doença.

Background Prenatal exposure to Cannabis is a worldwide growing problem. Although retina is part of the central nervous system, the impact of maternal Cannabis use on the retinal development and its postnatal consequences remains unknown. As the prenatal period is potentially sensitive in the normal development of the retina, we hypothesized that recreational use of Cannabis during pregnancy may alter retina structure in the offspring. To test this, we developed a murine model that mimics human exposure in terms of dose and use. Methods Pregnant BalbC mice were exposed daily for 5 min to Cannabis smoke (0.2 g of Cannabis) or filtered air, from gestational day 5 to 18 (N = 10/group). After weaning period, pups were separated and examined weekly. On days 60, 120, 200, and 360 after birth, 10 pups from each group were randomly selected for Spectral Domain Optical Coherence Tomography (SD-OCT) analysis of the retina. All retina layers were measured and inner, outer, and total retina thickness were calculated. Other 37 mice from both groups were sacrificed on days 20, 60, and 360 for retinal stereology (total volume of the retina and volume fraction of each retinal layer) and light microscopy. Means and standard deviations were calculated and MANOVA was performed. Results The retina of animals which mother was exposed to Cannabis during gestation was 17% thinner on day 120 (young adult) than controls (P = 0.003) due to 21% thinning of the outer retina (P = 0.001). The offspring of mice from the exposed group presented thickening of the IS/OS in comparison to controls on day 200 (P Conclusion Gestational exposure to Cannabis smoke may cause structural changes in the retina of the offspring that return to normal on mice adulthood. These experimental evidences suggest that children and young adults whose mothers smoked Cannabis during pregnancy may require earlier and more frequent clinical care than the non-exposed population.

There is a strong correlation between obesity and cancer. Here, we investigated the influence of IL-6 and gut microbiota of obese mice in melanoma development. We first evaluated B16F10 melanoma growth in preclinical models for obesity: mice deficient for leptin

Melanoma skin cancer is extremely aggressive with increasing incidence and mortality. Among the emerging therapeutic targets in the treatment of cancer, the family of transient receptor potential channels (TRPs) has been reported as a possible pharmacological target. Specifically, the ankyrin subfamily, representing TRPA1 channels, can act as a pro-inflammatory hub. These channels have already been implicated in the control of intracellular metabolism in several cell models, but little is known about their role in immune cells, and how it could affect tumor progression in a process known as immune surveillance. Here, we investigated the participation of the TRPA1 channel in the immune response against melanoma tumor progression in a mouse model. Using Trpa1+/+ and Trpa1-/- animals, we evaluated tumor progression using murine B16-F10 cells and assessed isolated CD8+ T cells for respiratory and cytotoxic functions. Tumor growth was significantly reduced in Trpa1-/- animals. We observed an increase in the frequency of circulating lymphocytes. Using a dataset of CD8+ T cells isolated from metastatic melanoma patients, we found that TRPA1 reduction correlates with several immunological pathways. Naïve CD8+ T cells from Trpa1+/+ and Trpa1-/- animals showed different mitochondrial respiration and glycolysis profiles. However, under CD3/CD28 costimulatory conditions, the absence of TRPA1 led to an even more extensive metabolic shift, probably linked to a greater in vitro killling ability of Trpa1-/- CD8+ T cells. Therefore, these data demonstrate an unprecedented role of TRPA1 channel in the metabolism control of the immune system cells during carcinogenesis.

Immunotherapy has improved patient survival in many types of cancer, but for prostate cancer, initial results with immunotherapy have been disappointing. Prostate cancer is considered an immunologically excluded or cold tumor, unable to generate an effective T-cell response against cancer cells. However, a small but significant percentage of patients do respond to immunotherapy, suggesting that some specific molecular subtypes of this tumor may have a better response to checkpoint inhibitors. Recent findings suggest that, in addition to their function as cancer genes, somatic mutations of PTEN, TP53, RB1, CDK12, and DNA repair, or specific activation of regulatory pathways, such as ETS or MYC, may also facilitate immune evasion of the host response against cancer. This review presents an update of recent discoveries about the role that the common somatic mutations can play in changing the tumor microenvironment and immune response against prostate cancer. We describe how detailed molecular genetic analyses of the tumor microenvironment of prostate cancer using mouse models and human tumors are providing new insights into the cell types and pathways mediating immune responses. These analyses are helping researchers to design drug combinations that are more likely to target the molecular and immunological pathways that underlie treatment failure.

OBJECTIVES: Ischemia and reperfusion (I/R) in the intestine could lead to severe endothelial injury, compromising intestinal motility. Reportedly, estradiol can control local and systemic inflammation induced by I/R injury. Thus, we investigated the effects of estradiol treatment on local repercussions in an intestinal I/R model. METHODS: Rats were subjected to ischemia via the occlusion of the superior mesenteric artery (45 min) followed by reperfusion (2h). Thirty minutes after ischemia induction (E30), 17β-estradiol (E2) was administered as a single dose (280 μg/kg, intravenous). Sham-operated animals were used as controls. RESULTS: I/R injury decreased intestinal motility and increased intestinal permeability, accompanied by reduced mesenteric endothelial nitric oxide synthase (eNOS) and endothelin (ET) protein expression. Additionally, the levels of serum injury markers and inflammatory mediators were elevated. Estradiol treatment improved intestinal motility, reduced intestinal permeability, and increased eNOS and ET expression. Levels of injury markers and inflammatory mediators were also reduced following estradiol treatment. CONCLUSION: Collectively, our findings indicate that estradiol treatment can modulate the deleterious intestinal effects of I/R injury. Thus, estradiol mediates the improvement in gut barrier functions and prevents intestinal dysfunction, which may reduce the systemic inflammatory response.

Alcohol use disorder is one of the most prevalent addictions, strongly influenced by environmental factors. Voluntary physical activity (VPA) has proven to be intrinsically reinforcing and we hypothesized that, as a non-drug reinforcer, VPA could mitigate ethanol-induced rewarding effects. The transcriptional factor cAMP response element binding protein (CREB), and deacetylases isozymes sirtuins 1 and 2 (SIRT-1 and SIRT-2) have a complex interplay and both play a role in the rewarding effects of ethanol. To test whether the exposure of mice to running wheels inhibits the development of ethanol-induced conditioned place preference (CPP), mice were assigned into four groups: housed in home cages with locked (“Sedentary”) or unlocked running wheels (VPA), and treated with saline or 1.8 g/kg ethanol during the conditioning phase. The groups were referred as Saline-Sedentary, Saline-VPA, Ethanol-Sedentary and Ethanol-VPA. The expression of CREB, SIRT-1 and SIRT-2 were evaluated in the prefrontal cortex (PFC) and nucleus accumbens (NAc). VPA prevented the development of ethanol-induced CPP. VPA, ethanol and the combination of both inhibited pCREB and pCREB/CREB ratio in the NAc, suggesting that both reward stimuli can share similar patterns of CREB activation. However, we have found that ethanol-induced increased CREB levels were prevented by VPA. Both VPA groups presented lower SIRT-1 levels in the NAc compared to the Sedentary groups. Thus, exposure to running wheels prevented ethanol-rewarding effects and ethanol-induced increases in CREB in the NAc. The molecular alterations underlying CPP prevention may be related to a lower expression of CREB in the NAc of Ethanol-VPA compared to the respective Sedentary group, given the positive correlation between CPP and CREB levels in the Ethanol-Sedentary group.