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Mutations in the mitochondrial genome (mtDNA) are ubiquitous in humans and can lead to a broad spectrum of disorders. However, due to the presence of multiple mtDNA molecules in the cell, co-existence of mutant and wild-type mtDNAs (termed heteroplasmy) can mask disease phenotype unless a threshold of mutant molecules is reached. Importantly, the mutant mtDNA level can change across lifespan as mtDNA segregates in an allele- and cell-specific fashion, potentially leading to disease. Segregation of mtDNA is mainly evident in hepatic cells, resulting in an age-dependent increase of mtDNA variants, including non-synonymous potentially deleterious mutations. Here we modeled mtDNA segregation using a well-established heteroplasmic mouse line with mtDNA of NZB/BINJ and C57BL/6N origin on a C57BL/6N nuclear background. This mouse line showed a pronounced age-dependent NZB mtDNA accumulation in the liver, thus leading to enhanced respiration capacity per mtDNA molecule. Remarkably, liver-specific atg7 (autophagy related 7) knockout abolished NZB mtDNA accumulat ion, resulting in close-to-neutral mtDNA segregation through development into adulthood. prkn (parkin RBR E3 ubiquitin protein ligase) knockout also partially prevented NZB mtDNA accumulation in the liver, but to a lesser extent. Hence, we propose that age-related liver mtDNA segregation is a consequence of macroautophagic clearance of the less-fit mtDNA. Considering that NZB/BINJ and C57BL/6N mtDNAs have a level of divergence comparable to that between human Eurasian and African mtDNAs, these findings have potential implications for humans, including the safe use of mitochondrial replacement therapy.Abbreviations: Apob: apolipoprotein B; Atg1: autophagy-related 1; Atg7: autophagy related 7; Atp5a1: ATP synthase, H+ transporting, mitochondrial F1 complex, alpha subunit 1; BL6: C57BL/6N mouse strain; BNIP3: BCL2/adenovirus E1B interacting protein 3; FCCP: carbonyl cyanide 4-(trifluoromethoxy)phenylhydrazone; GAPDH: glyceraldehyde-3-phosphate dehydrogenase; MAP1LC3A: microtubule-associated protein 1 light chain 3 alpha; MAP1LC3B: microtubule-associated protein 1 light chain 3 beta; mt-Atp8: mitochondrially encoded ATP synthase 8; MT-CO1: mitochondrially encoded cytochrome c oxidase I; MT-CO2: mitochondrially encoded cytochrome c oxidase II; mt-Co3: mitochondrially encoded cytochrome c oxidase III; mt-Cytb: mitochondrially encoded cytochrome b; mtDNA: mitochondrial DNA; MUL1: mitochondrial ubiquitin ligase activator of NFKB 1; nDNA: nuclear DNA; Ndufa9: NADH:ubiquinone oxireductase subunit A9; NDUFB8: NADH:ubiquinone oxireductase subunit B8; Nnt: nicotinamide nucleotide transhydrogenase; NZB: NZB/BINJ mouse strain; OXPHOS: oxidative phosphorylation; PINK1: PTEN induced putative kinase 1; Polg2: polymerase (DNA directed), gamma 2, accessory subunit; Ppara: peroxisome proliferator activated receptor alpha; Ppia: peptidylprolyl isomerase A; Prkn: parkin RBR E3 ubiquitin protein ligase; P10: post-natal day 10; P21: post-natal day 21; P100: post-natal day 100; qPCR: quantitative polymerase chain reaction; Rpl19: ribosomal protein L19; Rps18: ribosomal protein S18; SD: standard deviation; SEM: standard error of the mean; SDHB: succinate dehydrogenase complex, subunit B, iron sulfur (Ip); SQSTM1: sequestosome 1; Ssbp1: single-stranded DNA binding protein 1; TFAM: transcription factor A, mitochondrial; Tfb1m: transcription factor B1, mitochondrial; Tfb2m: transcription factor B2, mitochondrial; TOMM20: translocase of outer mitochondrial membrane 20; UQCRC2: ubiquinol cytochrome c reductase core protein 2; WT: wild-type.

Transient global ischemia is a leading cause of learning and memory dysfunction and induces a pattern of delayed neuronal death in the CA1 subfield of the hippocampus by down-regulating GluR2 mRNA AMPA receptors in this cerebral area. This study sought to investigate the neuroprotective effect of coumestrol against spatial memory impairment induced by global ischemia that leads to neural death by reducing the GluR2 receptors content in the hippocampal CA1 area. Our studies demonstrated that coumestrol administration prevented spatial memory deficits in mice. These findings suggest a cognitive enhancement role of coumestrol against cognitive impairment in ischemic events.

One of the consequences of the Western lifestyle and high-fat diet is non-alcoholic fatty liver disease (NAFLD) and its aggressive form, non-alcoholic steatohepatitis (NASH), which can progress to cirrhosis and hepatocellular carcinoma (HCC) and is rapidly becoming the leading cause of end-stage liver disease or liver transplantation. Currently, rodent NASH models lack significant aspects of the full NASH spectrum, representing a major problem for NASH research. Therefore, this work aimed to characterize a fast rodent model with all characteristic features of NASH. Eight-week-old male ApoE KO mice were fed with Western diet (WD), high fatty diet (HFD) or normal chow (Chow) for 7 weeks. Whole-body fat was increased by ~2 times in WD mice and HFD mice and was associated with increased glucose intolerance, hepatic triglycerides, and plasma ALT and plasma AST compared with Chow mice. WD mice also showed increased galectin-3 expression compared with Chow or HFD mice and increased plasma cholesterol compared with Chow mice. WD and HFD displayed increased hepatic fibrosis and increased F4/80 expression. WD mice also displayed increased levels of plasma MCP-1. Hepatic inflammatory markers were evaluated, and WD mice showed increased levels of TNF-α, MCP-1, IL-6 and IFN-γ. Taken together, these data demonstrated that the ApoE KO mouse fed with WD is a great model for NASH research, once it presents the fundamental parameters of the disease, including hepatic steatosis, fibrosis, inflammation, and metabolic syndrome.

Background The activation of the hypothalamic–pituitary–adrenal (HPA) axis is essential for metabolic adaptation in response to fasting. However, the neurocircuitry connecting changes in the peripheral energy stores to the activity of hypothalamic paraventricular corticotrophin-releasing factor (CRFPVN) neurons, the master controller of the HPA axis activity, is not completely understood. Our main goal was to determine if hypothalamic arcuate nucleus (ARC) POMC and AgRP neurons can communicate fasting-induced changes in peripheral energy stores, associated to a fall in plasma leptin levels, to CRFPVN neurons to modulate the HPA axis activity in mice. Results We observed increased plasma corticosterone levels associate with increased CRFPVN mRNA expression and increased CRFPVN neuronal activity in 36 h fasted mice. These responses were associated with a fall in plasma leptin levels and changes in the mRNA expression of Agrp and Pomc in the ARC. Fasting-induced decrease in plasma leptin partially modulated these responses through a change in the activity of ARC neurons. The chemogenetic activation of POMCARC by DREADDs did not affect fasting-induced activation of the HPA axis. DREADDs inhibition of AgRPARC neurons reduced the content of CRFPVN and increased its accumulation in the median eminence but had no effect on corticosterone secretion induced by fasting. Conclusion Our data indicate that AgRPARC neurons are part of the neurocircuitry involved in the coupling of PVNCRF activity to changes in peripheral energy stores induced by prolonged fasting.

Objective: the present study aims to evidence the most recurrent muscle injury techniques in the literature. Methodology: This is a bibliographical research of the integrative review type of literature. The articles were searched in 2021, from November 22 to 24, in the PubMed database, using the eligibility criteria. Results and Discussion: According to the search strategy used in this study, of all articles only 28 studies met the eligibility criteria. When analyzing the publication journals, it was observed that the most recurrent were PLoS One (7.14%), Int J Med Sci (7.14%), and J Trauma Acute Care with (7.14%). It was also evidenced that the number of publications on the subject has been growing over the years, when it compared the year 2016 (10.71%) with later years, except in 2019 with the same percentage of 10.71% and 2021 with zero publication. The breeds most used in the experiments were Sprague-Dawley (32.14%) and Wistar with 25%. There was a predominance of models due to contusion (35.71%), followed by excessive use injury (10.71%), and traumatic injury (10.71%), for induction of muscle injury in rodents. Conclusion: According to the results of this review, the most recurrent muscle injury induction models were injury by contusion, followed by excessive use injury, and traumatic injury. However, all the techniques addressed in the present study were able to reproduce with excellence the mechanism of muscle injury. Objetivo: el presente estudio tiene como objetivo destacar las técnicas de lesión muscular más recurrentes en la literatura. Metodología: Se trata de una investigación bibliográfica del tipo revisión integradora de literatura. Los artículos fueron buscados en el año 2021, del 22 al 24 de noviembre, en la base de datos PubMed, utilizando los criterios de elegibilidad. Resultados y Discusión: De acuerdo con la estrategia de búsqueda utilizada en este estudio, de todos los artículos, solo 28 estudios cumplieron con los criterios de elegibilidad. Al analizar las revistas de publicación, se observa que las más recurrentes fueron PLoS One (7,14%), Int J Med Sci (7,14%) y J Trauma Acute Care with (7,14%). También se evidenció que el número de publicaciones sobre el tema ha ido creciendo a lo largo de los años, al comparar el año 2016 (10,71%) con años posteriores, excepto en el 2019 con el mismo porcentaje de 10,71% y el 2021 con cero publicaciones. Las razas más utilizadas en los experimentos fueron Sprague-Dawley (32,14%) y Wistar con un 25%. Predominaron los modelos de contusión (35,71%), seguido de lesión por sobreuso (10,71%) y lesión traumática (10,71%), por inducir lesión muscular en roedores. Conclusión: Según los resultados de esta revisión, los modelos de inducción de lesiones musculares más recurrentes fueron las lesiones por contusión, seguidas de las lesiones por uso excesivo y las lesiones traumáticas. Sin embargo, todas las técnicas discutidas en el presente estudio pudieron reproducir con excelencia el mecanismo de lesión muscular. Objetivo: O presente estudo tem como objetivo evidenciar as técnicas de lesão muscular mais recorrentes na literatura. Metodologia: Trata-se de uma pesquisa bibliográfica do tipo revisão integrativa da literatura. Os artigos foram pesquisados no ano de 2021, no período de 22 a 24 de novembro, na base de dados PubMed, usando os critérios de elegibilidade. Resultados e Discussão: De acordo com a estratégia de busca utilizada neste estudo, da totalidade dos artigos apenas 28 estudos atenderam os critérios de elegibilidade. Ao analisar as revistas de publicação, observa-se que as mais recorrentes foram PLoS One (7.14%), Int J Med Sci (7.14%), e J Trauma Acute Care com (7.14%). Evidenciou-se também, que o número de publicações sobre a temática vem crescendo ao longo dos anos, quando compara o ano de 2016 (10.71%) com anos posteriores, exceto em 2019 com o mesmo percentual de 10.71% e 2021 com zero publicação. As raças mais utilizadas nos experimentos foram Sprague-Dawley (32.14%) e Wistar com 25%. Houve uma predominância dos modelos por contusão (35.71%), seguido da lesão por uso excessivo (10.71%), e por lesão traumática (10.71%), para indução de lesão muscular em roedores. Conclusão: Conforme os resultados desta revisão, os modelos de indução de lesão muscular mais recorrente foi a lesão por contusão, seguido da lesão por uso excessivo, e por lesão traumática. Porém, todas as técnicas abordadas no presente estudo conseguiram reproduzir com excelência o mecanismo de injúria muscular.

Obesity is mainly caused by excess energy intake and physical inactivity, and the number of overweight/obese individuals has been steadily increasing for decades. Previous studies showed that rodents fed westernized diets exhibit endocrine pancreas deterioration and a range of metabolic disorders. This study evaluated the effects of moderated aerobic treadmill exercise training on pancreatic islet cell viability and function in mice consuming a high-fat and sucrose diet. In the present study, 60-day-old male C57BL/6J mice were divided into four groups: control (C), fed a standard diet AIN-93M (3.83 kcal/g; 70% carbohydrate (cornstarch and dextrinized starch were chosen as the major source of carbohydrate for the AIN-93 diet. In addition, a small amount of sucrose), 20% protein (casein), and 10% fat (soybean) with no training (i.e., sedentary); C + training (CTR, fed the standard diet with eight weeks of exercise; high-fat diet + sucrose (HFDS), fed a high fat and sucrose diet (5.2 kcal/g; 20% carbohydrate (cornstarch and dextrinized starch were chosen as the major source of carbohydrate), 20% protein (casein), 60% fat (Lard was chosen as the major source of fat and a small amount of soybean) + 20% sucrose diluted in drinking water with no training; and HFDS + training (HFDSTR). After eight weeks, the HFDS mice displayed increased body weight (P

Hypophysiotropic somatostatin (SST) neurons in the periventricular hypothalamic area express growth hormone (GH) receptor (GHR) and are frequently considered as the key neuronal population that mediates the negative feedback loop controlling the hypothalamic–GH axis. Additionally, insulin-like growth factor-1 (IGF-1) may also act at the hypothalamic level to control pituitary GH secretion via long-loop negative feedback. However, to the best of our knowledge, no study so far has tested whether GHR or IGF-1 receptor (IGF1R) signaling specifically in SST neurons is required for the homeostatic control of GH secretion. Here we show that GHR ablation in SST neurons did not impact the negative feedback mechanisms that control pulsatile GH secretion or body growth in male and female mice. The sex difference in hepatic gene expression profile was only mildly affected by GHR ablation in SST neurons. Similarly, IGF1R ablation in SST neurons did not affect pulsatile GH secretion, body growth, or hepatic gene expression. In contrast, simultaneous ablation of both GHR and IGF1R in SST-expressing cells increased mean GH levels and pulse amplitude in male and female mice, and partially disrupted the sex differences in hepatic gene expression. Despite the increased GH secretion in double knockout mice, no alterations in body growth and serum or liver IGF-1 levels were observed. In summary, GHR and IGF1R signaling in SST neurons play a redundant role in the control of GH secretion. Furthermore, our results reveal the importance of GH/IGF-1 negative feedback mechanisms on SST neurons for the establishment of sex differences in hepatic gene expression profile.

O c?ncer de mama apresenta a maior incid?ncia mundial entre todos os tipos de tumores. Os tratamentos s?o direcionados conforme o subtipo molecular: luminais A e B, HER2+ e triplo negativo. Apesar das taxas de sucesso para os tipos hormonais e HER2+, os pacientes que apresentam recidivas para os tumores mais agressivos ou triplos negativos permanecem com baixa sobrevida geral e o progn?stico ? desfavor?vel devido, por exemplo, ? aus?ncia de um alvo terap?utico ou elevada agressividade. Ademais, altera??es na composi??o da microbiota e, consequentemente, dos ?cidos graxos de cadeia curta (SCFA) ? atrav?s das condi??es cl?nicas basais ou por efeitos colaterais dos tratamentos ? pode afetar o risco e o progn?stico para o c?ncer de mama. Neste sentido, este estudo avaliou os efeitos dos SCFA em diferentes modelos de c?ncer de mama com o prop?sito de identificar uma nova abordagem para o tratamento desses tumores. Os ensaios in vitro utilizando as linhagens celulares de c?ncer de mama de diferentes subtipos moleculares, humanas, MCF-7, SK-BR-3 e MDA-MB-231; ou murina, 4T1 exibiram efeitos contrastantes ap?s os tratamentos com os SCFA. Na an?lise da viabilidade celular, o acetato apresentou efeitos positivos em concentra??es acima de 100 mM. Enquanto o propionato apresentou efeito dual dependente da concentra??o. De maneira tempo e concentra??o-dependente, o butirato, propionato e valerato reduziram significativamente a viabilidade celular das linhagens humanas e, o valerato reduziu a viabilidade celular da linhagem murina sob o IC50 de 9,6 mM em 48 h. Por outro lado, os agonistas seletivos de FFA2 e FFA3, 4-CMTB e AR420626, respectivamente, reduziram modestamente a viabilidade celular apenas das linhagens mais agressivas MDA-MB-231 e 4T1. Da mesma forma que na avalia??o dos antagonistas de FFA2 e FFA3, CATPB e ?-hidroxibutirato, respectivamente, somente o CATPB produziu efeitos inibit?rios na linhagem 4T1. Esses dados sugerem que os SCFA apresentam efeitos antitumorais no c?ncer de mama provavelmente por mecanismos independentes ? liga??o aos receptores FFA2 e FFA3. A partir desse screening, os SCFA com melhores efeitos inibit?rios, butirato, propionato e valerato, foram selecionados para investigar seus efeitos na capacidade de malignidade nas c?lulas MDA-MB-231 e 4T1. Altera??es significativas na morfologia celular foram observadas, assim como na redu??o da forma??o de novas col?nias. Na avalia??o da ades?o celular, o valerato reduziu este par?metro na linhagem MDA-MB231, no entanto, aumentou em rela??o ? linhagem 4T1. Em rela??o ? avalia??o da migra??o celular, somente o tratamento com butirato (10 mM) preveniu a migra??o da linhagem 4T1 em 24 h. Apesar de discretos, estes resultados corroboram os efeitos antitumorais observados na redu??o da forma??o de col?nias e altera??es da morfologia celular ap?s a exposi??o com o butirato na linhagem 4T1, principalmente. Para investigar os efeitos desses SCFA na progress?o tumoral foi utilizado o modelo ortot?pico de c?ncer de mama metast?tico em camundongos f?mea Balb/CJ induzido pela inocula??o das c?lulas 4T1. Os resultados preliminares da administra??o por via oral dos SCFA tanto no protocolo terap?utico quanto no preventivo n?o demonstraram um efeito espelhado de acordo com a avalia??o in vitro, exceto pelo n?mero de met?stases pulmonares que foram significativamente reduzidas pelo butirato (600 mg/kg) no esquema terap?utico. Essas evid?ncias sugerem que a participa??o do microambiente tumoral est? envolvida na a??o dos SCFA, assim como seus efeitos celulares podem ser modulados por mecanismos alternativos aos receptores FFA2 e FFA3, como pela inibi??o de HDAC. Portanto, para estabelecer um tratamento promissor, a defini??o desses mecanismos pode contribuir na determina??o dos SCFA como um alvo terap?utico no manejo do c?ncer de mama. Breast cancer has the highest worldwide incidence of all types of tumors. Treatments are defined according to molecular subtypes: luminal A and B, HER2+, and triple-negative. Despite the success rates for hormonal and HER2+ types, patients who relapse for the most aggressive or triple-negative tumors remain with low overall survival and the prognosis is poor due, for example, to the absence of a therapeutic target or high aggressiveness. Furthermore, alterations in the microbiota composition, consequently, in short-chain fatty acids (SCFA) ? through baseline clinical conditions or side effects of treatments ? might affect the risk and prognosis for breast cancer. In this sense, this study evaluated the effects of SCFA in different models of breast cancer to identify a new approach for the treatment of these tumors. In vitro assays using breast cancer cell lines of different molecular subtypes, human (MCF-7, SK-BR-3, and MDA-MB-231) or murine (4T1) exhibited contrasting effects after treatments with SCFA. In the analysis of cell viability, acetate showed positive effects at concentrations above 100 mM. While propionate demonstrated a dual concentration-dependent effect. In a time and concentration-dependent manner, butyrate, propionate, and valerate significantly reduced the cell viability of human cells, and valerate reduced the viability of the murine cells under an IC50 of 9.6 mM in 48 h. On the other hand, selective FFA2 and FFA3 agonists 4-CMTB and AR420626, respectively, modestly reduced cell viability only of the more aggressive MDA-MB-231 and 4T1 cell lines. As in the evaluation of the FFA2 and FFA3 antagonists CATPB and ?hydroxybutyrate, respectively, only CATPB produced inhibitory effects in the 4T1 cells. These data suggest that SCFA have antitumor effects in breast cancer, probably through FFA2 and FFA3 receptor independent mechanisms. From this screening, SCFAs with best inhibitory effects, butyrate, propionate, and valerate, were selected to investigate their effects on the malignancy capacity of MDAMB-231 and 4T1 cells. Several changes in cell morphology were observed, as well as a reduction in the new colonies formation. In the evaluation of cell adhesion, valerate reduced this parameter in the MDA-MB-231 cells, however, it increased about the 4T1 cells. Regarding the evaluation of cell migration, only the treatment with butyrate (10 mM) prevented the migration of the 4T1 cells within 24 h. Although discreet, these results corroborate the antitumor effects observed in the reduction of colony formation and changes in cell morphology after exposure with butyrate in the 4T1 cells, mainly. To investigate the effects of these SCFAs on tumor progression, was used the orthotopic model of metastatic breast cancer in female Balb/CJ mice induced by inoculation of 4T1 cells. The preliminary results of oral administration of SCFA in both the therapeutic and preventive protocols did not demonstrate a mirror effect according to the in vitro evaluation, except for the number of lung metastases that were significantly reduced by butyrate (600 mg/kg) in the therapeutic scheme. These evidence suggests that the participation of the tumor microenvironment is involved in the action of SCFA, as well as their cellular effects can be modulated by alternative mechanisms to the FFA2 and FFA3 receptors, such as the HDACs inhibition. Therefore, to establish a promising treatment, the definition of these mechanisms might contribute to the determination of SCFA as a therapeutic target in the management of breast cancer. Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior - CAPES

The present study aims to compare the responses between male and female C57BL/6 mice to multiple metabolic challenges to understand the importance of sex in the control of energy homeostasis.Male and female C57BL/6 mice were subjected to nutritional and hormonal challenges, such as food restriction and refeeding, diet-induced obesity, feeding response to ghrelin and leptin, ghrelin-induced growth hormone secretion, and central responsiveness to ghrelin and leptin. The hypothalamic expression of transcripts that control energy homeostasis was also evaluated.Male mice lost more weight and lean body mass in response to food restriction, compared to females. During refeeding, males accumulated more body fat and exhibited lower energy expenditure and glycemia, as compared to females. Additionally, female mice exhibited a higher protection against diet-induced obesity and related metabolic imbalances in comparison to males. Low dose ghrelin injection elicited higher food intake and growth hormone secretion in male mice, whereas the acute anorexigenic effect of leptin was more robust in females. However, the sex differences in the feeding responses to ghrelin and leptin were not explained by variations in the central responsiveness to these hormones nor by differences in the fiber density from arcuate nucleus neurons. Female, but not male, mice exhibited compensatory increases in hypothalamic Pomc mRNA levels in response to diet-induced obesity.Our findings revealed several sexually differentiated responses to metabolic challenges in C57BL/6 mice, highlighting the importance of taking into account sex differences in metabolic studies.

Submitted by Vitor Barbosa Fialho Martins (vitor.fialho@unesp.br) on 2022-05-03T10:11:00Z No. of bitstreams: 1 Dissertação Mestrado - Vitor_Martins_Definitivo_.pdf: 2604031 bytes, checksum: dfd722070533b517c3b05f37fcfbc203 (MD5) Approved for entry into archive by Laudicélia Martins Arantes (lm.arantes@unesp.br) on 2022-05-03T12:06:23Z (GMT) No. of bitstreams: 1 martins_vbf_me_jabo.pdf: 2604031 bytes, checksum: dfd722070533b517c3b05f37fcfbc203 (MD5) Made available in DSpace on 2022-05-03T12:06:23Z (GMT). No. of bitstreams: 1 martins_vbf_me_jabo.pdf: 2604031 bytes, checksum: dfd722070533b517c3b05f37fcfbc203 (MD5) Previous issue date: 2022-03-04 O circovirus suíno Tipo 2 (PCV2) e as doenças associadas a ele (PCVAD) geram grandes perdas para a suinocultura mundial. Além de outras medidas de prevenção para reduzir a disseminação dessa doença infecciosa, a vacinação é um dos meios mais eficientes de se controlar a circovirose nos rebanhos suínos. Há diversos tipos de vacinas comercialmente disponíveis contra o circovirus, sendo que estas devem ser atualizadas com base na evolução que os genótipos prevalentes de PCV2 têm sofrido ao longo dos anos. Assim, o presente estudo propôs a realização de dois experimentos: o primeiro teve o objetivo de comparar a resposta imune humoral anti-PCV2 em camundongos isogênicos e heterogênicos induzidas por vacinas comerciais contra este vírus e o segundo teve o objetivo de se estabelecer um modelo de avaliação da potência de vacinas contra o Circovirus Suíno em camundongos. Para a realização do Experimento 1 foram testadas duas vacinas comerciais contra o PCV2, sendo uma baseada no genótipo “a” e outra no genótipo “b” de PCV2, nas linhagens C57BL/6 e Balb/C e no estoque heterogênico Swiss. Os soros dos animais vacinados foram submetidos à mensuração de anticorpos anti-PCV2 pelo método indireto de ELISA e as médias de DOs obtidas nesse ensaio foram avaliadas e submetidas à análise estatística. Já no Experimento 2, o estoque heterogênico de camundongos Swiss foi escolhido para a realização do delineamento que contou com o mesmo esquema vacinal utilizado no Experimento 1 e foi seguido por desafio com PCV2b em três momentos. Os animais tiveram o sangue colhido e foram então eutanasiados em três momentos para avaliação da resposta imune humoral por ELISA indireto e análise da carga viral de PCV2 em sangue total e tecidos (rim, baço, pulmões e linfonodo). Os resultados demonstraram a capacidade de camundongos isogênicos e heterogênicos vacinados em gerar anticorpos contra PCV2 baseada em soroconversão e títulos end-point avaliados, além de demonstrar não haver diferença estatística (p = 0,063) entre os níveis de anticorpos dos grupos testados. Quanto ao modelo de potência, foi possível ratificar os dados obtidos no Experimento 1, com relação à resposta imune humoral e identificar que camundongos Swiss não são capazes de desenvolver sinais clínicos e apresentar viremia e carga viral em pulmões analisados pela técnica de qPCR de acordo com o delineamento utilizado. Apesar disso, conclui-se que camundongos Swiss podem ser utilizados como modelo para avaliação de resposta imune humoral e estudos de potência baseados em perfil sorológico dos animais submetidos à vacinação contra PCV2. Porcine circovirus Type 2 (PCV2) and its associated diseases (PCVAD) generate large losses for a worldwide swine industry. In addition to other preventive measures to reduce the spread of this infectious disease, vaccination is one of the most efficient ways to control circovirus in swine herds. There are several types of vaccines commercially available against circovirus, and a current update is needed because of the evolution that the prevalent PCV2 genotypes have undergone over the years. Thus, the present study proposed the performance of two experiments, the first with the objective of comparing an anti-PCV2 humoral immune response in isogenic and heterogeneous mice induced by commercial vaccines against this virus and the second with the objective of establishing a model for evaluating the potency of vaccines against the Swine Circovirus in a mouse model. To carry out Experiment 1, two commercial vaccines against PCV2 were tested, one based on the “a” genotype and the other on the “b” PCV2 genotype, on the C57BL/6 and Balb/C lines and on the Swiss heterogeneous stock. Sera from vaccinated animals were subjected to measurement of anti-PCV2 antibodies by indirect ELISA method and means of ODs were evaluated and submitted to statistical analysis. In Experiment 2, Swiss mice was chosen to carry out the design that had the same vaccination schedule used in Experiment 1 and was followed by challenge with PCV2b at three times. . The animals had their blood collected and were then euthanized at three times to assess the humoral immune response by indirect ELISA and analysis of the viral load of PCV2 in whole blood and tissues (kidney, spleen, lungs and lymph node). The results demonstrated the ability of isogenic and heterogeneous vaccinated mice to generate antibodies against PCV2 based on seroconversion and evaluated end-point titers, in addition to demonstrating that there was no statistical difference (p = 0.063) between the antibody levels of the groups tested. As for the potency model, it was possible to confirm the data obtained in Experiment 1, regarding the humoral immune response and to identify that Swiss mice are not able to develop clinical signs, and present viremia and viral load in the lungs analyzed by qPCR technique according to the design used. Despite this, it is concluded that Swiss mice can be used as a model for evaluating the humoral immune response and potency studies based on the serological profile of the animals submitted to vaccination against PCV2, with a possibility of correlation of the data obtained in this model, when compared to the target species of the vaccine evaluated.

Dissertação (mestrado) — Universidade de Brasília, Faculdade de Medicina, Programa de Pós-Graduação em Ciências Médicas, 2021. INTRODUÇÃO: A Amicacina causa ototoxicidade irreversível, e a detecção precoce desse agravo é considerada uma tarefa difícil. Estudos clínicos dos efeitos da Amicacina no homem têm revelado que a droga pode produzir alterações que provocam zumbido, perda auditiva nas frequências altas e manifestações comportamentais. A equivalência anátomo-fisiológica do sistema auditivo periférico de humanos com os camundongos faz com que esse modelo de animal seja rotineiramente empregado em ensaios clínicos, pois contribuem para a prevenção, diagnóstico e tratamento das alterações causadas pelo uso desse fármaco. OBJETIVO: Este estudo teve por objetivo verificar a suscetibilidade do modelo Murino SWISS às lesões de células ciliadas externas causadas em virtude do uso do aminoglicosídeo Amicacina. MATERIAL E MÉTODO: Pesquisa experimental, prospectiva e de intervenção, aprovada pelo CEUA/UnB nº (63/2018). Os animais foram divididos em dois grupos: grupo controle(G1) e Amicacina Ototóxica (G2). O G1 recebeu solução de Cloreto de Sódio(soro) 10mg/kg/dia e o G2 recebeu Amicacina 400mg/kg/dia. As soluções foram ofertadas diariamente via intraperitoneal por 14 dias consecutivos. Foram realizados exames de emissões otoacústicas por produto de distorção nas frequências de 6 à 12kHz no T0 e T14 e estudo histológico das bulas timpânicas. As análises foram realizadas empregando-se o programa Prism5 ®. Foram consideradas significativas, diferenças com p

What is the central question of this study? What is the effect of an obesogenic diet on the expression of microRNAs (miRNAs) involved in cardiac hypertrophy in female mice? What is the main finding and its importance? Female mice fed an obesogenic diet exhibited cardiac hypertrophy associated with increased levels of miRNA-143-3p, decreased mRNA levels of Sox6 and increased mRNA levels of Myh7. Inhibition of miRNA-143-3p increased Sox6 mRNA levels and reduced Myh7 expression in cardiomyocytes, and prevented angiotensin II-induced cardiomyocyte hypertrophy. The results indicate that the miRNA-143-3p-Sox6-Myh7 pathway may play a key role in obesity-induced cardiac hypertrophy.Obesity induces cardiometabolic disorders associated with a high risk of mortality. We have previously shown that the microRNA (miRNA) expression profile is changed in obesity-induced cardiac hypertrophy in male mice. Here, we investigated the effect of an obesogenic diet on the expression of miRNAs involved in cardiac hypertrophy in female mice. Female mice fed an obesogenic diet displayed an increased body weight gain, glucose intolerance, insulin resistance and dyslipidaemia. In addition, obese female mice exhibited cardiac hypertrophy associated with increased levels of several miRNAs, including miR-143-3p. Bioinformatic analysis identified Sox6, regulator of Myh7 gene transcription, as a predicted target of miR-143-3p. Female mice fed an obesogenic diet exhibited decreased mRNA levels of Sox6 and increased expression of Myh7 in the heart. Loss-of-function studies in cardiomyocytes revealed that inhibition of miR-143-3p increased Sox6 mRNA levels and reduced Myh7 expression. Collectively, our results indicate that obesity-associated cardiac hypertrophy in female mice is accompanied by alterations in diverse miRNAs, and suggest that the miR-143-3p-Sox6-Myh7 pathway may play a key role in obesity-induced cardiac hypertrophy.

Diabetic neuropathy, often associated with diabetes mellitus, is a painful condition with no known effective treatment except glycemic control. Studies with neuropathic pain models report alterations in cannabinoid and opioid receptor expression levels; receptors whose activation induces analgesia. We examined whether interactions between CB(1)R and opioid receptors could be targeted for the treatment of diabetic neuropathy. For this, we generated antibodies that selectively recognize native CB(1)R-MOR and CB(1)R-DOR heteromers using a subtractive immunization strategy. We assessed the levels of CB(1)R, MOR, DOR, and interacting complexes using a model of streptozotocin-induced diabetic neuropathy and detected increased levels of CB(1)R, MOR, DOR, and CB(1)R-MOR complexes compared with those in controls. An examination of G-protein signaling revealed that activity induced by the MOR, but not the DOR agonist, was potentiated by low nanomolar doses of CB(1)R ligands, including antagonists, suggesting an allosteric modulation of MOR signaling by CB(1)R ligands within CB(1)R-MOR complexes. Because the peptide endocannabinoid, hemopressin, caused a significant potentiation of MOR activity, we examined its effect on mechanical allodynia and found that it blocked allodynia in wild-type mice and mice with diabetic neuropathy lacking DOR (but have CB(1)R-MOR complexes). However, hemopressin does not alter the levels of CB(1)R-MOR complexes in diabetic mice lacking DOR but increases the levels of CB(1)R-DOR complexes in diabetic mice lacking MOR. Together, these results suggest the involvement of CB(1)R-MOR and CB(1)R-DOR complexes in diabetic neuropathy and that hemopressin could be developed as a potential therapeutic for the treatment of this painful condition.

PurposeSome first-line cytotoxic chemotherapics, e.g. doxorubicin, paclitaxel and oxaliplatin, induce activation of the immune system through immunogenic cell death (ICD). Tumor cells undergoing ICD function as a vaccine, releasing damage-associated molecular patterns (DAMPs), which act as adjuvants, and neoantigens of the tumor are recognized as antigens. ICD induction is rare, however it yields better and long-lasting antitumor responses to chemotherapy. Advanced metastatic melanoma (AMM) is incurable for more than half of patients. The discovery of ICD inducers against AMM is an interesting drug discovery strategy with high translational potential. Here we evaluated ICD induction of four highly cytotoxic chromomycins A (CA5-8).MethodsICD features and DAMPs were evaluated using several in vitro techniques with metastatic melanoma cell line (B16-F10) exposed to chromomcins A5-8 such as flow cytometry, western blot, RT-PCR and luminescence. Additionally in vivo vaccination assays with CA5-treated cells in a syngeneic murine model (C57Bl/6) were performed to confirm ICD evaluating the immune cells activation and their antitumor activity.ResultsB16-F10 treated with CA5-8 and doxorubicin exhibited ICD features such as autophagy and apoptosis, externalization of calreticulin, and releasing of HMGB1. However, CA5-treated cells had the best profile, also inducing ATP release, ERp57 externalization, phosphorylation of eIF2α and altering expression of transcription of genes related to autophagy, endoplasmic reticulum stress, and apoptosis. Bona fide ICD induction by CA5 was confirmed by vaccination of C57BL/6 mice with CA5-treated cells which activated antigen-presenting cells and T lymphocytes and stimulated antitumor activity.ConclusionCA5 induces bona fide immunogenic cell death on melanoma.

In nature, confrontations between conspecifics are recurrent and related, in general, due to the lack of resources such as food and territory. Adequate defence against a conspecific aggressor is essential for the individual's survival and the group integrity. However, repeated social defeat is a significant stressor promoting several behavioural changes, including social defence per se . What would be the neural basis of these behavioural changes? To build new hypotheses about this, we here investigate the effects of repeated social stress on the neural circuitry underlying motivated social defence behaviour in male mice. We observed that animals re-exposed to the aggressor three times spent more time in passive defence during the last exposure than in the first one. These animals also show less activation of the amygdalar and hypothalamic nuclei related to the processing of conspecific cues. In turn, we found no changes in the activation of the hypothalamic dorsal pre-mammillary nucleus (PMD) that is essential for passive defence. Therefore, our data suggest that the balance between the activity of circuits related to conspecific processing and the PMD determines the pattern of social defence behaviour. Changes in this balance may be the basis of the adaptations in social defence after repeated social defeat.

Made available in DSpace on 2022-04-28T19:50:37Z (GMT). No. of bitstreams: 0 Previous issue date: 2022-02-01 Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) We compared the parameters related to glucose homeostasis, and liver and muscle proteomes in fluorosis-susceptible (A/J; S) and fluorosis-resistant (129P3/J; R) mice in response to fluoride (F) exposure and exercise. Ninety male mice (45 R-mice and 45 S-mice) were randomized into three groups: (SI; RI) No-F, No-Exercise, (SII; RII) 50 ppm F, No-Exercise, (SIII; RIII) 50 ppm F, Exercise. Overall, mean F concentrations in the plasma and femur were significantly higher in R-mice compared with S-mice. In R-mice, exercise resulted in an increase in F accumulation in the femur. In S-mice, the mean plasma glucose level was significantly higher in Group II compared with Groups I and III. There was an increase in liver proteins involved in energy flux and antioxidant enzymes in non-exercise groups (I, II) of S-mice in comparison with the corresponding groups of R-mice. The results also showed a decrease in muscle protein expression in Group I S-mice compared with their R-mice counterparts. In conclusion, the findings suggest an increased state of oxidative stress in fluorosis-susceptible mice that might be exacerbated by the treatment with F. In addition, fluorosis-susceptible mice have plasma glucose levels higher than fluorosis-resistant mice on exposure to F, and this is not affected by exercise. Bauru School of Dentistry University of São Paulo, SP Department of Physical Education School of Sciences São Paulo State University, SP School of Human and Health Sciences University of Huddersfield Centre for Oral Health Research School of Dental Sciences Newcastle University School of Health and Life Sciences Teesside University Department of Physical Education School of Sciences São Paulo State University, SP CAPES: 001 FAPESP: 2014/50798-1 FAPESP: 2015/05278-2 FAPESP: 2015/12109-2 FAPESP: 2015/50406-9 FAPESP: 2016/20020-4

The development of a Pressure Injury (PI) is a very recurrent outcome in long-stay patients in hospitals or even in-home care who are bedridden. Besides raising the costs of treatment and the length of stay of the user in health services, PIs also cause suffering. This wear generated by a PI has been suggested to establish its relationship with the development of depression, a severe and disabling psychiatric disorder. In this sense, substances with antioxidant and anti-inflammatory activity are being discussed as new forms of treatment and early healing of a PI. Thus, recent studies have sought to demonstrate the healing role of alpha-lipoic acid (ALA) in skin lesions. That way, the present study aimed to investigate the wound healing effect of the topical application of ALA in an animal model of pressure injury associated with behavioral and neurochemical changes. We aimed to investigate a new therapeutic perspective for PI healing and avoid its complications. For the development of the study, we used male Swiss mice (25-30g) and separated them into the following experimental groups: control, SHAM, vehicle, ALA, and Hydrogel with Alginate. All subjects were submitted to the ischemia/reperfusion cycle (I/R) for 4 consecutive days, by placing two magnets on the skin of the dorsal region. The animals had their PI treated for 5 days topically with ALA cream or Hydrogel with Alginate and polyurethane film in the association. The control group did not have lesion induction. The vehicle group received only the ALA vehicle, and the SHAM group had the lesion-induced but received no topical treatment. The animals were submitted to the following behavioral tests: open field, splash, tail suspension, and social interaction, six hours after application of the topical treatment on days 1 and 5 of treatment. Immediately after the behavioral tests, the animals were euthanized by decapitation and their prefrontal cortex (CPF), hippocampus (HC), lesion/scar were removed, and plasma was collected for the determination of Myeloperoxidase (MPO) enzyme activity. Only the lesion/scar was used to determine the concentration of the cytokine TNF-α. The area of the lesion or scar was also removed for histopathological evaluation. Photos of the lesion were taken on days 1 and 5 of treatment. The body weight was measured at the beginning of the protocol and the end of the treatments. In the open field test, the SHAM group demonstrated a reduction in the number of crossings. while all treatments reversed this alteration. SHAM animals demonstrated an increase in immobility time in the tail suspension test. All treatments reversed this alteration. In the sucrose spray test, the SHAM group showed increased latency for grooming and decreased number and time of grooming. On the other hand, ALA cream reversed the alteration in all parameters. The other treatments only had a reversal effect on the number and time of grooming. Furthermore, SHAM animals showed a significant reduction in social interaction while all treatments reversed this modification. Furthermore, SHAM animals showed a reduction in social interaction, while all treatments managed to minimize this behavioral alteration. Concerning the behavioral assessment, the depressive-like behaviors that were mitigated due to the healing/anti-inflammatory effect of the treatments can be highlighted by: the reduction of crossings in the open field, the increase of immobility in the tail suspension; increased latency, decreased number, and time of grooming when spraying sucrose in the mice’s back, and reduced social interaction. In terms of weight variation, it was revealed that PI induction leads to a reduction in the weight of animals, on the other hand, no treatment was able to reduce. Concerning the enzymatic activity of MPO, the SHAM group showed an increase in the parameter evaluated in CPF, HC, plasma, and lesion, and the treatments reversed this effect in all tissues investigated. When assessing the TNF-α in the lesion, the SHAM group caused an increase in the concentration. All the treatments reversed this effect. About the healing effects, treatments were able to reduce the area and increase the contraction of the lesion. About the histological parameters, all treatments reduced inflammatory scores caused by PI, but only ALA was able to induce angiogenesis. Altogether, these data show that the topical treatment with ALA has a wound healing activity on PI and that this local treatment can exert an indirect antidepressant action. Um desfecho muito recorrente em pacientes de longa permanência em hospitais ou mesmo no cuidado domiciliar que estão restritos ao leito é o aparecimento de lesões por pressão. Além de elevarem os custos do tratamento e o tempo de permanência do usuário nos serviços de saúde, as LP causam sofrimento físico e emocional. Esse desgaste gerado tem sido sugerido como um fator relacionado com o desenvolvimento de quadros depressivos, um transtorno psiquiátrico grave e incapacitante. Nesse sentido, substâncias com atividade antioxidante e anti-inflamatória estão sendo discutidas como novas formas de tratamento e cicatrização precoce de uma LP. Assim, estudos recentes tem buscado demonstrar o papel cicatrizante do ácido alfa-lipóico (ALA) baseado nessas ações. Dessa forma, o presente estudo se propôs a investigar o efeito cicatrizante da aplicação tópica de ALA em modelo de lesão por pressão animal associado às alterações comportamentais e neuroquímicas, objetivando desenvolver uma nova perspectiva terapêutica para a cicatrização de LP e assim evitar suas complicações. Para o desenvolvimento do estudo foram utilizados camundongos Swiss machos (25-30g), divididos nos seguintes grupos experimentais: controle, SHAM, creme base, creme ALA e Hidrogel com Alginato (fármaco referência), os quais, durante 4 dias consecutivos foram submetidos ao ciclo de isquemia/reperfusão (I/R), através da colocação de dois imãs na pele da região dorsal. Os animais tiveram suas LP tratadas durante 5 dias por via tópica com creme ALA ou com Hidrogel com Alginato e filme de poliuretano como cobertura secundária. Os animais do grupo controle não tiveram a indução da lesão, os do grupo creme base receberam somente o veículo do ALA e os do grupo SHAM tiveram a indução da lesão, mas não receberam nenhum tratamento tópico. Os animais foram submetidos aos testes comportamentais: campo aberto, borrifagem de sacarose, suspensão de cauda e interação social, seis horas após a aplicação do tratamento tópico nos dias 1 e 5 de tratamento. Imediatamente após os testes comportamentais os animais foram eutanasiados por rápida decapitação e seu córtex pré-frontal (CPF), hipocampo (HC), a lesão/cicatriz foram removidos e o plasma coletado para a determinação da atividade da enzima Mieloperoxidase (MPO). Somente a lesão/cicatriz foi utilizada para determinar a concentração da citocina TNF-α e para a realização da avaliação histopatológica. As fotos da lesão foram obtidas no dia 1 e 5 de tratamento. Já o peso corporal foi mensurado no início do protocolo e no fim dos tratamentos. No teste de campo aberto, o grupo SHAM demonstrou uma redução do número de travessias, enquanto todos os tratamentos reduziram esse efeito. Os animais do SHAM demonstraram um aumento no tempo de imobilidade no teste de suspensão da cauda, enquanto todos os tratamentos reduziram esse efeito. No teste de borrifagem de sacarose, o grupo SHAM mostrou aumento da latência para grooming e diminuição do número e tempo de grooming, enquanto o creme ALA minimizou o efeito em todos os parâmetros, os outros tratamentos só tiveram efeito de minimização sobre o número e tempo de grooming. Além disso, os animais do SHAM mostraram uma redução importante da interação social, enquanto todos os tratamentos conseguiram minimizar esse efeito. Em suma, os comportamentos depressivos símile que foram mitigados devido ao efeito cicatrizante/anti-inflamatório dos tratamentos foram a redução das travessias no campo aberto, o aumento de imobilidade na suspensão de cauda; aumento da latência, diminuição do número e tempo de grooming na borrifagem de sacarose e redução da interação social. Já no quesito de variação de peso, foi revelado que a indução da LP leva a redução do peso dos animais e nenhum tratamento foi capaz de reduzir. No que diz respeito a atividade enzimática de MPO, o grupo SHAM também obteve aumento do parâmetro avaliado em CPF, HC, plasma e lesão, e os tratamentos foram capazes de minimizar esse efeito em todos os tecidos investigados. Na avaliação de TNF-α da lesão, o grupo SHAM causou um aumento na concentração e os tratamentos foram capazes de reduzir esse efeito. Além disso, sobre os efeitos cicatriciais, houve a redução da área e o aumento da contração da lesão. Nos parâmetros histológicos, os tratamentos foram de capazes de reduzir os escores inflamatórios causados pela LP, mas somente o creme ALA foi capaz de induzir a angiogênese. Em conjunto esses dados evidenciam que o tratamento tópico com creme ALA possui ação cicatrizante em LP e que esse tratamento local parece contribuir para uma ação antidepressiva indireta.

Submitted by Michele Mologni (mologni@unoeste.br) on 2022-04-19T13:13:18Z No. of bitstreams: 1 Adriano Messias de Souza.pdf: 3076004 bytes, checksum: 5d06ef59734f236f32941c1b73a483c4 (MD5) Made available in DSpace on 2022-04-19T13:13:18Z (GMT). No. of bitstreams: 1 Adriano Messias de Souza.pdf: 3076004 bytes, checksum: 5d06ef59734f236f32941c1b73a483c4 (MD5) Previous issue date: 2021-10-28 Autism spectrum disorders (ASD) are a diverse group of conditions. They are characterized by some degree of difficulty with social interaction and communication and may be more vulnerable to developing chronic non-communicable conditions due to behavioral risk factors such as physical inactivity and poor food preferences, and are at increased risk of violence, injury and abuse. Health insurance covers some of these expenses, but families of autistic children have high out-of-pocket costs. ASD-like disorders are induced by propionic or valproic acids in an animal model, rats and mice, in the pre- and postnatal phases. The phenolic resveratrol (RSV) has demonstrated the ability to beneficially influence behavioral parameters similar to ASD, sociability, social preference for novelty, self-cleaning/self-care and reciprocal social interaction, but the results are controversial and scarce in animal and human models. The objective of this systematic review with meta-analysis was to compile the results of the included studies and analyze the effects of resveratrol on behavioral indicators similar to ASD in an animal model, rats and mice. The searches were carried out in the databases: “PubMed”, “ScienceDirect”, “Web of Science”, “Scielo” and Google in April 2021, and repeated in December 2021, using the keywords “resveratrol”,” autism spectrum disorder”, “autistic disorder”, “rats” and “mice”. 4 eligible articles were included in the meta-analyses. The results obtained demonstrate beneficial effects (P

The infection caused by Streptococcus equi, known as strangles, affects the respiratory system of horses, causing high morbidity and rapid spread among the herd. Bacterin vaccines, composed of inactivated whole cells of S. equi, have variable efficacy and duration. Infected animals produce specific antibodies against SeM, the immunodominant antigen of S. equi. This makes it a promising target for vaccine development. In this context, the objective of this work was to evaluate a vaccine combining S. equi bacterin and recombinant SeM protein. Mice were vaccinated with bacterin (S. equi ~1.2 × 108CFU/ml); rSeM protein (20μg); bacterin-rSeM combination; or PBS (Control Group) and challenged with a suspension of S. equi, containing 10 × LD50. All vaccinated mice survived the challenge and produced anti-rSeM and anti-S. equi antibodies, which were assessed by indirect ELISA. The Control Group reached endpoint criteria 96 h after infection. These results demonstrate that a vaccine combining the S. equi bacterin with rSeM protein protects mice against strangles. This combination vaccine could potentially protect horses and overcome the limitations of currently available strangle vaccines. RESUMO: A infecção causada por Streptococcus equi, denominada adenite, atinge o sistema respiratório de equinos, causando alta morbidade e rápida disseminação entre o rebanho. Vacinas bacterinas, compostas de células inteiras inativadas de S. equi apresentam eficácia e duração variáveis. Animais infectados apresentam anticorpos específicos à proteína SeM, antígeno imunodominante de S. equi, o que a torna um alvo promissor para o desenvolvimento de vacinas. Neste contexto, o objetivo deste trabalho foi avaliar uma vacina baseada na administração simultânea da bacterina e da proteína SeM recombinante. Camundongos foram vacinados com a bacterina (S. equi ~1.2 × 108CFU/ml); a proteína rSeM (20μg); a bacterina e rSeM simultaneamente; ou PBS (Grupo Controle) e, posteriormente, foram desafiados com uma suspensão de S. equi contendo 10 × LD50. Todos os animais vacinados apresentaram anticorpos anti-rSeM e contra S. equi, avaliados através de ELISA indireto, e mantiveram-se e sobreviveram ao desafio letal. O Grupo Controle atingiu critérios de endpoint 96 h após a infecção. Estes resultados demonstram que uma vacina constituída de células inteiras de S. equi com rSeM protege camundongos contra adenite, sugerindo a capacidade de proteção a equinos e, possivelmente, superando as limitações das vacinas contra adenite atualmente disponíveis.

The alveolar bone repair process may be influenced by multiple local and systemic factors, which include immune system cells and mediators. Macrophages allegedly play important roles in the repair process, and the transition of an initial inflammatory M1 profile into a pro-reparative M2 profile theoretically contributes to a favorable repair outcome. In this context, considering immunoregulatory molecules as potential targets for improving bone repair, this study evaluated the role of the immunoregulatory molecule FTY720, previously described to favor the development of the M2 phenotype, in the process of alveolar bone healing in C57Bl/6 (WT) mice. Experimental groups submitted to tooth extraction and maintained under control conditions or treated with FTY720 were evaluated by microtomographic (μCT), histomorphometric, immunohistochemical and molecular analysis to characterize healing and host response features at 0, 1, 3, 7 and 14 days. Our results demonstrated that the FTY720 group presented higher bone tissue density, higher bone tissue volume, greater tissue volume fraction, greater number and thickness of trabeculae and a higher number of osteoblasts and osteoclasts than the control group. Accordingly, the bone markers BMP2, BMP7, ALPL, SOST and RANK mRNA expressions increased in the FTY720 treated group. Furthermore, the levels of FIZZ, ARG2 and IL-10 mRNA increased in the FTY720 group together with the presence of CD206

Made available in DSpace on 2022-04-29T08:46:34Z (GMT). No. of bitstreams: 0 Previous issue date: 2022-01-01 Background Fluoride has become widely used in dentistry because of its effectiveness in caries control. However, evidence indicates that excessive intake interferes with the metabolic processes of different tissues. Thus, this study aimed to investigate the effects of long-term exposure to F on the parotid salivary gland of mice, from the analysis of oxidative, proteomic and genotoxic parameters. Materials and methods The animals received deionized water containing 0, 10 or 50 mg/L of F, as sodium fluoride, for 60 days. After, parotid glands were collected for analysis of oxidative biochemistry, global proteomic profile, genotoxicity assessment and histopathological analyses. Results The results revealed that exposure to fluoride interfered in the biochemical homeostasis of the parotid gland, with increased levels of thiobarbituric acid reactive species and reduced glutathione in the exposed groups; as well as promoted alteration of the glandular proteomic profile in these groups, especially in structural proteins and proteins related to oxidative stress. However, genotoxic assessment demonstrated that exposure to fluoride did not interfere with DNA integrity in these concentrations and durations of exposure. Also, it was not observed histopathological alterations in parotid gland. Conclusions Thus, our results suggest that long-term exposure to fluoride promoted modulation of the proteomic and biochemical profile in the parotid glands, without inducing damage to the genetic component. These findings reinforce the importance of rationalizing the use of fluorides to maximize their preventative effects while minimizing the environmental risks. Laboratory of Functional and Structural Biology Institute of Biological Sciences Federal University of Para, PA Laboratory of Clinical Immunology and Oxidative Stress Pharmacy Faculty Institute of Health Science Federal University of Para, PA Evandro Chagas Institute, PA Department of Biological Sciences Bauru Dental School University of São Paulo, SP Department of Chemistry University of Nebraska-Lincoln Department of Preventive and Restorative Dentistry School of Dentistry São Paulo State University (UNESP), SP Department of Preventive and Restorative Dentistry School of Dentistry São Paulo State University (UNESP), SP

Bisphenol A (BPA) is an endocrine disrupting chemical able to promote hormone-responsive tumors. The major route of BPA contamination being oral, the aim of the present study was to investigate BPA effects on oral cells. Here, we evaluated the impact of sub-chronic in vivo exposure to BPA and its in vitro effects on neoplastic and non-neoplastic oral cells. We evaluated the oral mucosa of mice chronically exposed to BPA (200 mg/L). The response of keratinocytes (NOK–SI) and Head and Neck (HN) Squamous Cell Carcinoma (SCC), HN12 and HN13 cell lines to BPA was examined. In vivo, BPA accumulated in oral tissues and caused an increase in epithelial proliferative activity. BPA disrupted the function of keratinocytes by altering pro-survival and proliferative pathways and the secretion of cytokines and growth factors. In tumor cells, BPA induced proliferative, invasive, pro-angiogenic, and epigenetic paths. Our data highlight the harmful effects of BPA on oral mucosa and, tumorigenic and non-tumorigenic cells. Additionally, BPA may be a modifier of oral cancer cell behavior by prompting a functional shift to a more aggressive phenotype.