The family of proteins known as heterogeneous nuclear ribonucleoproteins (hnRNPs) is large and diverse. Often, one and the same hnRNP will perform multiple cellular functions, leading to their description as “multifunctional proteins”. The two hnRNPs known as hnRNP A1 and hnRNP C1/C2 are multifunctional proteins found to affect the transcription, splicing, stability, and translation of specific genes’ mRNA. They are implicated in carcinogenesis, apoptosis, and DNA damage response mechanisms. The aims of this thesis were to study the hnRNP A1 and hnRNP C1/C2 dependent regulation of two highly stress responsive genes, the tumor suppressor p53 and the cytochrome P450 enzyme Cyp2a5/CYP2A6. We identified hnRNP C1/C2 as a DNA damage induced binding protein towards the coding region of p53 mRNA, and found that while a specific cis binding site appears to have a positive function in p53 expression, interaction of hnRNP C1/C2 with this site represses the expression. The data suggest that two distinct molecular mechanisms exist for the down-regulation of p53 by hnRNP C1/C2. One mechanism, active during transcriptional stress, is dependent upon the aforementioned site, and the other, independent. We discuss how hnRNP C1/C2 dependent repression of p53 may play a role in apoptosis. The data presented here further suggest that the transcriptional and post-transcriptional processes controlling the expression of the murine Cyp2a5 gene are linked via hnRNP A1, by performing functions in the nucleus as a transcription factor, or in the cytoplasmic compartment as a trans factor bound to the 3’UTR of the mRNA as needed. Our studies of the human ortholog of this gene, CYP2A6, suggest that this gene is regulated post-transcriptionally in a manner similar to that of its murine counterpart, via changes in mRNA stability and interaction of hnRNP A1 with its 3’ UTR.

TRPA1 channels are sensors for noxious stimuli in a subset of nociceptive neurons. TRPA1 channels are also expressed in cells of the mammalian inner ear, but their function in this tissue remains unknown given that Trpa1–/– mice exhibit normal hearing, balance and sensory mechanotransduction. Here we show that non-sensory (supporting) cells of the hearing organ in the cochlea detect tissue damage via the activation of TRPA1 channels and subsequently modulate cochlear amplification through active cellshape changes. We found that cochlear supporting cells of wild type but not Trpa1–/– mice generate inward currents and robust long-lasting Ca2+ responses after stimulation with TRPA1 agonists. These Ca2+ responses often propagated between different types of supporting cells and were accompanied by prominent tissue displacements. The most prominent shape changes were observed in pillar cells which here we show possess Ca2+-dependent contractile machinery. Increased oxidative stress following acoustic overstimulation leads to the generation of lipid peroxidation byproducts such as 4-hydroxynonenal (4-HNE) that could directly activate TRPA1. Therefore, we exposed mice to mild noise and found a longer-lasting inhibition of cochlear amplification in wild type than in Trpa1–/– mice. Our results suggest that TRPA1-dependent changes in pillar cell shape can alter the tissue geometry and affect cochlear amplification. We believe this novel mechanism of cochlear regulation may protect or fine-tune the organ of Corti after noise exposure or other cochlear injuries.

A corticoterapia tem sido utilizada frequentemente nas éguas suscetíveis. O uso de isuflupredona melhora a taxa de prenhez e altera o perfil proteico do líquido endometrial em relação a éguas não tratadas. A utilização de dexametasona diminui o acúmulo de líquido pós-cobertura, reduz o edema do útero, porém, desconhecem-se seus efeitos no perfil proteico do líquido endometrial. O objetivo do presente estudo foi analisar o efeito da dexametasona em éguas suscetíveis à endometrite persistente póscobertura sobre perfil proteico do líquido endometrial na presença ou ausência de infecção. Nove éguas suscetíveis foram utilizados, com idade entre 7 a 30 anos. Após a verificação dos sinais de estro as éguas foram submetidas a quatro tratamentos: (C) éguas não receberam nenhum tipo de tratamento e serviram como controle; (D) éguas receberam 40mg de dexametasona (IV), no momento da cobertura, com coleta da amostras após 6 horas, (I-6 e I-24) infusão intra-uterina de 1 x 109 de S. zooepidemicus/mL, com coleta da amostra após 6 e 24 horas; (I/D-6 e I/D-24) infusão intra-uterina de 1 x 109 S. zooepidemicus/mL e administração de 40mg de dexametasona (IV), com coleta da amostra após 6 e 24 horas. Todas as éguas foram submetidas a todos os tratamentos. As amostras foram coletadas e submetidas à eletroforese bidimensional para separação proteica e espectrometria de massa para a identificação das bandas proteicas relevantes. A corticoterapia provocou alteração na proteômica do líquido endometrial de éguas suscetíveis, caracterizada pelo aumento (TTR) e/ou diminuição (ApoA1) na densidade óptica de proteínas da fase aguda da inflamação. Conclui-se que a utilização da dexametasona em éguas com e sem presença de infecção altera a proteômica do fluido endometrial de éguas suscetíveis. Sugere-se que a dosagem ou a frequência de aplicação da dexametasona deva ser aumentada. / Corticotherapy has often been used in susceptible mares. The use of isuflupredona improves pregnancy rate and alters the protein profile of the endometrial fluid in relation to untreated mares. The use of dexamethasone decreases the post breeding fluid accumulation, reduces the uterine edema, however is unaware of its effects on the protein profile of endometrial fluid. The aim of the present study was analyze the effect of dexamethasone in mares susceptible to post-breeding persistent endometritis on the protein profile of endometrial fluid in the presence or absence of infection. Nine susceptible mares were used, aged 7-30 years old. After checking the signs of estrus, mares were subjected to four treatments: (C) mares received no treatment and served as controls; (D) mares received 40 mg of dexamethasone at breeding time, with collection of samples after 6 hours; (I-6 and I-24) intrauterine infusion of 1 x 109 S. zooepidemicus/ml and the sample was collected after 6 and 24 hours; (I/D-6 and I/D-24) intrauterine infusion of 1 x 109 S. zooepidemicus/ml and 40 mg of dexamethasone administration, collecting the sample after 6 and 24 hours. All of the mares were subjected to all treatments. Samples were collected and subjected to two-dimensional electrophoresis for protein separation and mass spectrometry for the identification of relevant protein bands. Corticotherapy resulted in alteration of the protein profile of the endometrial fluid of susceptible mares, characterized by an increase (TTR) and/or decrease (ApoA1) in optical density of the acute phase of proteins of inflammation. We conclude that the use of dexamethasone in mares with and without the presence of infection alters the protein profile of endometrial fluid of susceptible mares. It is suggested that the dosage or frequency of application of dexamethasone should be increased.

Orientador: Flávia Cristina Rodrigues Lisoni / Coorientador: Sonia Maria Oliani / Banca: Cristiane Damas Gil / Banca: Edson Guilherme Vieira / Resumo: INTRODUÇÃO: O câncer de colo de útero, também chamado de câncer cervical, é o segundo tipo de câncer mais frequente em mulheres mundialmente, sendo a quarta causa de morte por câncer em países em desenvolvimento. A carcinogênese de colo de útero está relacionada com alterações genéticas, infecção pelo Papilomavirus Humano (HPV), angiogênese e processos inflamatórios. A anexina-A1 (ANXA1), proteína de 37 kDa, que é expressa pelas células tumorais atua como moduladora do processo inflamatório. Os dados disponíveis sugerem que essa família de proteínas pode ter, além de seu importante papel no processo inflamatório, um envolvimento significativo no câncer, por meio de cascatas de sinalização que incluem genes relacionados com o ciclo celular, a diferenciação e a apoptose. OBJETIVOS: Em função da importância do efeito anti-inflamatório do peptídeo Ac2-26 nas células carcinogênicas, o presente trabalho teve como objetivo geral investigar a influência desse anti-inflamatório nas células SiHa. MATERIAIS E MÉTODOS: Utilizamos a linhagem celular SiHa (derivada das células de carcinoma epidermóide de cérvice), tratada com o peptídeo Ac2-26 na concentração de 10μg/mL por 2, 4, 24, 48 e 72 horas. Avaliamos morfologia; proliferação e migração celular; apoptose, necrose e apoptose tardia; citocinas pró inflamatórias; expressão protéica e dos genes COX-2, EP3, EP4, MMP2, MMP9, TIMP1 e TIMP2; nas amostras controle e tratadas com o peptídeo Ac2-26, pelas técnicas de microscopia de luz, Citometria de fluxo, Magipix, Western Blotting e PCR quantitativo em tempo real. RESULTADOS: Nas células de carcinoma de colo uterino observamos que não houve alteração na morfologia celular após o tratamento com peptídeo Ac2-26, entretanto houve a redução da proliferação e migração celular. No ensaio de citômetro de fluxo não houve indução de apoptose, apoptose... / Abstract: INTRODUCTION: Cervical cancer is the second most common cancer in women worldwide and is the fourth leading cause of cancer deaths in developing countries. The cervical carcinogenesis is related to genetic alterations, infection by the Human Papillomavirus (HPV), angiogenesis and inflammation. Annexin-A1 (ANXA1), 37 kDa protein, which is expressed by tumor cells acts as a modulator of the inflammatory process. Evidence suggests that this protein family may have, in addition to its important role in the inflammatory process, significant involvement in cancer, through signaling cascades that include genes related to cell cycle, differentiation and apoptosis. OBJECTIVES: Because of the importance of the anti-inflammatory effect of Ac2-26 peptide in carcinogenic cells, this study aimed to investigate the influence of anti-inflammatory in SiHa cells. MATERIALS AND METHODS: SiHa cell line (derived from squamous cell carcinoma of cervix), treated with Ac2-26 peptide at a concentration of 10μg/mL for 2, 4, 24, 48 and 72 hours. We evaluate morphology; cell proliferation and migration; apoptosis, late apoptosis and necrosis; pro-inflammatory cytokines; protein expression and COX-2 genes, EP3, EP4, MMP2, MMP9, TIMP1 and TIMP2; control and the samples treated with the peptide Ac2-26, by light microscopy techniques, flow cytometry, magipix, Western blotting and quantitative PCR in real time. RESULTS: In cervical carcinoma cells observed that there was no change in cell morphology after treatment with peptide Ac2-26, however, there was a reduction in cell proliferation and migration. In the test flow cytometry there was no induction of apoptosis, late apoptosis or necrosis, but the test Magpix decreased interleukin 6 (IL-6) in SiHa cells treated with the peptide Ac2-26. The effect of treatment carried out with the exogenous protein did not affect the expression of Annexin A1 endogenous protein, but showed... / Mestre

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Background: Exacerbated or prolonged inflammatory responses can be detrimental to the host, then any antiinflammatory mediators act to regulate the properties of pro-inflammatory factors and ensure the homeostasis of the organic systems. Objectives: Aims: This study has focused in the regulation of expression and the immunomodulatory properties of annexin A1, a protein of 37 KDa, originally known as the second messenger action of glucocorticoids, in inflammatory and infectious processes. Methods: Using molecular (qPCR), biochemical (luminometry) and immunological (flow cytometry, immunofluorescence) tools, we investigated the endogenous and exogenous of ANXA1 and its mimetic peptide (Ac2-26) in different conditions in vitro and in vivo: fungi infection, HIV/SIV infecion, bowel inflammatory disease and renal ischemic injury. Results: The Ac2-26 pre-treatment of PMN and PBMC of human peripheral blood stimulated with opsonized-zymosan inhibited the production of reactive oxygen species in a dose-dependent manner. In parallel, incubation of phagocytes with the peptide was shown to decrease expression of TLR2 receptor, responsible for the recognition of zymosan on phagocyte, suggesting a regulatory mechanism of surface receptor by ANXA1 peptide. Due to last data, the expression of CCR5, chemokine receptor and HIV co-receptor, was investigated in the same in vitro conditions. CCR5 transcription was down-regulated in PBMC from HIV-infected and healthy subjects, partially by ANXA1/FPR pathway. Consistently, both PBMC subject groups showed an impairment of percentage of CCR5+CD4+ T cells after Ac2-26 incubation. In contrast, Ac2-26 showed an up-regulation of CCR5 surface expression in monocytes, however the peptide switched the profile of monocyte subsets, increasing CD14highCD16- and decreasing CD14lowCD16+ populations. The last one is the most susceptible to HIV infection. In conclusion, the data suggest that the action of N-terminal ANXA1 is cell-dependent and may contribute indirectly modulating the HIV-1. Coherently, the study in vivo of ANXA1 expression in a non-human model of AIDS, showed that ANXA1 is regulated during the progression of the disease in the main compartments of infection, the blood and the gut. In comparison with samples of uninfected animals, the ANXA1 transcripts in peripheral blood increased from acute to chronic SIV infection, whereas in the intestinal mucosa it was down-regulated, reaching baseline levels only in chronic infection. Analysis of typical markers of AIDS progression, showed that increased ANXA1 transcripts was significantly correlated with the activation of T cells. However, the expression of ANXA1 had a positive correlation with antiinflammatory cytokines in the blood and in the intestine and a inverse dynamics of viral load and depletion of CD4 T cells, suggesting that activation of its transcription is related to an attempt to reduce the immune depletion during infection. In addition, based on the described differential modulation of the ANXA1 in the gut of primates, another goal was to analyze the expression of ANXA1 in patients with IBD (inflammatory bowel disease) or not treated with immunosuppressive drugs. The correlation among those factors and clinical data were analyzed. In IBD patients, the expression of ANXA1 is reduced in level of mRNA in peripheral blood and protein in the colonic mucosa. In these patients, immunotherapy with infliximab (anti-TNF-alpha) modulates the transcription of ANXA1 and TNF-α, and even systemic lymphocyte activation in accordance with the duration of treatment and the side effects of drugs, such as bacteremia. The dynamics of ANXA1 added to other clinical and immunological factors appear to be associated with the course of IBD. Finally, acute and chronic inflammatory process induced by ischemia/reperfusion (I/R) procedure, revealed that exogenous ANXA1 mimetic peptide granted a remarkable protection against kidney I/R injury, preventing glomerular filtration rate and urinary osmolality decreases and acute tubular necrosis development by affording striking structural protection due to the abortion of neutrophil extravasation, attenuation of macrophage infiltration and regulation of endogenous annexin A1 expression in renal epithelial cells. Conclusions: In a general analysis of all studies presented, we conclude that exogenous ANXA1 and its derived peptides act as key molecules in the modulation of specific activation, immunophenotype and distribution of phagocytes and lymphocytes participating as a line of defense or targeting of infectious agents, via FPRs or not. And, emphasizing the importance of ANXA1 in the defense of homeostasis, although it was found that the pathways related to the regulation of endogenous ANXA1 is differentially activated in tissues and fluids under acute and chronic inflammation caused by viral infection, autoimmunity or hypoxia. / Introdução: As respostas inflamatórias exacerbadas ou prolongadas podem ser prejudiciais para o hospedeiro, então muitos mediadores antiinflamatórios atuam para regular as propriedades dos fatores pró-inflamatórios e garantir a homeostasia dos sistemas. Objetivos: Assim, o foco do presente trabalho foi estudar a regulação da expressão e as propriedades imunomodulatórias da anexina A1, uma proteína de 37 KDa, inicialmente conhecida como a segunda mensageira da ação dos glicocorticóides, em processos inflamatórios e infecciosos. Metodologia: Para tanto, por meio de técnicas moleculares (qPCR), bioquímicas (luminometria) e imunológicas (citometria de fluxo, imunofluorescência), foi investigado o papel endógeno da ANXA1 e exógeno do peptídeo N-terminal da ANXA1 em diferentes condições in vitro e in vivo: infecção fúngica, infecção viral por HIV-1 e SIV, doença inflamatória intestinal e injúria isquêmica renal. Resultados: O peptídeo N-terminal da ANXA1, Ac2-26, quando usado como tratamento prévio de PMN e PBMC do sangue periférico humano estimuladas por zymosan-opsonizado, inibe a produção de espécies reativas de oxigênio de modo dose-dependente. Paralelamente, a incubação dos fagócitos com o peptídeo demonstrou diminuir a expressão do receptor TLR2, responsável pelo reconhecimento de zymosan nos fagócitos, sugerindo um mecanismo de regulação de receptores de superfície pelo Ac2-26. Frente a este dado e a sinalização de ANXA1 via FPRs, a expressão do receptor de quimiocina e coreceptor do vírus HIV-1, CCR5, também foi investigada em tais condições in vitro. Os transcritos de CCR5 diminuíram em PBMCs de indivíduos HIV+ e saudáveis, parcialmente pela via ANXA1/FPR. Consistentemente, Ac2-26 também diminuiu a expressão do CCR5 na superfície de células T CD4+, os principais alvos do vírus. Em contraste, nos monócitos o peptídeo aumentou a expressão de CCR5, no entanto, induziu uma mudança na distribuição dos subfenótipos, diminuindo a quantidade dos monócitos com maior susceptibilidade a infecção por HIV-1 (CD14lowCD16+). A investigação da ANXA1 endógena em um modelo não humano de AIDS, realizado pela infecção de macacos por SIV, mostrou que a expressão transcricional da ANXA1 é regulada durante a progressão da doença nos principais compartimentos de infecção, o sangue e o intestino. Em comparação com as amostras de animais não-infectados, os transcritos de ANXA1 aumentaram gradualmente no sangue periférico entre as fases aguda e crônica da infecção por SIV, enquanto que na mucosa intestinal houve uma regulação negativa, atingindo os níveis basais apenas na infecção crônica. A análise de marcadores típicos da progressão da AIDS mostrou que a expressão de ANXA1 está relacionada à ativação de células T. No entanto, a expressão da ANXA1 tem uma correlação positiva com o número de transcritos de citocinas antiinflamatórias e uma correlação negativa com a carga viral e depleção de células T CD4+ no sangue e no intestino, sugerindo que a ativação da sua transcrição está relacionada a tentativa de diminuir a exaustão imunológica durante a infecção. Frente aos dados da modulação diferencial da ANXA1 no intestino de primatas, o próximo objetivo foi analisar a expressão da ANXA1 nos portadores de IBD (doença inflamatória intestinal) tratados ou não com drogas imunossupressoras. Nos indivíduos com IBD, a expressão da ANXA1 é diminuída em nível de RNAm, no sangue periférico, e proteína, na mucosa colônica. Nestes pacientes, a imunoterapia com infliximab (anti-TNF-α) modulou sistemicamente a transcrição da ANXA1 e do TNF-α, e ainda a ativação linfocitária e a bacteremia. Em suma, a dinâmica da ANXA1 somada a outros elementos imunológicos e clínicos parecem estar associados a progressão do curso das IBDs. Finalmente, os processos inflamatórios agudo e crônico induzidos pelo procedimento de isquemia e reperfusão (I/R) revelou que o peptídeo mimético ANXA1 exógeno protegeu significativamente contra a lesão por I/R renal. Os animais pré-tratados com Ac2-26 mostraram menores taxas de filtração glomerular, osmolalidade urinária e desenvolvimento da necrose tubular aguda, possivelmente, por manter a integridade tecidual devido ao bloqueio total do extravasamento de neutrófilos, à atenuação de infiltração de macrófagos e à regulação da expressão protéica da ANXA1 endógena em células epiteliais renais. Estes resultados apontam um importante papel da ANXA1 na defesa das células epiteliais contra a lesão de I/R e indicam que os neutrófilos são mediadores-chave para o desenvolvimento da lesão do tecido renal após I/R. Conclusões: Em uma análise geral do conjunto de estudos apresentados, é possível concluir que a ANXA1 exógena e seus peptídeos derivados atuam como moléculas chave na modulação específica da ativação, imunofenótipo e distribuição dos fagócitos e linfócitos que participam como linha de defesa ou como alvo de agentes infecciosos, por meio da ligação com os FPRs ou por uma via ainda não elucidada. E, enfatizando a relevância da ANXA1 na defesa da homeostasia, ainda foi constatado que as vias relacionadas à regulação da ANXA1 endógena são ativadas diferencialmente em tecidos e fluidos sob inflamação aguda e crônica iniciadas quer seja por infecção viral, autoimunidade ou hipóxia. / Doutor em Genética e Bioquímica

A corticoterapia tem sido utilizada frequentemente nas éguas suscetíveis. O uso de isuflupredona melhora a taxa de prenhez e altera o perfil proteico do líquido endometrial em relação a éguas não tratadas. A utilização de dexametasona diminui o acúmulo de líquido pós-cobertura, reduz o edema do útero, porém, desconhecem-se seus efeitos no perfil proteico do líquido endometrial. O objetivo do presente estudo foi analisar o efeito da dexametasona em éguas suscetíveis à endometrite persistente póscobertura sobre perfil proteico do líquido endometrial na presença ou ausência de infecção. Nove éguas suscetíveis foram utilizados, com idade entre 7 a 30 anos. Após a verificação dos sinais de estro as éguas foram submetidas a quatro tratamentos: (C) éguas não receberam nenhum tipo de tratamento e serviram como controle; (D) éguas receberam 40mg de dexametasona (IV), no momento da cobertura, com coleta da amostras após 6 horas, (I-6 e I-24) infusão intra-uterina de 1 x 109 de S. zooepidemicus/mL, com coleta da amostra após 6 e 24 horas; (I/D-6 e I/D-24) infusão intra-uterina de 1 x 109 S. zooepidemicus/mL e administração de 40mg de dexametasona (IV), com coleta da amostra após 6 e 24 horas. Todas as éguas foram submetidas a todos os tratamentos. As amostras foram coletadas e submetidas à eletroforese bidimensional para separação proteica e espectrometria de massa para a identificação das bandas proteicas relevantes. A corticoterapia provocou alteração na proteômica do líquido endometrial de éguas suscetíveis, caracterizada pelo aumento (TTR) e/ou diminuição (ApoA1) na densidade óptica de proteínas da fase aguda da inflamação. Conclui-se que a utilização da dexametasona em éguas com e sem presença de infecção altera a proteômica do fluido endometrial de éguas suscetíveis. Sugere-se que a dosagem ou a frequência de aplicação da dexametasona deva ser aumentada. / Corticotherapy has often been used in susceptible mares. The use of isuflupredona improves pregnancy rate and alters the protein profile of the endometrial fluid in relation to untreated mares. The use of dexamethasone decreases the post breeding fluid accumulation, reduces the uterine edema, however is unaware of its effects on the protein profile of endometrial fluid. The aim of the present study was analyze the effect of dexamethasone in mares susceptible to post-breeding persistent endometritis on the protein profile of endometrial fluid in the presence or absence of infection. Nine susceptible mares were used, aged 7-30 years old. After checking the signs of estrus, mares were subjected to four treatments: (C) mares received no treatment and served as controls; (D) mares received 40 mg of dexamethasone at breeding time, with collection of samples after 6 hours; (I-6 and I-24) intrauterine infusion of 1 x 109 S. zooepidemicus/ml and the sample was collected after 6 and 24 hours; (I/D-6 and I/D-24) intrauterine infusion of 1 x 109 S. zooepidemicus/ml and 40 mg of dexamethasone administration, collecting the sample after 6 and 24 hours. All of the mares were subjected to all treatments. Samples were collected and subjected to two-dimensional electrophoresis for protein separation and mass spectrometry for the identification of relevant protein bands. Corticotherapy resulted in alteration of the protein profile of the endometrial fluid of susceptible mares, characterized by an increase (TTR) and/or decrease (ApoA1) in optical density of the acute phase of proteins of inflammation. We conclude that the use of dexamethasone in mares with and without the presence of infection alters the protein profile of endometrial fluid of susceptible mares. It is suggested that the dosage or frequency of application of dexamethasone should be increased.

Abstract A large body of animal studies indicates that reinforcement from alcohol is associated with dopaminergic neurotransmission in the mesocorticolimbic pathway. However, as most psychiatric phenomena cannot be studied with animals, human studies are needed. Furthermore, because of the fluctuating nature of phenomena regarding the status of abuse and withdrawal, repeated observations of the same study subjects under different situations can elucidate a variety of pathophysiological mechanisms. In this study 42 alcoholics were monitored during withdrawal and 30 alcoholics after four weeks of abstinence. 123I-β-CIT SPET was used as a method for the semi quantification of their striatal dopamine transporter (DAT) densities reflecting the function and structure of the dopaminergic system. DAT density was markedly lower during withdrawal among alcoholics as compared to control subjects, but it elevated during abstinence to the level of healthy volunteers. This increases in DAT density during withdrawal and afterwards correlated with the depression scores of alcoholics. DAT density correlated with the Novelty Seeking (NS) personality trait, especially among abstinent alcoholics. After four weeks of controlled abstinence alcoholics with an A1 allele of dopamine receptor D2 were found to have higher DAT densities than alcoholics without it. The results indicate that striatal DAT density is associated with mood, personality, A1 genotype and the length of the abstinence period after heavy alcohol drinking.

This diploma thesies is focused on design of a sawing machine for profiles of the aluminium profile system. In the beginning the basic types of sawing maschines are described, followed by the analysis of alternative solutions, selecting the best option, design of the final solution and evaluation. The resulting solution should be ergonomically acceptable, safe, affordable and efficient.

No description available.

Background: Mild Cognitive Impairment (MCI) is seen as a state between normal aging and dementia, with patients having an increased risk of developing Alzheimer’s disease (AD) and other sorts of dementia. MCI has been linked to a change in gut microbiota which impacts the microbiota-gut-brain axis (MGBA), consequently affecting neurological functions. A way of altering microbiota and thereby promoting cognitive health is through the administration of prebiotics, probiotics, and synbiotics. Aim: This systematic literature review aims to assess the impact of prebiotics, probiotics, and synbiotics on MCI by compiling existing data on the matter. Methods: Three databases - Web of Science, Cochrane, and PubMed - were searched and articles were included based on the following inclusion criteria: (1) randomized clinical trials (RCTs), (2) conducted on adults evaluated with MCI during the study, (3) including a prebiotic, probiotic, or synbiotic intervention of any kind, (4) comparing the intervention with a placebo or control group, (5) written in English, (6) reporting the main outcome of cognitive function using any neuropsychological evaluation test. Results: Five studies were included in the final selection. These studies showed that cognitive function improved after probiotic intervention, significantly affecting several cognitive domains: attention, calculation, orientation in time, and delayed memory. Two studies showed that subjects with low cognitive scores at baseline benefited more from probiotic supplementation compared to high-scoring subjects. Conclusions: Probiotics appear to improve cognition in MCI subjects; however, further research is needed to conclude the effects of prebiotics and synbiotics.