Os parabenos utilizados como conservantes nas indústrias de cosméticos, alimentos e fármacos não são removidos por completo nas estações de tratamento de água e esgoto, além disso, podem causar danos a biota aquática. O presente estudo teve como finalidade aplicar um método analítico novo para quantificar o metil (MP), etil (EP), propil (PP), butil (BP), benzilparabeno (BzP) e a mistura (metil e propilparabeno) em amostras de água dos aquários com tilápias do Nilo (Oreochromis niloticus). A técnica analítica usada foi a cromatografia líquida com detector de arranjo de diodo (HPLC-DAD). Avaliou-se a toxicidade dos parabenos em tilápias e os efeitos nos biomarcadores bioquímicos dos animais após 6 e 12 dias dos testes de exposição e por administração via injeção intraperitoneal. A concentração dos parabenos utilizada em todos os testes foi de 4,0 mg L-1 (de cada parabeno individualmente) e de 6,0 mg L-1 do metil e de 1,7 mg L-1 do propilparabeno para a mistura. Foram feitas análises nos biomarcadores superóxido dismutase (SOD), catalase (CAT), glutationa peroxidase (GPx), glutationa redutase (GR), glutationa reduzida(GSH-t) e peroxidação lipídica (MDA). O limite de detecção dos parabenos foi de 0,03 mg L-1 (MP e EP), 0,05 mg L-1 (PP) e 0,10 mg L-1 (BP e BzP), e o limite de quantificação foi de 0,13 mg L-1 (MP, EP, PP), 0,18 mg L-1 (BP) e 0,25 mg L-1 (BzP). Foi possível quantificar somente o PP e a mistura (MP + PP) nas amostras de água dos aquários que continham peixe, no máximo 30 h após a exposição. Nas amostras de água sem a presença dos peixes, foi possível quantificar o BP e a mistura, metil e propilparabeno, durante os 12 dias de exposição. Os testes de toxicidade mostraram que a concentração letal para 50% dos indivíduos após 48 h de exposição foi de 67,11 mg L-1 do MP, 24,08 mg L-1 do EP, 17,34 mg L-1 do PP, 7,98 mg L-1 do BzP e 7,80 mg L-1 do BP, sendo que estes dois últimos compostos podem ser considerados os mais tóxicos da classe. Outro modo de ação tóxica também observada dos parabenos foi a narcose, ou seja, a perda temporária da consciência e da mobilidade. À medida que aumenta o comprimento da cadeia, aumenta a lipofilicidade destas substâncias, que está relacionada com o coeficiente de partição octanol/água (Kow) das mesmas e consequentemente aumenta a toxicidade. Estes dados indicaram que quanto mais lipofílico mais tóxico é o composto. Relacionando as atividades enzimáticas testadas com os níveis de peroxidação lipídica, o metilparabeno foi o único composto capaz de provocar danos aos tecidos testados por meio das espécies reativas de oxigênio. Isso foi comprovado através da inibição da atividade das enzimas analisadas com o aumento nos níveis de MDA. Por outro lado, mesmo com as enzimas antioxidantes apresentando atividades elevadas isso não foi suficiente para impedir a redução nos níveis de GSH-t. Tais resultados indicam que os parabenos podem agir negativamente nas tilápias. / Parabens, used as preservatives in cosmetics, foodstuffs and pharmaceuticals are not completely removed from water in sewage treatments, which may cause damage to aquatic biota. The present study addresses a new methodology to measure the quantity of methyl (MP), ethyl (EP), propyl (PP), butyl (BP), benzyl (BzP) parabens and a mixture of methyl and propylparaben in water. Aquarium water of experiments with tilapia samples was analyzed for 12 days by liquid chromatography with a doide array detector (HPLC-DAD). The toxicity of parabens in Nile tilapia (Oreochromis niloticus) and its effects on biochemical biomarkers were also evaluated after 6 and 12 days of exposure and intraperitoneal injection. The concentrations of parabens used in all tests were 4.0 mg L-1 (alone) and to mixture was 6.0 mg L-1 of methyl and 1.7 mg L-1 of propylparaben. Biomarkers, superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), glutathione reductase (GR), glutathione reduced (GSH-t) and lipid peroxidation (MDA) were analyzed. The results show the detection limits for the analysis of parabens were 0.03 mg L-1 (MP and EP), 0.05 mg L-1 (PP) and 0.10 mg L-1 (BP and BzP), and the quantification limits were 0.13 mg L-1 (MP, EP, PP), 0.18 mg L-1 (BP) and 0.25 mg L-1 (BzP). In the water sample with fish, the compounds PP and the mixture (MP + PP) could be quantified up to 30h after exposure. In the water sample without fish, the compounds BP and the mixture were quantified for 12 days of exposure. Toxicity test revealed the lethal concentrations for 50% of individuals after 48 h of exposure were 67.11 mg L-1 for MP, 24.08 mg L-1 for EP, 17.34 mg L-1 for PP, 7.98 mg L-1 for BzP and 7.80 mg L-1 for BP. Therefore, BzP and BP can be considered the most toxic of the class. As the chain length grows, the lipophilicity of the substances increases. Such an increase is related to their octanol/water (Kow) partition coefficient and, consequently, increases toxicity. Another toxic action observed for the parabens was the temporary loss of consciousness and mobility of the organisms. According to the enzymatic activity tested and the lipid peroxidation levels, the methylparaben was the only compound that caused damage by reative oxidative species, supported by the inhibition of the activities of the enzymes and the increase in the MDA levels. However, the high activity of the antioxidant enzymes to exposure and intraperitoneal injections could not prevent the reduction in the levels of GSH-t. Such results indicate parabens can cause negative effects on tilapia.

Ma, Buig Yue Brigette. / Thesis M.D. Chinese University of Hong Kong 2014. / Includes bibliographical references (leaves 232-270). / Title from PDF title page (viewed on 18, November, 2016).

Os meningiomas são os tumores primários do Sistema Nervoso Central (SNC) mais frequentes e representam 35,5% dos casos considerando-se todas as faixas etárias. Apesar dos progressos ocorridos nas últimas décadas, a tumorigênese dos meningiomas ainda permanece como um desafio. Há um consenso da necessidade de ferramentas moleculares para ajudar tanto no diagnóstico quanto no prognóstico dos meningiomas. Neste contexto, alguns trabalhos demonstram a importância do papel dos receptores de estrógeno e progesterona, assim como o entendimento das alterações nos níveis de expressão dos microRNAs (miRNAs) na tumorigênese dos meningiomas. Alguns estudos demonstram que o perfil de expressão sérico dos miRNAs tem correlação com a classificação e evolução clínica, sendo de grande interesse o uso desse material por se tratar de um procedimento não-invasivo, ou seja, como biomarcadores. Objetivos: avaliar o perfil de expressão tecidual e sérica de microRNAs associados as vias dos receptores de estrógeno e progesterona em meningiomas grau I, II e III. Pacientes e métodos: foram utilizadas amostras de tecido e plasma de 40 pacientes com meningiomas grau I, II e III. Para a análise da expressão dos miRNAs miR-34a, miR-143, miR-145 e miR-335 foi utillizada a técnica de PCR em tempo real. Resultados: os miRNAs: miR-34a e miR-145 apresentaram diferença estatística significativa nas amostras de tecido tumoral entre os grupos estudados com menor expressão nas amostras de meningiomas grau II quando comparadas as amostras grau I e III. Não observamos diferença estatística estatística significativa na expressão dos miRNAs nas amostras de plasma. Conclusão: os miRNAs selecionados não apresentaram correlação com a progressão tumoral em meningiomas. / Meningiomas are the most common primary Central Nervous System (CNS) tumors, accounting for 35.5% of the cases, considering all age groups. Despite the progress made in recent decades, the tumorigenesis of meningiomas still remains a challenge. There is a consensus of the need for molecular tools to assist both diagnosis and prognosis of meningiomas. In this context, some studies demonstrate the importance of the role of estrogen and progesterone receptors, as well as the understanding of alterations in microRNA (miRNAs) expression levels in the tumorigenesis of meningiomas. Some studies have shown that the serum expression profile of the miRNAs correlates with the classification and clinical evolution, being of great interest the use of this material because it is a non-invasive procedure, i.e., as biomarkers. Objectives: To evaluate the tissue and serum expression profile of microRNAs associated with the estrogen and progesterone receptor pathways in meningiomas grade I, II and III. Patients and methods: tissue and blood samples from 40 patients with grade I, II and III meningiomas were used. For analysis of miRNA expression miR-34a, miR-143, miR-145 and miR-335 was used the real-time PCR technique. Results: miRNAs: miR-34a and miR-145 presented a significant statistical difference in the tumor tissue samples between the groups with lower expression in the samples of grade II meningiomas when compared to samples I and III. We did not observe statistically significant statistical difference in miRNA expression in blood samples. Conclusion: the selected miRNAs showed no correlation with tumor progression in meningiomas.

Background: Psoriatic arthritis (PsA) is a chronic heterogenous inflammatory arthritis with five phenotypes. The two least studied phenotypes are investigated in this thesis, including: psoriatic spondyloarthropathy (PsSpA) and psoriatic arthritis mutilans (PAM). The aims of this thesis were to determine the prevalence, clinical characteristics and radiographic characteristics of PsSpA and PAM in a cohort of PsA patients, and serum-soluble bone- turnover biomarkers of these phenotypes. Aims: Comparisons were made with PsA patients without axial disease (pPsA), and ankylosing spondylitis (AS) patients. Methods: A prospective single-centre cross-sectional study was conducted of PsA and AS patients. Serum on psoriasis-only patients (PsC) and healthy controls (HC) were also obtained. Multivariate clinical, radiographic, genetic and serum biomarker comparisons were made between these five groups of subjects. Results: The study enrolled 201 PsA and 201 AS patients, who were then reclassified as 118 PsSpA, 127 pPsA and 157 AS cases, alongside 200 PsC and 50 HC subjects. Several clinical biomarkers, imaging biomarkers, serum-soluble biomarkers and genetic biomarkers were identified that differentiate PsSpA from pPsA and AS. PsSpA affected a significant proportion of PsA patients, and was not a milder version of AS. PsSpA involvement was as disabling and clinically impactful as AS. PAM was found to be associated with PsSpA, and clinical biomarkers of PAM occurrence and radiographic progression were identified. Conclusions: In conclusion, this thesis indicates that PsSpA is on a spectrum of musculoskeletal disease, in between pPsA and AS; with PsSpA comprising a continuum itself, and with a phenotype expression related to disease duration. These findings may prompt the inception of an international-consensus classification system for PsSpA, for which there is a great clinical need. Given that PsSpA has its own discrete clinical and biomarker signature, its clinical management and research should be tailored from that of pPsA and AS. Ultimately this may further the effort for stratified and personalised medicine.

Technologies for early detection of tumors is critical for better therapy outcome and overall change in cancer survival. These assays must be capable of detecting tumors at early stages in order to prevent metastasis of the tumor and help reduce mortality. Biological molecules can serve as markers that can indicate the presence of cancerous cells. Current biomarkers approved by the FDA include CA 125, which is a tumor associated antigen (TAA). However, the sensitivities of these TAAs is not high enough to detect at early stages of disease. Recent technologies have found that antibodies that recognize these TAAs, also known as autoantibodies, provide more sensitive means to screen for tumors. This review aims to present recent literature data relative to the field of cancer diagnosis and treatment. However, one should note that this article covers only fraction of the broad science behind this subject.

A periodontite é uma doença infecciosa caracterizada pela inflamação dos tecidos de suporte dos dentes, perda de inserção e perda óssea. O diagnóstico convencional da periodontite é realizado por meio da avaliação dos parâmetros clínicos e radiográficos. Entretanto, novas modalidades diagnósticas para identificação do início e progressão da periodontite estão sendo estudadas, o que possibilitaria o diagnóstico precoce da progressão da doença, assim como avaliação da resposta ao tratamento periodontal. Este estudo se propôs a monitorar a atividade da doença periodontal e descrever as características clínicas e moleculares de sítios periodontais ativos em progressão em pacientes com periodontite crônica, a partir da avaliação da expressão gênica de sítios periodontais e da avaliação de proteínas inflamatórias salivares e do fluido gengival crevicular, antes e após a terapia periodontal básica. Foram selecionados 27 indivíduos e classificados em dois grupos: grupo Controle (n=9) - pacientes saudáveis; grupo DP (n=18) - pacientes com periodontite crônica. O exame clínico foi realizado por um único examinador experiente em três tempos: (i) quinze dias antes da terapia periodontal, (ii) no dia da terapia periodontal e (iii) 60 dias após a terapia periodontal. A coleta de fluido gengival foi realizada no baseline, 15 e 60 dias após a terapia; a coleta de saliva foi realizada no baseline e 60 dias após a terapia e a coleta de tecido gengival apenas no baseline. Para a coleta de fluido gengival e tecido gengival, os sítios foram classificados em: sítios com inflamação (PS &ge; 5 mm e presença de sangramento à sondagem nos dois exames clínicos iniciais); sítios sem inflamação (PS &le; 3 mm e ausência de sangramento à sondagem nos dois exames clínicos iniciais); e sítios controle (PS &le; 3 mm e ausência de sangramento à sondagem). Os sítios com e sem inflamação pertenciam ao mesmo paciente do grupo DP. A análise de expressão gênica de VEGF, MMP-8, IL-10, RANK-L, OPG e TGF-&beta;1 foi realizada através da Reação em Cadeia da Polimerase em Tempo Real (RT-PCR). As análises de marcadores na saliva e fluido gengival foram realizadas pelo imunoensaio Multiplex Cytokine Profiling, exceto para MMP-8 que foi feito através do método ELISA. Os sítios com inflamação apresentaram maior expressão de RANK-L, OPG, IL-10 e TGF-&beta;1 (p < 0,05) e maior quantidade (pg) de VEGF (p=0,009) e IL-10 (p < 0,05) no fluido gengival. Os pacientes do grupo DP apresentaram maior quantidade (pg) de RANK-L (p=0,03) e OPG (p=0,0002) na saliva, antes da terapia. A atividade da doença em sítios periodontais a após terapia básica é um evento de baixa ocorrência, mas apenas uma pequena fração dos sítios permaneceu com perda de inserção progressiva. Os biomarcadores utilizados neste estudo podem ser úteis para detectar inflamação, mas não para identificar sítios ativos em progressão. / Periodontal disease is a chronic microbial infection characterized by inflammation of supportive tissues and alveolar bone loss. Because of the increasing prevalence of the disease, diagnostic modalities for the early identification of periodontitis initiation and progression are being studied. Cytokines associated to host defense has been identified in saliva, gingival crevicular fluids and gingival tissues of periodontal patients. In this study, we aimed to monitor periodontal disease activity and investigate clinical and molecular features of active sites through saliva, gingival crevicular fluid and gingival tissue samples. Fifty-seven subjects were enrolled for this study, 18 with chronic periodontitis (PD group) and 9 subjects that were periodontal and systemically healthy (control group). The patients underwent clinical examination and collection of saliva before and two months after the non-surgical periodontal therapy; collection of gingival crevicular fluid at baseline, 15 days and 2 months after therapy; and collection of gingival tissue samples at baseline. For collection of gingival crevicular fluid and gingival tissue samples, sites were classified as: inflamed (probing depth &ge; 5 mm and bleeding on probe); non-inflamed (probing depth &le; 3 mm without bleeding on probe); and control sites (probing depth &le; 3 mm without bleeding on probe from control group). Inflamed and non-inflamed sites belongs to the same patient PD group. Samples of whole saliva were collected for assessment of the levels of MMP-8, VEGF, IL-10, RANKL, OPG and TGF-&beta;1 using the multiplex cytokine profiling assay. Samples of gingival crevicular fluid were collected for assessment of the levels of MMP-8, VEGF and IL-10 using the multiplex cytokine profiling assay, except for MMP-8 (ELISA assay). Inflamed and non-inflamed lesion in each patient underwent biopsy for the Real Time PCR gene expression analysis for MMP-8, VEGF, IL-10, RANKL, OPG and TGF-&beta;1. At baseline, higher expression of mRNA for RANK-L, OPG, IL-10 e TGF-&beta;1 were found in inflamed sites (p < 0.05) and higher total amount of IL-10 (0,29 pg/sample) compared to non-inflamed (0,21 pg/sample) and control sites (0.21 pg/sample) (p < 0.05). 15 days after therapy, total amount of VEGF were higher in inflamed sites (11.43 pg/sample), compared to non-inflamed sites (7.38 pg/sample) (p=0.009). PD group showed higher total amount (pg) of RANKL (p=0.03) and OPG (p=0.0002) after periodontal therapy. Thus, we concluded that disease activity seems to be an event of relative low probability of occurrence, however, a small percentage of sites keeps showing progressive attachment loss even after basic periodontal therapy. The specific biomarkers used in this study may be helpful to detect inflammation but not to discriminate progressive active sites.

A associação entre o diabetes e a periodontite produz uma descarga de proteínas inflamatórias que pode ser refletida na saliva. Com base nisso, o objetivo desse estudo foi mensurar as concentrações salivares de interleucina-6 (IL-6), metaloproteinase da matriz-8 (MMP-8) e osteoprotegerina (OPG) em pacientes com periodontite crônica associada ou não ao diabetes mellitus tipo 2. Um total de 90 indivíduos foi dividido em quatro grupos: saudáveis (Controle, n=22), pacientes com doença periodontal (DP, n=24), apenas com diabetes mellitus (DM, n=20) e com doença periodontal e diabetes mellitus (DP+DM, n=24). Dados clínicos e metabólicos foram registrados. Amostras de saliva não-estimulada foram analisadas pelo ensaio imunoenzimático (ELISA). Diferenças estatisticamente significantes foram detectadas entre os grupos para todos marcadores (p<0,05). Em relação à IL-6, diferenças estatisticamente significantes foram encontradas comparando os grupos Controle com DP e com DP+DM, e entre os grupos DP+DM com DM (p<0,005). Para MMP-8, a média das concentrações do grupo Controle foi significativamente menor que em todos os grupos doentes (p<0,01) e nenhuma diferença entre os grupos doentes foi detectada. Para OPG, diferenças estatisticamente significantes foram encontradas entre os grupos Controle com DP e entre Controle com DM (p<0,05). Todos os parâmetros clínicos foram estatisticamente significantes entre os grupos (p<0,001), exceto supuração. No grupo DP, SS (sangramento à sondagem) mostrou uma correlação positiva com as concentrações de IL-6 (r=0,48; p<0,05), PS>=7 (profundidade de sondagem>=7mm) correlacionou-se positivamente com as concentrações de MMP-8 (r=0,46; p<0,05), e os níveis de HbA1c também correlacionaram-se positivamente com as concentrações de IL-6 (r=0,54; p<0,000). Concluindo, a saliva é um adequado substrato para identificação de biomarcadores inflamatórios em pacientes periodontais com ou sem diabetes. A IL-6 é um biomarcador candidato para periodontite e periodontite associada ao diabetes na saliva. Além disso, conclui-se que a super-expressão de MMP-8 e OPG pode acionar o aumento do colapso periodontal observado em indivíduos com diabetes tipo 2. / Background: Association of diabetes and periodontitis produces an inflammatory proteins discharge that can be reflected in saliva. The aim of this study was to measure salivary concentrations of interleukin-6 (IL-6), matrix metalloproteinase-8 (MMP-8) and osteoprotegerin (OPG) in patients with chronic periodontitis associated or not to type 2 diabetes mellitus. Methods: A total of 90 subjects was divided in four groups: healthy (Control, n=22), patients with Periodontal Disease (PD, n=24), Diabetes Mellitus only (DM, n=20), Periodontal Disease and Diabetes Mellitus (PD+DM, n=24). Clinical and metabolic data were recorded. Non-stimulated saliva samples were analyzed by enzyme-linked immunosorbent assay (ELISA). Results: Significant differences were detected between groups for the biomarkers (p<0.05). Regarding IL-6, significant differences were found comparing Control with PD and with PD+DM, and comparing PD+DM with DM groups (p<0.005). For MMP- 8, the concentrations mean of the Control group was significantly lower than all the diseased groups (p<0.01), and no differences between diseased groups were detected. For OPG, significant differences were found between Control and PD, and between Control and DM groups (p<0.05). All clinical parameters were significant between groups (p<0.001), except suppuration. In PD group, BOP (bleeding on probing) showed positive correlation with IL-6 concentrations (r=0.48; p<0.05), PPD>=7 (pocket depth>=7mm) correlated positively with MMP-8 concentrations (r=0.46; p<0.05), also HbA1c levels correlated positively with IL-6 concentrations (r=0.54; p<0.000). Conclusion: Saliva is an adequate substrate for inflammatory biomarkers identification in periodontal patients. IL-6 is a candidate biomarker for periodontitis and periodontitis associated to diabetes in the saliva.

A esquizofrenia (SZ) é uma doença mental crônica e grave, que compromete o funcionamento psicossocial do indivíduo nos mais variados graus. Atinge 1% da população mundial, considerando todo seu espectro de sintomas (DSM-IV). O pobre funcionamento cognitivo é um dos principais fatores que explicam as elevadas taxas de prejuízos e encargos associados à esquizofrenia. A etiologia da SZ é desconhecida, tendo muitas hipóteses etiológicas como fatores genéticos, epidemias virais durante a gestação, época de nascimento, traumatismos de parto, infecções perinatais, condições neurológicas ou neuropsiquiátricas que geram sintomas tipo esquizofrênicos ou desenvolvimento anormal (avaliados por testes psicológicos, estudos de neuroimagem e neuropatológicos que sugerem alterações no desenvolvimento cerebral). A fisiopatologia da SZ pode ser resultante de uma desregulação na plasticidade sináptica por alterações de neurotofinas, radicais livres e processos inflamatórios. Existe uma larga evidência que os radicais livres podem ter um papel importante na fisiopatologia da SZ, podendo induzir danos na membrana celular, em proteínas e DNA. Problemas com estresse oxidativo, como o aumento da peroxidação lipídica foram relatados previamente em pacientes com SZ em primeiro episódio, virgens de tratamento e naqueles cronicamente medicados. As citocinas inflamatórias têm sido estudadas como importantes participantes na etiologia e desenvolvimento das doenças psiquiatricas. Seu papel ainda não é bem estabelecido, porém diversas alterações têm sido vistas nas doenças psiquiátricas. Dentre as citocinas, destacam-se as interleucinas (IL) e o fator de necrose tumoral alfa (TNF-α), que podem ter ação inflamatória e anti-inflamatória. Dentre as próinflamatórias, podemos destacar a IL-6 e o TNF-α. Alteração de IL na SZ tem sido relatada nesses últimos anos, relacionada à etiologia e à atividade da doença. Pacientes em episódio agudo da doença apresentaram aumento dos níveis séricos de IL pró-inflamatórias sugerindo atividade inflamatória sistêmica. Identificar, além dos sintomas clínicos, possíveis alterações bioquímicas e de neuroimagem em pacientes com SZ pode ajudar em futuras intervenções tanto para identificar, como para prevenir ou atenuar o curso da SZ. Estudos que permitam avançar no entendimento da psicopatologia deste grupo de pacientes são de grande importância, na medida em que proporcionarão futuras abordagens terapêuticas. Está bem estabelecido que a matéria cinzenta cortical e o volume de córtex préfrontal estão diminuídos em pacientes com SZ. Entretanto, os fatores que levam à perda de tecido não estão claras. Uma hipótese para esse fato é que o estado próinflamatório aumentado em SZ está relacionado com a diminuição volumétrica da massa cinzenta. O objetivo deste estudo piloto foi correlacionar os níveis séricos de IL-6 com o volume cortical total de pacientes com SZ e controles. Foram selecionados 36 pacientes com SZ (28 do sexo masculino, com idade média de 37,17 ± 12,05; anos de doença 15,56 ± 11,75), 35 controles pareados idade (21 do sexo masculino, idade média= 36,97 ± 13,04). As imagens foram adquiridas por um equipamento de ressonância magnética Philips Achieva 1.5T no Hospital de Clínicas de Porto Alegre, Brasil. Todas as imagens foram processadas usando o pipeline automatizado de FreeSurfer v5.1. Concluímos que a IL-6 está negativamente correlacionada com o volume cortical total (p= 0,027; rho= -0,370) nos pacientes com esquizofrenia, tal correlação não foi vista nos controles (p= 0,235, rho= -0,206). Nosso resultado sugere que a ativação inflamatória crônica em pacientes com SZ pode estar relacionada com a diminuição volumétrica total do córtex. / Schizophrenia (SZ) is a chronic and severe mental illness, which affects the psychosocial functioning of the individual in many degrees. It reaches 1% of the population, considering all its spectrum of symptoms (DSM-IV). Poor cognitive functioning is one of the main factors responsible for the high rates of disability and costs associated with schizophrenia. The etiology of SZ is unknown, and many etiological assumptions are taken, as genetic factors, viral epidemics during pregnancy, time of birth, birth trauma, perinatal infections, neuropsychiatric or neurological conditions that produce symptoms like schizophrenia or unnatural development (assessed by psychological tests, neuroimaging and neuropathological studies that suggest changes in brain development). The pathophysiology of SZ may be due to a deregulation in synaptic plasticity caused by changes in neurotrophins, free radicals and inflammatory processes. There is a wide evidence that free radicals may have a main role in the pathophysiology of SZ, and can induce damage into the membrane cell, in proteins and DNA. Problems with oxidative stress, such as increased lipid peroxidation have been previously reported in treatment virgem patients with SZ in first episodes and in those chronically treated. And inflammatory cytokines have been studied as important parts in the etiology of psychiatric diseases’ development. Its role is not well established, however a number of changes have been noticed in psychiatric illnesses. Among the cytokines, the Interleukins (IL) and the tumor necrosis factor alpha (TNF-α) stand out, these two may have inflammatory and anti-inflammatory action. Among the pro-inflammatory, we can highlight IL-6 and TNF-α. IL change in the SZ has been reported in these last few years, related to the etiology and disease activity. Patients with acute episode of the disease showed increased serum levels of IL proinflammatory suggesting systemic inflammatory activity. Identify not only the clinical symptoms, possible biochemical and neuroimaging abnormalities in patients with SZ can help in future interventions both to identify and prevent or slow down the course of SZ. Studies to enable progress in the understanding of psychopathology this group of patients are of great importance to the extent that provide future therapeutic approaches. It is well established that cortical gray matter and the prefrontal cortex volume are reduced in patients with SZ. However, the factors that lead to tissue loss are unclear. One possible explanation is that the increased proinflammatory state in SZ is related to the volumetric reduction of the gray matter. The objective of this pilot study was to correlate serum levels of IL-6 in the hole cortex volume of schizophrenic patients and controls. We selected 36 patients with SZ (28 male, average age 37.17±12.05; years of illness 15.56±11.75), 35 matched controls (21 male, average age= 36.97±13.04). Images were obtained by an MRI equipment, brand Philips Achieva 1.5T at Hospital de Clinicas de Porto Alegre, Brazil. All images were processed using automated pipeline FreeSurfer v5.1. We concluded thatIL-6 is negatively correlated with the total cortical volume in patients (p= 0.027, rho= -0.370), this correlation was not seen in controls (p= 0.235; rho= -0.206). Our results suggest that chronic inflammatory activation in patients with SZ can be related to the total volumetric reduction of the cortex.

Cette thèse a pour objectif d’identifier et caractériser des biomarqueurs liés aux traitements d’immunothérapie allergénique (ITA). En particulier, le travail s’est porté sur les biomarqueurs prédictifs de l’efficacité du traitement : leur estimation avant la prise médicamenteuse permettrait d’évaluer les bénéfices cliniques à l’issue de l’ITA. À partir d’échantillons de sérum collectés avant ITA et provenant de patients inclus dans une étude clinique contrôlée, randomisée et dirigée contre les pollens de graminées, une méthode d’analyse protéomique différentielle (la 2D-DIGE) a permis d’identifier une protéine comme candidat biomarqueur prédictif de l’efficacité de l’ITA. Dans un premier temps, les variants moléculaires de la protéine identifiée ont été caractérisés. L’analyse en spectrométrie de masse de la protéine a permis d’identifier les modifications post-Traductionnelles associées aux isoformes plus abondants dans le sérum des patients pour lesquels une réponse positive à l’ITA est observée. Une approche protéomique de quantification relative a été mise en œuvre par LC-MS/MS sans marquage, et a permis d’identifier des peptides de la protéine d’intérêt, portant des modifications post-Traductionnelles spécifiques, associés à un bénéfice clinique accru en fin d’ITA. Dans un deuxième temps, l’implication de la fétuine-A dans la physiopathologie de l’allergie a été étudiée. Des modèles cellulaires humains ont mis en évidence le caractère essentiel des acides sialiques portés par la protéine pour l’activation de la voie TLR4, dans les mécanismes de l’immunité innée initiant la réaction allergique. In vivo, les modèles murins d’asthme allergique développés ont en revanche montré la fonction anti-Inflammatoire de la protéine. La fétuine-A a donc un rôle ambivalent, mais est indubitablement liée à la régulation de l’inflammation allergique. La validation des candidats biomarqueurs peptidiques de la protéine dans d’autres cohortes cliniques représente une future étape clé, qui permettrait d’envisager l’usage de ces marqueurs en clinique, pour améliorer la sélection des patients les plus susceptibles de répondre à l’ITA. / This thesis aimed at identifying and characterizing predictive biomarkers associated with allergen-Specific immunotherapy (AIT) efficacy. In particular, we were focused on biomarkers predictive of AIT efficacy: quantifying such markers before treatment would allow estimating the final clinical benefit. For this purpose, serum samples were collected before AIT from patients included in a double-Blind, placebo controlled clinical study against grass pollen allergy. Their analysis by means of differential proteomics (2D-DIGE) pointed out a protein, named fetuin-A, as a candidate biomarker predictive of AIT efficacy. First, the protein fetuin-A isoforms were extensively characterized by mass spectrometry, and specific post-Translational modifications (PTMs) were associated to the isoforms more abundant in sera from patients positively responding to AIT. A second proteomic approach allowed identifying fetuin-A peptides, differentially expressed among groups of patients. Some peptides from the candidate protein, bearing specific post-Translational modifications (PTMs), are associated with an increased clinical benefit at the end of the treatment. In a second part, we studied the involvement of the fetuin-A during the course of allergic inflammation. Human cellular assays highlighted that sialic acids on the protein PTMs are essential to activate the TLR4 pathway, and inducing innate immune mechanisms linked to allergy. In vivo, murine models of allergic asthma showed an opposite anti-Inflammatory function of the protein. Thus, the protein fetuin-A is still ambivalent, but is undoubtedly linked to the regulation of allergic inflammation. Validation of peptides candidate biomarkers in larger clinical cohorts is of utmost interest, since using such biomarkers in clinics would improve patients’ selection and therefore clinical benefit from the treatment.

La caractérisation moléculaire multidimensionnelle des tumeurs et des tumeurs gliales en particulier est une étape importante pour l’identification de biomarqueurs (diagnostique, pronostique, théranostique et/ou de prédisposition), pour l’identification de cibles thérapeutiques et pour une meilleure compréhension de l’oncogénèse moléculaire.Nos travaux ont permis de confirmer et de consolider certaines données de la littérature comme par exemple : (i) la valeur pronostique favorable de la codélétion 1p/19q, (ii) la valeur pronostique favorable de la mutation IDH, (iii) le caractère mutuellement exclusive des mutations TP53 et de la codélétion 1p/19q et (iv) la rareté des altérations génétiques du PDGFRA dans les gliomes de bas grade (GDBG). De manière plus originale, nous avons identifié plusieurs sous-groupes génomiques de GDBG pertinents sur le plan clinico-biologique, notamment au sein des GDBG non 1p/19q codélétés : (i) 19q-délété ; (ii) 11p-délété, (iii) 7-gagné, (iv) 19-gagné et (v) inclassés. La perte du bras chromosomique 19q annule la valeur pronostique favorable de la mutation IDH dans les GDBG non 1p/19q codélétés. Nous avons également identifié des mutations géniques originales dans les GDBG (i.e. mutation TEP1 et RNF40) qui renforcent le rôle des télomères et du remodelage de la chromatine au sein des GDBG.Enfin, nous nous sommes concentrés sur la caractérisation des GDBG 11p-délétés qui sont de phénotype majoritairement astrocytaire et de moins bon pronostic. Ces GDBG surexpriment des gènes des cellules immunitaires (les GIM -Glioma infiltrating microglia-, les macrophages de type 1, les macrophages de type 2) et sont infiltrés par des cellules macrophagiques et microgliales. Ce microenvironnement dérégulé peut constituer une cible thérapeutique au sein des GDBG 11p-délétés. En conclusion, nos travaux participent à la dissection clinico-moléculaire des GDBG et à préciser la biologie d’un sous-type de GDBG caractérisé la perte du bras chromosomique 11p. / Multildimensional molecular characterization of tumors and more specifically of gliomas is of pivotal importance to identify: (i) new biomarkers (i.e. diagnostic, prognostic, theranostic or predisposing), (ii) new therapeutic targets and (iii) to improve our understanding of molecular oncogenesis.Our work has confirmed and consolidated previous data published in the literature, for example that: (i) 1p/19q co-deletion is associated with better prognosis, (ii) IDH mutation is associated with better prognosis, (iii) TP53 mutations and 1p/19q codeletion are mutually exclusive and (iv) PDGFRA is rarely altered, at genomic level, in low-grade gliomas (LGG).More originally, we have identified several genomic groups, with clinical and biological relevances, in LGG and more specifically in LGG without 1p/19q co-deletion: (i) 19q-deleted, (ii) 11p-deleted, (iii) 7-gained, (iv) 19-gained and (v) unclassified. Interestingly, 19q deletion abrogates the positive prognostic value of IDH mutation in LGG without 1p/19q codeletion.We have also identified new recurrent somatic gene mutations in LGG (i.e. TEP1 and RNF40 mutations), supporting the critical role of telomeres and chromatin remodelling in LGG.Finally, we have characterized further 11p-deleted LGG that exhibit mostly astrocytic phenotype and poor prognosis. This subgroup includes LGG overexpressing genes of inflammatory/immune cells (GIM -Glioma infiltrating microglia-, M1 macrophages and M2 macrophages) and infiltrated by macrophagic/microglial cells. This peculiar microenvironment detected in 11p-deleted LGG might be used as a therapeutic target. In conclusion, our work participates to characterize clinico-biological portrait of LGG and to describe a singular genomic subgroup of LGG characterized by 11p loss.