The proteome of the corpus callosum in schizophrenia

Orientador: Daniel Martins de SouzaDissertação (mestrado) Universidade Estadual de Campinas, Instituto de BiologiaResumo: A esquizofrenia (SCZ) é um transtorno mental incurável que está relacionado ao neurodesenvolvimento e afeta cerca de 1% da população mundial. A SCZ pode ser considerada a principal forma de psicose devido a sua grande frequência e importância clínica, sendo devastadora tanto para o paciente quanto para os seus familiares. No presente trabalho foram analisados e comparados o proteoma e fosfoproteoma do corpo caloso provindos de pacientes com esquizofrenia e controles pareados coletados post-mortem. Foram usados amostras previamente enriquecida para as proteínas solúveis (citoplasma) e proteoma total, nessa última análise verificamos também o conjunto de proteínas diferencialmente fosforiladas. A análise desses conjuntos de amostras foram feitas por nano-cromatografia líquida seguida de espectrometria de massas em tandem (nano LC-MS/MS). As proteínas encontradas diferencialmente expressas em ambos os estudos foram submetidas a análise de vias metabólicas no programa Ingenuity Pathway Analysis. Foi encontrado que vias de sinalização celular estão desreguladas na maior região de substância branca cerebral, com destaque para as vias Efrina e 14-3-3. Os dados gerados nessa dissertação auxiliaram em uma melhor compreensão das bases moleculares da esquizofrenia, através da integração das vias bioquímicas e na identificação de moléculas-chave na patologiaAbstract: Schizophrenia (SCZ) is an incurable mental disorder that is related to neurodevelopment and affects about 1% of world population. SCZ can be considered the main form of psychosis given its high frequency and clinical significance, and it is devastating for both patients and family. This study aimed to analyze and compare the proteome and phosphoproteome of the corpus callosum stemmed from schizophrenia patients and matched controls collected post-mortem. We used previously enriched samples to analyze soluble proteins (cytoplasm) and total proteome, in which were also identified the set of differentially phosphorylated proteins. The analysis of these sets of samples were made by nano liquid chromatography followed by tandem mass spectrometry (nano LC-MS / MS). The proteins found differentially expressed in both studies were subjected to analysis of metabolic pathways in the Ingenuity Pathway Analysis software. It was found that cell signaling pathways are deregulated in most of cerebral white matter, especially the Ephrin and 14-3-3 way. The data generated in this work aided in better understanding the molecular basis of schizophrenia, through the integration of biochemical pathways and the identification of key molecules in the pathologyMestradoBioquimicaMestra em Biologia Funcional e Molecular151787/2F2014-0CNP

Human cerebral organoids and fetal brain tissue share proteomic similarities

The limited access to functional human brain tissue has led to the development of stem cell-based alternative models. The differentiation of human pluripotent stem cells into cerebral organoids with self-organized architecture has created novel opportunities to study the early stages of the human cerebral formation. Here we applied state-of-the-art label-free shotgun proteomics to compare the proteome of stem cell-derived cerebral organoids to the human fetal brain. We identified 3,073 proteins associated with different developmental stages, from neural progenitors to neurons, astrocytes, or oligodendrocytes. The major protein groups are associated with neurogenesis, axon guidance, synaptogenesis, and cortical brain development. Glial cell proteins related to cell growth and maintenance, energy metabolism, cell communication, and signaling were also described. Our data support the variety of cells and neural network functional pathways observed within cell-derived cerebral organoids, confirming their usefulness as an alternative model. The characterization of brain organoid proteome is key to explore, in a dish, atypical and disrupted processes during brain development or neurodevelopmental, neurodegenerative, and neuropsychiatric diseases7CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO - CNPQCOORDENAÇÃO DE APERFEIÇOAMENTO DE PESSOAL DE NÍVEL SUPERIOR - CAPESFINANCIADORA DE ESTUDOS E PROJETOS - FINEPFUNDAÇÃO CARLOS CHAGAS FILHO DE AMPARO À PESQUISA DO ESTADO DO RIO DE JANEIRO - FAPERJFUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESPNão temNão temNão temNão tem14/21035-0; 16/07332-7; 13/08711-3; 14/10068-4JN, VS-C, and DM-D-S are supported by the São Paulo Research Foundation (FAPESP) grants 14/21035-0, 16/07332-7, 13/08711-3, and 14/10068-4. CS was recipient of a CAPES-FAPERJ Postdoc fellowship. Other funds are provided by the National Council for Scientific and Technological Development (CNPq), the Instituto Nacional de Ciência e Tecnologia de Neurociência Translacional (INCT-INNT), Foundation for Research Support in the State of Rio de Janeiro (FAPERJ), Coordination for the Improvement of Higher Education Personnel (CAPES), Brazilian Funding Authority for Studies and Projects (FINEP), and Brazilian Development Bank (BNDES

Impacto de um novo benchmark no mercado de balcão brasileiro

This paper can be divided in two main parts. First, we analyze the effects of the publication of a new benchmark on the Brazilian stock lending market. In the second part, we use the findings provided by the first analysis to create an instrument to assess the relation between short-selling and two characteristics of the spot market: price efficiency and stock liquidity. The study is based on a unique data set of all the loan contracts for a 60-day window around the introduction of the benchmark, containing the information of 162,195 deals of 279 tickers. We present three main findings. First, the publication of the new benchmark caused an overall reduction in average loan fees, and a narrowing of loan and brokerage fee dispersion. Second, the impact on loan fees was higher for tickers with a history of high volatility, high average loan fee and low average number of borrowers per day. Third, short-selling constraints - measured by the exogenous variation of loan fees - are associated with lower price efficiency, and an inconclusive result for market liquidityEsta dissertação pode ser dividida em duas principais partes. Na primeira, nós analisamos os efeitos da publicação de um novo benchmark no mercado de aluguel de ações. Na segunda, nós utilizamos as descobertas obtidas na primeira parte para criar um instrumento que nos permite avaliar a relação entre a venda a descoberto e duas características do mercado à vista: eficiência de preço e liquidez. O estudo é baseado em uma base de dados com todas as negociações de empréstimo de ações em um período de 60 dias em torno da implementação do novo benchmark, contendo 162.195 negócios de 279 ações. Nós apresentamos três principais achados. Primeiro, a publicação do benchmark resultou em uma queda geral das taxas de empréstimos, e uma diminuição da dispersão da taxa de empréstimo e de corretagem. Segundo, o impacto nas taxas de empréstimo foi mais alto para ações com histórico de alta volatilidade, alta taxa de empréstimo média, e baixo número de tomadores por dia. Terceiro, restrições às operações de venda a descoberto - mensuradas pela variação exógena das taxas de empréstimo - estão associadas com menor eficiência de preços, e a um resultado inconclusivo em relação à liquidez de mercad

O papel dos bancos públicos no desenvolvimento da economia brasileira (1950-2010)

O presente trabalho tem como intenção analisar as funções desempenhadas pelos principais agentes do Sistema Financeiro Público brasileiro, durante o período de 1950 até 2010, destacando ¿ em uma perspectiva keynesiana ¿ o papel desempenhado por eles no que tange a sua contribuição ao desenvolvimento econômico e social do Brasil, analisando a gestão dos recursos públicos a eles destinados. As três principais instituições desse Sistema são: o BNDES, a Caixa Econômica Federal e o Banco do Brasil. O BNDES ¿ criado em 1952 com a intenção de ser um braço direito do governo na implantação de políticas desenvolvimentistas, como agente planejador, coordenador e financiador de projetos de investimento de longo prazo ¿ é hoje reconhecido como grande realizador de investimentos de longo prazo e principal banco de fomento brasileiro, figurando entre os maiores do mundo, dentre seus congêneres (ARAÚJO & CINTRA, 2011). O Banco do Brasil foi fundado em 1808 em meio a um conjunto de medidas de organização econômica e financeira associadas à instalação da coroa portuguesa no Brasil, com o objetivo de criar um instrumento que pudesse comandar e centralizar as operações de circulação monetária e de financiamentos públicos e privados (CARDOSO, 2009). Atualmente, o Banco do Brasil figura entre os maiores e mais importantes bancos brasileiros e se destaca como maior instituição de crédito rural brasileira (HERMANN, 2009). Em 1861 foi fundada, por Dom Pedro II, a Caixa Econômica Federal com o intuito de ser o órgão responsável pela captação de modestos recursos, provenientes das classes mais baixas, garantindo as famílias uma opção segura de poupança individual e familiar (ENDRES, 2003). Hoje, pertencente ao grupo dos cinco maiores bancos nacionais, a Caixa se diferencia como grande responsável pelos financiamentos habitacionais do BrasilThis paper is intended to analyze the roles played by key players in the Brazilian Public Financial System during the period 1950 to 2010, highlighting - in a Keynesian perspective - the role played by them in relation to their contribution to economic and social development of Brazil, through the analysis of the management of public resources allocated to them (funding); The three main institutions of that system are: BNDES, Caixa Econômica Federal and Banco do Brasil. BNDES - created in 1952 with the intention of being a right arm of implementation of government development policies, as agent planner, coordinator and funder of projects to long-term investment - is now recognized as a high achiever of long-term investments and principal Brazilian development bank, ranks among the largest in the world, among their counterparts (ARAÚJO e CINTRA, 2011). Banco do Brasil was founded in 1808 in the midst of a set of measures of economic and financial organization associated with the installation of the Portuguese crown in Brasil, aiming to create an instrument that could control and centralize operations monetary and public finance and private (CARDOSO, 2009). Currently, the Bank of Brasil is among the largest and most important Brazilian banks and stands out as the largest institution of Brazilian rural credit (HERMANN, 2009). In 1861, Caixa Econômica Federal was founded by Dom Pedro II, in order to be the agency responsible for attracting modest resources from the lower classes. It guarantees families a safe option for individual and family savings (ENDRES, 2003). Today, being one of the five largest domestic banks, Caixa differentiates itself as largely responsible for housing finance in Brazi

A new drug delivery system for the local anesthetic prilocaine in lipossomes : preparation, characterization and biological tests

Orientadores: Eneida de Paula, Daniele Ribeiro de AraujoTese (doutorado) - Universidade Estadual de Campinas, Instituto de BiologiaResumo: O controle da dor é de extrema importância em odontologia, já que a maioria dos procedimentos odontológicos envolve estímulos dolorosos. Com a descoberta dos anestésicos locais, que se caracterizam pela capacidade de abolir a dor sem a perda da consciência, deu-se um grande passo para o avanço da odontologia. Um anestésico local ideal deve apresentar ação anestésica duradoura (suficiente para os procedimentos operatórios) e baixa toxicidade. Várias pesquisas têm sido desenvolvidas com anestésicos locais objetivando prolongar a duração de seu efeito e diminuir sua toxicidade. Um caminho muito promissor foi aberto com o desenvolvimento de formulações anestésicas de liberação prolongada, utilizando carreadores como lipossomas, que mantêm o fármaco por mais tempo e em maior concentração no sítio de ação. Este trabalho teve por finalidade preparar formulações de liberação prolongada do anestésico local prilocaína, muito usado em odontologia. Objetivouse: (i) preparar prilocaína encapsulada em lipossomas; (ii) caracterizar físicoquimicamente essa formulação, (iii) avaliar a atividade terapêutica, com testes de nocicepção em animais e (iv) avaliar a formulação quanto à estabilidade e à toxicidade local. Após a preparação, testes de caracterização físico-química demonstraram a interação da prilocaína com os lipossomas. Com a técnica de Ressonância Paramagnética Eletrônica pôde-se observar um decréscimo de 11% no parâmetro de ordem da membrana lipossomal, em presença do anestésico local. Com a análise por espalhamento de luz quase-elástico (light scattering) observou-se que as vesículas lipossomais apresentaram diâmetro médio de 382nm (±30), que não sofreu variação significativa (p>0,05) após encapsulação da prilocaína. Ensaios de liberação in vitro evidenciaram taxa de liberação mais lenta para a prilocaína lipossomal (equilíbrio em 90 min) que para prilocaína em solução (60 min). Análises por light scattering (p>0,05), de peroxidação lipídica (p>0,05) e ressonância magnética nuclear de prótons (H1-NMR) mostraram que, após processo de esterilização em autoclave, a prilocaína lipossomal permaneceu estável por um período de 30 dias após a preparação. Esses experimentos não mostraram diferenças na estabilidade físico-química da prilocaína ou da preparação lipossomal, estéreis ou não. Testes de bloqueio do nervo infraorbital em ratos e de tail-flick em camundongos revelaram maior efeito analgésico para a prilocaína lipossomal em relação à prilocaína em solução (p0,05) para o efeito da prilocaína lipossomal. A avaliação da toxicidade local em ratos mostrou que a prilocaína lipossomal não provocou edema de pata, quando comparada com soluções controle: salina, tampão, prilocaína em solução e lipossoma (p>0,05). Na avaliação histológica da mucosa oral dos ratos, não houve diferença significativa (p>0,05) entre os animais tratados com prilocaína lipossomal e seus controles. Com esses resultados, pôde-se concluir que a encapsulação de prilocaína em lipossomas aumentou a duração do bloqueio nervoso sensorial sem induzir o aumento na toxicidade local, podendo ser considerada mais uma opção no arsenal já disponível para a anestesia local em odontologiaAbstract: Pain control is an extremely important issue in dentistry since most of the dentistry procedures involve painful stimuli. The discovery of local anesthetics, which have the capability of abolishing the pain without loosing the consciousness, meant a major step in dentistry advance. An ideal local anesthetic must present lasting anesthetic action (long enough for surgery procedures) and low toxicity. Many researches have been developed with local anesthetics aiming at the prolongation of their anesthetic effect duration and decrease of their toxicity. A very promising path was open with the development of long-acting local anesthetics formulations, using carriers as liposomes that are able to enhance the bioavailability, to reduce the systemic toxicity and to increase the local anesthetic half-life in vivo. This present study comprised (i) the preparation of liposomal prilocaine ( a local anesthetic widely used in dentistry) formulation, (ii) the physicochemical characterization of the formulation, (iii) the assessment of its anesthetic efficacy and (iv) the evaluation of its physicochemical stability, as well as of its toxic effects. After preparation, the physicochemical characterization showed the prilocaine-liposome interaction. Electron spin resonance results showed a decrease in the order parameter of liposomal membrane, in presence of prilocaine. Laser light-scattering analysis revealed a vesicle population of liposomes with 382nm (± 30) diameter, without size changes after prilocaine incorporation. In the in vitro drug release assay, the liposomal formulation led to a slower release rate of prilocaine compared to its plain formulation. Equilibrium was delayed from 60 min (prilocaine in solution) to 90 min with the drug delivery system. Liposomal prilocaine was found to be stable up to 30 days after preparation, according to the analysis by laser light scattering (p > 0.05), thiobarbituric acid reactions (p > 0.05) and H1-nuclear magnetic resonance, once these assays did not show differences on physicochemical stability of prilocaine in solution or prilocaine liposomal, sterilized or not. Rat infraorbital nerve blockade and mice tail-flick tests revealed that a prolonged anesthetic effect was produced by liposomal prilocaine in comparison to prilocaine in solution (p0,05). Local toxicity evaluation in rats showed that the liposomal prilocaine did not evoke rat paw edema when compared to the control groups: saline, Hepes buffer, prilocaine in solution and liposome (p > 0.05). There were no statistical differences (p > 0.05) between lipossomal prilocaine and their controls, in histological evaluation of rat oral mucous. In conclusion, the prilocaine encapsulation in liposomes enhanced the nerve sensorial blockade, without increasing local toxicity. Liposomal prilocaine can, therefore, be considered an option to local anesthesia in dentistryDoutoradoBioquimicaDoutor em Biologia Funcional e Molecula

Psychiatric Disorders Biochemical Pathways Unraveled By Human Brain Proteomics

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Approximately 25 % of the world population is affected by a mental disorder at some point in their life. Yet, only in the mid-twentieth century a biological cause has been proposed for these diseases. Since then, several studies have been conducted toward a better comprehension of those disorders, and although a strong genetic influence was revealed, the role of these genes in disease mechanism is still unclear. This led most recent studies to focus on the molecular basis of mental disorders. One line of investigation that has risen in the post-genomic era is proteomics, due to its power of revealing proteins and biochemical pathways associated with biological systems. Therefore, this review compiled and analyzed data of differentially expressed proteins, which were found in postmortem brain studies of the three most prevalent psychiatric diseases: schizophrenia, bipolar disorder and major depressive disorders. Overviewing both the proteomic methods used in postmortem brain studies, the most consistent metabolic pathways found altered in these diseases. We have unraveled those disorders share about 21 % of proteins affected, and though most are related to energy metabolism pathways deregulation, the main differences found are 14-3-3-mediated signaling in schizophrenia, mitochondrial dysfunction in bipolar disorder and oxidative phosphorylation in depression.2671317Sao Paulo Research Foundation (FAPESP) [13/08711-3, 14/21035-0, 14/14881-1]Brazilian National Council for Scientific and Technological Development (CNPq) [460289/2014-4]CNPq [151787/2F2014-0]Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq

The Energy Metabolism Dysfunction in Psychiatric Disorders Postmortem Brains: Focus on Proteomic Evidence

Psychiatric disorders represent a great medical and social challenge and people suffering from these conditions face many impairments regarding personal and professional life. In addition, a mental disorder will manifest itself in approximately one quarter of the world's population at some period of their life. Dysfunction in energy metabolism is one of the most consistent scientific findings associated with these disorders. With this is mind, this review compiled data on disturbances in energy metabolism found by proteomic analyses of postmortem brains collected from patients affected by the most prevalent psychiatric disorders: schizophrenia (SCZ), bipolar disorder (BPD), and major depressive disorder (MDD). We searched in the PubMed database to gather the studies and compiled all the differentially expressed proteins reported in each work. SCZ studies revealed 92 differentially expressed proteins related to energy metabolism, while 95 proteins were discovered in BPD, and 41 proteins in MDD. With the compiled data, it was possible to determine which proteins related to energy metabolism were found to be altered in all the disorders as well as which ones were altered exclusively in one of them. In conclusion, the information gathered in this work could contribute to a better understanding of the impaired metabolic mechanisms and hopefully bring insights into the underlying neuropathology of psychiatric disorders

Psychiatric disorders biochemical pathways unraveled by human brain proteomics

Approximately 25 % of the world population is affected by a mental disorder at some point in their life. Yet, only in the mid-twentieth century a biological cause has been proposed for these diseases. Since then, several studies have been conducted toward a better comprehension of those disorders, and although a strong genetic influence was revealed, the role of these genes in disease mechanism is still unclear. This led most recent studies to focus on the molecular basis of mental disorders. One line of investigation that has risen in the post-genomic era is proteomics, due to its power of revealing proteins and biochemical pathways associated with biological systems. Therefore, this review compiled and analyzed data of differentially expressed proteins, which were found in postmortem brain studies of the three most prevalent psychiatric diseases: schizophrenia, bipolar disorder and major depressive disorders. Overviewing both the proteomic methods used in postmortem brain studies, the most consistent metabolic pathways found altered in these diseases. We have unraveled those disorders share about 21 % of proteins affected, and though most are related to energy metabolism pathways deregulation, the main differences found are 14-3-3-mediated signaling in schizophrenia, mitochondrial dysfunction in bipolar disorder and oxidative phosphorylation in depression2671317CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO - CNPQFUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESP460289/2014-4; 151787/2F2014-013/08711-3; 14/21035-0; 14/14881-

Liposomal Formulations Of Prilocaine, Lidocaine And Mepivacaine Prolong Analgesic Duration.

A laboratory investigation was undertaken to compare the in vivo antinociceptive effects of 2% liposomal formulations of prilocaine (PLC), lidocaine (LDC) and mepivacaine (MVC) compared to plain solutions of each of these three local anesthetics. Large unilamellar vesicles were prepared by extrusion (400 nm), at pH 7.4. The membrane/water partition coefficients were obtained from encapsulation efficiency values, after incorporation of each local anesthetic to the vesicles. The anesthetic effect of each liposomal formulation was compared to the respective local anesthetic solution in water, using the infraorbital nerve-blockade test, in rats. The partition coefficients were: 57 for PLC, 114 for LDC and 93 for MVC. In vivo results showed that local anesthetic-free liposomes, used as control, had no analgesic effect. In contrast, the encapsulated formulations induced increased intensities of total anesthetic effect (35.3%, 26.1% and 57.1%) and time for recovery (percentage increases of 30%, 23.1% and 56%), respectively, for PLC, LDC and MVC when compared to the plain solutions (P < 0.01). These results indicate that liposomes provide effective drug-delivery systems for intermediate-duration local anesthetics. Mepivacaine was affected to the greatest extent, while LDC benefited least from liposome encapsulation, possibly due to greater vasodilatory properties of LDC.531092-