Thyroid hormones regulate anxiety in the male mouse

Thyroid hormone levels are implicated in mood disorders in the adult human but the mechanisms remain unclear partly because, in rodent models, more attention has been paid to the consequences of perinatal hypo and hyperthyroidism. Thyroid hormones act via the thyroid hormone receptor (TR) alpha and beta isoforms, both of which are expressed in the limbic system. TR's modulate gene expression via both unliganded and liganded actions. Though the thyroid hormone receptor (TR) knockouts and a transgenic TRalpha1 knock-in mouse have provided us valuable insight into behavioral phenotypes such as anxiety and depression, it is not clear if this is because of the loss of unliganded actions or liganded actions of the receptor or due to locomotor deficits. We used a hypothyroid mouse model and supplementation with tri-iodothyronine (T3) or thyroxine (T4) to investigate the consequences of dysthyroid hormone levels on behaviors that denote anxiety. Our data from the open field and the light-dark transition tests suggest that adult onset hypothyroidism in male mice produces a mild anxiogenic effect that is possibly due to unliganded receptor actions. T3 or T4 supplementation reverses this phenotype and euthyroid animals show anxiety that is intermediate between the hypothyroid and thyroid hormone supplemented groups. In addition, T3 but not T4 supplemented animals have lower spine density in the CA1 region of the hippocampus and in the central amygdala suggesting that T3-mediated rescue of the hypothyroid state might be due to lower neuronal excitability in the limbic circuit.

Aluminium and the human breast

The human population is exposed to aluminium (Al) from diet, antacids and vaccine adjuvants, but frequent application of Al-based salts to the underarm as antiperspirant adds a high additional exposure directly to the local area of the human breast. Coincidentally the upper outer quadrant of the breast is where there is also a disproportionately high incidence of breast cysts and breast cancer. Al has been measured in human breast tissues/fluids at higher levels than in blood, and experimental evidence suggests that at physiologically relevant concentrations, Al can adversely impact on human breast epithelial cell biology. Gross cystic breast disease is the most common benign disorder of the breast and evidence is presented that Al may be a causative factor in formation of breast cysts. Evidence is also reviewed that Al can enable the development of multiple hallmarks associated with cancer in breast cells, in particular that it can cause genomic instability and inappropriate proliferation in human breast epithelial cells, and can increase migration and invasion of human breast cancer cells. In addition, Al is a metalloestrogen and oestrogen is a risk factor for breast cancer known to influence multiple hallmarks. The microenvironment is established as another determinant of breast cancer development and Al has been shown to cause adverse alterations to the breast microenvironment. If current useage patterns of Al-based antiperspirant salts contribute to causation of breast cysts and breast cancer, then reduction in exposure would offer a strategy for prevention, and regulatory review is now justified.

Transcriptomics and proteomics show that selenium affects inflammation, ctoskeleton, and cancer pathways in human rectal biopsies

Epidemiologic studies highlight the potential role of dietary selenium (Se) in colorectal cancer prevention. Our goal was to elucidate whether expression of factors crucial for colorectal homoeostasis is affected by physiologic differences in Se status. Using transcriptomics and proteomics followed by pathway analysis, we identified pathways affected by Se status in rectal biopsies from 22 healthy adults, including 11 controls with optimal status (mean plasma Se = 1.43 μM) and 11 subjects with suboptimal status (mean plasma Se = 0.86 μM). We observed that 254 genes and 26 proteins implicated in cancer (80%), immune function and inflammatory response (40%), cell growth and proliferation (70%), cellular movement, and cell death (50%) were differentially expressed between the 2 groups. Expression of 69 genes, including selenoproteins W1 and K, which are genes involved in cytoskeleton remodelling and transcription factor NFκB signaling, correlated significantly with Se status. Integrating proteomics and transcriptomics datasets revealed reduced inflammatory and immune responses and cytoskeleton remodelling in the suboptimal Se status group. This is the first study combining omics technologies to describe the impact of differences in Se status on colorectal expression patterns, revealing that suboptimal Se status could alter inflammatory signaling and cytoskeleton in human rectal mucosa and so influence cancer risk.

Establishment of a Brazilian Line of Human Embryonic Stem Cells in Defined Medium: Implications for Cell Therapy in an Ethnically Diverse Population

Pluripotent human embryonic stem (hES) cells are an important experimental tool for basic and applied research, and a potential source of different tissues for transplantation. However, one important challenge for the clinical use of these cells is the issue of immunocompatibility, which may be dealt with by the establishment of hES cell banks to attend different populations. Here we describe the derivation and characterization of a line of hES cells from the Brazilian population, named BR-I, in commercial defined medium. In contrast to the other hES cell lines established in defined medium, BR-I maintained a stable normal karyotype as determined by genomic array analysis after 6 months in continuous culture (passage 29). To our knowledge, this is the first reported line of hES cells derived in South America. We have determined its genomic ancestry and compared the HLA-profile of BR-I and another 22 hES cell lines established elsewhere with those of the Brazilian population, finding they would match only 0.011% of those individuals. Our results highlight the challenges involved in hES cell banking for populations with a high degree of ethnic admixture.

Human Stem Cell Cultures from Cleft Lip/Palate Patients Show Enrichment of Transcripts Involved in Extracellular Matrix Modeling By Comparison to Controls

Nonsyndromic cleft lip and palate (NSCL/P) is a complex disease resulting from failure of fusion of facial primordia, a complex developmental process that includes the epithelial-mesenchymal transition (EMT). Detection of differential gene transcription between NSCL/P patients and control individuals offers an interesting alternative for investigating pathways involved in disease manifestation. Here we compared the transcriptome of 6 dental pulp stem cell (DPSC) cultures from NSCL/P patients and 6 controls. Eighty-seven differentially expressed genes (DEGs) were identified. The most significant putative gene network comprised 13 out of 87 DEGs of which 8 encode extracellular proteins: ACAN, COL4A1, COL4A2, GDF15, IGF2, MMP1, MMP3 and PDGFa. Through clustering analyses we also observed that MMP3, ACAN, COL4A1 and COL4A2 exhibit co-regulated expression. Interestingly, it is known that MMP3 cleavages a wide range of extracellular proteins, including the collagens IV, V, IX, X, proteoglycans, fibronectin and laminin. It is also capable of activating other MMPs. Moreover, MMP3 had previously been associated with NSCL/P. The same general pattern was observed in a further sample, confirming involvement of synchronized gene expression patterns which differed between NSCL/P patients and controls. These results show the robustness of our methodology for the detection of differentially expressed genes using the RankProd method. In conclusion, DPSCs from NSCL/P patients exhibit gene expression signatures involving genes associated with mechanisms of extracellular matrix modeling and palate EMT processes which differ from those observed in controls. This comparative approach should lead to a more rapid identification of gene networks predisposing to this complex malformation syndrome than conventional gene mapping technologies.

p38 alpha MAP Kinase Controls IL-17 Synthesis in Vogt-Koyanagi-Harada Syndrome and Experimental Autoimmune Uveitis

PURPOSE. Interleukin (IL)-17, which is responsible for the initial influx of leukocytes into the target tissue, was recently described as the main cytokine involved in autoimmune diseases. Vogt-Koyanagi-Harada (VKH) syndrome is a significant cause of noninfectious blindness in the world. Herein the authors aimed at unraveling the involvement of IL-17 in VKH and in experimental autoimmune uveitis, focusing on the signaling pathways involved in IL-17 synthesis. METHODS. Mice were immunized with 161-180 peptide and pertussis toxin. Draining lymph node cells, harvested 21 days after immunization, were cultured in the presence or absence of p38 alpha mitogen-activated protein kinase (MAPK) inhibitor (SB203580) and assayed for cytokine production and quantification of CD4(+)IL-17(+) cells. Mice received intraocular injections of SB203580, and disease severity was evaluated by histologic examination of the enucleated eyes at day 21. CD4(+) lymphocytes from MSK-1/2-deficient mice, human CD4(+) cells silenced with MSK1 siRNA, or peripheral blood mononuclear cells (PBMCs) from VKH patients were cultured in the presence or absence of p38 alpha MAPK inhibitor and then assayed for IL-17, IFN-gamma, and IL-4 production. RESULTS. The inhibition of p38 alpha MAPK fully blocked the synthesis of IL-17 by PBMCs from VKH patients and lymphocytes from EAU mice. The absence of the msk1/2 gene resulted in failure to produce IL-17 by murine and human lymphocytes. Interestingly, intraocular injections of SB203580 in EAU mice did not suppress development of the disease. CONCLUSIONS. These data show that p38 alpha MAPK-MSK1/2 is involved in the control of IL-17 synthesis by CD4(+) T cells and that inhibition of p38 alpha MAPK in vitro suppresses IL-17 synthesis but that inhibition of this kinase in vivo did not protect from EAU. (Invest Ophthalmol Vis Sci. 2010;51:3567-3574) DOI: 10.1167/iovs.09-4393

Putative outer membrane proteins of Leptospira interrogans stimulate human umbilical vein endothelial cells (HUVECS) and express during infection

Cell adhesion molecules (CAMs) are surface receptors present in eukaryotic cells that mediate cell-cell or cell-extracellular matrix interactions. Vascular endothelium stimulation in vitro that lead to the upregulation of CAMs was reported for the pathogenic spirochaetes, including rLIC10365 of Leptospira interrogans. In this study, we report the cloning of LIC10507, LIC10508, LIC10509 genes of L interrogans using Escherichia coli as a host system. The rational for selecting these sequences is due to their location in L. interrogans serovar Copenhageni genome that has a potential involvement in pathogenesis. The genes encode for predicted lipoproteins with no assigned functions. The purified recombinant proteins were capable to promote the upregulation of intercellular adhesion molecule 1 (ICAM-1) and E-selectin on monolayers of human umbilical vein endothelial cells (HUVECS). In addition, the coding sequences are expressed in the renal tubules of animal during bacterial experimental infection. The proteins are probably located at the outer membrane of the bacteria since they are detected in detergent-phase of L interrogans Triton X-114 extract. Altogether our data suggest a possible involvement of these proteins during bacterial infection and provide new insights into the role of this region in the pathogenesis of Leptospira. (C) 2008 Elsevier Ltd. All rights reserved.

Human endogenous RNAs: Implications for the immunomodulation of Toll-like receptor 3

Toll-like receptors (TLRs), a family of mammalian receptors, are able to recognize nucleic acids. TLR3 recognizes double-stranded (ds)RNA, a product of the replication of certain viruses. Polyinosinic-polycytidylic acid, referred to as poly(I:C), an analog of viral dsRNA, interacts with TLR3 thereby eliciting immunoinflammatory responses characteristic of viral infection or down-regulating the expression of chemokine receptor CXCR4. It is known that dsRNA also directly activates interferon (IFN)-induced enzymes, such as the RNA-dependent protein kinase (PKR). In the present study, the mRNA expression of TLR3, CXCR4, IFN gamma and PKR was investigated in a culture of peripheral blood mononuclear cells (PBMCs) stimulated with poly(I:C) and endogenous RNA from human PBMCs. No cytotoxic effect on the cells or on the proliferation of CD3(+), CD4(+) and CD8(+) cells was observed. TLR3 expression in the PBMCs in the presence of poly(I:C) was up-regulated 9.5-fold, and TLR3 expression in the PBMCs treated with endogenous RNA was down-regulated 1.8-fold (p=0.002). The same trend was observed for IFN gamma where in the presence of poly(I:C) an 8.7-fold increase was noted and in the presence of endogenous RNA a 3.1-fold decrease was observed. In the culture activated with poly(1:C), mRNA expression of CXCR4 increased 8.0-fold and expression of PKR increased 33.0-fold. Expression of these genes decreased in the culture treated with endogenous RNA when compared to the culture without stimulus. Thus, high expression of mRNA for TLR3, IFN gamma, CXCR4 and PKR was observed in the presence of poly(I:C) and low expression was observed in the cells cultured with endogenous RNA. In conclusion, TLR3 may play major physiological roles that are not in the context of viral infection. It is possible that RNA released from cells could contain enough double-stranded structures to regulate cell activation. The involvement of endogenous RNA in endogenous gene expression and its implications in the regulation thereof, are still being studied, and will have significant implications in the future.

Human regulatory protein Ki-1/57 has characteristics of an intrinsically unstructured protein

The human protein Ki-1/57 was first identified through the cross reactivity of the anti-CD30 monoclonal antibody Ki-1; in Hodgkin lymphoma cells. The expression of Ki-1/57 in diverse cancer cells and its phosphorylation in peripheral blood leukocytes after mitogenic activation suggested its possible role in cell signaling. Ki-1/57 interacts with several other regulatory proteins involved in cellular signaling, transcriptional regulation and RNA metabolism, suggesting it may have pleiotropic functions. In a previous spectroscopic analysis, we observed a low content of secondary structure for Ki-1/57 constructs. Here, Circular dichroism experiments, in vitro RNA binding analysis, and limited proteolysis assays of recombinant Ki-1/57(122-413) and proteolysis assays of endogenous full length protein from human HEK293 cells suggested that Ki-1/57 has characteristics of an intrinsically unstructured protein. Small-angle X-ray scattering (SAXS) experiments were performed with the C-terminal fragment Ki-1/57(122-413). These results indicated an elongated shape and a partially unstructured conformation of the molecule in solution, confirming the characteristics of an intrinsically unstructured protein. Experimental curves together with ab initio modeling approaches revealed an extended and flexible molecule in solution. An elongated shape was also observed by analytical gel filtration. Furthermore, sedimentation velocity analysis suggested that Ki-1/57 is a highly asymmetric protein. These findings may explain the functional plasticity of Ki-1/57, as suggested by the wide array of proteins with which it is capable of interacting in yeast two-hybrid interaction assays.

Reliability of GPS based traffic data: an experimental evaluation

GPS tracking of mobile objects provides spatial and temporal data for a broad range of applications including traffic management and control, transportation routing and planning. Previous transport research has focused on GPS tracking data as an appealing alternative to travel diaries. Moreover, the GPS based data are gradually becoming a cornerstone for real-time traffic management. Tracking data of vehicles from GPS devices are however susceptible to measurement errors – a neglected issue in transport research. By conducting a randomized experiment, we assess the reliability of GPS based traffic data on geographical position, velocity, and altitude for three types of vehicles; bike, car, and bus. We find the geographical positioning reliable, but with an error greater than postulated by the manufacturer and a non-negligible risk for aberrant positioning. Velocity is slightly underestimated, whereas altitude measurements are unreliable.

Modelo experimental de lesões intra-epiteliais e de adenocarcinoma ductal pancreático induzidos por 7,12-dimetilbenzantraceno em camundongos

INTRODUÇÃO: O adenocarcinoma de pâncreas apresenta um mau prognóstico. A utilização de modelos experimentais é necessária para a compreensão do comportamento biológico tumoral, principalmente das lesões precoces (neoplasias intra-epiteliais pancreáticas - NIPan) e para o desenvolvimento de opções terapêuticas. OBJETIVO: Avaliar a carcinogênese pancreática induzida por 7,12-dimetilbenzantraceno (DMBA), em camundongos, aplicando a classificação das neoplasias intra-epiteliais pancreáticas. MÉTODOS: 90 camundongos machos, mus musculus, da cepa CF1, foram submetidos à laparotomia mediana e 1 mg de DMBA foi implantado na porção cefálica do pâncreas. Os animais foram divididos em dois grupos, com eutanásia em 30 e 60 dias. Em seguida, o pâncreas foi retirado, fixado em formalina e foram confeccionadas lâminas coradas com hematoxilina eosina. Os cortes histológicos foram avaliados por dois patologistas de acordo com os seguintes critérios: pâncreas normal, hiperplasia reacional, NIPan 1A, NIPan 1B, NIPan 2, NIPan 3 e carcinoma. As alterações inflamatórias também foram analisadas. RESULTADOS: A avaliação patológica evidenciou, no grupo de 30 dias: 4 (16,7%) animais com hiperplasia reativa, 16 (66,6%) com NIPan e 4 (16,7%) com adenocarcinoma. No grupo de 60 dias: 10 (27,1%) animais com hiperplasia reativa, 13 (35,1%) com NIPan e 14 (37,8%) com adenocarcinoma. A diferença entre os grupos apresentou significância estatistística (P < 0,05 – teste exato de Fisher). A prevalência de alterações inflamatórias em 30 dias foi: pancreatite aguda (n=11), pancreatite crônica (n=5) e inflamação dependente da bolsa (n=8). No grupo de 60 dias 11 espécimes apresentavam pancreatite aguda e 26 pancreatite crônica. CONCLUSÕES: O modelo experimental com DMBA em camundongos, induz neoplasia intra-epitelial pancreática e adenocarcinoma ductal histologicamente semelhantes ao carcinoma pancreático em humanos. Este modelo pode ser utilizado na investigação da carcinogênese com enfoque na progressão molecular das lesões precursoras até o adenocarcinoma.

Ansiedade e desempenho: uma análise experimental

A ansiedade é um dos conceitos mais importantes dentro da Psicologia. Uma grande quantidade de trabalhos experimentais tratando sobre os efeitos da ansiedade-de-teste, tem sido conduzida. A discussão sobre ansiedade-de-teste e sua relação com o desempenho acadêmico deve ser precedida por uma elucidação do conceito de ansiedade. Este conceito tem sido interpretado de diferentes formas pelas várias escolas de pensamento na Psicologia e Filosofia. Os dois primeiros capítulos deste trabalho são dedicados à uma apresentação dos vários conceitos de ansiedade e de teorias que consideramos importantes que abordam o assunto. A seguir, passamos a descrever a análise experimental que foi realizada com o objetivo de detectar os efeitos da ansiedade-de-teste no desempenho e verificar os resultados da manipulação da variável tempo limitado e da magnitude do "stress", na relação estudada. Grande parte da literatura sobre ansiedade tem sublinhado que a influência desta sobre o comportamento é bastante complexa. A pesquisa aqui reportada se baseia nas investigações de Spielberger que focalizam a relação entre o desempenho do estudante e seu nível de ansiedade. Como Spielberger, julgamos ser necessário controlar as variáveis dificuldade da tarefa e inteligência dos sujeitos. Com base nos resultados encontrados, concluímos que uma série de outros aspectos devem ser considerados, tais como elaboração da prova, matéria envolvida, experiências anteriores dos alunos, imagem do professor etc. Busca-se conhecer a natureza da ansiedade-de-teste e encontrar formas efetivas para controlar este tipo específico de ansiedade, de maneira a aparelhar melhor os alunos para lidarem com ele. É necessário também que os educadores tomem conhecimento das características e implicações da ansiedade-de-teste, de forma que sejam capazes de tirar partido dela e consigam minimizar seus efeitos negativos no processo ensino-aprendizagem.

O efeito do desempenho de um papel de ajuda na ansiedade de falar em público: um estudo experimental

O objetivo principal desta pesquisa foi testar o ”princípio da pessoa que ajuda”, fenômeno que ressalta os benefícios que recebemos quando ajudamos alguém. Para a consecução desse objetivo, sujeitos com alta ansiedade de falar em público desempenharam um papel de ajuda: serviram de "terapeutas comportamentais" para outros sujeitos com alta ansiedade de falar. Depois de selecionado, os sujeitos -de alta ansiedade de falar em público -foram distribuídos por estes quatro grupos experimentais: (1) grupo cujos sujeitos aprenderam, com terapeutas, técnicas comportamentais para a redução da ansiedade de falar, e, posteriormente, ensinaram essas técnicas a outros sujeitos; (2) grupo em que os sujeitos aprenderam, com os terapeutas, as técnicas para a redução da ansiedade de falar, mas não as ensinaram a outros sujeitos; (3) grupo que aprendeu as técnicas com os sujeitos do grupo 1; (4) grupo cujos sujeitos apenas participaram do pré e pós-testes. Previu-se que os sujeitos do grupo 1 apresenta riam maior redução da ansiedade de falar do que os sujeitos do grupo 2, porque aqueles exerceriam um papel de ajuda. Porém, os testes estatísticos indicaram que a hipótese substantiva foi rejeitada: o grupo 1 não teve a sua ansiedade significativamente mais reduzida do que o grupo 2, embora os grupos que receberam o programa de tratamento para a ansiedade de falar (grupos 1, 2 e 3) tenham melhorado significativamente mais do que o grupo 4. São discutidas possíveis explicações para esses resultados e feitas sugestões para novas pesquisas.

Ansiedade infantil em contexto cirúrgico: estudo experimental

Na infância, a hospitalização, em especial por circunstâncias cirúrgicas, constitui uma experiência potencialmente ameaçadora e causadora de ansiedade, medo e stress, que não se circunscreve apenas à criança, mas também aos pais e à família. A presença de níveis elevados de ansiedade infantil pode ter consequências nefastas e comprometedoras do desenvolvimento psicológico. A preparação psicológica da criança é crucial para a redução da ansiedade e das alterações do comportamento pós-hospitalização, razão porque considerámos fundamental colmatar a inexistência de um programa similar destinado ao utente pediátrico do HDESPDL. Desta forma, concebemos o Programa Infantil de Preparação para a Cirurgia (PIPCirurgia), um instrumento de preparação psicológica destinado a crianças entre os 6-11 anos propostas para cirurgia eletiva com internamento programado. O PIPCirurgia pretende preparar criança e pais, trabalhar estratégias de coping e reduzir a ansiedade e as alterações comportamentais pós-hospitalização. Validámos a sua utilidade através de um plano metodológico experimental aplicado a uma amostra de 60 crianças, equitativamente subdividida em Grupo Experimental (submetido ao PIPCirurgia) e Grupo de Controlo (recebeu o procedimento atualmente disponibilizado pela instituição). A investigação efetivou-se através de dois estudos, Estudo I: Ansiedade infantil e Caraterísticas Sociodemográficas e Estudo II: Efeitos do PIPCirurgia na redução da Ansiedade Infantil e das Alterações do Comportamento Pós-Hospitalização. Utilizaram-se métodos de recolha de dados quantitativos. Aplicou-se um Questionário de Caraterização Sociodemográfica, a versão traduzida e validada para a população portuguesa do State-Trait Anxiety Inventory for Children, e uma versão por nós traduzida e testada do Post-Hospitalization Behavior Questionnaire. Concluímos que, para as amostras em estudo, existiu uma relação entre a Ansiedade Infantil e a presença de antecedentes familiares de doença, assim como entre a Ansiedade-Estado pré-operatória e a situação de emprego materno. Confirmou-se a utilidade do PIPCirurgia para reduzir a Ansiedade Infantil relacionada com a cirurgia e minimizar as alterações comportamentais pós-hospitalização.

análise experimental do efeito da diversificação em carteiras de ações: a relação entre risco e quantidade de ativos sob a ótica da estratégia ingênua

The objective is to analyze the relationship between risk and number of stocks of a portfolio for an individual investor when stocks are chosen by "naive strategy". For this, we carried out an experiment in which individuals select actions to reproduce this relationship. 126 participants were informed that the risk of first choice would be an asset average of all standard deviations of the portfolios consist of a single asset, and the same procedure should be used for portfolios composed of two, three and so on, up to 30 actions . They selected the assets they want in their portfolios without the support of a financial analysis. For comparison we also tested a hypothetical simulation of 126 investors who selected shares the same universe, through a random number generator. Thus, each real participant is compensated for random hypothetical investor facing the same opportunity. Patterns were observed in the portfolios of individual participants, characterizing the curves for the components of the samples. Because these groupings are somewhat arbitrary, it was used a more objective measure of behavior: a simple linear regression for each participant, in order to predict the variance of the portfolio depending on the number of assets. In addition, we conducted a pooled regression on all observations by analyzing cross-section. The result of pattern occurs on average but not for most individuals, many of which effectively "de-diversify" when adding seemingly random bonds. Furthermore, the results are slightly worse using a random number generator. This finding challenges the belief that only a small number of titles is necessary for diversification and shows that there is only applicable to a large sample. The implications are important since many individual investors holding few stocks in their portfolios