Two functional forms of the Xenopus laevis estrogen receptor translated from a single mRNA species.

Steroid receptors are nuclear proteins that regulate gene transcription in a ligand-dependent manner. Over-expression of the Xenopus estrogen receptor in a vaccinia virus-derived expression system revealed that the receptor localized exclusively in the nucleus of the infected cells, irrespective of the presence or absence of the ligand. Furthermore, two forms of the receptor were produced, a full-length and a N-terminal truncated version, which are translated from a single mRNA species by the use of two AUG within the same reading frame. These 66- and 61-kDa receptors were also observed after in vitro translation of the mRNA as well as in primary Xenopus hepatocytes. Both forms are potent estrogen-dependent transcriptional activators in transient transfection experiments, as well as in in vitro transcription assays.

Expression of T cell receptor genes in the thymus: localization of transcripts in situ and comparison of mature and immature subsets.

We have analyzed the expression of T cell receptor (TcR) genes in the thymus using in situ RNA hybridizations with probes to the constant regions of the TcR alpha, beta, gamma and delta chains. Localization of transcripts revealed low TcR alpha mRNA levels in the thymus cortex and very low levels in the subcapsular region. In contrast, TcR beta message was very abundant in the cortex. TcR gamma or delta mRNA+ thymocytes showed a scattered, predominantly cortical localization. In contrast to gamma, TcR delta transcripts were abundant in the subcapsular region. Control experiments with sorted TcR alpha/beta or gamma/delta cells revealed a detection efficiency of 75%-85% for the respective TcR mRNA and data on TcR gene expression in mature, CD3+ thymocytes were consistent with previous reports. The analysis of immature, CD3- thymocyte subsets, however, revealed a virtual absence of TcR alpha transcripts and an unexpectedly high proportion of cells (14%-29%) expressing the gene for the TcR delta chain. The data are discussed in view of current models of lineage relationships in the thymus.

IL-2/anti-IL-2 antibody complexes show strong biological activity by avoiding interaction with IL-2 receptor alpha subunit CD25.

IL-2 is crucial to T cell homeostasis, especially of CD4(+) T regulatory cells and memory CD8(+) cells, as evidenced by vigorous proliferation of these cells in vivo following injections of superagonist IL-2/anti-IL-2 antibody complexes. The mechanism of IL-2/anti-IL-2 antibody complexes is unknown owing to a lack of understanding of IL-2 homeostasis. We show that IL-2 receptor alpha (CD25) plays a crucial role in IL-2 homeostasis. Thus, prolongation of IL-2 half-life and blocking of CD25 using antibodies or CD25-deficient mice led in combination, but not alone, to vigorous IL-2-mediated T cell proliferation, similar to IL-2/anti-IL-2 antibody complexes. These data suggest an unpredicted role for CD25 in IL-2 homeostasis.

Natural killer cell receptor-repertoire and functions after induction therapy by polyclonal rabbit anti-thymocyte globulin in unsensitized kidney transplant recipients.

Polyclonal rabbit anti-thymocyte globulin (rATG) is widely used in solid organ transplantation (SOT) as induction therapy or to treat corticosteroid-resistant rejection. In vivo, the effect of rATG on natural killer (NK) cells has not been studied. These cells are of particular relevance after SOT because classical immunosuppressive drugs do not inhibit or even can activate NK cells. A cohort of 20 recipients at low immunological risk, that had been receiving rATG as induction therapy, was analyzed for receptor repertoire, cytotoxicity and capacity of NK cells to secrete IFN-γ before kidney transplantation and at different time points thereafter. NK cells expressed fewer killer-cell immunoglobulin-like receptors (KIR), fewer activating receptors NKG2D, but more inhibitory receptor NKG2A compatible with an immature phenotype in the first 6 months post transplantation. Both cytotoxicity of NK cells and the secretion of IFN-γ were preserved over time after transplantation.

Association between variants of the leptin receptor gene (LEPR) and overweight: a systematic review and an analysis of the CoLaus study.

BACKGROUND: Three non-synonymous single nucleotide polymorphisms (Q223R, K109R and K656N) of the leptin receptor gene (LEPR) have been tested for association with obesity-related outcomes in multiple studies, showing inconclusive results. We performed a systematic review and meta-analysis on the association of the three LEPR variants with BMI. In addition, we analysed 15 SNPs within the LEPR gene in the CoLaus study, assessing the interaction of the variants with sex. METHODOLOGY/PRINCIPAL FINDINGS: We searched electronic databases, including population-based studies that investigated the association between LEPR variants Q223R, K109R and K656N and obesity- related phenotypes in healthy, unrelated subjects. We furthermore performed meta-analyses of the genotype and allele frequencies in case-control studies. Results were stratified by SNP and by potential effect modifiers. CoLaus data were analysed by logistic and linear regressions and tested for interaction with sex. The meta-analysis of published data did not show an overall association between any of the tested LEPR variants and overweight. However, the choice of a BMI cut-off value to distinguish cases from controls was crucial to explain heterogeneity in Q223R. Differences in allele frequencies across ethnic groups are compatible with natural selection of derived alleles in Q223R and K109R and of the ancient allele in K656N in Asians. In CoLaus, the rs10128072, rs3790438 and rs3790437 variants showed interaction with sex for their association with overweight, waist circumference and fat mass in linear regressions. CONCLUSIONS: Our systematic review and analysis of primary data from the CoLaus study did not show an overall association between LEPR SNPs and overweight. Most studies were underpowered to detect small effect sizes. A potential effect modification by sex, population stratification, as well as the role of natural selection should be addressed in future genetic association studies.

O Papel das Agências de Viagens e Turismo no Desenvolvimento do Turismo Emissor e Receptor

Devido a mudanças tecnológicas e da globalização, o turismo vem passando por grandes transformações, permitindo um maior acesso às informações e uma maior aproximação entre o consumidor e o mercado turístico. O papel das agências de viagens e turismo (AVT’s) é de fundamental importância para o desenvolvimento da actividade turística de um país. Face a isto, as AVT’s devem aderir ao novo cenário, procurando acompanhar a evolução, renovando e actualizando sempre os seus serviços. O presente estudo tem por objectivo avaliar o papel das AVT’s da ilha de São Vicente, no âmbito do turismo emissor e receptor. Foi realizado um estudo de caso, incluindo os doze representantes das AVT’s da ilha de S. Vicente, com excepção das empresas exclusivamente marítimas. Foi usado um guião de entrevistas para a obtenção de dados relacionados aos responsáveis dos serviços, a empresa, aos colaboradores internos, clientes, serviços oferecidos, sazonalidade e sua problemática, parceiros, as tecnologias de informação e as perspectivas futuras. As informações foram analisadas sob um enquadramento teórico e prático. Constatou-se que sete agências dedicam-se ao turismo emissor e receptor, três ao turismo emissor, e duas ao receptor. Foi possível observar que as AVT’s apresentam semelhanças na prestação de serviços e a maioria tem a preocupação de acompanhar as exigências actuais do turismo. Pode-se concluir que o turismo emissor e receptor estão bem representados pelas AVT’s de São Vicente. Technological changes and globalization has allowed big transformations in Tourism, greater access to information and a closer relationship between the consumer and tourist market. The role of the Travel and Tourism Agencies (TTA) has fundamental importance for the tourism development in a country, and they must adhere to the new environment, trying to follow the evolution, always renewing and updating their services. The present study aims to evaluate the role of TTA’s in São Vicente Island, in the field of inbound and outbound tourism. It was conducted a case study, including 12 representatives of TTA's in São Vicente, with exception of shipping companies. It was used a scripted interview to obtain data about the service delegate, the company, internal employees, customers, offered services, seasonality and its problems, partners, information technology and future prospects. Data were analyzed from a theoretical and practical view. It was found that seven agencies dedicated to inbound and outbound tourism, three to outbound and two to inbound. The TTA’s have similarities in service delivery and most have the concern to keep up with current demands of tourism. It can be concluded that inbound and outbound tourism are well represented by TTA’s in São Vicente.

Assinatura Digital Foco Smart Card

A assinatura digital é um processo de assinatura electrónica baseado no sistema criptográfico assimétrico composto por um algoritmo ou série de algoritmos, mediante o qual é gerado um par de chaves assimétricas exclusivas e complementares. Tal como a assinatura manuscrita, a assinatura digital não pode ser falsificada, pelo que se adiciona uma função hash ao valor da chave gerada, para garantir a segurança do sistema. A assinatura digital permite, ao mesmo tempo, a identificação positiva do autor de uma mensagem (ou do signatário de um documento) e a verificação da integridade da mesma. Refira-se, porém, que a assinatura digital pode ser anexa a qualquer mensagem, seja esta decifrada ou não, apenas para que o receptor tenha a certeza da identidade do emissor e de que a mensagem chegou intacta ao destino. Baseia-se na criptografia da chave pública, que usa um algoritmo de duas chaves, a privada e a pública, diferentes mas matematicamente associadas: a primeira cria a assinatura digital e decifra os dados; a segunda verifica a assinatura e devolve-a ao formato original. O autor da mensagem mantém secreta a sua chave privada e divulga a chave pública, e vice-versa. A identidade do dono da chave pública é atestada pelo certificado digital emitido por uma entidade certificadora (uma espécie de notário). Independentemente da forma que assume, a assinatura electrónica, e a digital em particular, deve cumprir três funções: a autenticação (tal como acontece com o habitual gatafunho a que chamamos assinatura, apenas o indivíduo deve ser capaz de a reproduzir); não repudiação (quando o indivíduo assina não pode voltar atrás); e integridade (é necessário assegurar que os documentos assinados não são alterados sem o consentimento dos signatários).

The aldosterone-mineralocorticoid receptor pathway exerts anti-inflammatory effects in endotoxin-induced uveitis.

We have previously shown that the eye is a mineralocorticoid-sensitive organ and we now question the role of mineralocorticoid receptor (MR) in ocular inflammation. The endotoxin-induced uveitis (EIU), a rat model of human intraocular inflammation, was induced by systemic administration of lipopolysaccharide (LPS). Evaluations were made 6 and 24 hours after intraocular injection of aldosterone (simultaneous to LPS injection). Three hours after onset of EIU, the MR and the glucocorticoid metabolizing enzyme 11-beta hydroxysteroid dehydrogenase type 2 (11β-HSD2) expression were down-regulated in iris/ciliary body and the corticosterone concentration was increased in aqueous humor, altering the normal MR/glucocorticoid receptor (GR) balance. At 24 hours, the GR expression was also decreased. In EIU, aldosterone reduced the intensity of clinical inflammation in a dose-dependent manner. The clinical benefit of aldosterone was abrogated in the presence of the MR antagonist (RU26752) and only partially with the GR antagonist (RU38486). Aldosterone reduced the release of inflammatory mediators (6 and 24 hours: TNF-α, IFN-γ, MIP-1α) in aqueous humor and the number of activated microglia/macrophages. Aldosterone partly prevented the uveitis-induced MR down-regulation. These results suggest that MR expression and activation in iris/ciliary body could protect the ocular structures against damages induced by EIU.

The chemokine receptor CXCR4 strongly promotes neuroblastoma primary tumour and metastatic growth, but not invasion.

Neuroblastoma (NB) is a heterogeneous, and particularly malignant childhood neoplasm in its higher stages, with a propensity to form metastasis in selected organs, in particular liver and bone marrow, and for which there is still no efficient treatment available beyond surgery. Recent evidence indicates that the CXCR4/CXCL12 chemokine/receptor axis may be involved in promoting NB invasion and metastasis. In this study, we explored the potential role of CXCR4 in the malignant behaviour of NB, using a combination of in vitro functional analyses and in vivo growth and metastasis assessment in an orthotopic NB mouse model. We show here that CXCR4 overexpression in non-metastatic CXCR4-negative NB cells IGR-NB8 and in moderately metastatic, CXCR4 expressing NB cells IGR-N91, strongly increased tumour growth of primary tumours and liver metastases, without altering the frequency or the pattern of metastasis. Moreover shRNA-mediated knock-down experiments confirmed our observations by showing that silencing CXCR4 in NB cells impairs in vitro and almost abrogates in vivo growth. High levels of CXCL12 were detected in the mouse adrenal gland (the primary tumour site), and in the liver suggesting a paracrine effect of host-derived CXCL12 on NB growth. In conclusion, this study reveals a yet unreported NB-specific predominant growth and survival-promoting role of CXCR4, which warrants a critical reconsideration of the role of CXCR4 in the malignant behaviour of NB and other cancers.

An European Organisation for Research and Treatment of Cancer phase I study of lapatinib and docetaxel as neoadjuvant treatment for Human Epidermal Growth Factor Receptor 2 (HER2) positive locally-advanced/inflammatory or large operable breast cancer.

BACKGROUND: Lapatinib is an effective anti-HER2 therapy in advanced breast cancer and docetaxel is one of the most active agents in breast cancer. Combining these agents in pre-treated patients with metastatic disease had previously proved challenging, so the primary objective of this study aimed to determine the maximum tolerated dose (MTD) in treatment-naive patients, by identifying acute dose-limiting toxicities (DLT) during cycle 1 in the first part of a phases 1-2 neoadjuvant European Organisation for Research and Treatment of Cancer (EORTC) trial. PATIENTS AND METHODS: Patients with large operable or locally-advanced HER2 positive breast cancer were treated with continuous lapatinib, and docetaxel every 21days for 4 cycles. Dose levels (DLs) were: 1000/75, 1250/75, 1000/85, 1250/85, 1000/100 and 1250/100 (mg/day)/(mg/m(2)). RESULTS: Twenty-one patients were included. Two DLTs occurred at dose level 5 (1000/100); one grade 4 neutropenia ⩾7days and one febrile neutropenia. A further 3 patients were therefore treated at the same dose with prophylactic granulocyte-colony stimulating factor (G-CSF), and 3 patients at dose level 6. No further DLTs were observed. CONCLUSIONS: Our recommended dose for phase II is lapatinib 1000mg/day and docetaxel 100mg/m(2) with G-CSF in HER2 positive non-metastatic breast cancer. The dose of lapatinib should have been 1250mg/day but we were mindful of the high rate of treatment discontinuation in GeparQuinto with lapatinib 1250mg/day combined with docetaxel. No grade 3-4 diarrhoea was observed. Pharmacodynamics analysis suggests that concomitant medications altering P-glycoprotein activity (in addition to lapatinib) can modify toxicity, including non-haematological toxicities. This needs verification in larger trials, where it may contribute to understanding the sources of variability in clinical toxicity and treatment discontinuation.

Importância / Função do Enfermeiro Na prestação de Cuidados Paliativos ao Doente Oncológico Terminal

Cabo Verde está vivenciando uma transição epidemiológica caracterizada pelo aumento da incidência das doenças crónicas não transmissíveis, nomeadamente as oncológicas, em detrimento das infecto-contagiosas. Neste cenário verifica-se a necessidade de adequação dos cuidados de saúde aos doentes oncológicos terminal de modo a dar resposta as necessidades físicas, psicológicas e sociais deste doentes e dos seus familiares. Durante muito tempo persistiu a ideia de que não existe mais nada a fazer por um doente quando é diagnosticada uma doença oncológica (cronica e evolutiva). Mas o advento da filosofia dos cuidados paliativos veio provar que essa ideia é errónea, pois quando a cura não é possível existe ainda um campo de cuidado amplo a ser explorado. O cuidar faz parte da existência humana desde o nascimento e deve acompanha-lo até a sua finitude. O cuidar é a essência da Enfermagem, logo os cuidados paliativos, termo que denomina o cuidado em fim da vida é inerente a prática de Enfermagem.Partindo desses pressupostos surge o presente trabalho com o objectivo de compreender qual a Importância / função do enfermeiro na prestação de cuidados paliativos ao doente terminal oncológico. Para dar reposta ao nosso objectivo optámos pela metodologia qualitativa e método de revisão da literatura. Sendo assim realizamos uma pesquisa de documentos relacionados com a problemática, publicadas no intervalo de tempo entre 2005-2012, em livros, revistas, artigos científicos e bases de dados online, nomeadamente EBSCO, SCIELO, B-on e no RCAAP, orientadas pelas palavras-chave: cuidados paliativos, doente terminal, doente oncológico, cuidados de enfermagem.Desta pesquisa podemos constatar que o enfermeiro tem um papel primordial na equipa de cuidados paliativos, pois ele é o elo de ligação entre o doente, a família e a equipa; é o elemento da equipa que está mais próximo do doente e seus familiares. Os cuidados paliativos visam aliviar o sofrimento não só físico mas também psicológico, espiritual, social, emocional, religioso do doente e seus familiares e nesse processo a função do enfermeiro consiste em avaliar as necessidades em cada uma destas áreas e planear, implementar e avaliar as intervenções apropriadas com o objectivo de melhorar a qualidade de vida e possibilitar uma morte digna. Para dar resposta a essas necessidades o enfermeiro deve desenvolver as suas competências e habilidades nos quatro pilares básicos da prática de cuidados paliativos que são controlo de sintomas, comunicação, apoio a família e trabalho em equipa.

Characterization of functional role of α-ketoglutarate receptor in the kidney : role of the intrinsic circadian timing system in renin expressing cells

Le rein joue un rôle essential dans le maintien de l'homéostasie des fluides extracellulaires (FEC) et la pression artérielle. L'objectif de notre groupe est d'identifier de nouveaux mécanismes impliqués dans le contrôle de l'homéostasie des FEC et de la pression artérielle par le rein. Projet 1) Caractérisation du rôle fonctionnel du récepteur à l'a-cétogluatarate Oxgrl dans le rein Oxgrl est le récepteur spécifique de l'a-cétogluatarate, une moléule intermédiaire du cycle de l'acide citrique, filtrée par le rein et réabsorbée ou secrétée au niveau des tubules proximaux. Le rôle fonctionnel de ces deux récepteurs reste inconnu. Nos résultats montrent qu'Oxgrl est localisé au niveau des cellules intercalaires du tube collecteur (CCD). Des souris (Oxgrr/_) montrent une diminution du pH urinaire ,une augmentation de la concentration de l'acide urinaire titrable et une augmentation des niveaux d'a-cétoglutarate. Le traitement au Na-bicarbonate provoque une augmentation plus prononcée de l'alcalose métabolique chez les souris Oxgrl"7"' accompagnée d'une augmentation de la concentration de bicarbonate et une diminution du niveau de chlore plasmatique. En parallèle, des études de microperfusion ont montré que a-cétoglutarate stimule la réabsorption éléctroneutre de NaCl dans le CCD des souris de type sauvage mais pas des souris Oxgrl"7". En résumé, ces résultats montrent que l'a-cétoglutarate joue un rôle de molécule messagère du tubule proximal jusqu'au tube collecteur au niveau du rein et qu'Oxgrl pourrait être impliqué dans la régulation de l'échange Cl/bicarbonate et la réabsorption du NaCl dans les cellules intercalées. Projet 2) Rôle du système circadien dans les cellules productrices de rénine. Le système chronologique circadien est un mécanisme moléculaire ubiquitaire qui permet à l'organisme de coordonner ses fonctions principales en fonction du temps géophysique. Comme l'activité de la rénine plasmatique montre une rythmicité circadienne nette chez l'homme et la souris ; dans ce projet, nous avons abordé la question à savoir dans quelle mesure le système circadien est impliqué dans cette variabilité circadienne. Pour cela, le gène Bmall, élément principal de l'horloge moléculaire, a été perturbé dans les cellules granulaires productrices de rénine par le système Cre-LoxP. Nos résultats montrent que les souris Renld- Cre/Bmalllox/lox (cKO) présentent de faibles taux d'ARNm de Reni, altèrent la dynamique d'expression de la protéine rénine, mais il le niveau de concentration plasmatique de la rénine reste le même. Cependant, les souris cKO montrent une réduction significative de la concentration plasmatique de l'aldostérone. Nos analyses de l'urine récupérée dans des intervalles de temps de 24 et 1 heure montrent une augmentation du volume urinaire, une tendance à une hypercalciurie, ainsi qu'une altération de la dynamique d'excrétion urinaire de sodium chez les souris cKO. Plusieurs gènes impliqués dans la production/sécrétion de la rénine et dans le contrôle de la fonction rénale montrent une altération de l'expression circadienne d'ARNm. Par ailleurs, les souris cKO montrent une baisse significative de la pression artérielle. Nos résultats suggèrent que l'horloge intrinsèque des cellules productrices de la rénine joue un rôle important dans le control des FEC et l'homéostasie de la pression artérielle via régulation de la fonction rénale.