Epitope map for a growth hormone receptor agonist monoclonal antibody, MAb 263

Monoclonal antibody (MAb) 263 is a widely used monoclonal antibody that recognizes the extracellular domain (ECD) of the GH receptor. It has been shown to act as a GH agonist both in vitro and in vivo, and we report here that it must be divalent to exert its effect on the full-length receptor. To understand the mechanism of its agonist action, we have determined the precise epitope for this antibody using a novel random PCR mutagenesis approach together with expression screening in yeast. A library of 5200 clones of rabbit GH receptor ECD mutants were screened both with MAb 263 and with an anticarboxy-tag antibody to verify complete ECD expression. Sequencing for clones that expressed complete ECD but were not MAb 263 positive identified 20 epitope residues distributed in a discontinuous manner throughout the ECD. The major part of the epitope, as revealed after mapping onto the crystal structure model of the ECD molecule, was located on the side and upper portion of domain 1, particularly within the D - E strand disulfide loop 79 - 96. Molecular dynamics docking of an antibody of the same isotype as MAb 263 was used to dock the bivalent antibody to the 1528-Angstrom(2) epitope and to visualize the likely consequences of MAb binding. The minimized model enables the antibody to grasp two receptors in a pincer-like movement from opposite sides, facilitating alignment of the receptor dimerization domains in a manner similar to, but not identical with, GH.

Seeking SOCS and sex steroids

In seeking to explain why oral estrogen inhibits the GH-IGF-I axis, a recent study has unearthed a new way that steroid hormones can influence growth hormone action. This involves suppressors of cytokine signalling (SOCS), which offer a new level of understanding in signal control and crosstalk.

High rate of detection of primary aldosteronism, including surgically treatable forms, after 'non-selective' screening of hypertensive patients

Background Wide testing of the aldosterone: renin ratio among hypertensive individuals has revealed primary aldosteronism to be common, with most patients normokalaemic. Some investigators, however, have reported aldosterone-producing adenoma to be rare among patients so detected. Objective To test the hypothesis that differences among reported studies in the rate of detection of aldosterone-producing adenoma (as opposed to bilateral adrenal hyperplasia) reflect differences in the procedures used for diagnosis of primary aldosteronism, and the methods used to identify aldosterone-producing adenomas. Methods In the newly established Princess Alexandra Hospital Hypertension Unit (PAHHU), we used procedures developed by Greenslopes Hospital Hypertension Unit (which reports that more than 30% of patients with primary aldosteronism have aldosterone-producing adenomas) to diagnose primary aldosteronism and determine the subtype. All patients with an increased aldosterone: renin ratio (measured after correction for hypokalaemia and while the patient was not receiving interfering medications) underwent fludrocortisone suppression testing to confirm or exclude primary aldosteronism; if they were positive, they underwent genetic testing to exclude glucocorticoid-remediable aldosteronism before adrenal venous sampling was used to differentiate lateralizing from bilateral primary aldosteronism. Results This approach allowed PAHHU to diagnose, within 2 years, 54 patients [only seven (13%) hypokalaemic] with primary aldosteronism. All tested negative for glucocorticoid-remediable aldosteronism. Aldosterone production was lateralized to one adrenal in 15 patients (31%; only six hypokalaemic) and was bilateral in 34 (69%; all normokalaemic) of 49 patients who underwent adrenal venous sampling. Among patients with lateralizing adrenal hyperplasia, computed tomography revealed an ipsilateral mass in only six and a contralateral lesion in one. Fourteen patients underwent unilateral adrenalectomy, which cured the hypertension in seven and improved it in the remainder. In patients with bilateral primary aldlosteronism, hypertension responded to spironolactone (112.5-50 mg/ day) or amiloride (2.5-10 mg/day). Conclusion When performed with careful regard to confounding factors, measurement of the aldosterone: renin ratio in all hypertensive individuals, followed by fludrocortisone suppression testing to confirm or exclude primary aldosteronism and adrenal venous sampling to determine the subtype, can result in the detection of significant numbers of patients with specifically treatable or potentially curable hypertension. (C) 2003 Lippincott Williams Wilkins.

Primary aldosteronism

Approaching the fiftieth year since its original description, primary aldosteronism is now thought to be the commonest potentially curable and specifically treatable form of hypertension. Correct identification of patients with primary aldosteronism requires that the effects of time of day, posture, dietary sodium intake, potassium levels and medications on levels of aldosterone and renin be carefully considered. Accurate elucidation of the subtype is essential for optimal treatment, and adrenal venous sampling is the only reliable means of differentiating aldosterone-producing adenoma from bilateral adrenal hyperplasia. With genetic testing already available for one inherited form, making more cumbersome biochemical testing for that subtype virtually obsolete and bringing about improvements in treatment approach, an intense search is underway for genetic mutations causing other, more common familial varieties of primary aldosteronism.

Primary aldosteronism: From genesis to genetics

The fact that a debate concerning the unexpectedly high prevalence of normokalaemic primary aldosteronism (PAL) attracted a large audience at the 2002 Scientific Meeting of the International Society of Hypertension makes it timely to address this issue. The affirmative case argues that PAL is the most common potentially curable and specifically treatable form of hypertension, itself the most common chronic disorder in Western societies, with significant morbidity and mortality, consuming large proportions of health budgets. Recent discoveries about the genetics of aldosterone production and of its unexpectedly broad effects on the cardiovascular system need to be placed in clinical context.

E@D: projeto Serpro de educa????o a dist??ncia

O projeto refere-se ?? experi??ncia do Servi??o Federal de Processamento de Dados (Serpro) na implanta????o de uma solu????o completa de educa????o a dist??ncia, em que, considerando a necessidade de capacita????o de grande contingente de pessoas em curto espa??o de tempo e com baixa disponibilidade de recursos, ocorrida em 1999, surgiu a criatividade de um grupo de t??cnicos para criar uma alternativa que viria a se transformar em solu????o de grande sucesso e de utilidade tanto para o Serpro quanto para seus clientes. Ap??s seis anos do in??cio da experi??ncia, j?? foram desenvolvidos mais de 50 cursos com atendimento a aproximadamente 35 mil pessoas, demonstrando a viabilidade dessa modalidade de aprendizagem na educa????o corporativa

Cheques indevidos: corrigindo um erro de v??rios milh??es de d??lares

O caso trata das dificuldades de tomada de decis??o por parte de uma autoridade federal ao serem constatados erros no repasse de subs??dios para agricultores em um programa governamental. O texto relata o conflito entre as posi????es da auditoria e da ??rea t??cnica a respeito das a????es a serem desenvolvidas; a primeira, considerando a quest??o sob o ponto de vista legal e do interesse p??blico, e a segunda, priorizando o interesse dos agricultores e os impactos pol??ticos de uma decis??o contr??ria ?? categoria. O caso tem sido usado em cursos para subsidiar discuss??es sobre ??tica, accountability, conflito de interesses e gest??o de erros n??o previstos

Emiss??o de certid??o negativa de d??bitos de tributos e contribui????es federais via internet

Obter uma Certid??o Negativa de D??bitos de Tributos e Contribui????es Federais da Pessoa Jur??dica, at?? dezembro de 1997, implicava na presen??a do contribuinte em uma Delegacia da Receita Federal. Com a implanta????o do sistema certid??o negativa na Internet isso mudou; o projeto foi desenvolvido, objetivando permitir que o contribuinte obtenha a Certid??o Negativa sem a necessidade de deslocamento, propiciando-lhe maior conforto e economia financeira, conseq??entemente, diminuindo o fluxo de contribuintes ??s Unidades da SRF. Se o contribuinte est?? em dia com suas obriga????es tribut??rias, ?? poss??vel emiti-la via Internet, no site da SRF. A Certid??o Negativa ?? imprescind??vel para que as empresas participem de licita????es, vendam im??veis ou obtenham cr??dito banc??rio. Pode ficar a cargo da entidade que exige a certid??o, verificar a situa????o fiscal do contribuinte na Internet

Sucess??o gerencial para chefia geral de centros de pesquisa (P&D)

A iniciativa surgiu quando constatada grande defasagem entre o n??vel de qualifica????o do grupo t??cnico-cient??fico (pesquisadores) e do grupo gerencial na EMBRAPA. Foi ent??o implementada nova metodologia de sele????o de chefes das Unidades Descentralizadas. A nova modalidade de sele????o conseguiu combinar participa????o democr??tica dos empregados com a avalia????o objetiva do m??rito dos candidatos e com o compromisso com uma proposta de trabalho compat??vel com os objetivos e a miss??o da UD. Al??m disso, oferece ?? sociedade a oportunidade de participar do gerenciamento de suas unidades de pesquisa

Decreto n?? 6.563: aprova o Estatuto e o Quadro Demonstrativo dos Cargos em Comiss??o e das Fun????es Gratificadas da Funda????o Escola Nacional de Administra????o P??blica - ENAP, e d?? outras provid??ncias.

O PRESIDENTE DA REP??BLICA, no uso das atribui????es que lhe confere o art. 84, incisos IV e VI, al??nea ???a???, da Constitui????o, e tendo em vista o disposto no art. 50 da Lei n. 10.683, de 28 de maio de 2003, decreta: ficam aprovados o Estatuto e o Quadro Demonstrativo dos Cargos em Comiss??o e das Fun????es Gratificadas da Funda????o Escola Nacional de Administra????o P??blica - ENAP, na forma dos Anexos I e II a este Decreto.