Molecular diagnosis in the clinical practice: an endocrinologist’s perspective

A endocrinologia é uma de muitas especialidades médicas que vem sendo transformada pelo maior conhecimento das bases moleculares das doenças. O teste genético com o propósito de definir o diagnóstico molecular vem ganhando espaço na avaliação rotineira de pacientes com endocrinopatias, e o advento do sequenciamento paralelelo em larga escala (SPLE) está ampliando o potencial de incorporação da informação molecular na prática clínica. O principal benefício do teste genético é a precisão diagnóstica, resultando em melhora e individualização do tratamento de pacientes e seus familiares, e da prevenção de doenças. Entretanto, testes genéticos não são infalíveis e podem trazer alguns riscos potenciais, estando portanto indicados quando a suspeita clínica é forte e o benefício do diagnóstico molecular é claro. Nesta revisão serão explorados conceitos genéticos em evolução e as atuais indicações de diagnóstico molecular em endocrinologia. Ferramentas moleculares serão revisadas, incluindo aquelas visando a identificação de alterações no número de cópias gênicas (cariótipo, FISH, MLPA, aCGH, SNParray) ou na sequencia nucleotídica do DNA (PCR alelo-específica, RFLP, sequenciamento Sanger, SPLE ou NGS). Ainda, serão discutidos os percalços da difícil atribuição de impacto funcional e relevância biológica a variantes de DNA identificadas, e os desafios atuais das tecnologias de análise molecular de larga escala. A endocrinologia molecular vive um momento transformador e empolgante, com o potencial de, em breve, se traduzir em múltiplos benefícios no cuidado de nossos pacientes.Endocrinology is one of several medical specialities that have been gradually transformed by a deeper understanding of the molecular bases of disorders. Genetic testing with the purposes of defining a precise molecular diagnosis has increasingly gained space in the routine assessment of patients with endocrinopathies, and the advent of massive parallel sequencing (MPS) is boosting the incorporation of molecular information in the clinic. The main benefit of genetic testing is diagnostic precision, resulting in improved and individualized care for patients and family members, and better disease prevention. However, genetic tests are not infallible and may bear several potential risks, being thus indicated when clinical suspicion is strong and the benefit of determining a molecular diagnosis is unambiguous. In this review, these evolving concepts and current indications for molecular diagnosis in endocrinology will be explored. Molecular tools will be revised and contextualised, including those aimed at identifying changes in gene dosage (karyotpe, FISH, MLPA, aCGH, SNParray) or in the DNA nucleotide sequence (allele-specific PCR, RFLP, Sanger sequencing, MPS or NGS). Finally, matters surrounding the complex attribution of biologically relevant functional impact to identified DNA variants will be explored, together with the challenges brought by high throughput molecular analysis. These are exciting times for molecular endocrinology, and hopefully soon a translation to multiple benefits for patients will be self-evident

ChIP-on-chip analysis reveals angiopoietin 2 (Ang2, ANGPT2) as a novel target of steroidogenic factor-1 (SF-1, NR5A1) in the human adrenal gland

The nuclear receptor steroidogenic factor-1 (SF-1, NR5A1) is a key regulator of adrenal and gonadal biology. Disruption of SF-1 can lead to disorders of adrenal development, while increased SF-1 dosage has been associated with adrenocortical tumorigenesis. We aimed to identify a novel subset of SF-1 target genes in the adrenal by using chromatin immunoprecipitation (ChIP) microarrays (ChIP-on-chip) combined with systems analysis. SF-1 ChIP-on-chip was performed in NCI-H295R human adrenocortical cells using promoter tiling arrays, leading to the identification of 445 gene loci where SF-1-binding regions were located from 10 kb upstream to 3 kb downstream of a transcriptional start. Network analysis of genes identified as putative SF-1 targets revealed enrichment for angiogenic process networks. A 1.1-kb SF-1-binding region was identified in the angiopoietin 2 (Ang2, ANGPT2) promoter in a highly repetitive region, and SF-1-dependent activation was confirmed in luciferase assays. Angiogenesis is paramount in adrenal development and tumorigenesis, but until now a direct link between SF-1 and vascular remodeling has not been established. We have identified Ang2 as a potentially important novel target of SF-1 in the adrenal gland, indicating that regulation of angiogenesis might be an important additional mechanism by which SF-1 exerts its actions in the adrenal gland

Diversidade de besouros aquáticos (Insecta: Coleoptera) em um trecho do Rio Marambaia, Mangaratiba, Rio de Janeiro, Brasil

Coleoptera é a ordem de insetos mais diversa, incluindo mais de 400 mil espécies popularmente conhecidas como besouros. Pelo menos 13 mil espécies de 37 famílias são consideradas aquáticas, das quais 16 famílias são registradas no Brasil. O presente estudo teve como objetivo realizar o levantamento dos besouros aquáticos ocorrentes em um trecho do Rio Marambaia, Mangaratiba, Rio de Janeiro, Brasil. Um total de 456 espécimes foi obtido na área de estudo, representando os seguintes 12 gêneros e seis famílias: Copelatus, Cybister, Desmopachria, Hydaticus e Rhantus (Dytiscidae); Heterelmis (Elmidae); Gyretes (Gyrinidae); Derallus, Chasmogenus e Tropisternus (Hydrophilidae); Pheneps (Psephenidae); e Iapir (Torridincolidae). Dentre eles, os gêneros Copelatus e Cybister, e as espécies de Hydrophilidae, Chasmogenus fluminensis Clarkson & Ferreira-Jr; Derallus angustus Sharp; Tropisternus apicipalpis Chevrolat; e Tropisternus variolosus Hansen, são aqui registrados pela primeira vez no município de Mangaratiba- RJ. Os registros de Cybister e T. apicipalpis também são os primeiros no Estado do Rio de Janeiro

ENGAJAMENTO NO TRABALHO E CLIMA PARA SERVIÇOS EM UMA INSTITUIÇÃO FEDERAL DE ENSINO SUPERIOR

O objetivo desta pesquisa foi analisar em que medida o clima para serviços é fator determinante para o engajamento no trabalho em uma Instituição Federal de Ensino Superior. A pesquisa, quantitativa, analisou dados coletados por survey e foi aplicada a 59 servidores técnico-administrativos da Universidade Federal do Ceará (UFC), participantes do curso de atendimento estratégico e que trabalham diretamente com o público. Verificou-se que os servidores possuem engajamento moderado, com maior ênfase na absorção. Igualmente, o clima para serviços foi avaliado como moderado, com mediana influência positiva para o engajamento no trabalho e predomínio das diretrizes estratégicas voltadas ao atendimento ao cliente. Entretanto, condições facilitadoras e a avaliação do cliente podem ampliar essa relação, embora em menor proporção. O compromisso institucional com a qualidade do serviço e a delegação de autoridade estão entre as principais ações que podem ser elaboradas a fim de elevar o clima para serviços e o engajamento

Perfil das pacientes com doença Trofoblástica Gestacional atendidas em centros de saúde suplementar da Amazônia: Profile of patients with Gestational Trophoblastic disease seen at supplementary health centers in the Amazônia

A Doença Trofoblástica Gestacional (DTG) é uma complicação rara na gravidez que divide-se em dois grupos: Mola Hidatiforme (MH) e Neoplasia Trofoblástica Gestacional (NTG). O objetivo da pesquisa foi calcular a frequência de DTG em pacientes atendidas em dois centros de referência oncológica de saúde suplementar da cidade de Manaus de 2011 até dezembro de 2019, especificamente, descrever dados sociodemográficos e clínicos, calcular a evolução para NTG classificando as mesmas em alto e baixo risco, assim como descrever formas de tratamento e calcular mortalidade. Trata-se de um estudo observacional retrospectivo, que incluiu pacientes com diagnóstico de DTG que foram atendidas em dois centros suplementares de Manaus de 2011 até dezembro de 2019. Os dados foram obtidos através de entrevistas e preenchimento de protocolo do estudo, após a assinatura do TCLE, depois foram tabulados e analisados estatisticamente. A frequência de DTG foi de cinco pacientes, a idade média foi 32,2 anos. Em sua maioria eram brancas (80%) e possuíam ensino superior completo (80%). A frequência de NTG foi de três pacientes (60%), 100% de baixo risco, 33% apresentaram metástase pulmonar e 100% realizaram monoterapia com Metotrexato. Não ocorreram óbitos nas pacientes da cauística, ficando abaixo da média brasileira de 4% 5

The role of enteric hormone GLP-2 in the response of bone markers to a mixed meal in postmenopausal women with type 2 diabetes mellitus

Abstract\ud \ud Background\ud Type 2 diabetes mellitus (T2D) is a complex disease associated with several chronic complications, including bone fragility and high fracture risk due to mechanisms not yet fully understood. The influence of the gastrointestinal tract and its hormones on bone remodeling has been demonstrated in healthy individuals. Glucagon-like peptide 2 (GLP-2), an enteric hormone secreted in response to nutrient intake, has been implicated as a mediator of nutrient effects on bone remodeling. This study aimed to analyze the dynamics of bone resorption marker C-terminal telopeptide of type I collagen (CTX), bone formation marker osteocalcin, and GLP-2 in response to a mixed meal in diabetic postmenopausal women.\ud \ud \ud Methods\ud Forty-three postmenopausal women with osteopenia or osteoporosis (20 controls – group CO – and 23 diabetic – group T2D) were subjected to a standard mixed meal tolerance test, with determination of serum CTX, plasma osteocalcin and serum GLP-2 concentrations at baseline and 30, 60, 120 and 180 minutes after the meal.\ud \ud \ud Results\ud T2D women had higher body mass index as well as higher femoral neck and total hip bone mineral density. At baseline, luteinizing hormone, follicle-stimulating hormone, osteocalcin and CTX levels were lower in group T2D. In response to the mixed meal, CTX and osteocalcin levels decreased and GLP-2 levels increased in both groups. The expected CTX suppression in response to the mixed meal was lower in group T2D.\ud \ud \ud Conclusions\ud Bone turnover markers were significantly reduced in T2D women at baseline. Confirming the role of nutrient intake as a stimulating factor, GLP-2 increased in response to the mixed meal in both groups. Importantly, CTX variation in response to the mixed meal was reduced in T2D women, suggesting abnormal response of bone remodeling to nutrient intake in T2D.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Coordenação de Aperfeiçoamento de Pessoal de nível superior (CAPES

The influence of seasonality on the content of goyazensolide and on anti-inflammatory and anti-hyperuricemic effects of the ethanolic extract of Lychnophora passerina (Brazilian arnica).

Ethnopharmacological relevance: Lychnophora passerina (Mart ex DC) Gardn (Asteraceae), popularly known as Brazilian arnica, is used in Brazilian folk medicine to treat pain, rheumatism, bruises, inflammatory diseases and insect bites. Aim of the study: Investigate the influence of the seasons on the anti-inflammatory and anti-hyperuricemic activities of ethanolic extract of L. passerina and the ratio of the goyazensolide content, main chemical constituent of the ethanolic extract, with these activities. Materials and methods: Ethanolic extracts of aerial parts of L. passerina were obtained from seasons: summer (ES), autumn (EA), winter (EW) and spring (EP). The sesquiterpene lactone goyazensolide, major metabolite, was quantified in ES, EA, EW and EP by a developed and validated HPLC-DAD method. The in vivo antihyperuricemic and anti-inflammatory effects of the ethanolic extracts from L. passerina and goyazensolide were assayed on experimental model of oxonate-induced hyperuricemia in mice, liver xanthine oxidase (XOD) inhibition and on carrageenan-induced paw edema in mice. Results: HPLC method using aqueous solution of acetic acid 0.01% (v/v) and acetonitrile with acetic acid 0.01% (v/v) as a mobile phase in a gradient system, with coumarin as an internal standard and DAD detection at 270 nm was developed. The validation parameters showed linearity in a range within 10.0?150.0 ?g/ml, with intraday and interday precisions a range of 0.61?3.82. The accuracy values of intraday and interday analysis within 87.58? 100.95%. EA showed the highest goyazensolide content. From the third to the sixth hour after injection of carrageenan, treatments with all extracts at the dose of 125 mg/kg were able to reduce edema. Goyazensolide (10 mg/kg) showed significant reduction of paw swelling from the second hour assay. This sesquiterpene lactone was more active than extracts and presented similar effect to indomethacin. Treatments with ES, EA and EP (125 mg/kg) and goyazensolide (10 mg/kg) reduced serum urate levels compared to hyperuricemic control group and were able to inhibit liver XOD activity. One of the mechanisms by which ES, EA, EP and goyazensolide exercise their anti-hyperuricemic effect is by the inhibition of liver XOD activity. Goyazensolide was identified as the main compound present in ES, EA, EW and EP and it is shown to be one of the chemical constituents responsible for the anti-inflammatory and anti-hyperuricemic effects of the ethanolic extracts. Conclusion: The anti-inflammatory and anti-hyperuricemic activities of the ethanolic extracts from L. passerina were not proportionally influenced by the variation of goyazensolide content throughout the seasons. The involvement of goyazensolide on in vivo anti-inflammatory and anti-hyperuricemic activities of L. passerina extracts was confirmed, as well as the possibility of participation of other constituents on these effects. This study demonstrated that the aerial parts of L. passerina may be collected in any season for use as anti-inflammatory agent. For use in hyperuricemia, the best seasons for the collection are summer, autumn and spring. The ethanolic extract of L. passerina and goyazensolide can be considered promising agents in the therapeutic of inflammation, hyperuricemia and gout