RESUMO
Germline inactivating variants in BRCA1 lead to a significantly increased risk of breast and ovarian cancers in carriers. While the functional effect of many variants can be inferred from the DNA sequence, determining the effect of missense variants present a significant challenge. A series of biochemical and cell biological assays have been successfully used to explore the impact of these variants on the function of BRCA1, which contribute to assessing their likelihood of pathogenicity. It has been determined that variants that co-localize with structural or functional motifs are more likely to disrupt the stability and function of BRCA1. Here we assess the functional impact of 37 variants chosen to probe the functional impact of variants in phosphorylation sites and in the BRCT domains. In addition, we perform a meta-analysis of 170 unique variants tested by the transcription activation assays in the carboxy-terminal domain of BRCA1 using a recently developed computation model to provide assessment for functional impact and their likelihood of pathogenicity.
Assuntos
Proteína BRCA1/química , Proteína BRCA1/metabolismo , Domínios e Motivos de Interação entre Proteínas , Relação Estrutura-Atividade , Alelos , Substituição de Aminoácidos , Proteína BRCA1/genética , Teorema de Bayes , Sequência Conservada , Humanos , Mutação de Sentido Incorreto , Domínios e Motivos de Interação entre Proteínas/genética , TermodinâmicaRESUMO
Bowdichia virgilioides Kunth (Family Fabaceae) is a plant that is distributed widely in the tropical and subtropical regions of the world. In the northeast region of Brazil, where B. virgilioides is called "sucupira-preta," the stem bark is used in folk medicine to treatment of inflammatory and painful diseases. This study aimed to evaluate the antinociceptive activity of the aqueous extract of the dried stem bark of B. virgilioides. The aqueous extract of B. virgilioides in doses of 50, 100, 200, and 400 mg/kg was administered orally 1 hour prior to pain induction. Only the doses of 200 and 400 mg/kg produced an inhibition by 61% and 74%, respectively, in the number of abdominal writhings induced by acetic acid. This antinociceptive effect was not reversed by pretreatment with naloxone, indicating that the effect is not associated with the activation of opioid receptors. In the formalin test, using the two highest doses, the extract had no effect in the first phase but produced an analgesic effect on the second phase with the inhibition of licking time (P < .001). In the hot plate test, no effect was seen at the dose of 400 mg/kg p.o. Our findings show that B. virgilioides contains pharmacologically active constituents that possess antinociceptive activity justifying its popular therapeutic use in treating conditions associated with the painful conditions.
Assuntos
Analgésicos/uso terapêutico , Fabaceae , Dor/tratamento farmacológico , Fitoterapia , Extratos Vegetais/uso terapêutico , Ácido Acético , Analgésicos/farmacologia , Animais , Comportamento Animal/efeitos dos fármacos , Fabaceae/química , Formaldeído , Masculino , Camundongos , Naloxona/farmacologia , Antagonistas de Entorpecentes/farmacologia , Dor/etiologia , Casca de Planta , Extratos Vegetais/farmacologia , Caules de PlantaRESUMO
We studied bone marrow stromal cell cultures from patients with childhood myelodysplastic syndromes (MDS, refractory anemia with excess of blasts, RAEB) and from matched normal donors. Stromal cell monolayers were characterized as myofibroblasts by the expression of smooth muscle alpha-actin, collagen IV, laminin and fibronectin. When normal cord blood cells were plated onto myelodysplastic stromas, a pathologic cell differentiation was observed, indicating altered myelosupportive properties. cDNA array analysis showed that patient stromas expressed increased levels of thrombospondin-1, collagen-I alpha2-chain, osteoblast-specific factor-2 and osteonectin, indicating the presence of increased osteoblast content, as confirmed by enhanced alkaline phosphatase synthesis. Alterations in the myelodysplastic stroma environment might contribute to abnormal hematopoiesis in this pathology.
Assuntos
Medula Óssea/patologia , Regulação Neoplásica da Expressão Gênica , Hematopoese , Músculo Liso/patologia , Síndromes Mielodisplásicas/patologia , Células Estromais/patologia , Actinas/metabolismo , Fosfatase Alcalina/metabolismo , Anemia Refratária com Excesso de Blastos , Medula Óssea/metabolismo , Estudos de Casos e Controles , Diferenciação Celular , Criança , Pré-Escolar , Colágeno Tipo IV/metabolismo , Feminino , Sangue Fetal/química , Sangue Fetal/metabolismo , Fibroblastos/patologia , Fibronectinas/metabolismo , Perfilação da Expressão Gênica , Humanos , Técnicas In Vitro , Lactente , Laminina/metabolismo , Masculino , Músculo Liso/metabolismo , Síndromes Mielodisplásicas/genética , Síndromes Mielodisplásicas/metabolismo , Análise de Sequência com Séries de Oligonucleotídeos , Osteoblastos/metabolismo , Pré-Leucemia , Células Estromais/metabolismoRESUMO
The apparently dormant breast cancer micrometastases in haemopoietic marrow are correlated with distant metastatic carcinoma dissemination. We studied in vitro interactions of carcinoma cells with adjacent stromata, using connective tissue cell cultures from breast and bone marrow samples of normal donors, comparing them to the pericancerous breast tissue and bone marrows of 12 selected patients with invasive breast carcinomas. Cancer cells were detected by immunocytochemistry and RT-PCR in all the bone marrows and in most blood samples of the studied patients. We monitored the growth and interaction of carcinoma MCF-7 cells with the stromata. The normal breast stroma sustained typical massive cancer growth. The pericancerous breast stroma induced the invasive mesenchymal pattern of growth. Normal bone marrow stroma induced the same conversion and was highly adhesive, retaining the cells in the stroma, but carcinoma patients' bone marrow stromata underwent low adhesive interactions with cancer cells, releasing them potentially into the circulation. The semi-quantitative RT-PCR indicated an enhanced expression of the hepatocyte growth factor and its receptor c-met in breast and bone marrow stromata of cancer patients. The input of cancer cells into the normal bone marrow may induce modifications of the local microenvironment, favourable for growth and release of carcinoma cells into the systemic circulation, which correlate with the poor prognosis of patients with bone marrow micrometastases.