RESUMO
We report a direct DNA sequencing analysis of the MECP2 gene undertaken on a further 64 Italian patients with Rett syndrome by using a LICOR 4200 Automated Sequencer. All of the girls entering the study had a consistent clinical diagnosis for this disorder. All coding regions and the flanking intronic splice site sequences were amplified as three non-overlapping fragments by using both forward and reverse primers. The results were then compared to the MECP2 reference sequences published in GenBank. Mutations of the MECP2 gene were identified in 64 of 75 (85.33%) unrelated sporadic Rett syndrome girls. Genotype/phenotype correlation studies, in particular in groups of patients with the same mutation, did not offer definitive and interesting data.
Assuntos
Proteínas Cromossômicas não Histona , Análise Mutacional de DNA , Proteínas de Ligação a DNA/genética , Testes Genéticos , Mutação/genética , Proteínas Repressoras , Síndrome de Rett/genética , Adolescente , Adulto , Sequência de Bases , Criança , Pré-Escolar , Códon sem Sentido/genética , DNA/genética , Feminino , Mutação da Fase de Leitura/genética , Genótipo , Humanos , Itália , Proteína 2 de Ligação a Metil-CpG , Mutação de Sentido Incorreto/genética , Fenótipo , Síndrome de Rett/fisiopatologiaRESUMO
We used an infrared (IR) automated fluorescence monolaser sequencer for the analysis of 13 autosomal short tandem repeat (STR) systems (TPOX, D3S1358, FGA, CSF1PO, D5S818, D7S820, D8S1179, TH01, vWA, D13S317, D16S359, D18S51, D21S11) and the X-Y homologous gene amelogenin system. These two systems represent the core of the combined DNA index systems (CODIS). Four independent multiplex reactions, based on the polymerase chain reaction (PCR) technique and on the direct labeling of the forward primer of every primer pair, with a new molecule (IRDye800), were set up, permitting the exact characterization of the alleles by comparison with ladders of specific sequenced alleles. This is the first report of the whole analysis of the STRs of the CODIS core using an IR automated DNA sequencer. The protocol was used to solve paternity/maternity tests and for population studies. The electrophoretic system also proved useful for the correct typing of those loci differing in size by only 2 bp. A sensibility study demonstrated that the test can detect an average of 10 pg of undegraded human DNA. We also performed a preliminary study analyzing some forensic samples and mixed stains, which suggested the usefulness of using this analytical system for human identification as well as for forensic purposes.
Assuntos
Corantes Fluorescentes , Repetições Minissatélites , Paternidade , Polimorfismo Genético , Análise de Sequência de DNA/métodos , Automação , DNA/análise , Corantes Fluorescentes/química , Humanos , Indóis/química , Raios Infravermelhos , Estrutura Molecular , Técnicas de Amplificação de Ácido Nucleico , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Moldes GenéticosRESUMO
Holter's test was performed in eight patients with advanced breast cancer and paraneoplastic hypercalcemia. In spite of several reports of cardiac arrhythmias, none were found in these patients.
Assuntos
Arritmias Cardíacas/etiologia , Hipercalcemia/complicações , Adulto , Neoplasias da Mama/complicações , Eletrocardiografia Ambulatorial , Feminino , Humanos , Hipercalcemia/etiologia , Hipercalcemia/fisiopatologia , Pessoa de Meia-IdadeRESUMO
An empirical approach to the detection of groups of women at high risk of developing breast carcinoma by a score-system is presented. A comparison between the diffusion of breast self-examination and Pap-test, and a comparison between medical breast checks and gynecologic ones are also presented.