Characterization of the c.190T>C missense mutation in BRCA1 codon 64 (Cys64Arg).

In the Milan area (Northern Italy), we identified a family characterized by a high prevalence of ovarian and breast cancer cases (5 out of 6 subjects, over 3 generations), and a predominant prevalence of ovarian lesions (4 out of 5 patients). Analysis of BRCA1 and BRCA2 genes allowed the identification of the missense c.190T>C mutation in codon 64 (Cys64Arg) of BRCA1. The aims of the present investigation were to characterize the functional implications of the c.190T>C mutation at the molecular level, and to search whether additional polymorphisms might be linked to the peculiar phenotypic features observed in the Italian pedigree. Molecular modelling studies suggested that substitution of the cysteine 64 with an arginine likely disrupts the architecture of the BRCA1 RING finger domain, responsible for the interaction with BARD1, essential for the tumor-suppressor activity of the BRCA1-BARD1 complex. By splicing site information analysis, exonic splicing enhancer site characterization, and analysis of transcript fragment length and sequence, we showed that the c.190T>C mutation was able to modulate the splicing of exon 5 in a fashion opposite to the c.190T>G transversion, responsible for the functionally-related Cys64Gly amino acid substitution. Genotyping of BRCA1 and BRCA2 in the Italian family revealed the presence of two significant polymorphisms: the cancer-associated c.2612C>T SNP in BRCA1, and the c.-26G>A SNP in the BRCA2 gene, acting as an ovarian cancer risk modifier in carriers of deleterious BRCA1 mutations. Analysis of these SNPs in a genotypically-unrelated Polish family, characterized by prevalent breast neoplasms in carriers of the c.190T>C mutation, revealed a genetic profile consistent with the hypothetic role of both polymorphisms.

The effect of methionine-enkephalin on nitric oxide release in mice is age and gender related.

Gender- and age-related differences in nitric oxide (NO) release and in response to drugs of abuse has been reported in both humans and experimental animals. So far, we have demonstrated in vivo methionine-enkephalin- (MENK-) modulated NO release in mice. However, no data on the influence of age and gender on this immunomodulatory effect of MENK have been reported. In this study we examined the influence of age (2, 4, 8 month old mice) and gender (male and female mice) on MENK-induced NO release of mouse peritoneal macrophages (PEMs) of the CBA strain of mice. NO release was not age but was gender related in that males generally produced more NO than females. The effect of MENK on NO release was age (demonstrated only in mature 4 and 8 month old mice) and gender related in that it could be observed only in female mice. Apoptotic cells that paralleled the increase of NO in MENK-treated female mice were, however, observed also in male mice although MENK was in males without effect. These data provide evidence that some immunomodulatory properties of MENK are age and gender related which may be relevant to the potential use of MENK in adjuvant therapy for immunocompromised status.