[A new DNA diagnostic system for the detection of human CYP21 gene mutations associated with adrenal cortex hyperplasia].

Congenital Adrenal Hyperplasia (CAH) is one of the most widespread severe autosomal recessive hereditary diseases. CAH is caused by the impaired biosynthesis of the key human hormones cortisol and aldosterone and is accompanied by the excess synthesis of androgens. Over 90% of CAH cases are caused by a deficiency of the steroid 21-hydrohylase (P450c21). The degree of damage in this enzyme is responsible for the severity of the clinical manifestation of CAH from potentially lethal to mild symptoms. Various mutations of the gene encoding this enzyme are the main source of the reduced activity of the 21-hydrolase. The location of the highly homological pseudogene CYP21P in close proximity to the functional gene impedes the DNA diagnostics of CAH. To detect the eight most frequent CYP21 gene mutations associated with CAH, we developed a new real-time PCR-based system of DNA diagnostics using new allele-specific primers and TaqMan probes for the analyzed mutations. The method was primarily tested on artificial DNA templates, where the analyzed mutations were introduced by site-directed mutagenesis. Then, it was tested on DNA samples from 43 patients with clinical and biochemical manifestations of CAH; seven patients were used as a control. Two mutant alleles were detected in two different individuals: the nonsense Q318X and the missense V281L mutations.

Ovarian function and disease activity in patients with systemic lupus erythematosus.

OBJECTIVE: Menstrual cycle disturbances frequently occur during the onset or in exacerbation periods of systemic lupus erythematosus (SLE), suggesting a possible relationship. The aim of the study is to assess the ovarian function in SLE patients with active disease before the treatment with high doses of glucocorticoids (GC) and cytotoxic agents. METHODS: We evaluated 94 female SLE patients (mean age of 29.2+/-7.0 years). The mean SLEDAI score was 11.4+/-8.1. Seventy-nine patients had a current use of GC with a median dose of 10 mg/day (8-15). The other 15 patients were untreated. After examination and blood sample collection 40% of the patients were treated and high doses of GC (>30 mg/day); 68% from this group of patients were treated GC in combination with cyclophosphamide (CYC). Forty healthy women with the same mean age were evaluated as controls. A careful gynecological history and a gynecological examination were carried out in patients and controls. Hormonal serum levels of follicle-stimulating hormone (FSH), luteinizing hormone (LH), prolactin, estradiol (E2) and progesterone in SLE patients and controls were measured by enzyme-linked immunosorbent assay (ELISA). RESULTS: Menstrual cycle disorders with oligomenorrhea as dominant aspect were observed in 54% of SLE patients. The hormonal studies showed decreased progesterone level in 52% of patients, reduced E2 concentration in 25% of patients; increased levels of LH, FSH and prolactin were observed with the lower frequency (13%, 9%, 10% respectively). Menstrual cycle disorders and the hormonal unbalance such as decreased progesterone level and hyperprolactinemia were found related significantly to high SLEDAI score (p<0.05, p=0.001, p<0.05). In the group of non-treated SLE patients the menstrual and hormonal disorders were observed in the same spectrum and with the same frequency as in all the examined SLE patients. SLEDAI score was found correlated significantly with the frequency of menstrual cycle disorders in non-treated SLE patients (p<0.05). CONCLUSION: The reported study shows the disease activity as a major factor associated with menstrual cycle disorders in SLE patients before treatment with alkylating agents and high doses of GC. Therefore, SLE women might be considered as a risk group for altered ovarian function.