Relationship between expansion of the CAG repeat in exon 1 of the androgen receptor gene and idiopathic male infertility.

OBJECTIVE: To determine whether expansion of CAG repeats in exon 1 of the androgen receptor is correlated with impaired spermatogenesis in patients with male idiopathic infertility. DESIGN: A retrospective study. SETTING: Medical school in Besançon, France. PARTICIPANT(S): Thirty-seven infertile patients with azoospermia or oligospermia and 50 fertile controls. INTERVENTION(S): History, physical, hormonal assays, semen analysis, and collection of blood samples in order to study the androgen receptor's gene. MAIN OUTCOME MEASURE(S): Blood samples were collected from each infertile patient and control. The length of the CAG repeat segment was evaluated by using polymerase chain reaction (PCR) electrophoresis in exon 1 and PCR single-strand conformation polymorphism in exons 2-8. RESULT(S): The mean length of the CAG repeats was significantly different between infertile and fertile patients (23.91 +/- 0.5 vs. 22.20 +/- 0.4). No mutation was detected in exons 2-8 of the androgen receptor gene in infertile patients. CONCLUSION(S): Expansion of the CAG repeat segment of the androgen receptor is correlated with male idiopathic infertility. The number of CAG repeats may therefore have a modulatory effect on normal androgen receptor function.

A sulfone group-labeled TEM-DNA probe: comparison with a 32P-labeled probe in dot-hybridization.

A non-radioactive DNA probe for the TEM-type beta-lactamase gene was obtained by using the 'Chemiprobe' system. It was used along with a 32P-labeled TEM probe to screen for TEM beta-lactamase gene in 107 bacterial isolates representing 7 Gram-negative genera and previously classified as TEM-positive or negative. The DNA to be tested was extracted from these bacterial isolates by the Birnboim-Doly method and, after blotting into charged nylon membranes, it was submitted to hybridization with either the TEM 'Chemiprobe' or the 32P-TEM probe. The TEM 'Chemiprobe' could detect as few as 25 pg specific DNA if it was used at a concentration of 5 ng per cm2 of membrane. The results obtained by both probes were concordant in 93.5% of the entire sample. The TEM 'Chemiprobe' was specific since only one false positive was observed. Furthermore, it appeared at least as sensitive as the 32P-labeled TEM probe. As the dot-hybridization with the sulfone-labeled probe was sensitive, simple and easy to perform, it will be useful for large-scale screening in clinical laboratory.

Steroid sulfatase activities in normal and cirrhotic livers and plasma levels of estrone sulfate, estrone and estradiol-17 beta in men.

Estrone and dehydroepiandrosterone (DHEA) sulfatases were studied in livers of normal and cirrhotic men. Their Km were 3.2 microM and 1.2 microM respectively. The microsomal sulfatases were solubilized by Miranol H2M and ultrasound. After gel filtration, the soluble material gave a single peak of activity for both substrates with a molecular weight of approximately 330,000. In terms of pmol of product.min-1 per mg of fresh tissue, the mean (+/- SD) values of estrone and DHEA sulfatase activities were lower in cirrhotic livers [(n = 7) (4.09 +/- 2.90 and 0.38 +/- 0.20)] than in normal livers [(n = 13)(8.29 +/- 4.00 and 0.69 +/- 0.20)]. The differences were statistically significant : p less than 0.03 for estrone sulfatase and p less than 0.01 for DHEA sulfatase. In cirrhotic men, the mean level of plasma estrone is increased whereas that of estrone sulfate is decreased. The variations may be related to the decrease of serum albumin in cirrhotic subjects.

Estrone sulfate concentrations in plasma of normal individuals, postmenopausal women with breast cancer, and men with cirrhosis.

Estrone sulfate is quantitatively the most important estrogen in plasma. A method for its determination in human plasma is described, and the precision, accuracy, sensitivity, and specificity are defined. Free steroids were extracted from plasma with diethyl ether and steroid sulfates were isolated with use of Vlitos' reagent (methylene blue in dilute H2SO4/Na2SO4 solution). After enzymic hydrolysis, estrone was isolated by chromatography on Celite and measured by radioimmunoassay. The mean concentrations (nmol/L +/- 1 SD) of estrone sulfate were 2.51 +/- 0.90 for plasma from 13 women in follicular phase, 5.33 +/- 1.55 for 17 women in luteal phase, 0.89 +/- 0.60 for 44 postmenopausal women, and 0.96 +/- 0.43 for 24 postmenopausal women with breast cancer. Results for postmenopausal women with or without breast cancer did not differ significantly. For 13 normal men, estrone sulfate concentrations were 2.62 +/- 0.79 nmol/L, and for a group of 19 cirrhotic men the mean value was 1.43 +/- 0.95 nmol/L, significantly lower than normal.