Fluorescent in situ hybridization evaluation of human Y-bearing spermatozoa separated by albumin density gradients.

OBJECTIVE: Fluorescent in situ hybridization of decondensed sperm nuclei was employed to evaluate the effectiveness of the Ericsson albumin gradient enrichment technique of Y-bearing sperm. DESIGN: Mature human spermatozoa were separated from donor- and randomly selected patient semen using three different albumin density layers. Sperm nuclei, in the albumin layer with highest density, were decondensed and the percentage Y-bearing sperm was determined by fluorescent in situ hybridization. RESULTS: A slight, although statistically significant, increase in percentage Y-bearing sperm was observed. CONCLUSION: Although statistically significant, the clinical relevance of the small increase in Y-bearing sperm remains uncertain. A randomized controlled clinical study should help to clarify the above laboratory results.

Intrafamilial variability in the clinical expression of familial hypercholesterolemia: importance of risk factor determination for genetic counselling.

A specific mutation in the low-density lipoprotein receptor (LDLR) gene causes familial hypercholesterolemia (FH) in about 60% of Afrikaner FH heterozygotes. Molecular diagnosis of this so-called FH Afrikaner-1 mutation was performed in a family with the disease. One individual did not develop coronary heart disease (CHD) by age 84, despite having the FH Afrikaner-1 mutation, while his son who inherited the same gene, developed CHD before age 50 and had to undergo bypass surgery. All the sibs in the third generation inherited the defective LDLR gene allele. This variation in clinical presentation creates a counselling dilemma. It also raises questions about the effect of diet and life style, and the possibility of other genes either contributing to the severity of the disease, or protecting against high lipid levels in plasma. An investigation of the influence of selected factors on the clinical expression of the FH Afrikaner-1 mutation in this family indicated that it was especially the elevated apolipoprotein (a) levels, in addition to low levels of high density lipoprotein cholesterol and raised triglyceride and apolipoprotein B levels, that were associated with a greater risk of developing CHD. These findings are thus in accordance with the view that the severity of CHD in FH patients is not only determined by nature of the gene defect, but is also influenced by other risk factors.

Immunocytochemical identification of nuclear estrogen-receptors in human acute myeloid leukemia.

The peripheral and bone marrow blast cells of twenty patients newly-diagnosed as acute myeloid leukemia (AML), subtypes M1-M5, were analysed for the presence of estrogen-receptors by ER-ICA immunocytochemistry and dextran-coated charcoal cytosolic (DCC) assay. Two patients demonstrated myeloblasts with nuclear staining patterns consistent with the presence of estrogen-receptors. The positive immuno-staining results were confirmed by DCC in one patient with AML, subtype M4. This patient demonstrated an unusual myeloblast karyotype with a reciprocal translocation t(6;11)(q27;q23.3) involving the designated locus of the ESR gene on chromosome 6 (6q24-qter). The abnormal juxtaposition of DNA control elements close to the promoter of the ESR gene may be one mechanism by which aberrant synthesis of estrogen-receptor protein can occur to provide a growth advantage for leukemia cell clones.

Deletion mapping of 39 random isolated Y-chromosome DNA fragments.

Thirty-nine recombinants isolated from a Y chromosome-specific library were deletion mapped. Seven deletion intervals were defined by hybridization of probes to DNA of eight individuals with aberrant Y chromosomes. Extreme cytogenetic limits of the deletion intervals were determined by in situ hybridization of one probe per deletion interval. Five intervals, with a total of twenty-five probes, were allocated to the long-arm euchromatic region. The probes described will be useful for characterization of aberrant Y chromosomes, in searching for expressed sequences on the Y chromosome, and for further study of the evolutionary relationship between the Y chromosome and other chromosomes.

Severe mental retardation in six generations of a large South African family carrying a translocation t(6;10)(q27;q25.2).

Partial monosomy 10q25.2----qter, detected in a newborn baby with multiple congenital abnormalities, was found to be derived from a balanced maternal translocation t(6;10)(q27;q25.2). The pedigree of six generations of the family is presented. In an extensive cytogenetic study of this family, the chromosome complements of 57 subjects, potentially capable of carrying some form of this translocation, were analysed. A total of 14 male carriers (four obligatory) and 14 female carriers (three obligatory) of this translocation was found. Partial trisomy 10q25.2----qter, associated with severe mental retardation, occurred in nine cases, eight males and one female. Two of these eight males were detected prenatally and subsequently therapeutically aborted. The phenotypes of the family members with partial trisomy 10q25.2----qter are compared to each other and to those reported in publications. No further cases of partial monosomy 10q25.2----qter were encountered. A review of published reports of partial monosomy and partial trisomy 10qter is given. The apparent absence of infertility, the occurrence of many first trimester miscarriages, and the marked sex ratio are discussed.

Balanced chromosome translocations and abnormal phenotypes. A report of 5 cases.

Five cases in which phenotypic abnormalities were found in association with apparent balanced chromosomal translocations are described. In 3 patients, one of the parents was found to be a carrier of the same translocation. In a further patient, the translocation was shown to be de novo and in the remaining patient the father was not available for chromosome studies. In a review of the literature the breakpoints in 36 familial balanced translocations were compared with 40 de novo translocations (including the present cases) all associated with phenotypic abnormalities. No common translocation was found in these groups, but it was observed that chromosomes 4 and 5 were significantly more involved in de novo translocations than in familial translocations. The possible aetiology and implications for prenatal diagnoses are discussed.

Chromosome studies in 496 infertile males with a sperm count below 10 million/ml.

Chromosome studies were performed on 106 men with azoospermia and 390 men with oligozoospermia (consistent sperm count below 10 million/ml). Constitutional chromosome abnormalities were found in 14.1% of the azoospermia group and in 5.1% of the oligozoospermia group. An overall incidence of 7.1% constitutional abnormalities indicates that this criterion of selection may be advisable for routine chromosome analysis of infertile men. A reduction of 25% in the workload increases the yield of chromosome abnormalities in the group of infertile men to 10-14 times above that expected in the normal population.