Method for rapid separation of 3,5,3'-triiodothyroacetic acid in human serum by fast protein liquid chromatography

The 3,5,3'-triiodothyroacetic acid (TRIAC) has been approved as a valuable agent in the management of hyperthyroidism secondary to inappropriate secretion of thyrotropin. We have developed a fast protein liquid chromatography (FPLC) method for separation and quantification of TRIAC. Serum samples charged with TRIAC were extracted with methanol/ammonium acetate, the supernatants were evaporated to dryness, reconstituted in NaOH and injected on a reversed phase column for chromatography. For separation an isocratic elution method (methanol water; 0.1% trifluoroacetic acid) was used. The area under the curve (ml%) was compared with those of the calibration curves. Recoveries were 70 +/- 10.8%. TRIAC was eluted in 2.33 ml. Conclusively, the present method shows that TRIAC can be measured by FPLC and may be applied to the measurement of TRIAC in pharmacological studies.     

Acrosomal status evaluation in human ejaculated sperm with monoclonal antibodies

An important question in mammalian gamete physiology concerns how capacitation and the occurrence of acrosome reactions in motile sperm relate to fertility. Evaluation of these relationships has been restricted by practical limitations because rapid, quantitative assays are unavailable. We have developed a rapid, reproducible assay for the evaluation of acrosomal status utilizing monoclonal antibodies specific to antigens localized in the acrosomal cap region of the sperm head. Mice were immunized with human ejaculated sperm preparations and the resultant hybridomas producing antisperm antibody were selected by solid-phase radioimmunoassay and indirect immunofluorescence (IIF). Two monoclonal antibodies (HS-19, HS-21) recognized target antigens restricted to the acrosomal cap by IIF, and 87 +/- 8.5% of the sperm in fresh ejaculates from 10 different sperm donors showed positive cap fluorescence with these reagents. Loss of HS-21 binding as measured by IIF was correlated with disappearance of the acrosomal cap as observed directly by transmission electron microscopy. Acrosomal disappearance, artificially induced in vitro using the calcium ionophore A23187, also resulted in a loss of HS-21 binding. The induction of acrosomal loss by ionophore was dependent upon extracellular calcium. The data presented suggest that specific monoclonal antibodies can be used for the rapid evaluation of acrosomal status in mammalian sperm.     

Maternal factors in onset of Huntington disease.

Analyses of father-offspring and mother-offspring similarity in onset age suggest that nuclear genes account for a significant portion of the modification of onset age in Huntington disease. The effects of non-nuclear modifiers are supported by the finding that the offspring of affected women have significantly older mean ages of onset than offspring of affected men irrespective of the onset age in the parent. The absence of increased father-daughter similarity indicates that modification is not X-linked. The absence of reproductive advantage for late-onset individuals and the absence of a multigenerational maternal-lineage effect suggest that the modifying effect of the sex of the affected parent occurs in a single parental generation. Offspring of affected women with onset between ages 35 and 49 had a significantly older mean onset age than their mothers. This suggests that a protective effect may be conferred upon the offspring of affected women.     

Histochemical demonstration of peptidases in the human kidney

The localization of several peptidases in the human kidney was investigated histochemically. The membrane-bound peptidases, aminopeptidase A (APA), aminopeptidase M, gamma-glutamyltransferase (gamma-GT) and dipeptidylpeptidase IV, were mainly demonstrable in the brush border of the proximal tubule. In addition, APA was found in the glomeruli, while gamma-GT was found in the basal labyrinth of the proximal tubule. The lysosomal peptidases, dipeptidylpeptidase I and cathepsin B, were most strongly concentrated in the different-sized lysosomes of the proximal tubule, but they were also found in the small lysosomes of the distal tubule. Dipeptidylpeptidase II showed only a weak reaction in lysosomes of the proximal tubule. It is concluded that, in comparison with other previously studied species, the human kidney has a well-developed equipment with membrane-bound and lysosomal peptidases.     

Sperm parameters and testicular volumes in Saguinus mystax

Adult male Saguinus mystax tamarins were evaluated for sperm parameters and testicular volumes. Sperm concentrations average 195.5 X 10(6)/cc with 41.7% motile sperm. Semen specimens were classified as normal, relative to sperm morphology, when 95% or more of the sperm in the specimen had normal morphology; 76% of the animals evaluated had normal semen specimens using this criterion. Testicular volumes averaged 726.9 mm3. A total of 50 infants were sired by 16 of these males during the period covered by this report.     

Yeast pheromone alpha-factor is synthesized as a high molecular weight precursor

The sex pheromone alpha-factor of Saccharomyces cerevisiae, a tridecapeptide of approx. 1,700 molecular weight, was found to be synthesized in vivo as a high molecular weight precursor of Mr = 28,000. Inhibition of N-linked glycosylation by tunicamycin leads to three precursor species of lower molecular weight indicating three carbohydrate residues linked to the alpha-factor precursor molecule. A molecular weight of 18,000 was determined for the unglycosylated molecule.