Familial effects on plasma sex-steroid content in man: testosterone, estradiol and Sex-hormone-binding globulin.

We investigated whether familial factors influence the plasma content of sex-steroids and sex-hormone-binding globulin (SHBG) in 98 adult males of 66 families. They had no apparent endocrine dysfunction, The 0800-1100 hr plasma levels of testosterone, 5 alpha-dihydrotestosterone (DHT), estradiol-17 beta (E2) and estrone (E1) were measured by radioimmunoassay. The free fractions of E2 and testosterone were determined by equilibrium dialysis, and binding capacity of SHBG was also calculated. The data were analyzed by analysis of variance. We observed that the differences in the plasma content of testosterone (p = .02). SHBG binding capacity (p = 0.1), and E2 (p = 0.3), free E2 index (p = .05) were all substantially less variable within groups of brothers than among non-brothers. The variability of the plasma concentration of DHT, free testosterone and E1 was not significantly less within brothers than among non-brothers. The correlation between either plasma testosterone content (r = .14) or SHBG binding capacity (r = .12) and percent of ideal body weight was not significant statistically. Age had no effect on the results. Our data suggest that genetic and/or environmental factors may affect the plasma content of testosterone, E2 and SHBG binding capacity.

The preparation of carbohydrate-protein conjugates: cyanuric trichloride coupling of 2-aminoethyl glycosides, and mixed-anhydride coupling of 8-carboxyoctyl glycosides to bovine serum albumin.

Preparation of the following glycosides is described: 2-aminoethyl beta-D-glycosides of (A) 2-acetamido-3,4,6-trio-O-acetyl-2-deoxy-D-glucopyranose, (B) 2-acetamido-4-O-(2-acetamido-3,4,6-trio-O-acetyl-2-deoxy-beta-D-glucopyranosyl)-3,6-di-O-acetyl-2-deoxy-beta-D-glucopyranose (N,N'-diacetylchitobiose pentaacetate), (C) 4-O-(2,3,4,6-tetra-O-acetyl-beta-D-glucopyranosyl)-2,3,6-trio-O-acetyl-beta-D-glucopyranose (cellobiose heptaacetate); 8-carboxyoctyl glycosides of (D) cellobiose, and (E) N,N'-diacetylchitobiose. Conjugates were prepared from (A), (B), and (C) by coupling to bovine serum albumin by cyanuric trichloride and subsequent deacetylation; (D) and (E) were coupled to bovine serum albumin by the mixed-anhydride reaction. Conjugates (A) and (B) were insoluble; conjugates (C), (D), and (E) functioned as artificial antigens and gave rise to precipitating antibodies in rabbits. Specificities of the antisera were determined by inhibition studies.

Electrospray analysis of biological samples for trace amounts of trichloroacetic acid, dichloroacetic acid, and monochloroacetic acid.

Trichloroethylene (TCE) has been identified as a widespread groundwater contaminant. Trichloroacetic acid (TCA) and dichloroacetic acid (DCA) are toxicologically relevant metabolites of TCE that produce tumors in B6C3F1 mice. A sensitive method for measuring these metabolites in plasma has been developed to obtain pharmacokinetic data from TCE exposure. This is particularly important because DCA is more potent at producing hepatoproliferative lesions than TCA. At present, it is unclear whether DCA is produced by humans. Existing gas chromatographic methods cannot detect DCA at low nanogram-per-milliliter levels. A Finnigan TSQ 700 mass spectrometer (MS) with electrospray ionization was used to measure TCA, DCA, and monochloroacetic acid (MCA) in plasma. The MS was operated in negative ion tandem MS mode. The limit of detection for TCA and DCA was 4 ng/ml, and the limit of detection for MCA was 25 ng/mL. Plasma samples from human subjects exposed to 100 ppm TCE for 4 h contained TCA at concentrations as high as 10 micrograms/mL. DCA concentrations were less than 5 ng/mL, and MCA was not detected (less than 25 ng/mL).

Conversion of trichloroacetic acid to dichloroacetic acid in biological samples.

Trichloroethylene (TCE) has been identified as an environmental contaminant in groundwater. Trichloroacetic acid (TCA), dichloroacetic acid (DCA), chloral hydrate (CH), trichloroethanol (TCOH), and trichloroethanol glucuronide (TCOG) have been identified as metabolites of TCE. Studies have shown that TCA and DCA are toxicologically significant metabolites that can induce liver tumors in B6C3F1 mice. Methods for the analysis of these metabolites are important for conducting pharmacokinetic studies. In this study, TCA and DCA were derivatized to their methyl esters by dimethyl sulfate under acidic conditions and analyzed by gas chromatography with electron capture detection. In developing a method for esterifying TCA and DCA, the conversion of TCA to DCA was observed in freshly drawn blood upon the addition of acid. After blood was drawn from the animals, the amount of TCA converted to DCA by the addition of acid decreased with time. This conversion could be prevented by freezing blood samples overnight prior to the addition of acid. Further experiments demonstrated that this activity could be restored by the addition of dithionite to inactivate blood samples or the addition of dithionite to methemoglobin prior to the addition of acid. The results reported here show that reduced hemoglobin may be involved in the acid-catalyzed conversion of TCA to DCA.

Immunological studies on dermatophytes. I. Serological reactivities of neutral polysaccharides with rabbit antiserum to Microsporum quinckeanum.

Antiserum produced in the rabbit to autoclaved mycelial suspensions of Microsporum quinckeanum reacted with three neutral polysaccharides isolated from each of five species of dermatophytes, M. quinckeanum, Trichophyton granulosum, T. interdigitale, T. rubrum, and T. schönleinii. The serological reactivities of these polysaccharides, grouped as galactomannans I, galactomannans II, and glucans, were compared by qualitative precipitation analyses in gel and quantitative complement-fixation analyses. Significant differences were found among the glucans and galactomannans II but not among the galactomannans I of these species.

Immunological studies on dermatophytes. II.

The contribution of terminal galactofuranose residues to the antigenic specificity and to cross-reactivity of galactomannans isolated from five species of dermatophytes, Microsporum quinckeanum, Trichophyton granulosum, T. interdigitale, T. rubrum, and T. schoenleinii, was investigated. Galactofuranose units were removed from galactomannans I and galactomannans II by mild acid hydrolysis. The resulting mannans were tested for serological reactivity with rabbit antiserum to M. quinckeanum by qualitative precipitation in gel and by quantitative complement-fixation analyses. Our results showed that, with this antiserum, the galactofuranose residues contributed greatly to the antigenic specificity and to cross-reactivity of the galactomannans II, but these residues were less significant as antigenic determinants in the galactomannans I. We have shown that mannans isolated from three Candida species reacted with rabbit antiserum to M. quinckeanum.

Immunological studies on dermatophytes. IV. Chemical structures and serological reactivities of polysaccharides from Microsporum praecox, Trichophyton ferrugineum, Trichophyton sabouraudii, and Trichophyton tonsurans.

The chemical structures and serological specificities of polysaccharides isolated from four species of dermatophytes, Microsporum praecox, Trichophyton ferrugineum, T. sabouraudii, and T. tonsurans, were investigated. Each of these species yielded a mixture of crude polysaccharides which could be separated into three water-soluble, neutral polysaccharides free of nitrogen. These were grouped as galactomannan I, galactomannan II, and glucan. The galactomannans I were quite similar in chemical structure. When measured by complement fixation, their serological cross-reactivities were similar with rabbit antisera to each of these species except T. sabouraudii. The differences in their relative reactivities with this antiserum could be correlated with differences in structure and specificity of this antiserum for galactofuranose end groups. The galactomannans II differed both in chemical structure and in their serological reactivities with antisera to each of these species. The galactomannan II from T. ferrugineum differed most in chemical structure and was the least reactive serologically. The glucans also differed in both structure and serological reactivities.

Immunological studies on dermatophytes. 3. Further analyses of the reactivities of neutral polysaccharides with rabbit antisera to Microsporum quinckeanum, Trichophyton schoenleinii, Trichophyton rubrum, Trichophyton interdigitale, and Trichophyton granulosum.

The serological reactivities of polysaccharides isolated from five species of dermatophytes, Microsporum quinckeanum, Trichophyton granulosum, T. interdigitale, T. rubrum, and T. schoenleinii, with rabbit antisera to these species were studied qualitatively by precipitation in gel and quantitatively by complement-fixation analyses. Significant differences in the serological reactivities of the galactomannans I were detected with antisera to T. schoenleinii and T. interdigitale. The differences appeared to be related to the specificity of these antisera for the galactofuranose residues in the polysaccharides. Antisera to M. quinckeanum, T. granulosum, or T. rubrum did not detect differences between the galactomannans I. The serological reactivities of the galactomannans II were different with each of the five antisera. The reactivities of the glucans could be correlated with the amount of alpha 1 --> 6 linked glucopyranose residues when antisera to T. schoenleinii and M. quinckeanum were used.

Cerebrovascular disease in Utah, 1968--1971.

Utah mortality rates for cerebrovascular disease (ICD numbers 430--438) are 13% below U.S. rates. About 70% of Utahns are members of the Church of Jesus Christ of Latter-day Saints, commonly called Mormons of LDS, which proscribes use of tobacco and alcohol. Other studies on this group have found significantly lower occurrence of many cancers and ischemic heart disease. We tested the hypothesis that Utah's lower cerebrovascular disease (CBVD) mortality was contributed by the LDS population. We classified by religion all CBVD deaths (2,521) (except subarachnoid hemorrhage and cerebral embolism) occurring in the state in 1968--1971. No significant difference was found between LDS and non-LDS, but both groups had mortality rates below U.S. expectation. Although recent studies have reported smoking to be a risk factor for CBVD, we found no consistent difference between the LDS and non-LDS, even in the younger age groups. The results do not support the hypothesis that tobacco is an important etiologic agent in CBVD mortality.

Free radical damage to cultured porcine aortic endothelial cells and lung fibroblasts: modulation by culture conditions.

Culture conditions modulating cell damage from xanthine plus xanthine oxidase-derived partially reduced oxygen species were studied. Porcine thoracic aorta endothelial cells and porcine lung fibroblasts were maintained in monolayer culture. Cells were prelabeled with 51Cr before xanthine plus xanthine oxidase exposure. Endothelial cells showed 30 to 100% more lysis than fibroblasts and thus seemed more sensitive to this oxidant stress. The effect of cell culture age, as indicated by population doubling level (PDL), was examined. Response of low PDL endothelial cells and fibroblasts subjected to oxidant stress was compared with the response of PDL 15 cells. Both low PDL endothelial cells and fibroblasts responded differently to the lytic effect of xanthine oxidase-derived free radicals than did higher PDL cells. Specific activities of the antioxidant enzymes catalase, manganese superoxide dismutase, copper-zinc superoxide dismutase, glutathione peroxidase, and glucose-6-phosphate dehydrogenase were measured in both low and high PDL fibroblasts and endothelial cells. Antioxidant enzyme specific activities could only partially explain the differences in response to oxidant stress between fibroblasts and endothelial cells and between low and high PDL cells. Cell culture medium composition modulated the rate of production, and relative proportions of xanthine plus xanthine oxidase-derived partially reduced species of oxygen, i.e. superoxide, hydrogen peroxide, and hydroxyl radical. Serum content of medium was important in modulating free radical generation; superoxide production rates decreased 32%, H2O2 became undetectable, and hydroxyl radical generation decreased 54% in the presence of 10% serum. The medium protein and iron content also modulated free radical generation. The data suggest that cell culture media constituents, cell type, and cell culture age greatly affect in vitro response of cells subjected to oxidant stress.