Molecular genetics and pathophysiology of 17 beta-hydroxysteroid dehydrogenase 3 deficiency.

Autosomal recessive mutations in the 17 beta-hydroxysteroid dehydrogenase 3 gene impair the formation of testosterone in the fetal testis and give rise to genetic males with female external genitalia. Such individuals are usually raised as females, but virilize at the time of expected puberty as the result of increases in serum testosterone. Here we describe mutations in 12 additional subjects/families with this disorder. The 14 mutations characterized to date include 10 missense mutations, 3 splice junction abnormalities, and 1 small deletion that results in a frame shift. Three of these mutations have occurred in more than 1 family. Complementary DNAs incorporating 9 of the 10 missense mutations have been constructed and expressed in reporter cells; 8 of the 9 missense mutations cause almost complete loss of enzymatic activity. In 2 subjects with loss of function, missense mutations testosterone levels in testicular venous blood were very low. Considered together, these findings strongly suggest that the common mechanism for testosterone formation in postpubertal subjects with this disorder is the conversion of circulating androstenedione to testosterone by one or more of the unaffected 17 beta-hydroxysteroid dehydrogenase isoenzymes.

Male pseudohermaphroditism caused by mutations of testicular 17 beta-hydroxysteroid dehydrogenase 3.

Defects in the conversion of androstenedione to testosterone in the fetal testes by the enzyme 17 beta-hydroxysteroid dehydrogenase (17 beta-HSD) give rise to genetic males with female external genitalia. We have used expression cloning to isolate cDNAs encoding a microsomal 17 beta-HSD type 3 isozyme that shares 23% sequence identity with other 17 beta-HSD enzymes, uses NADPh as a cofactor, and is expressed predominantly in the testes. The 17 beta HSD3 gene on chromosome 9q22 contains 11 exons. Four substitution and two splice junction mutations were identified in the 17 beta HSD3 genes of five unrelated male pseudohermaphrodites. The substitution mutations severely compromised the activity of the 17 beta-HSD type 3 isozyme.

Allergenic activity of short ragweed pollen (Ambrosia elatior) from different years and/or suppliers: criteria for the selection of an in-house allergen reference preparation.

We have investigated dialyzed extracts (A-F) of short ragweed pollens (Ambrosia elatior) from six consecutive production years and four different companies by protein nitrogen units, high performance liquid chromatography, isoelectric focusing, SDS-PAGE, RAST inhibition, crossed radioimmunoelectrophoresis, and antigen E contents. Extract A showed lowest yields in all quantitative assays. Isoelectric focusing, SDS-PAGE, crossed immunoelectrophoresis, and crossed radioimmunoelectrophoresis showed comparable patterns for all extracts except for extract A. Apparently, the activity of short ragweed pollens depends on the year of production and/or the skill of the producer. These findings are in contrast to similar examinations of other pollen species.

High-performance liquid chromatographic molecular weight determination of allergen extracts. Examination of the influence of the column material on allergenic activity and allergen patterns.

To investigate whether the column material employed in size-exclusion high-performance liquid chromatography (HPLC) influences the allergenic activity and antigen/allergen patterns of allergen extracts, the molecular weights of a mite and a birch pollen allergen extract were determined using a Bio-Sil TSK 250 column with a guard column. The allergenic activities were measured by RAST inhibition and the antigen/allergen patterns were determined by crossed (radio)immunoelectrophoresis. The original extracts and the corresponding eluates after HPLC runs showed the same allergenic activity and the same number of antigen/allergen precipitation lines. Only slight differences in the peak heights of some precipitates were observed.

Quality of timothy pollen (Phleum pratense) from different pollen seasons and different suppliers.

We investigated seven batches of timothy (Phleum pratense) pollen from four different pollen seasons and three different manufacturers in the USA and Europe by several biochemical and immunochemical methods. By measuring the contents of hexoses, proteins and the total allergenic activity using RAST inhibition we could not find any significant differences in quality between pollen of different seasons, different geographical origin and different manufacturers. There were only slight differences in the staining of some protein bands and in peak height of some antigens in isoelectric focusing (IEF), SDS-PAGE and crossed immunoelectrophoresis. These differences in IEF and SDS-PAGE patterns seem to be specific for the pollen samples from the USA and Europe, respectively. We also observed minor differences in IEF immunoprint patterns, mainly for some basic proteins. The allergen patterns in crossed radioimmunoelectrophoresis were very similar, with only minor differences in concentration of the basic allergens. It appears that timothy pollen from different years can be produced in different geographical areas by different manufacturers with fairly constant allergenic activity.

Comparison of classical Lowry, modified Lowry and a dye-binding assay for the estimation of protein in allergen extracts and influence of different parameters on the modified Lowry assay.

Protein values of dialysed allergen extracts determined by Lowry, modified Lowry (trichloroacetic acid precipitation of the proteins) and dye-binding assay were compared. The influence of different parameters on the modified Lowry was examined. The reproducibility of the modified Lowry was checked with three independent measurements. For the examination of recovery a constant amount of 6-grass pollen allergen proteins was added to the samples of the standardized human serum albumin prepared for the calibration curve. The samples were measured by modified Lowry. The mean of the ratio between the protein values of the dialysed allergen extracts obtained by modified Lowry and those obtained by classical Lowry was 3.59 (coefficient of variation Cv = 45%). The mean of the ratio between the protein values of the allergen extracts obtained by modified Lowry and dye-binding assay was 1:0.71 (Cv = 31%). Phenol interfered with the modified Lowry. Phenolic allergen extracts showed higher "protein values" than non-phenolic allergen extracts. This influence could be reduced by a second precipitation of the dissolved precipitate. The precipitation of non-phenolic dialysed aqueous allergen extracts was complete after the first trichloroacetic acid precipitation. By incubating samples with the Folin-Ciocalteu's reagent at 55 degrees C in a waterbath, the time necessary for developing the colour could be reduced from 45 min to 5 min. Protein measurements by modified Lowry of a 6-grass pollen allergen extract in three different laboratories showed good reproducibility. For these extract 785 micrograms protein/ml (Cv = 4%) could be measured.(ABSTRACT TRUNCATED AT 250 WORDS)

Leukocyte histamine release and humoral changes during oral and subcutaneous hyposensitization of grass pollen allergic children.

Thirty-one children with seasonal hay fever, sensitive to rye grass by skin test, RAST, and leukocyte histamine release, received preseasonal hyposensitization with a rye grass pollen extract either subcutaneously (cumulative dose 40,000 PNU) or orally (cumulative dose 400,000 PNU). While there was a significant (p less than 0.005) increase in specific serum IgG-antibodies and a decrease (p less than 0.005) of leukocyte sensitivity in the subcutaneously treated group, oral treatment with an aqueous pollen extract could not be demonstrated to be immunologically effective.

Comparison of three variations of the radioallergosorbent test-inhibition assay for measuring allergenic activity of grass pollen extracts.

Three variations of the radioallergosorbent (RAST)-inhibition assay (RI) have been compared for measuring the allergenic activity of grass pollen extracts. The main difference between the three assays consisted in the solid supports to which the allergens were coupled. These supports were paper disks, microcrystalline cellulose, and Matrex. It could be demonstrated that all three variations of RI yielded almost identical results as expressed by their F value (F0). Correlations between the three methods were highly significant (P less than 0.001). Three independent valid assays showed an excellent reproducibility of each test system (P less than 0.01). Three different preparations of each of the supports yielded highly reproducible F0 (0.76 +/- 3% SD). Evaluation of the 50% inhibition values achieved with the different assays based on protein showed differences between the three supports. Provided the same allergen extract is coupled to the different solid supports and the results are related to a reference preparation, different laboratories using different forms of RI will be able to compare their results.

[Experiences in implementing a dynamically oriented group therapy in myocardial infarct patients and their spouses]. / Erfahrungen bei der Durchführung dynamisch orientierter Gruppengespräche bei Herzinfarktpatienten und ihren Ehepartnern.

It is reported on the performance of dynamically orientated group talks with 5 patients with myocardial infarction and their wives within the psychic rehabilitation in phase II. 30 group meetings of a duration of 2 hours each were performed in the outpatient department in a weekly rhythm. At the beginning the implementation of a relaxation method, the autogenic training, for furthering the motivation of the patient for an independent elaboration of the problem in the group as well as for reducing the inability to relaxation often existing in patients with myocardial infarction proved to be favourable. Before the beginning and after the end of the group talks psychodiagnostic tests were carried out. Though scarcely significant differences were the results in the test values, the course of the group talks is to be estimated as successful. The aim reflexion and analysis especially of the interactions between the partners could be obtained after a relatively long initial phase. Conclusions were made for further work in this field.