Willard R. Starnes

Extraction and characterization of a thyrotropic material from the human placenta

A material with thyrotropic activity was extracted from fresh human placentas. After chromatography of the extract on carboxymethyl cellulose, thyroid-stimulating activity ranged from 0.12 to 1.06 mU of thyroid-stimulating hormone (TSH) per mg of protein in bioassays of eight preparations. The amount of TSH per placenta varied from 113 to 2200 mU and approximated the content of the pituitary gland. Additional purification by gel filtration on Sephadex G-100 gave a maximum activity of 8 mU/mg. The most active portion was eluted in the same position as (125)I-labeled bovine or human TSH, a fact suggesting that the molecular size of this thyrotropic substance was similar to that of pituitary TSH. Another placental fraction with weaker activity was eluted earlier indicating that the placental material was heterogeneous. In the McKenzie mouse bioassy, the response of the placental thyrotropin paralleled that of the beef TSH standard. There was no long-acting thyroid stimulator effect. Antibodies to both human and bovine pituitary TSH neutralized the biologic activity of the placental TSH. Placental thyrotropin cross-reacted very weakly in a sensitive radioimmunoassay for human pituitary TSH; it cross-reacted completely in a radioimmunoassay for bovine pituitary TSH, and this assay was used for following the purification. The role of this thyrotropic material as a possible cause of thyroid hypertrophy and hyperfunction in pregnancy and in patients with trophoblastic tumors remains to be investigated.

Quantitative chromatographic estimation of urinary C19 and C21 steroids

An extensive methodological study has been undertaken with a view to developing a reproducible procedure which would permit a quantitative estimation of a broad spectrum of C19 and C21 steroids, with minimum chromatography involved. The method described in this paper utilizes one partition column chromatography and seven consecutive paper-chromatographic systems for the simultaneous isolation and the quantitative estimation of 30 urinary steroid metabolites. The technique is relatively simple but reliable and reproducible. Its feasibility is not beyond the capacity of a well-equipped clinical research laboratory, and its sensitivity permits detection and quantitation of compounds normally present in such minute quantities that their isolation hitherto required use of radioisotope techniques.

An improved method for the determination of urinary myoinositol by gas-liquid chromatography

Abstract Previous methods for the determination of urinary myoinositol by gas-liquid chromatography of its trimethylsilyl derivatives have required preincubation of urine samples with urease. Since we have observed that commercially available urease preparations contain significant amounts of myoinositol, a method for the separation of urinary myoinositol and urea has been developed which avoids the use of urease.

Purification of Bovine Thyroid-Stimulating Hormone by Solubility Fractionation with Ammonium Sulfate∗:

Summary Solubility fractionation of relatively crude TSH with ammonium sulfate was investigated as a means of purifying the hormone. The results obtained with three variations in solubility fractionation techniques using decreasing concentrations of ammonium sulfate demonstrated that each method yielded a 5- to 7-fold purification of the 0.49 to 0.84 U/mg starting material. Total recovery of TSH was 90-100%. Recoveries in the high potency fractions were 60-80% when TSH was extracted with ammonium sulfate or recovered by fractional precipitation and 25-35% when batch eluted from diethylaminoethyl (DEAE) cellulose. Solubility fractionation appears to be a simple and rapid method for the purification of TSH to 4-6 U/mg which may be adequate for most purposes. Highly purified TSH may then be obtained by standard column chromatography with DEAE-cellulose.

Isolation and identification of 3α,21-dihydroxy-5α-pregnane-11,20-dione from urine

An unknown steroid encountered in patterning C21 corticosteroids in urine has been isolated and identified as 3α,21-dihydroxy-5α-pregnane-11,20-dione (allo-THA) and a reaction scheme for its synthesis from 3α,11β,21-trihydroxy-5α-pregnane-20-one (allo-THB) has been presented. Studies on the mobility of 3α,21-dihydroxy-5α-pregnane-11,20-dione in several paper chromatographic systems have also been presented.

Non-specificity of biotin activity for Leuconostoc.

Summary It has been found that L. dextranicum elai is unique in that the dl-forms of oxybiotin and desthiobiotin are equal in activity to d-biotin in allowing maximum growth in an aspartate-free fructose medium. Since no increased lag period was observed the results suggest that these biotin analogs are utilized directly and without prior conversion to the vitamin. It was also found that γ-(3,4-ureylenecyclohexyl) butyric acid was not a blocking agent for this organism, but instead possessed biotin activity.

Carbohydrate Content of Euglobulins of Normal and Rheumatoid Sera

Summary 1. Euglobulins were prepared by 3 different methods from“latex positive” pooled serum. 2. Total protein bound hexoses, glucosamine, and sialic acid were measured on each euglobulin preparation. All were somewhat similar with the exception of the euglobulin prepared with hydrochloric acid, which had a higher per cent of hexose to protein. All showed marked increase in content when compared with euglobulins isolated from a pooled “latex negative” normal serum. 3. Paper electrophoretic examination reveals proteins that migrate as gamma globulins and are highly PAS positive. 4. A uronic acid-containing mucopolysaccharide was isolated from the cold agglutinating euglobulin.