Masahiko Endo

Werner's syndrome as "hyaluronuria".

Abstract Although excretion of acid glycosaminoglycans into urine of five patients with Werners syndrome were within normal limits, the quantity of hyaluronic acid increased in this disease. To this novel finding, a term “Hyaluronuria” was proposed.

Chemistry of organosilicon compounds: CC. Allyl coupling reactions of allylsilanes and allylstannanes with allylic halides, ethers and acetates promoted by a Lewis acid☆

Abstract Facile allyl couplings between allylsilanes or allylstannanes and various allylic substrates such as halides, ethers or acetates promoted by a Lewis acid, leading to 1,5-dienes with regiospecific transposition in the allylic part of allylmetallics have been observed. The combination of allylstannanes with allylic acetates promoted by bis(diethylaluminum) sulfate gives the most satisfactory results. The factors influencing the regioselectivity with respect to allylic substrates are discussed.

Hormonal effect on glycoproteins and glycosaminoglycans in rabbit uteri.

Abstract The effect of the female hormones on glycoproteins and glycosaminoglycans in uteri has been studied. The uteri taken from the ovariectomized rabbits treated with estrogen, estrogen plus progesterone, and sham administration (control) were incubated in vitro with [U- 14 C]glucose. Subsequently, the tissues were digested extensively with pronase, yielding crude glycan fractions. The amount and radioactivity of the crude glycan fraction increased by the treatment with estrogen, but reduced to certain level with progesterone. Separation of glycoproteins and glycosaminoglycans was achieved by stepwise elution from Dowex 1 (X2, chloride form) with increasing concentration of NaCl. The yield and radioactivity, together with the results of chemical, enzymatic, and electrophoretic studies on the resulting fractions indicated that the metabolism of a slightly acidic glycoprotein, hyaluronic acid, sulfated glycoproteins, low-sulf ated chondroitin sulfate, heparan sulfate, chondroitin 4-sulfate, and dermatan sulf ate were stimulated remarkably with estrogen, but the stimulation was restored to certain level with progesterone. The degree of the change with these hormones was, however, found to be different from each other. It was noticed that sulfated glycoproteins were the most sensitive to the hormones. On the other hand, the estrogenic stimulation of the metabolism of a neutral glycoprotein and oversulfated chondroitin sulfates was not restored with progesterone.

Glycosaminoglycans and acidic glycoproteins in rabbit uterus under estrogenic conditions

Uterine slices obtained from the estrogen-treated rabbits were digested with pronase. Glycosaminoglycans and acidic glycopeptides were then isolated by Dowex 1 column chromatography and preparative electrophoresis on cellulose acetate membrane (Separax), in succession. Each subfraction thus obtained was identified by the mobility on Separax electrophoresis and the digestibility with mucopolysaccharidases (Streptomyces hyaluronidase, testicular hyaluronidase, chondroitinase AC, chondroitinase ABC and heparinase). The resulting data showed that each complex saccharide (hyaluronic acid, heparan sulfate, chondroitin sulfate A, chondroitin sulfate C, dermatan sulfate, sulfated glycopeptide and sialoglycopeptide) was separated into 2-5 fractions, indicating charge and/or molecular heterogeneity of each complex saccharide.

Histochemical localization of estrogen induced sulfated glycoprotein in rabbit uterus.

SummaryHistochemical localization of the estrogen-induced sulfated glycoproteins was made in the estrogen-treated rabbit uterus. Biochemical studies by a group of Endoet al. affirmed these particular glycoproteins were PAS-positive and metachromatic as stained with TB. No sign of digestion, however, has been detected in a series of tests with α-amylase, testicular hyaluronidase, streptomyces hyaluronidase, chondroitinase AC and chondroitinase ABC, and heparinase.The apical portions of the epithelial and glandular cells, obviously expanded by the estrogen treatment, display strong β-metachromasia with TB (pH 4.0), saliva-resistant PAS-positive reactions, and also alcianophilia with AB (pH 2.5). These reactions are not reduced after the treatment with the enzymes above-mentioned. Meanwhile, in the stromal matrix, the same enzymes give an influence to diminish the reactions to various extent. Our results suggest that the estrogen-induced sulfated glycoprotein is definitely localized in the apical portions of the epithelial and glandular cells. The identity is emphasized between the substance that is elucidated in the histochemical sections and the sulfated glycoproteins that have been specified solely by means of biochemical assays.

A method for quantitative recovery of acidic glycosaminoglycans after electrophoresis on cellulose acetate strip.

Abstract Acidic glycosaminoglycans after electrophoretic separation on a strip of cellulose acetate membrane were stained with toluidine blue or azure I. The basic dye was removed with 0.2 m NH4OH in 80% ethyl alcohol. Then elution with water was effected for quantitative recovery of glycosaminoglycans.

Hormonal effect on sugar-nucleotides in rabbit uteri☆☆☆

Abstract GDP-Man, UDP-Glc and UDP-Gal, UDP-GlcNAc with UDP-GalNAc and UDP-GlcUA were isolated from uteri after in vitro incubation with glucose-U-14C of estrogen-treated (ES), estrogen-progesterone-treated (PG) and nontreated (C) ovariectomized rabbits. The amounts of the sugar-nucleotides increased 1.4–1.9 times with estrogen; except for UDP-GlcNAc, the control levels were restored with progesterone. The relative contents per unit weight of dry tissue of the nucleotides were shown to be lower than the control levels due to enhanced tissue growth. The nucleotides were labeled with 14C 5–7 times more in ES and 2–4 times more in PG compared with the controls and the specific activity was 4–4.5 times more in ES and 2–3.7 times more in PG. Of the nucleotides, UDP-Glc plus UDP-Gal and UDP-GlcUA showed identical behavior differing from GDP-Man and UDP-GlcNAc. The level of UDP-GlcNAc was not reversed with progesterone, and GDP-Man changed differently from the other nucleotides. The data suggest that estrogen activates the enzymes involved in the metabolism of these sugar-nucleotides similarly, but that progesterone suppresses the estrogenic effect on most of the nucleotides.

Biochemical Characterization of Hyaluronic Acid from a Case of Benign, Localized, Pleural Mesothelioma

The tissue of a benign, localized, pleural mesothelioma was digested with protease, and the crude polysaccharide was fractionated by a column of Dowex-1 (Cl-form). The eluate from the column was electrophoresed and incubated with various mucopolysaccharide lysases. Based on the results of column chromatography, electrophoresis, and enzymatic digestion, it was found that hyaluronic acid was the major constituent of the glycosaminoglycans in pleural mesothelioma. The hyaluronic acid from pleural mesothelioma seemed to be identical in structure with that from human umbilical cord; however, the hyaluronic acid from mesothelioma was eluted before that from human umbilical cord when fractionated by the column of Sepharose 4B, suggesting a differnence in molecular size between the two. Also, evidence was obtained for the presence of dermatan sulfate and heparan sulfate.