Distribution of sulphate and iduronic acid residues in heparin and heparan sulphate

1. A method was developed for determination of the uronic acid composition of heparin-like glycosaminoglycans. Polymers or oligosaccharides are degraded to monosaccharides by a combination of acid hydrolysis and deamination with HNO₂. The resulting uronic acid monosaccharides (accounting for about 70% of the uronic acid contents of the starting materials) are isolated and converted into the corresponding aldono-1,4-lactones, which are separated by g.l.c. The calculated ratios of glucuronic acid/iduronic acid are reproducible within 5%. 2. Samples of heparin from pig intestinal mucosa (molar ratio of sulphate/disaccharide unit, 2.40) and heparan sulphate from human aorta (sulphate/disaccharide ratio, 0.46) were subjected to uronic acid analysis. l-Iduronic acid constituted 77% and 19% respectively of the total uronic acid contents. 3. The correlation between the contents of sulphate and iduronic acid indicated by this finding also applied to the fractionated deamination products of the two polymers. The sulphated fragments varied in size from disaccharide to octasaccharide (or larger) and showed sulphate/disaccharide molar ratios in the range of 0.05–2.0. The proportion of iduronic acid increased with increasing ester sulphate contents of the oligosaccharides. 4. Previous studies on the biosynthesis of heparin in a cell-free system have shown that l-iduronic acid residues are formed by C-5 epimerization of d-glucuronic acid units at the polymer level; the process requires concomitant sulphation of the polymer. The results obtained in the present structural study conform to these findings, and suggest further that similar mechanisms may operate in the biosynthesis of heparan sulphate. The epimerization reaction appears to be linked to the sulphation of hydroxyl groups but does not seem to require sulphation of the target uronic acid residues. The significance of sulphamino groups in relation to the formation of iduronic acid is unknown.     

The Ultrastructure of a Cyanophage Attack on Anabaena variabilis

Cyanophages multiplying on the nitrogen fixing blue-green alga Anabaena variabilis Kütz. were revealed by electron microscopy. Severe ultrastructural changes have been observed in the vegetative cells, whereas the heterocysts appeared resistant to the cyanophage. A lytic cycle was observed from adsorption to lysis.     

Cyclical changes in the fine structure of bovine endometrial gland cells

Summary Cyclical changes in bovine endometrial gland cells were investigated in six heifers, three at estrus and three at day twelve of the estrous cycle in the luteal phase. The epithelium is generally low at estrus but high in the luteal phase. There are ciliated and non-ciliated cells. The ciliated cells are fewer and lighter and show inconspicuous cyclical changes.The secretory cells show more prominent changes. At estrus, their free border is flat with short microvilli. The conspicuous rough-surfaced endoplasmic reticulum may synthesize protein for later secretion. The Golgi complex seems inactive. The high number of cytosegresomes and dense bodies might express cell regression caused by endocrine changes.In the luteal phase, the cells are lighter with long microvilli. The Golgi apparatus shows vacuoles and immature secretory droplets. Secretory vacuoles with light contents occur in the apical cytoplasm. Some of them appear to discharge their contents into the lumen. This is interpreted as evidence of merocrine secretion. Accumulations of tubular, smooth-surfaced endoplasmic reticulum, masses of glycogen granules, and several fat droplets are present.Some lymphocytes and degranulated granulocytes are seen near the basement membrane, more frequently at estrus.Financial support for this study was received from Anslaget för främjande av medicinsk forskning vid Veterinärhögskolan.The authors express their gratitude to Prof. A. Bane and Dr. J.-E. E. Ringmar, Department of Obstetrics and Gynecology, for their help with the selection and clinical control of the animals and for keeping them in good condition.Post doctoral fellow, No. 43-KO-52 (1968) from the Educational Ministry of Japan.     

Ernährungsbiologie einer rhipidoglossen Kiemenschnecke

Zusammenfassung Die Arbeitsweise und die Leistungen der Rhipidoglossenradula von Theodoxus fluviatilis werden durch die kräftigen 4. Zwischenzähne und die Randbürstenzähne bestimmt. Die 4. Zwischenzähne lockern im wesentlichen die dem Fre\grund anhaftenden oder aufliegenden ein- bis wenigzelligen Algen (Diatomeen, chlorococcale und konjugate Grünalgen), die Randzähne fegen das gelockerte Nahrungsgut quantitativ zusammen. Fädige Grünalgen (z. B. Cladophorales) und Gewebeteile höherer Pflanzen werden nicht abgebissen bzw. abgeschabt.Die Diatomeen werden nur verdaut, wenn die Kieselschalen bereits bei der Nahrungsaufnahme mechanisch zerkleinert werden. Diese Zerkleinerung erfolgt allein auf einem Substrat mit rauher Oberfläche; sie wird durch die während des Bisses zwischen 4. Zwischenzähnen und Substrat auftretenden Reibungskräfte erzielt. Theodoxus wurde bei refiner Diatomeenernährung über mehrere Generationen gezüchtet. Tierisches Eiwei\ ist als Zusatzkost nicht erforderlich. Mit besonderen Hilfsma\nahmen kann Theodoxus im Laboratorium auch mit Cyanophyceen oder besonders mit Flagellaten (Chlamydomonas), die den Schnecken an den im Freiland besiedelten Standorten nicht zur Verfügung stehen, ernährt werden.Sämtliche Grünalgen mit stärkerer Cellulosewandung (Chlorococcales, Conjugatae) sind unverdaulich. Die Unverdaulichlichkeit beruht vermutlich auf einem Fehlen von Cellulasen im Verdauungstraktus. Die Zellmembranen und extrazellularen Scheiden der Cyanophyceen, die aus Hemicellulosen und Pektinen aufgebaut sind, werden im Magen aufgelöst. Theodoxus ist ein reiner Diatomeenfresser. Die ökologische Einnischung in die litorale Steinregion ist vorwiegend ernährungs-physiologisch begründet und erklärt das Vorkommen in Flie\gewassern und an Brandungsufern stehender Sü\gewässer sowie der Ostsee.     

Morphologische und experimentelle untersuchungen über die variabilität per farbmuster auf der schale von theodoxus fluviatilis L.

Ohne ZusammenfassungMit Unterstützung der Deutschen Forschungsgemeineshaft.     

Purification of thymocyte growth peptide (TGP) from sheep thymus. Relationship to FTS/thymulin

Thymocyte growth peptide (TGP) initiates DNA synthesis in immature thymocytes and has previously been characterized as an acidic peptide isolated from calf thymus. We now report the isolation of TGP from sheep thymus and show it to be a nonapeptide with a large N-terminal blocking moiety characterized by high UV absorbance. The amino acid composition is identical to FTS, consisting of 2 Gly, 2 Ser, 2 Glx, 1 Ala, 1 Lys, 1 Asx. In contrast to FTS, TGP is acidic with an apparent isoelectric point of 4.2 and a high UV absorbance at 270–280 nm. Reverse phase chromatography of TGP at an acidic pH results in a change of the molecule and the appearance of two new compounds TGP-A and TGP-B, both with less than 50% of the original TGP activity. Full activity could be restored by the addition of ZnCl₂ to TGP-A. Both TGP-A and B have some amino acid composition and high UV absorbance as native TGP. We propose that TGP consists of a non-peptide moiety bound to the N-terminal of the nonapeptide Glu-Ala-Lys-Ser-Gln-Gly-Gly-Ser-Asn and that the active molecule is stabilized by Zn²⁺.