Otto Wolfgang Thiele

The Effect of Magnesium on the Hemolysis of Calf and Adult Cattle Erythrocytes

Zusammenfassung1.Bei der hypotonen Hämolyse wird ein Stroma-Präparat mit minimalem Hämoglobin-Gehalt erzielt, wenn eine Pufferkonzentration von 20 imOsm bei pH 7,4 verwendet wird. Menschliche und Kälber-Erythrozyten retinieren mehr Hämoglobin als Erythrozyten erwachsener Rinder, speziell bei hoheren Puffer-Konzentrationen.2.Während der hypotonischen Hämolyse geht ein Teil der Membran-bestandteile der Erythrozyten erwachsener Rinder verloren; bei Kälber-Erythrozyten tritt ein solcher Verlust kaum ein.3.Durch Zusatz von Magnesium-Ionen (3 mM MgCl2) während der hypotonischen Hämolyse läßt sich der Verlust von Membranbestandteilen der Erythrozyten vermeiden; dabei erfolgt allerdings eine geringfügige Erhöhung der Hämoglobin-Retention bei Erythrozyten erwachsener Rinder. Bei jungen Kälbern ist die Hämoglobin-Retention unter diesen Bedingungen wesentlich größer. Sie nimmt mit zunehmendem Alter der Kälber ab.4.Der Magnesium-Gehalt der Erythrozyten erwachsener Rinder ist signifikant geringer (1,56μVal/ml gepackter Zellen) als der von Kälbern (2,47μVal/ml).5.Durch Inkubation von Erythrozyten erwachsener Rinder mit Magnesium-Ionen und Adenosin läßt sich der zelluläre Magnesium-Gehalt beträchtlich steigern, so daß er annähernd die Werte von Kälber-Erythrozyten erreicht.6.Das Ausmaß der Hämoglobin-Retention während der hypotonen Hämolyse von Erythrozyten erwachsener Rinder, die mit Magnesium-Ionen und Adenosin inkubiert worden waren, erreicht ähnlich hohe Werte wie bei unbehandelten Kälber-Erythrozyten. Dies zeigt, daß die zelluläre Magnesium-Konzentration der Erythrozyten wesentlich ist für das Ausmaß der Retention von Hämoglobin bei der hypotonen Hämolyse.Summary1.In hypotonic hemolysis, the hemoglobin removal is optimum at pH 7.4 and 20 imOsm buffer concentration. Calf erythrocytes retain more hemoglobin than adult cattle erythrocytes, particularly at higher buffer concentrations.2.During hypotonic hemolysis, adult cattle erythrocytes do not quantitatively retain their membrane constituents. The release of membrane constituents is much less pronounced in calf erythrocytes.3.Addition of magnesium ions (3 mM MgCl2) during hypotonic hemolysis prevents the disruption of adult cattle erythrocyte membranes; this is accompanied by a slightly increased hemoglobin retention. In young calves, the hemoglobin retention is much more pronounced on hemolysis in the presence of magnesium ions. It declines with increasing age of calf.4.The magnesium content of adult cattle erythrocytes is lower (1.56μequ/ml packed cells) than that of young calves (2.47μequ/ml packed cells).5.Storage of adult cattle erythrocytes decreases their magnesium content. Subsequent incubation with magnesium ions and adenosine increases the magnesium content considerably, thus approaching the magnesium level of calf erythrocytes.6.The hemoglobin retention of adult cattle erythrocytes incubated with magnesium ions and adenosine becomes similar to that of untreated calf erythrocytes indicating that the cellular magnesium level is essential for the extent of release of hemoglobin by hemolysis.

Lipid patterns of various hydrogen oxidizing bacterial species

Abstract Strains of hydrogen-oxidizing bacteria, Pseudomonas facilis, Nocardia opaca 1 b, Paracoccus denitrificans and Corynebacterium autotrophicum 7 C , were grown both under chemolithoautotrophic (referred to as autotrophic) and under chemoorganoheterotrophic (referred to as heterotrophic) conditions and harvested during the phase of stationary growth. The lipid patterns of all strains were studied by quantitative chemical analyses, UV spectra, TLC and GLC of fatty acid methyl esters. In general, there were no essential qualitative differences between lipid patterns of autotrophically and those of heterotrophically grown bacteria of the same strain. The conditions of growth apparently influence only the quantitative composition of lipids. In particular, Nocardia opaca is comparatively rich in triglycerides. Ubiquinones were found in Pseudomonas facilis and in Corynebacterium autotrophicum . The latter finding is quite unusual in a Corynebacterium . Carotenoid glycosides are probably present in Corynebacterium autotrophicum , thus explaining the orange color of this organism. The bacteria studied contain a C 19 cyclopropane acid, Pseudomonas facilis and Corneynebacterium autotrophicum also

The lipids of hydrogen-oxidizing bacteria: Occurrence ofcis-9,10-methylene hexadecanoic acid inHydrogenomonas H 16

Knallgasbakterien (Hydrogenomonas H 16) wurden unter Speicherbedingungen in Submerskultur herangezogen. Die dabei gespeicherte Poly-β-hydroxybuttersäure wurde zusammen mit anderen «lokker gebundenen » Lipiden mit Chloroform extrahiert. Die daraus abgetrennten polaren Lipide bestanden ausschliesslich aus Phosphatidyläthanolamin. Die Analyse der Fettsäurezusammensetzung dieses Phosphatids ergab einen Gehalt von 42% (bezogen auf das Gewicht der Gesamtfettsäuren) einer C17-Cyclopropansäure. Durch spektroskopische Untersuchungen und oxydativen Abbau wurde letztere alscis-9,10-Methylenhexadecansäure identifiziert.

Transfer of blood-group determinant from bovine serum nonlipids to erythrocyte lipids of various mammals.

Received July 15, 1977 1. Heinrich, H.C., Bartels, H.: Ktin. Wschr. 45, 553 (1967); Hausmann, K., et al. : Acta Haemat. 42, 193 (1969) 2. Hoechst Pharmaceutical Research Laboratories, Behring Institute, 1977 3. Miles, L.E.M., et al.: Anal. Biochem. 6I, 209 (1974) 4. Heinrich, H.C., et al. : Klin. Wschr. 44, 827 (1966); Heinrich, H.C., Gabbe, E.E., Whang, D.H. : Atompraxis 11, 430, 660 (1965) 5. Heinrich, H.C., Gabbe, E.E., Briiggemann, J.: Z. Naturforsch. (in press)

Transfer of Bovine J Blood-Group Activity to Human Erythrocytes in vitro

THe bovine J determinant is transferred from a bovine serum nonlipid fraction to a human erythrocyte membrane lipid by an incubation procedure. The transferred J determinant is detected in the total lipids extracted from transformed human erythrocytes by an inhibition test in the bovine J system. It is also detected by cross-reacting human anti-A sera after treatment of human cells with papain prior or subsequent to transformation.

Distribution of the A Blood Group Activity on Lipid and Nonlipid Fractions of Pig Organs

Lipids and protein-containing nonlipids of pig erythrocytes, serum and several organs were tested for A blood group activity. Both lipids and nonlipids of brain, myocardium, skeletal muscle and adipose tissue show no A activity. The way of distribution of A activity on lipids and nonlipids of other tissues and serum differs among individual pigs. With respect to the acquisition of A activity of erythrocytes in a postnatal period, it seems likely that the lipidic A substance Tor part of it containing the A determinant) is transferred from plasma to the erythrocyte membrane.

Ubiquinones (Coenzyme Q) in hydrogen oxidizing bacteria.

The isoprenoid quinone content and composition of several strains of the following hydrogen oxidizing bacteria were studied: Xanthobacter autotrophics, Xanthobacter flavus, Paracoccus denitrificans, Pseudomonas pseudoflava, P. carboxydovorans, P. carboxydo-hydrogena. All strains studied contained ubiquinone (UQ) in concentrations between 0.7 and 2.8 μmol/g bacterial dry weight. No menaquinones were found. UQ of all strains (except one) contained side chain lengths of 10isoprenoid units (UQ-10), two strains contained in addition UQ-8. UQ-9 was found in Pseudomonas carboxydohydrogena only.

Bovine J blood-group activity in the lipids of erythrocytes of different age

ZusammenfassungJ-positive Rinder-Erythrozyten wurden durch Ultrazentrifugation in einem diskontinuierlichen Dichtegradienten von Ficoll-400 in Populationen verschiedenen Alters getrennt. Mit zunehmendem Alter wurde eine Abnahme der Phosphatid- und Cholesterin-Konzentration gefunden. Die J-Aktivität der aus den Erythrozyten isolierten Lipide nahm mit zunehmendem Zellalter erheblich zu. Demnach erfolgt der Transfer der J-Determinanten vom Blutplasma auf die Erythrozyten-Membran ziemlich langsam. Es wird vermutet, daß die zunehmende J-Aktivität der älteren Zellen auf einer Abnahme der Membran-gebundenen Phosphatide beruht, die, wie früher nachgewiesen, den Transer der J-Determinanten hemmen.SummaryJ-positive cattle erythrocytes were separated according to age by ultracentrifugation in a discontinuous density gradient of Ficoll-400 (Pharmacia) in isotonic buffer solution. A decrease in phospholipid and in cholesterol content with increasing age was detected. The J activity was found to increase markedly with increasing cell age. This fact supports the view that the in vivo transfer of the J determinant from plasma to erythrocytes is a rather slow process. It is suggested that the increasing J activity of older cells is probably due to a decrease of erythrocyte membrane bound phospholipids which inhibit the transfer of the J determinant to the red cells.

Über die Speicherung von Lipiden durch Knallgasbakterien

The ability to accumulate lipids was investigated in two strains of hydrogen oxidizing bacteria (Hydrogenomonas H 16 and strain 11/x). Along with the deposition of poly-β-hydroxybutyrate the amount of other lipids is shown to increase 1.8 times in strain H 16. It is suggested that the increase of the latter lipids is due to the formation of membrane lipids that are needed for the formation of membranes around the intracellular globules of poly-β-hydroxybutyrate. In strain 11/x the amount of lipids increases 7 times along with the storage of carbohydrates. In this case, the majority of lipids consists of triglycerides. It is suggested that there is a true storage of neutral fat in strain 11/x.

Distribution of the a blood-group activity in porcine serum

The A blood-group activity of pig serum is bound to a lipid, in some cases also to a nonlipid fraction. The major lipidic A activity (roughly 50%) is carried with the HDL class, while the VLDL and LDL classes contain roughly 25% each.