A search for the indianapolis-variant of human alcohol dehydrogenase in liver autopsy samples from North Germany and Japan

Summary Human liver alcohol dehydrogenase (ADH) variants were screened in random autopsy specimens from 53 North German and 34 Japanese individuals. Based on pH-activity profile and electrophoretic pattern, only ADH₂ and ADH₃ variants were detected. In relatively fresh specimens, an anodic band or -ADH band was also observed. The recently reported new molecular forms collectively called ADH_Indianapolis (Bosron et al. 1980) could not be demonstrated and therefore may be confined hitherto only to the American black population.This paper is dedicated to Professor Dr. Dr. H. Baitsch on his 60th birthday     

Studies on Thin-Layer Chromatography after Incubation with [35S]Methionine as an Aid to Identification of Mycobacteria

Thin-layer chromatography (TLC) of ethyl ether-ethanol extracts of mycobacteria obtained after incubation with [³⁵S] methionine is useful for differentiation among mycobacterial species, as the distribution of radioactive spots in TLC shows a characteristic pattern except for a few species, including M. intracellulare and M. gordonae. Some supplementary studies have been carried out in the present investigation and the following results have been obtained.

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Ly-m19: The Lyb-2 region of mouse chromosome 4 controls a new surface alloantigen

Spleen cells from an SJL mouse immunized with 70'/3 cells, an established pre-B cell line, were fused with cells of the nonsecretor myeloma line NS.1. One established hybridoma cell line (clone K10.6) continuously secreted antibody that recognized a new antigenic specificity tentatively named Ly-m19. This newly found antigen is detectable on both T and B cells. Cytotoxicity assays reveal that 75 percent of the spleen and lymph-node cells, 35 percent of bone-marrow cells, and 15 percent of thymus cells reacted with antibody of clone K10.6. Strains expressing the specificity Ly-m19.1 are characterized by negative reactions and include the strains AKR, CE/J, RF/J, GR/A, SJL, P/J, BDP/J, and LG/J. All other strains so far tested are Ly-m19.2. This strain distribution pattern distinguishes Ly-m19 from any known murine lymphocyte alloantigen, but it parallels the Lyb-2 ^c haplotype. Linkage test of a set of AKXL recombinant inbred strains revealed close linkage of Ly-m19 and Lyb-2 loci on mouse chromosome 4.Abbreviations used in this paper LPS lipopolysaccharide - B6 C57BL/6 - Con-A concanavalin A - MLC mixed-lymphocyte culture The prefix m (monoclonal) is used following a suggestion by Klein and co-workers (1979).     

Phlegmacins and anhydrophlegmacinquinones: Dimeric hydroanthracenes from seedlings of Cassia torosa

From seedlings of Cassia torosa four dimeric hydroanthracenes have been isolated. Two, a pair of atropisomeric dimers consisting of two molecules o     

Dammarane saponins of leaves of Panax pseudo-ginseng subsp. himalaicus

From dried leaves of Panax pseudo-ginseng subsp. himalaicus collected in Eastern Himalaya, new dammarane saponins, named pseudo-ginsenosides-F₁₁ and -F₈ were isolated along with the known Ginseng-root saponins, ginsenosides-Rb₃, Rd and -Re. Pseudo-ginsenoside-F₈ was proved to be a mono-acetyl-ginsenoside-Rb₃ and the location of its acetyl group was established mainly by ¹³C NMR spectroscopy. Pseudo-ginsenoside-F₁₁, was identified as the 6-O-α-rhamnopyransyl(1 → 2)-β-glucopyranoside of 3β,6α,12β,25-tetrahydoxy-(20S,24R)-epoxy-dammarane. The C-24 configuration of ocotillone and its related triterpenes was confirmed to be 24R excluding the recent comment by Lavie et al.     

Synthesis of diastereomeric bis-unsaturated prostaglandins

With the report given herein all diastereomers of PGF₂, PGE₂, and PGD₂ which bear the naturally recognized 15-S hydroxylated center, whether in the natural or $\text{ent}$-prostanoic acid skeleton, have been prepared by a route involving initial introduction of the carboxyl (α) chain (1). A major advantage of the initial α-ylation⁺ route is the facile reduction of the 13,14-en-15-one system with methanolic NaBH₄ which proceeds without competing 1,4-reduction. The products are thus free of 13,14-dihydro-PG₂ contaminants (2). The initial pharmaco logical evaluation of these diastereomers will be submitted for publication in this journal (3).     

A multiplicity of erythrocyte glycolipids of the neolacto series revealed by immuno-thin-layer chromatography with monoclonal anti-I and anti-i antibodies

The thin-layer-chromatography immunostaining procedure was applied to human erythrocyte glycolipids using monoclonal anti-i and anti-I antibodies which are directed against epitopes on linear and branched carbohydrate chains of the neolacto (poly-N-acetyl-lactosamine) series. An examination of native and mild-acid-treated glycolipids from normal adult (I_{adul t} antigen type), neonatal (i_cord) , and I-antigen-deficient adult (i_adult) erythrocytes enabled certain structural inferences to be made as follows: (a) cells of both I and i phenotypes contain a multiplicity of glycolipids of the neolacto series whose backbones consist of 8 or more sugar residues; (b) the octasaccharide backbones are predominantly linear in cells of i phenotype and branched in those of I type; and (c) more complex glycolipids having decasaccharide and larger backbones with both linear and branched sequences occur in erythrocytes of both phenotypes, 0     

Effects of an inhibitor of glucosylceramide synthase on glycosphingolipid synthesis and neurite outgrowth in murine neuroblastoma cell lines.

1-Phenyl-2-decanolyamino-3-morpholino-1-propanol (PDMP), an effective inhibitor of UDP-glucose:ceramide glucosyltransferase, caused inhibition of cell growth in murine neuroblastoma cell lines. Metabolic labeling of glycosphingolipids with [14C]galactose in NS-20Y, Neuro2a, and N1E-115 cells showed reduced incorporation of radioactivity into gangliosides and neutral glycosphingolipids when threo-PDMP was present in the medium. Treatment of NS-20Y cells with threo-PDMP resulted in a time-dependent decrease in mass levels of gangliosides and neutral glycosphingolipids. After 24 h in the presence of 50 microM threo-PDMP, neutral glycosphingolipid mass was reduced to 32%, where glucosylceramide was the most affected (90% decrease). The ganglioside mass was reduced to 57% of the original content. Neurite outgrowth from neuroblastoma cells in serum-free medium was significantly inhibited by threo-PDMP in a dose-dependent manner. Threo-PDMP also caused retraction of neurites which had been induced to extend in serum-free medium. Pretreatment of cells with GM1 partially restored the ability of NS-20Y cells for neurite outgrowth in the medium containing threo-PDMP. These results suggest a possible role for glycosphingolipids in neurite outgrowth of murine neuroblastoma cells.     

A coenzyme A-independent transacylase is linked to the formation of platelet-activating factor (PAF) by generating the lyso-PAF intermediate in the remodeling pathway

The remodeling pathway for the biosynthesis of platelet-activating factor (PAF) consists of the following reaction sequence: alkylacylglycerophosphocholine----lyso-PAF----PAF. Results presented in this article describe a novel transacylase activity that generates the lyso-PAF intermediate, which can then be acetylated to form PAF. Ethanolamine-containing lysoplasmalogens, 1-acyl-2-lyso-sn-glycero-3-phosphoethanolamine, alkyllysophosphoethanolamine, unlabeled lyso-PAF, 1-acyl-2-lyso-GPC, where GPC is sn-glycero-3-phosphocholine, and choline-containing lysoplasmalogens were all able to stimulate the formation of [3H]lyso-PAF from a [3H]alkylacyl-GPC precursor pool associated with HL-60 cell (granulocytic type) membranes. Other glycerolipids containing free hydroxyl groups (3-alkyl-2-lyso-sn-glycero-1-phosphocholine, lysophosphatidylserine, lysophosphatidylinositol, diacylglycerols, alkylglycerols, and monoacylglycerols), cholesterol, phosphatidylcholine, and phosphatidylethanolamine had no stimulatory effect on the release of [3H]lyso-PAF from the prelabeled membranes under identical incubation conditions. The observed transacylase reaction is directly coupled to PAF production, since the addition of a lysoethanolamine plasmalogen preparation to HL-60 membranes in the presence of [14C]acetyl-CoA stimulated PAF formation; under these conditions the lysoethanolamine plasmalogen was acylated. The transacylase responsible for the release of lyso-PAF from the membrane-associated alkylacyl-GPC was not affected by Ca2+, EGTA, or a known phospholipase A2 inhibitor, p-bromophenacyl bromide. The fact that the unnatural analog of lyso-PAF, lysophosphatidylserine, and lysophosphatidylinositol did not influence transacylase activity, whereas detergents such as deoxycholate and Triton X-100 inhibited the activity, demonstrated the observed stimulatory effects of the choline- and ethanolamine-containing lysophospholipids on the formation of [3H]lyso-PAF from [3H]alkylacyl-GPC were not due to any detergent property of these lysophospholipids. Thus, we conclude a CoA-independent transacylase (possessing phospholipase A2/acyltransferase activities) can be responsible for the formation of the lyso-PAF intermediate in the remodeling route of PAF biosynthesis.