Glycosphingolipids were purified from porcine erythrocytes and plasma. Two minor glycolipids with human blood group A and H antigenicities were found in both sources as components. The two antigenic glycolipids were identified as a hexaglycosylceramide (IV3 alpha GalNAc,IV2 alpha Fuc-Lc4Cer) for the A antigen and pentaglycosylceramide (IV2 alpha Fuc-Lc4Cer) for the H antigen and belonged to lactoseries (type 1 sugar chain) in contrast to those with neolacto core (type 2 sugar chain) in human erythrocytes, thereby endorsing biochemically the previous serological observations that the A antigen on porcine erythrocytes is uptake from plasma, probably the H antigen being the case. In addition to major glycolipids of globoseries in red cells and plasma, a variety of acidic glycolipids including two classes of sulphatides (sulphated galactosylceramide and sulphated lactosylceramide) and five classes of gangliosides (GM3, GD3, GM1, fucosyl GM1 and GD1a) containing N-acetylneuraminic acid and N-glycolylneuraminic acid were obtained from plasma.

Two glycosphingolipids with human blood group A and H antigenicity were isolated from porcine erythrocyte membranes which were obtained from the pooled blood. The yield of the A- and H-antigenic glycolipids was approximately 0.2 and 0.1% of total neutral glycolipids, respectively. No B antigen was detected. Through several methods the porcine erythrocyte antigens were all found to belong to lactoseries (type 1 chain), IV2Fuc alpha, IV3GalNAc alpha Lc4Cer for type A and IV2-Fuc alpha Lc4Cer for type H, in contrast to the antigenic glycolipids in human erythrocytes, which mostly belong to neolactoseries (type 2 chain). The constituent fatty acids of the A antigen were 75% normal acids and 25% 2-hydroxy acids, and the long chain base was 95% sphingenine. This is the first demonstration of the A- and H-antigenic glycolipids on erythrocytes of pig in whose gastric mucin the human blood group A and H substances have been demonstrated.

Human blood group A- and H-antigenic glycosphingolipids were isolated from pooled porcine plasma. The structures of the A-active hexa- and H-active pentaglycosylceramides of lactoseries (type 1 sugar chain) were the same as those in porcine erythrocytes. These results endorse biochemically the previous observations that the A and H antigens on porcine erythrocytes are taken up from plasma.

Five neutral glycosphingolipids (GSLs) (Gb4Cer, Gb3Cer, LacCer, GlcCer and GalCer) were isolated from porcine erythrocyte membranes. Gb4Cer was the most abundant (over 70% of total neutral GSLs). In addition, a minor GSL fraction (less than 1% of the total), which migrated more slowly than Gb5Cer on TLC, composed of five GSLs. Two of them exhibited blood group A and O (H) activity, respectively, and were studied for their structures by spectrometries with NMR and FAB-mass, sequential hydrolysis by exoglycosidases, methylation and immunostaining. Chemical structure of the blood group A-antigen was proposed to be GalNAc alpha 1-3 (Fuc alpha 1-2) Gal beta 1-3 Gal-4Glc beta 1NAc beta 1-3 Gal beta 1-4 Glc beta 1-1Cer and GalNAc alpha 1-3 (Fuc alpha 1-2) Gal beta 1-4 GlcNAc beta 1-3 Gal beta 1-4Glc beta 1-1Cer. The blood group O (H)-antigen showed the structure lacking non-reducing terminal alpha-linked N-acetylgalactosamine from the A-antigen, or a biosynthetic precursor from of the A-antigen.

1. Six neutral GSL fractions were purified from porcine erythrocyte membranes. 2. They were identified to be LacCer (14% of total neutral GSLs), 2-hydroxy acid-rich and -poor Gb3Cer (3 and 7%, respectively) and Gb4Cer (71%) by means of NMR spectrometry. 3. Monohexosylceramides (5%) were composed of GlcCer and GalCer with near amount. 4. All these GSL classes contained a high concentration (more than 20% of total acids in each class) of 2-hydroxy fatty acids. 5. GalCer and GlcCer contained considerable amounts of C16- and C18-acids, and of C18-phytosphingosine, whereas C24-acids and C18-sphingosine were predominant in the other GSLs. 6. A minor GSL fraction (less than 1% of total neutral GSLs) which migrated more slowly than Gb5Cer on a thin layer plate and composed of several GSL components contained L-fucose.

A phosphotyrosine phosphatase was enriched 9-fold from human placenta homogenates using ammonium sulfate fractionation and chromatographies on P-cellulose and Phenyl Sepharose. The enzyme preparation hydrolyzed phosphotyrosine, p-nitrophenyl phosphate, as well as phosphoenol-pyruvate. The enzyme was unstable, and its Km value for phosphotyrosine increased during the purification procedure. The optimum pH for phosphotyrosine was found to be pH 7.0. The phosphotyrosine phosphatase activity was completely inhibited at 2.6 mM Cu2+. Zn2+ was slightly inhibitory to the phosphatase activity. The effects of several inhibitors on the phosphotyrosine phosphatase activity were different from those on the p-nitrophenyl phosphatase activity. These results suggest that the phosphotyrosine phosphatase activity from human placenta is different from previously reported acid and alkaline phosphatases.

A sensitive enzyme immunoassay method for rat gamma-glutamyl transpeptidase was developed that was able to quantitate between 5 to 500 ng of the enzyme. The enzyme contents of fetal liver, adult liver, hyperplastic nodules, and various tissues including hepatoma were determined. Normal rat liver contained very little enzyme, while fetal liver and hepatoma tissue contained high levels. The gamma-glutamyl transpeptidase content of the liver was assayed during the process of 3'-methyl 4-dimethyl-aminoazobenzene-induced hepatocarcinogenesis. The content was increased at an early stage, then decreased, and again increased at the stage when hepatoma developed. The change of the enzyme content determined by an enzyme immunoassay was highly consistent with that determined by enzymatic assay. This indicates that the increased activity of the enzyme as previously reported was accompanied by an increased immunoreactive enzyme protein. Immunohistochemical studies on the hyperplastic nodule revealed the presence of gamma-glutamyl transpeptidase in the hepatocyte membrane.

The cytotoxicity of 8 natural dyes, commercially available as food additives in Japan, was studied on cultured fetal rat hepatocytes. Laccaic acid, one of the carminic acid samples and monascus pigments were found to be very toxic to cultured hepatocytes. Laccaic acid caused an increase in γ-glutamyl transpeptidase activity. An 11-fold increase was seen 4 days after such addition, a significantly greater elevation than that produced by either the water or acetone solvents employed, or other dyes which have no toxic effects. On the other hand, monascus pigments had an increasing effect on both γ-glutamyl transpeptidase and glutathione S-transferase, the later enzyme being elevated approximately 9 times greater than control values within 2–4 days. The increases in γ-glutamyl transpeptidase and glutathione S-transferase activities by laccaic acid and monascus pigments could be inhibited by the simultaneous addition of either actinomycin D or cycloheximide. This suggests that the induction of these enzymes requires transcription and translation processes.

The cytotoxicity of 11 dyes, used as food dyes in Japan, on cultured fetal rat hepatocytes was studied. Xanthene dyes containing halogen atoms in their molecules such as phloxin, rose bengal, and erythrosine were more toxic than other groups of food dyes. In the present paper the effect of food dyes on the cell growth of hepatocytes was also examined. Phloxin was especially toxic to the cell growth and a dose-response relation was observed between the concentration of phloxin and the cell growth of hepatocytes when the dye was added 3 days after plating.

The toxicity of 14 commercial natural dyes which are widely used as food additives in Japan was studied on Paramecium caudatum. Laccaic acid and capsanthin were found to be very toxic to Paramecium caudatum. Some of the commercially available carminic acid and crocin were also toxic. The inhibitory effect of natural food dyes on leucine aminopeptidase, acid phosphatase and esterase in vitro was proportional to the toxic effect of the dyes on the survival time of Paramecium caudatum. Analyses of the commercial natural food dyes by high performance liquid chromatography failed to identify the toxic components.

The toxic effect of 14 food dyes was studied in Paramecium caudatum. It was found that xanthene dyes containing halogen atoms in their molecules were more toxic than other groups of food dyes. Phloxin and rose bengal containing chlorine were especially toxic. The effect of food dyes on leucine aminopeptidase, acid phosphatase, and γ-glutamyl transpeptidase activity in P. caudatum was studied in order to investigate the mechanism of toxicity. Phloxin and rose bengal inhibited leucine aminopeptidase remarkably. The inhibitory effect of food dyes on leucine aminopeptidase in vitro is consistent with the toxic effect of the dyes on the survival time of P. caudatum. A possible correlation between toxicity and inhibition of the activity of enzymes involved in the digestive process is discussed.

Summary In this paper, the vibration response of the satelliteequipment panel during the launch phase is predicted in consideration of the robustness. By using a simplified model consistedof a panel and an acousticcavity, MonteCarlo simulationin which analyticalparameters are changed at randomintheconceivablerangeofvariationis done. From the result,the acceleration power spectral density(PSD) of the satellite equipment panel is obtained. After statistical processing, the effect of variation on the vibration response is estimated and the proper design criteria of the equipment are defined.