Separation of lac dye components by high-speed counter-current chromatography.

High-speed counter-current chromatography has been successfully applied to the separation of the lac dye components. A 25-mg quantity of the sample was separated using a two-phase solvent system composed of tert.-butyl methyl ether-n-butanol-acetonitrile-water (2:2:1:5). The fractions were analyzed by high-performance liquid chromatography and electrospray tandem mass spectrometry. The separation yielded 2.6 mg of 97.2% pure laccaic acid C, 9.5 mg of 98.1% pure laccaic acid A, 3.6 mg of 98.2% pure laccaic acid B, and 0.5 mg of a 95.0% pure anthraquinonedicarboxylic acid with a molecular mass of 360.

Effect of maternal lifestyle on cord blood IgE factor.

During recent decades much interest has been focused on the possibility of predicting and preventing atopic diseases during pregnancy. The idea of being able to detect a predisposition early and take suitable environmental measures in order to avoid overt allergy is an attractive position. Elevated cord IgE of around 1.0 IU/ml has been proposed as a predictor in western children. However, there remains no information about the effect of maternal lifestyle during pregnancy on these levels. Total IgE levels were therefore determined using Pharmacia CAP system and PRIST, with sensitivities of 0.01 kU/l and 0.25 kU/l, respectively, from serum samples taken from 1138 Japanese pairs of cord blood and pregnant women responding to a questionnaire regarding 17 health practices, intake of 32 food allergens and 5 environmental factors. Of these, 28 (2.5%) pairs of samples were excluded from further analysis because of high contamination of IgA (> 15.4 mg/ml) in cord blood. Median cord blood IgE was 0.286 kU/l and geometric mean IgE was 66.25 kU/l in maternal sera using CAP system; there was no significant correlation between maternal log (IgE) and cord blood IgE. Similar results were obtained from PRIST, whose correlation with CAP system was significant (r = 0.884, p < 0.001 for maternal and r = 0.765, p < 0.001 for cord blood). Multiple logistic analysis demonstrated that avoidance of simultaneous exposure to hens' eggs and cow's milk (relative risk = 1.3, p < 0.05) as well as soy beans (relative risk = 2.8, p < 0.01) should be advised to mothers with positive allergic histories and/or high total IgE (> 400 IU/ml), especially in women aged more than 35 years who are pregnant with a male child. However, maintenance of healthy lifestyles, especially taking proper exercise and sleeping, and avoidance of inhalant allergens during late pregnancy may be a more important strategy for the reduction of cord blood IgE levels.

Concentration and uptake of taurine in umbilical blood platelets.

The concentration and uptake of taurine in the umbilical and adult blood platelets were studied. Taurine was the most abundant free amino acid in both umbilical and adult blood platelets. The taurine concentration in umbilical blood platelets (2.30 pmoles/10(4) cells) was significantly lower than that of adult blood platelets (3.27 pmoles/10(4) cells) in contrast to the reverse relationship in taurine concentrations in umbilical and adult blood plasma. No other amino acid showed such significant difference in the concentrations between umbilical and adult blood platelets. Taurine uptake into umbilical blood platelets was temperature sensitive and sodium-dependent in a manner similar to that of adult blood platelets. The uptake conformed well to Hanes-plot. The Vmax of the uptake into adult blood platelets was about 3.6 times higher than that of umbilical blood platelets, but no significant difference was seen in the Km value between the two groups.

Transamination of L-cysteine sulfinate in the growing rat.

The enzyme activities involved in the transamination of L-cysteine sulfinate (L-alanine 3-sulfinic acid), L-aspartate and L-cysteine were examined in fetal, neonatal and maternal rat liver and placenta. In fetal and neonatal rat liver, aminotransferase activity was most active with L-cysteine sulfinate as a substrate and was also active with L-aspartate, while activity with L-cysteine was very low. The activity of transamination of L-cysteine sulfinate in rat liver developed in parallel with that of L-aspartate and L-cysteine. The aminotransferase activity markedly increased after the 19th day of gestation, reaching the same value as adult liver on the 3rd day after birth. The ratios of transamination of L-cysteine sulfinate to that of L-aspartate and to that of L-cysteine were constant during development. These observations suggest that L-cysteine sulfinate, L-aspartate and L-cysteine are transaminated by the same enzyme in the rat liver during development. Since placental aminotransferase activity was extremely low compared with that of the liver, it was suggested that the placenta did not play an important role in the transamination of these amino acids during pregnancy.

Effect of guanidinoethyl sulfonate on taurine concentrations and fetal growth in pregnant rats.

Guanidinoethyl sulfonate (GES), a transport antagonist of taurine, was given to pregnant rats from day 11 to 21 of gestation as a 1% solution in drinking water. On day 21 of gestation in GES-treated pregnant rats, the concentration of taurine markedly decreased in the fetal whole body (54% of the control), the fetal liver (37%), the fetal whole brain (87%), the placenta (32%), the maternal liver (33%), the maternal whole brain (32%), and the maternal plasma (46%). The wet weight of fetal whole body, liver and brain of fetus, and placenta also showed a significant drop. But there were no differences of weight gain, in the liver and brain weights of the mother of the control and GES-treated pregnant rats. The urinary excretion of taurine in pregnant rats treated with GES was much higher than that of the controls. These results suggest that the administration of GES to pregnant rats induces much urinary taurine excretion with a resulting decrease in the tissue taurine content and readily produces taurine-deficient fetal rats.

Taurine concentrations in fetal, neonatal and pregnant rats.

The concentrations of taurine in the fetal and neonatal organs, and the maternal organs, plasma and urine of rats between the 15th day of gestation and the 21st day after birth were determined using an automatic amino acid analyzer. In the fetal liver and brain and in the placenta, the taurine concentration was the highest of all ninhydrin positive compounds. In the fetal liver and placenta, the concentrations of taurine increased significantly with the gestational days. Concentrations of taurine in the brain were much higher in the fetus and neonate than that in the adult. Moreover, the total amount of taurine per fetus increased markedly after the 15th day of gestation, and near term, reached almost the same amount as in the adult rat liver. In contrast to this, a significant decrease was observed in the taurine concentration in the maternal liver and muscle near term. The concentration of taurine in the urine of pregnant rats decreased near term, but in the plasma of pregnant rats the concentration of taurine did not change during pregnancy.

[Developmental changes in taurine and other sulfur-containing amino acid concentrations in fetal and neonatal rats].

The concentrations of taurine and other sulfur-containing amino acids in the fetal, neonatal and maternal liver, placenta, and whole fetal body between the 15th day of gestation and the 14th day after birth were determined using an automatic amino acid analyzer. In the fetal liver and placenta, the taurine concentration was the highest among all ninhydrin positive compounds. In these tissues the concentration of taurine increased significantly with the number of gestational days. Moreover, the total amount of taurine per fetus increased markedly up to term after the 15th day of gestation, and reached almost the same value as the total amount for adult rat liver. A striking difference in the changes in taurine concentrations during gestation between the fetal and maternal rat liver was observed: In contrast to the increase in the fetal liver, a significant decrease in the maternal liver was observed near term. But in these organs, no significant change in methionine, cystathionine and cysteine concentrations was observed during the perinatal period. These results suggest that taurine is supplied to the developing rat fetus from the maternal liver throughout gestation.

Estimation of gestosis of pregnancy (EPH-gestosis), relationship between fetal development and gestosis index.

Gestosis index score at delivery was compared with factors reflecting the fetal development in a series of 95 patients with EPH-gestosis collected in Okayama University Medical School in 1975-1979. The prediction of small for date (SFD) was performed by means of multivariate analysis of 10 variables, i.e. gestosis index, maternal body weight, height, uterine fundal length, abdominal circumference, maternal age at delivery and urinary estriol within one week prior to delivery. The result was that gestosis index was as useful as uterine fundal length in predicting SFD. The more gestosis index score increased, the more markedly the fetal development ws disturbed. Especially in the cases with scoring above 4 and in those with hypertension and proteinuria, the incidence of SFD increased obviously. No correlation between gestosis index and neonatal asphyxia was noticed. From the growth pattern of uterine fundal length and BPD, intrauterine growth retardation (IUGR) in pregnancy with EPH-gestosis occurred mainly within the third trimester of pregnancy. The functional development of the fetus with EPH-gestosis was evaluated with the use of urinary estriol level and fetal heart rate (FHR) monitoring. In the cases with EPH-gestosis scoring above 4, extreme disturbance of functional development of the fetus was observed.