Age-related changes of lipid fractions and total fatty acids in liver lipids and heart lipids of female and male rats aged 37-1200 days (liver) and 331-1200 days (heart).

1. Total lipids and the lipid fractions cholesterol ester, triacylglycerol, free cholesterol, free fatty acids and phospholipids, as well as the fatty acid patterns of total lipids, were measured in liver homogenates of female and male rats (Wistar SPF, strain Hannover) aged 37-1213 days. 2. The same parameters were measured in the apex of the heart in female and male rats aged 331-1213 days. 3. All parameters were monitored every 49th day. Five female and five male animals were used in each experiment. 4. The lipid fractions in liver showed a positive linear regression vs age, whereas all lipids in rat heart showed a negative regression vs age in both sexes. 5. The significance of regression vs age of fatty acids was much less than that in the lipid fractions of liver and heart of these animals.

Lipid analysis in bovine lens parts after in vitro incubation in the presence of an HMG-CoA-reductase inhibitor.

Normal bovine lenses and bovine lenses after incubation in TC 199 in the presence of a lipid lowering drug were divided into four parts: the equatorial ring, the nucleus, the anterior cortex and posterior cortex. The lipids were extracted according to Egge et al. (1). Total lipids were determined gravimetrically, the lipid fractions phospholipids (PL), cardiolipin (CL), free fatty acids (FFA) and cholesterol (CH) were separated by thin layer chromatography and determined quantitative by densitometry after charring with 10% sulfuric acid. There are differences in lipid distribution between the four lens compartments, but there are no differences between normal and drug treated lenses in total lipids and lipid fractions. This result could be explained in different ways: either during this incubation time of 24 hours at 37 degrees C there will be no effect on the lens epithelial cells or there is no de-novo synthesis of lipids, especially of cholesterol in the bovine lens epithelium which could be influenced by lipid lowering drugs like HMG-CoA reductase inhibitors.

Age-related changes of lipid fractions and total fatty acids in the serum of female and male rats aged from 37 to 1200 days.

1. Total lipids and the lipid fractions cholesterol ester, triacylglycerols, free cholesterol, free fatty acids and phospholipids as well as the fatty acid patterns of total lipids were measured in the serum of female and male rats (Wistar SPF, strain Hannover) aged 37-1213 days. 2. All parameters were monitored every 49th day. Five female and five male animals were used in each experiment. 3. All lipid fractions showed a significant positive linear regression vs age in both sexes. There were multiple significant differences in lipid fractions between the sexes. 4. Among the fatty acids only docosahexaenoic acid shows a significant correlation with age in both sexes.

Concentrations of serum cortisol, progesterone, estradiol-17 beta, cholesterol and cholesterol ester in the doe during the reproductive stadium, in the fetal serum, in the amniotic fluid and in the milk of rabbits, as well as correlations between these parameters.

1. Serum progesterone, estradiol-17 beta and cortisol, as well as cholesterol and cholesterol ester concentrations in pregnant and lactating rabbits (New Zealand white hybrids, n = 9), were measured. These parameters were also studied in the amniotic fluid, the milk and the fetal serum (28-day old fetuses). 2. Serum progesterone and estradiol-17 beta were significantly enhanced during gestation, while the content of cortisol showed a marked elevation at the end of pregnancy. The concentrations of these hormones decreased before parturition. 3. Serum cholesterol and cholesterol ester concentrations markedly decreased in the second half of gestation (74 and 76%, respectively) and elevated after parturition and in the first week of lactation.

Haematological values during normal reproduction of the maternal and the fetal rabbit.

1. Haematological values of non-pregnant/non-lactating, pregnant as well as lactating rabbits and 28-day-old fetuses were measured. 2. The haemoglobin content in does decreased during the observed periods from 122 +/- 8 g/l to 100 +/- 11 g/l. In 28-day-old fetuses it was 85 +/- 0 g/l. 3. The erythrocyte count in 28-day-old fetuses was 2.4 X 10(12)/l. In the does, the erythrocyte count was 5.2 X 10(12)/l in week 4 of gestation. The erythrocyte volume in fetuses was about 45% higher than that of the doe. 4. In fetuses the leucocyte count was approximately one ninth that of the mother in week 4 of gestation (0.41 +/- 0.08 X 10(9)/l vs 3.8 +/- 0.4 X 10(9)/l).

Iron and copper concentrations in the maternal and fetal serum, placenta and amniotic fluid during the reproductive stadium as well as in the milk of rabbits.

Serum concentrations of iron and copper from rabbits (New Zealand White hybrids; N = 12) were determined during the reproductive stadium (gestation and four weeks of lactation). Samples of serum from fetuses, placental tissue and amniotic fluid were also examined. Iron: a decrease of iron in the maternal serum during the second half of gestation was observed, whilst a significant rise occurred in the first week of lactation. The content of iron in the fetal serum dropped from day 21 to day 28 of gestation. The iron concentration in the placental tissue decreased during this time. A rise of the iron level in the amniotic fluid was determined from day 21 to day 28 of gestation. The iron content in the milk was about 33 mumol/l (first and second day of lactation). Copper: in the first half of pregnancy the copper level diminished slightly compared with the content of non-pregnant, non-lactating rabbits, while a rise was observed in the fourth week of this period. The copper concentration decreased in the first week of lactation and then reached the peak level in the second week of this phase. The copper level in the fetal serum declined from day 21 to day 28 of gestation, while the copper content in the amniotic fluid increased significantly on day 28, in comparison with day 21 of gestation. In contrast, a decline of the copper concentration in the placental tissue was noticed from day 21 to day 28 of this period. The copper content in the milk was nearly 25 mumol/l (first and second day of lactation).

6,7,4'-Trihydroxyisoflavan: a potent and selective inhibitor of 5-lipoxygenase in human and porcine peripheral blood leukocytes.

The effect of 6,7,4'-trihydroxyisoflavan on human platelet 12-lipoxygenase and human and porcine PMNL 5-lipoxygenase activities has been studied. 6,7,4'-Trihydroxyisoflavan was found to inhibit 5-lipoxygenase more strongly than 12-lipoxygenase; its concentration for 50% inhibition (IC50) was 1.6 microM for human and porcine 5-lipoxygenase and 22 microM for human platelet 12-lipoxygenase. Inhibition of microsomal cyclooxygenase from ram seminal vesicles is exhibited at much higher concentrations of 6,7,4'-trihydroxyisoflavan (IC50 = 200 microM).

[The microanalysis of serum lipoprotein lipids (author's transl)]. / Mikrobestimmung von Lipoproteinlipiden im Serum.

A method is described which allows the determination of phospholipids, free and esterified cholesterol, triglycerides and free fatty acids in lipoprotein fractions starting from 50 microliter of serum. Lipoproteins were separated by successive precipitation: VLDL with Heparin/Mg++, LDL with Dextran sulfate/Mg++ and finally HDL with Dextran sulfate/Mn++. Lipids extracted from the precipitated lipoproteins were determined gravimetrically and by densitometry after thin layer chromatography and charring (van Gent, C.M. (1968), Z. Anal. Chem. 236, 344--350; Egge, H. et al. (1970) Z. Klin. Chem. Klin. Biochem. 8, 488--491). The results obtained from the serum of 12 adult healthy persons were compared with those from lipoprotein fractions separated by preparative ultracentrifugation (Havel, R. J. et al. (1955) J. Clin. Invest. 34, 1345--1353). The distribution of lipids in beta-lipoproteins (d less than 1.063 g/ml) and HDL (1.063 less than d less than 1.21 g/ml) prepared by both methods showed good agreement. Some differences were observed between VLDL (d less than 1.006 g/ml) and VHDL (d greater than 1.21 g/ml) prepared either by precipitation or ultracentrifugation. Compared to the total lipid of the sera, recovery rates were 95--105%. Variation coefficients were in the range of 15--20% for VLDL lipids, 5--10% for LDL and HDL lipids and 10--15% for VHDL lipids. Gravimetrically determined total lipids had a variation coefficient of 4 and 6% for LDL and HDL respectively.

Positional isomers of unsaturated fatty acids in rat liver lipids.

The fatty acids of liver lipids from rats raised on a fat free diet from the 30th to the 90th day after birth were analyzed with special regard to the detection of positional isomers of mono-, di-, tri-, and tetraenoic fatty acids. The methyl esters obtained after transesterification of total lipids were separated by argentation chromatography into five fractions: I saturated, II monoenoic, III dienoic, IV dienoic nonmethylene interrupted, V tri- and tetraenoic fatty acid esters. After hydroxylation of the double bonds with osmium tetroxide, the analysis of the poly-O-trimethylsilyl derivatives by gas liquid chromatography on S.C.O.T. columns combined with mass spectrometry revealed the presence of 19 monoenoic, 15 dienoic, and 9 trienoic as well as 3 tetraenoic fatty acid isomers including the normally occurring representatives of the (n-3), (n-6), (n-7), and (n-9) fatty acid families. The majority of the identified isomers can be coordinated to one of these families like 7-16:1; 11-20:1; 6,9-18:2; 8,11-20:2; 5,11-20:2; 5,8,11-20:3; 7,10,13-22:3 to the (n-9) family, 11-18:1; 13-20:1; 5,11-18:2; 7,13-20:2; 6,11-18:2; 6,9-16:2; 8,11-18:2; 10,13-20:2; 5,8,11-18:3; 7,10,13-20:3; 4,7,10,13-20:4 to the (n-7) family and 11,14-20:2; 5,11,14-20:3; 6,9,12-18:3; 8,11,14-20:3; 5,8,11,14-20:4; 7,10,13,16-22:4 to the (n-6) family. All these naturally occuring isomers can be placed into a network of desaturation and chain elongation steps which allows certain conclusions about the substrate specificity of the delta6-, delta5- and delta4-desaturase systems. The great number of isomers found in the (n-7) family indicates that the members of this family are actively metabolized in partial essential fatty acid deficiency.

Technology for the identification of unusual cis, cis octadecadienoic fatty acids.

In this article methods are presented for the separation and identification of unusual cis, cis dienoic and polygnoic long chain fatty acids. Special emphasis has been laid on the identification of cis, cis octadecadienoic acids. The steps followed are: after transesterification the fatty acid methyl esters are separated by preparative gas chromatography according to chain length followed by argentation chromatography on thin-layer plates. After hydroxylation of the double bonds with osmium tetroxide the polyhydroxy compounds are derivatized to the per-O-trimethylsilyl-ethers. Separation and identification of individual compounds are achieved by combined gas chromatography-mass spectrometry using SCOT columns and low ionization energy.