Some aspects of amino acid metabolism in the rat fetus

• 1.1. In spite of an eventual catabolic phase during the last third of pregnancy, nitrogen retention seems to increase in pregnant rats. Furthermore, the high uterine blood flow and the high placental transfer of amino acids maintains an adequate nutrient supply to the fetuses.
• 2.2. The terminal rat fetus has a high circulating plasma amino acid level, as well as an increased free amino acid tissue pool when compared to its mother's.
• 3.3. In the rat fetus the development of enzymatic capabilities shows a sudden emergence (also denomined clustering) in late fetal life. In a general trend, the activities of enzymes related with amino acid metabolism are not well developed during rat fetal life.
• 4.4. The rate of amino nitrogen excertion in rat fetus is low, mainly due to the low development of urea cycle enzyme activities.
• 5.5. The rates of protein synthesis in many tissues are high in the rat fetus and they show a progressive decrease until delivery. On the other hand, the rates of protein breakdown are also higher during fetal life than in the adult.

     

Some new aspects of 17alpha-estradiol metabolism in man

17Alpha-estradiol (1,3,5(10)-estratriene-3,17alpha-diol) together with a tracer dose of the tritium-labeled compound was administered orally and sublingually to male volunteers. The serum concentrations of 17alpha-estradiol (free and liberated by enzymatic hydrolysis) were quantified by GC/MS, and the serum total radioactivity and urinary radioactivity excretion were determined. After oral administration, 17alpha-estradiol was rapidly and intensively conjugated; only tiny quantities of the free steroid (<1% of total) appeared in serum. Sublingual administration resulted in temporary (up to 3 h p.a.) higher serum levels of the free compound. The metabolite patterns obtained by TLC of extracts from serum and urine demonstrated that 17alpha-estradiol is the subject of a poor phase I metabolism in man. A great discrepancy was found in the serum concentrations of 17alpha-estradiol (free + conjugated) determined by GC/MS and the serum radioactivity expressed in 17alpha-estradiol equivalents. By TLC analysis of the steroid conjugates extracted from serum, various 17alpha-estradiol conjugate peaks were found. By enzymatic hydrolysis with beta-glucuronidase/aryl sulfatase from Helix pomatia they were only partially cleaved. Thus, the difference between the serum radioactivity and the 17alpha-estradiol levels determined by GC/MS had to be attributed to an incomplete conjugate hydrolysis. It has been shown with the synthesized 17alpha-estradiol sulfate conjugates that only the 3-sulfate is cleaved by enzymatic hydrolysis, whereas the 17-sulfate group resists enzymatic hydrolysis. The methanolysis procedure (acetyl chloride in MeOH) has proved to be an efficient method for cleaving both the 3-sulfate group and the 17-sulfate group. In contrast to the 17alpha-estradiol conjugates in serum, the urinary conjugates were intensively split by the enzyme preparation. From this, it has to be concluded that the serum conjugates were deconjugated and newly reconjugated before urinary excretion.     

Some new aspects in biosensors

This paper reviews recent advances in biosensors contributed mainly by our laboratory. The biosensors, based on the new immobilization materials - sol-gel organic-inorganic hybrid materials, cryohydrogel (or organohydrogel) and bilayer lipid membranes, are presented. The analytical performances of the biosensors are discussed. Applications of the biosensors in extreme environment are emphasized. A new generation of biosensors - surface plasmon resonance biosensors and capacitance biosensors, are also described.     

Some aspects of glutathione metabolism in ataxia-telangiectasia fibroblasts

Levels of glutathione (GSH) and two enzymes involved in GSH metabolism, glutathione reductase (GR) and glutathione-S-transferase(s) (GST), were measured in four SV40-transformed human fibroblast cell lines. MRC5-V1 and GM0637, derived from normal individuals, had mean GSH levels of 4.2 and 6.5 nmoles/10(6) cells, respectively. TAT2SF and AT5BIVA, both from ataxia-telangiectasia (A-T) patients, respectively had 6.5 and 4.2 nmol/10(6) cells, indicating that basal GSH levels were similar in A-T and normal cells. There was some variation in GST activity among the four cell lines but deficiency in this enzyme cannot be associated with radiosensitivity in A-T. When GR activity was measured, A-T cells had approximately 82 per cent of the mean normal activity. Though statistically significant, (P = 0.05), this small deficiency could be due to chance and is unlikely to be responsible for the radiosensitive phenotype of A-T.