Covalent binding to proteins of reactive intermediates resulting from prostaglandin H synthase-catalyzed oxidation of stilbene and steroid estrogens

Prostaglandin H synthase (PSH) is known to metabolically activate a variety of xenobiotics in vitro by means of its peroxidase activity. Recently, stilbene and steroid estrogens have been found to be cooxidized by ram seminal vesical microsomes, a rich source of PHS, to nonextractable metabolites bound to microsomal protein. To investigate further the nature of this protein binding, different radiolabeled estrogens were incubated with purified PHS, holoenzyme in the presence of various amounts of albumin (BSA), and radioactivity bound to protein was determined after gel electrophoretic separation. Diethylstilbestrol (DES), its analog hexestrol, and the steroid estrogens estrone and 2-hydroxy-estrone were cooxidized by PHS in vitro to metabolites that bound covalently to PHS and to BSA. Although a preferential binding of DES to PHS was found in the presence of excess BSA, reactive intermediates derived from DES, or from the other estrogens, were sufficiently stable to react with the competing nucleophile BSA as well. With respect to the metabolic reactions catalyzed by PHS, in addition to one-electron oxidation of phenolic functions, PHS catalyzed the aromatic hydroxylation of synthetic and steroid estrogens as shown by 3H2O release from regiospecifically labeled compounds and confirmed by product identification. Although DES was extensively metabolized by PHS, its aromatic hydroxylation was minor by comparison to estradiol, a difference possibly related to the compounds' redox potentials. Thus, cooxidation of estrogens in vitro resulted in phenoxy radicals, semiquinones and quinones, reactive intermediates capable of protein binding that may contribute to the adverse effects of stilbene and steroid estrogen observed in vivo and in short-term assays.     

Energy requirements of fat sand rats (Psammomys obesus) and their efficiency of utilization of the saltbush Atriplex halimus for maintenance

The fat sand rat (Psammomys obesus; Gerbillinae), a diurnal gerbillid rodent, is herbivorous and able to thrive while consuming only the saltbush Atriplex halimus (Chenopodiaceae), a plant relatively low in energy content and high in ash and water. We measured the basal metabolic rate offat sand rats, their energy requirements in captivity when they were offered only A. halimus, and their efficiency of utilization of this diet for maintenance.
Before consuming A. halimus leaves, the fat sand rats scraped off the surface layers with their teeth. This behavioural activity removed 6.4% to 8.5% of the dry matter of the leaf, but increased the gross energy and organic matter content of the leaf by only approximately 3.1%. Basal metabolic rate of the fat sand rats was 167.9 kJ.kg-O 7s d-', approximately 57% of that expected for an eutherian mammal of its body mass, and energy requirements for maintenance, or average daily metabolic rate, were 498.7 kJ.kg-0'7s d-', approximately 90% of that expected for a rodent of its body mass. Dry matter digestibility of the consumed A. halimus averaged 67% and apparent digestible energy and apparent metabolizable energy averaged 65.3% and 63.4% of the gross energy, respectively. The efficiency of utilization of A . halimus for maintenance energy (k,) by the fat sand rats was 0.32 and the heat increment of feeding (HIF) was 0.68. The k, of A. halimus appeared to be low compared to other feeds, and this characteristic plus its low energy value and high water content forced the fat sand rats to consume large quantities of forage for maintenance.
It was concluded that although A. halimus has a low energy and high ash content, there are several advantages for fat sand rats consuming mainly this diet. Among them are: (1) it provides a more stable diet throughout the year than do seeds; (2) fat sand rats have no competition for this food resource from other rodents; and (3) their burrows are at the base of the     

Prenatal and postnatal expression of mRNA coding for rat prothrombin.

The levels of prothrombin mRNA in prenatal and postnatal rat tissues were analyzed in order to determine tissue distribution of prothrombin expression and to determine if increases in liver prothrombin mRNA during development correlated with previously documented developmental increases in plasma prothrombin levels. Maternal tissues were also analyzed in order to determine if prothrombin mRNA levels varied due to gestational or postpartum influences. Northern analysis demonstrated that rat liver prothrombin mRNA levels increased several-fold late in gestation and reached maximal levels by 13 days after birth. Prothrombin mRNA was also expressed in diaphragm, stomach, intestine, kidney, spleen and adrenal tissues during development. In maternal tissues during pregnancy, prothrombin mRNA was expressed in liver, diaphragm, stomach, uterus and placenta. Prothrombin mRNA levels in each of these tissues that were positive by Northern analysis were quantitated by solution hybridization analysis. Between gestational day 18 and postnatal day 13, liver prothrombin mRNA levels increased from approx. 600 to 2100 molecules per cell (a 3.5-fold increase). In maternal liver during pregnancy, between day 18 and day 22, prothrombin mRNA levels increased from approx. 1800 to 2100 molecules per cell. Immediately after delivery, maternal liver prothrombin mRNA levels decreased to approx. 50% of preparturition levels. Prothrombin mRNA levels in placental tissue ranged from approx. 100 to 250 molecules per cell. In other fetal, postnatal and maternal tissues, prothrombin mRNA expression was less than 100 molecules per cell. These results demonstrate that the level and tissue-type expression of prothrombin mRNA varies in response to prenatal and postnatal influences.     

Diet selection and energy and water budgets of the common spiny mouse Acomys cahivinus

The common spiny mouse, Acomys cahirinus (body mass=47 g), is widely distributed in Israeli deserts where it inhabits natural crevices on rocky slopes. This omnivorous rodent consumes a varied diet, and in particular snails. We determined diet selection and energy and water balances of spiny mice when they were offered snails and seeds. The spiny mice maintained steady state body mass. Dry matter consumption of snails was 0.014 g* g⁻¹.-d⁻¹ and of seeds was 0.049g*.g⁻¹d⁻¹ for a total of 0.063 g*g⁻¹*d⁻¹. Total water intake was 0.101ml-g⁻¹.d⁻¹ and metabolizable energy intakewas 0.990 kJ. gxs^-1.d⁻¹ for a ratio (ml: kJ) of 0.102. This ratio was similar to that reported in a previously published study on free-living spiny mice. We concluded that snails and seeds allowed spiny mice to fulfil their energy and water requirements with minimal dry matter and fresh matter intakes. Furthermore, spiny mice selected a diet that provided them with a water (ml) to energy (kJ) ratio of approximately 0.1, although it appeared that they are able to survive on a much drier diet.     

Average daily metabolic rate of gerbils of two species: Gerbillus pyramidum and Gerbillus allenbyi

Gerbillus pyramidum and G. allenbyi are primarily granivorous, nocturnal rodents that are sympatric over many sandy areas of the Negev Desert. However, in their overall distribution, G. pyramidum occurs in extreme desert areas whereas G. allenbyi does not. We measured the average daily metabolic rate (ADMR) of gerbils of each species when they were offered pelleted diet. Given the difference in their distribution, we reasoned that the more xeric G. pyramidum would have lower ADMR than G. allenbyi; however, given the similarity in their diets, we reasoned that their ADMRs would be similar. The latter alternative was supported. ADMR of G. pyramidum (body mass = 31·9 ± 5·4 g) was 427·1 kJ·kg^−0.75·d⁻¹, 58% of that predicted for a rodent of its body mass; whereas ADMR of G. allenbyi (body mass = 22·3±2·3 g) was 387·7 kJ·kg^−0.75·d⁻¹, 49%) of that predicted. On the basis of these results, we suggest that factors other than their ADMRs are important in determining their geographical distribution.     

Interaction of phytoestrogens and other environmental estrogens with prostaglandin synthase in vitro

The phytoestrogens daidzein, genistein, equol and coumestrol were found to stimulate microsomal prostaglandin H synthase (PHS) in vitro in a concentration-dependent manner when PHS-activity was measured by arachidonic acid-dependent oxygen uptake. These compounds were co-oxidized by PHS and the conversion of parent compounds was measured by HPLC analysis. The stimulation of PHS-cyclooxygenase by these compounds was partially reversed at high concentrations probably due to their antioxidant properties causing inhibition. In contrast, the monomethyl ethers of daidzein and genistein, formononetin and biochanin A, had little or weakly inhibitory effect on PHS, and appear to be no or poor co-substrates for PHS. Compared to the equine estrogen equilin, its metabolite d-equilenin was poorly metabolized by PHS and inhibited rather than stimulated PHS-cyclooxygenase activity in vitro. The resorcylic acid lactones zearalenone and zeranol, on the other hand, were surprisingly good inhibitors of PHS-cyclooxygenase. Furthermore, zeranol inhibited both the arachidonic acid and the hydrogen-peroxide-dependent oxidation of DES in contrast to indomethacin which inhibited only cyclooxygenase-dependent co-oxidation of DES. The results of this in vitro study are discussed in the context of data on synthetic and steroidal estrogens and support the idea that PHS-activity may be modulated by interaction with certain estrogenic compounds.     

Enzymatic conversion of N carbamoyl-d-amino acids to d-amino acids

An enzyme isolated from Agrobacterium radiobacter was shown to catalyse the following reaction: H₂O + N-carbamoyl-d-amino acid→ d-amino acid + NH₃ + CO₂ Some properties of this new enzyme, N-carbamoyl-d-amino acid amidohydrolase, are presented in this paper. The potential application of this enzyme for the preparation of some d-amino acids used as pharmaceutical intermediates is discussed.     

Microbial degradation of oil spills enhanced by a slow-release fertilizer.

The improved cleanup of marine oil spills by stimulating biodegradation through the use of a slow-release fertilizer is reported. A paraffin-supported fertilizer containing MgNH4PO4 as active ingredient was developed and evaluated in laboratory and field experiments using quantitative infrared spectrometry and chromatographic techniques. The biodegradation of Sarir crude oil in the sea was considerably enhanced by paraffin-supported fertilizer. After 21 days 63% had disappeared as compared to 40% in the control area.