Reductive biotransformations of organic compounds by cells or enzymes of yeast.

Saccharomyces cerevisiae catalyses the asymmetric reductive biotransformation of a variety of compounds containing a carbonyl group or carbon-carbon double bond. Oxidoreductases participating in these reactions which have commercial potential in biotransformation processes are likely to have relatively broad substrate specificity. Important carbonyl reductases falling into this category include YADH- and yeast NADP-dependent beta-ketoester reductases. The enoyl reductase component of the FAS complex may have a role in asymmetric yeast reduction of carbon-carbon double bonds of unnatural substrates. Other nicotinamide-requiring oxidoreductases of yeast are also surveyed to rationalize observed biotransformations of whole yeast cells in terms of specific enzymes. Genetic and protein engineering may enable enzymes to be tailored to accept new substrates. A greater understanding of the enzymes and reactions involved will facilitate further optimization and exploitation of these catalytic systems in industrial processes.     

3H]L-657,743 (MK-912): a new, high affinity, selective radioligand for brain alpha 2-adrenoceptors

L-657,743 (MK-912), a highly potent and selective alpha 2-adrenoceptor antagonist was tritiated to a high specific activity and its binding characteristics to brain tissue were determined. The specific binding of [3H]L-657,743 to rat cerebrocortex was saturable, reversible, and dependent on tissue concentration. In saturation studies, [3H]L-657,743 binding was resolved into two high affinity components exhibiting Kd values of 86 pM and 830 pM with densities of 82 fmol/mg protein and 660 fmol/mg protein, respectively. Based on the binding potencies of a variety of compounds with differing receptor selectivities, the sites labeled by [3H]L-657,743 were characteristic of alpha 2-adrenoceptors. In contrast to alpha 2-antagonists, alpha 2-agonists displayed shallow competition curves. In the presence of 100 microM GTP, Gpp(NH)p or 150 mM NaCl, the competition curve for epinephrine was shifted to the right, whereas that for yohimbine was unaffected. In studies utilizing human cerebrocortical tissue, [3H]L-657,743 also bound with high affinity to sites characteristic of alpha 2-adrenoceptors.     

Chromogenic redox assay for beta-lactamases yielding water-insoluble products. II. Heterogeneous sandwich assay for hCG.

A very sensitive and rapid heterogeneous sandwich enzyme immunoassay for human chorionic gonadotropin (hCG) is described. The assay is based on the application of the novel chromogenic redox substrate system for beta-lactamase which is used as label. The chromogen system consists of a thioacetylcephalosporin beta-lactamase substrate, which upon turnover by the enzyme label releases the thiolate with the concomitant reduction of the tetrazolium salt to a colored formazan. The concentration of the formazan is directly related to the amount of the hormone in the sample and is read spectrophotometrically. The enzyme-antibody conjugates, produced through use of heterobifunctional maleimide crosslinker, maintain 90% of the enzyme activity after 30 days at 25 degrees C. Concentrations of the hormone as low as 5 mIU/ml, equivalent to 25 fmol/ml, are detectable in 3 h.     

Embodying illness,embodying cancer

Individuals and societies embody illnesses in different ways, in part determined by the way a person knows and lives his or her diagnosis and prognosis. Based on research in Northern Italy, on the experiences and meanings of cancer and on the practice of nondisclosure of the diagnosis, we find nondisclosure reflects a world divided - life/death, good/bad, mind/body — with the unwanted converted to other. The strong association of cancer with death, suffering, and hopelessness in much of Italy, coupled with the tremendous power attributed to naming and sentencing makes nondisclosure a major mechanism for keeping the condemned in this social world, and keeping death, decay, and suffering in the other. It is the social reality that is dominant here, such that informing a patient of cancer can be tantamount to social death.

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Résumé Les individus et les sociétés incorporent la maladie de façon différente, déterminée en partie de comment une personne connait et vit son diagnostic et prognostic. A partir de la recherche des experiences des significations du cancer et de la pratique de ne pas dire la diagnostic au Nord de l'Italie, on a remarqué que l'habitude de ne rien dire reflète un monde séparé entre la vie et la mort, entre le bon et le mal, entre l'esprit et le corps, de sorte que ce qui West pas voulu soit transfomé en l'autre. L'association forte du cancer à la mort, à la souffrance, au désespérance en toute l'Italie, unie au grand pouvoir donne au fait de dénommer et de donner une sentence, rend ne pas dire un méchanisme important pour garder le condamnd dans ce monde social et pour garder la mort, la décadence et la souffrance dans l'autre. C'est la réalité sociale qui est ici dominante, tel que le fait d'informer un patient de cancer soit comme une mort sociale.

     

Validation of age readings from otoliths of juvenile roundnose grenadier, Coryphaenoides rupestris, a deep-water macrourid fish

Validation of the ageing of deep-water fish is difficult and there are only a few instances where the rings on the otoliths have been shown to be laid down annually. Roundnose grenadier have been fished commercially in the North Atlantic since the 1960s and the adult fish have frequently been aged by counting the rings in otoliths or scales. All the ageing was done on the assumption that the rings in the otoliths or scales were annual. Between 1975 and 1992, the Scottish Association for Marine Science carried out seasonal trawling surveys in the Rockall Trough using a fine-meshed trawl, and collected otoliths from a wide size range of roundnose grenadier. An examination of the growing edge of otoliths from juvenile fish from these collections suggests that the rings in the otoliths are laid down annually. The broader, opaque zones which represent the growth phase were dominant between September and March. The thinner, hyaline zones were dominant between April and July. The apparent delay in the growth phase compared with most shallow-water species is discussed in relation to the availability of mesopelagic prey.     

Control of the Amiloride-Sensitive Na+ Current in Salivary Duct Cells by Extracellular Sodium

We have previously reported that intralobular salivary duct cells contain an amiloride-sensitive Na⁺ conductance (probably located in the apical membranes). Since the amiloride-sensitive Na⁺ conductances in other tight epithelia have been reported to be controlled by extracellular (luminal) Na⁺, we decided to use whole-cell patch clamp techniques to investigate whether the Na⁺ conductance in salivary duct cells is also regulated by extracellular Na⁺. Using Na⁺-free pipette solutions, we observed that the whole-cell Na⁺ conductance increased when the extracellular Na⁺ was increased, whereas the whole-cell Na⁺ permeability, as defined in the Goldman equation, decreased. The dependency of the whole-cell Na⁺ conductance on extracellular Na⁺ could be described by the Michaelis-Menten equation with a K _m of 47.3 mmol/1 and a maximum conductance (G _max) of 2.18 nS. To investigate whether this saturation of the Na⁺ conductance with increasing extracellular Na⁺ was due to a reduction in channel activity or to saturation of the single-channel current, we used fluctuation analysis of the noise generated during the onset of blockade of the Na⁺ current with 200 μmol/l 6-chloro-3,5-diaminopyrazine-2-carboxamide. Using this technique, we estimated the single channel conductance to be 4 pS when the channel was bathed symmetrically in 150 mmol/l Na⁺ solutions. We found that Na⁺ channel activity, defined as the open probability multiplied by the number of available channels, did not alter with increasing extracellular Na⁺. On the other hand, the single-channel current saturated with increasing extracellular Na⁺ and, consequently, whole-cell Na⁺ permeability declined. In other words, the decline in Na⁺ permeability in salivary duct cells with increasing extracellular Na⁺ concentration is due simply to saturation of the single-channel Na⁺ conductance rather than to inactivation of channel activity. Received: 27 July 1995/Revised: 7 December 1995     

Floristic diversity and co-occurrences in a subtropical broad-leaved forest and two contrasting regrowth stands in central-west Yunnan Province,China

Many of the natural forested ecosystems that still remain in mainland China are being cleared with potentially detrimental effects on woody plant species diversity on both local and regional scales. The most extensive stand of subtropical broad-leaved forest remaining in China is located in Yunnan Province. In an effort to document the influence of human-induced disturbance on Yunnan's woody flora, floristic inventories were conducted in a stand of primary forest and in regrowth stands located in its interior and along its outer margin in the Xujiaba Nature Sanctuary in the Ailao Mountain Range. Of particular interest was the location of the disturbance relative to the primary forest source area. A total of 134 woody plant species representing 74 genera and 43 families were recorded. The floristics of the two regrowth stands were significantly different from each other, with < 10% of their respective floras comprised of co-occurring species. The interior regrowth stand had a higher number of co-occurring species with the primary forest; however, > 40% were still non-co-occurring.The principal families represented in the primary forest and the interior regrowth stand were Aquifoliaceae, Berberidaceae, Fagaceae, Lauraceae, Rosaceae, Smilacaceae, Symplocaceae, Theaceae, and Vacciniaceae. The three dominant species with relative importance values ranging from > 5% to 18% in both the primary forest and the interior regrowth stand were Castanopsis wattii, Lithocarpus jingdongensis, and Symplocos sumuntia. The edge regrowth stands had the lowest species diversity and were dominated by the native pine Pinus yunnanensis, with a relative importance of 24%. The principal families represented in the edge regrowth stand were Betulaceae, Ericaceae, Fagaceae, Myricaceae, Pinaceae, and Theaceae. Only the Fagaceae and Theaceae were well-represented in all three stands. The results of the study document the low species diversity in post-cutting regrowth on the margins of the primary forest as compared with post-cutting regrowth in the forest interior.