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41.
Mouse protoporphyrinogen oxidase. Kinetic parameters and demonstration of inhibition by bilirubin. 总被引:4,自引:1,他引:3
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The penultimate step of haem biosynthesis, the oxidation of protoporphyrinogen to protoporphyrin, was examined with purified murine hepatic protoporphyrinogen oxidase (EC 1.3.3.4) in detergent solution. The kinetic parameters for the two-substrate (protoporphyrinogen and oxygen) reaction were determined. The limiting Km for protoporphyrinogen when oxygen is saturating is 6.6 microM, whereas the Km for oxygen with saturating concentrations of protoporphyrinogen is 125 microM. The kcat. for the overall reaction is 447 h-1. The ratio of kcat. to the Km for protoporphyrinogen is approx. 20-fold greater than the kcat./Km,O2 ratio. The ratio of protoporphyrin formed to dioxygen consumed is 1:3. Ubiquinone-6, ubiquinone-10 and dicoumarol stimulate protoporphyrinogen oxidase activity at low concentrations (less than 15 microM), whereas coenzyme Q0 and menadione show no activation at these concentrations. Above 30 microM, all five quinones inhibit the enzyme activity. FAD does not significantly affect the activity of the enzyme. Bilirubin, a product of haem catabolism, is shown to be a competitive inhibitor of the penultimate enzyme of the haem-biosynthetic pathway, protoporphyrinogen oxidase, with a calculated Ki of 25 microM. The terminal enzyme of haem-biosynthetic pathway, namely ferrochelatase, is not inhibited by bilirubin at concentrations over double the Ki value for the oxidase. In contrast with other enzymic systems, the toxicity of bilirubin is not reversed by binding to albumin. 相似文献
42.
P R Post C N Santos R Carvalho O S Lopes R Galler 《Memórias do Instituto Oswaldo Cruz》1991,86(2):239-246
The Oswaldo Cruz Foundation produces most of the yellow fever (YF) vaccine prepared worldwide. As part of a broader approach to determine the genetic variability in YF 17D seeds and vaccines and its relevance to viral attenuation the 17DD virus was purified directly from chick embryo homogenates which is the source of virus used for vaccination of millions of people in Brazil and other countries for half a century. Neutralization and hemagglutination tests showed that the purified virus is similar to the original stock. Furthermore, radioimmune precipitation of 35S-methionine-labeled viral proteins using mouse hyperimmune ascitic fluid revealed identical patterns for the purified 17DD virus and the YF 17D-204 strain except for the 17DD E protein which migrated slower on SDS-PAGE. This difference is likely to be due to N-linked glycosylation. Finally, comparison by northern blot hybridization of virion RNAs of purified 17DD with two other strains of YF 17D virus revealed only genome-sized molecules for all three viruses. These observations suggest that the vaccine phenotype is primarily associated with the accumulation of mutations. 相似文献
43.
To study breed differences in ewes induced to parturition with dexamethasone (DXM), 15 Corriedale and 16 Polwarth ewes were injected with 15 mg of DXM four days before the expected lambing date (Days 144 and 145, respectively) in Experiment 1. Interval from treatment to parturition was twice as long for the Polwarth than for the Corriedale breed (57.9 vs 28.8 hours, P = 0.001). In Experiment 2, the effect of DXM on the interval from parturition to expulsion of the placenta and on biological parameters of the lambs (birth weight, ability to stand after birth and daily weight gain during the first week of life) was studied in 30 Corriedale ewes. On Day 144 of gestation, ewes were given 15 mg of DXM (n= 22) or 15 ml of saline (controls, n= 8). The DXM group lambed 33.8 +/- 2.7 hours after injection. Treatment caused an increase in the time of placental expulsion (4.3 vs 3.4 hours, P>/= 0.05) and on the percentage of ewes retaining the placenta beyond 6 hours (18% vs 0%, P>/= 0.05). Birth weight of lambs was lower in the induced group (3.9 vs 4.5 kg, P= 0.03), but this did not affect the other parameters of the lambs studied. 相似文献
44.
R. A. Santos J. C. G. Caldeira H. M. Pinheiro J. M. S. Cabral 《Biotechnology letters》1991,13(5):349-354
A novel aqueous two-phase system, based on polyethyleneglycol (PEG) and monosodium glutamate, was tested for the 1-dehydrogenation of hydrocortisone-based substrates. This system led to higher substrate solubilities and biocatalyst/steroid separation levels when compared with alternative systems. The addition of short-chain monohydric alcohols resulted in higher solubilities and more favourable partition coefficients for the tested substrates. Bioconversion activities in PEG/glutamate systems with 2,5% (v/v) methanol were comparable to those measured in monophasic buffer-methanol medium. 相似文献
45.
Candida albicans and three other Candida species contain an elongation factor structurally and functionally analogous to elongation factor 3. 总被引:3,自引:0,他引:3
A cell-free poly(U)-dependent translation elongation system from Candida albicans is ATP-dependent due to the presence of an elongation factor 3 (EF3)-like activity. Saccharomyces cerevisiae ribosomes added to a C. albicans postribosomal supernatant (PRS) supported poly(U)-dependent elongation, suggesting that the C. albicans lysate contained a soluble translation factor functionally analogous to the S. cerevisiae translation factor EF-3. The presence of EF-3 in C. albicans was confirmed by Western blotting using an antibody raised against S. cerevisiae EF-3. This antibody was also used to screen a selection of Candida species, all of which possessed EF-3 with molecular mass in the range of 110-130 kDa. 相似文献
46.
Nonaspiration fine needle cytology. Application of a new technique to nodular thyroid disease 总被引:2,自引:0,他引:2
The basic principle underlying fine needle aspiration (FNA) is the aspiration of cellular material from target masses, often utilizing fairly high suction pressures. The procedure requires a needle and a syringe, advisedly held in a syringe holder, enabling single-handed suction to be exercised. Mastery of the technique is variable, with few operators acquiring consistent skill. A new technique, pioneered in France but essentially unpublicized, eliminates active aspiration, replacing it by the principle of capillary suction of fluid or semifluid material into a thin channel (a fine needle). This nonaspiration sampling method was tested in a consecutive series of 50 solid thyroid nodules. Simultaneously performed conventional FNAs served as controls. Cell samples were cytologically assessed as unsuitable, diagnostic/adequate or diagnostic/superior, without knowledge of the sampling method employed. Diagnostically superior specimens were obtained significantly more frequently by the nonaspiration technique in 36 benign lesions and 13 neoplasms. The method of nonaspiration fine needle cytology ("cytopuncture") is described and illustrated, and the implications for its use in other sites are discussed. 相似文献
47.
Effects of t-butyl-4-hydroxyanisole and other phenolic antioxidants on tumoral cells and Trypanosoma parasites 总被引:1,自引:0,他引:1
J Ferreira L Coloma E Fones M E Letelier Y Repetto A Morello J Aldunate 《FEBS letters》1988,234(2):485-488
The antioxidant food additives 2(3)-tert-butyl-4-hydroxyanisole (BHA), 2,6-di(tert-butyl)-p-cresol (BHT) and the methyl and propyl esters of gallic acid inhibited Trypanosoma cruzi culture growth and oxygen consumption. The I50 values for growth and oxygen uptake with BHA were 0.284 and 0.400 and for BHT 0.083 and 0.235 mM, respectively. Moreover, BHA inhibited the respiration of several tumor cells, as well as of the procyclic and bloodstream trypomastigote forms of T. brucei brucei, with I50 in the range 0.29-0.52 mM. Inhibition of the parasites' oxygen uptake by BHA was not of the pure Michaelis-Menten type, but may be of a mixed form. It is postulated that these compounds are inhibitors because they resemble ubiquinone. 相似文献
48.
49.
Summary A pot experiment with lettuce involving three N forms each at six application levels, showed that lettuce can be grown satisfactorily
with a very low nitrate content when supplied with ammonium sulphate and a nitrification inhibitor. For plants growing on
nitrate N, the optimum midrib sap nitrate concentration as maturity approached was about 1400 mg/1 NO3-N. Large losses of mineral N were observed from the peat medium, even in the absence of plants.
A relationship is presented which would enable a lettuce grower to estimate whole-shoot nitrate concentration from a quick
test of midrib sapi.e. NO3-N (mg/kg in fresh shoot) =0.14×NO3-N (mg/l in sap).
Tipburn was worst at intermediate levels of applied N, and was less serious with pure ammonium nutrition than with nitrate. 相似文献
50.
The termite Incisitermes schwarzi has multiple sex chromosomes that have arisen by repeated translocations between autosomes and previously existing sex chromosomes. Two sex-linked allozyme loci--Acp-1 and Est-3--are holozygous, not hemizygous, in males (the heterogametic sex). Both loci show less than 1% crossing-over between X and Y chromosomes, and alleles of both are in marked disequilibrium with respect to X vs Y linkage. The two loci assort independently in female meiosis, indicating that they lie on different sex chromosomes. But they are tightly linked in male meiosis because of nonrandom assortment of the multiple X and Y chromosomes in males of this species. The findings of holozygosity and strong linkage disequilibrium suggest that differential selection in the two sexes at or near these loci may be responsible for the establishment of the translocations in this species. The existence of active Y-linked alleles also suggests that the translocations may have occurred recently. 相似文献