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41.
42.
Direct analysis of CYP21B genes in 21-hydroxylase deficiency using polymerase chain reaction amplification 总被引:8,自引:0,他引:8
Steroid 21-hydroxylase deficiency is the leading cause of impaired cortisol synthesis in congenital adrenal hyperplasia (CAH). We have studied the structure of the CYP21B gene in 30 unrelated CAH patients using the polymerase chain reaction (PCR) to differentiate the active CYP21B gene from its highly related CYP21A pseudogene. The PCR approach obviates the need to distinguish the CYP21A and CYP21B genes by restriction endonuclease digestion and electrophoresis before analysis with labeled probes. Furthermore, direct nucleotide sequence analysis of CYP21B genes is demonstrated on the PCR-amplified DNA. Gene deletion of CYP21B, gene conversion of the entire CYP21B gene to CYP21A, frame shift mutations in exon 3, an intron 2 mutation that causes abnormal RNA splicing, and a mutation leading to a stop codon in exon 8 appear to be the major abnormalities of the CYP21B gene in our patients. These mutations appear to account for 21-hydroxylase deficiency in 22 of 26 of our salt-wasting CAH patients. 相似文献
43.
The iron chelators desferrioxamine and 1-alkyl-2-methyl-3-hydroxypyrid-4-ones inhibit vascular prostacyclin synthesis in vitro. 下载免费PDF全文
The iron chelators desferrioxamine (DFO), 1,2-dimethyl(L1)-, 1-ethyl-2-methyl(L1NEt)- and 1-propyl-2-methyl(L1NPr)-3-hydroxypyrid-4-ones inhibited rat aortic prostacyclin (PGI2) synthesis in vitro (rank order of potency: DFO greater than L1 greater than L1NEt greater than L1NPr) when stimulated with adrenaline, arachidonate and the Ca2+ ionophore A23187. The inhibitory action of the chelators was blocked by Fe3+ and Al3+ and reversed by washing and H2O2, but not by ascorbate. These data suggest that iron chelators inhibit prostanoid synthesis in intact tissue through the removal or binding of Fe3+ linked to cyclo-oxygenase. These iron chelators may be of therapeutic value in the treatment of inflammatory and other diseases via two mechanisms: (1) the inhibition of pro-inflammatory prostanoid synthesis and (2) the inhibition of toxic-free-radical generation by cyclo-oxygenase. 相似文献
44.
Inka Brockhausen Arthur A Grey Henrianna Pang Harry Schachter Jeremy P Carver 《Glycoconjugate journal》1988,5(4):419-448
Sixteen asparagine-linked oligosaccharides ranging in size from (Man)2(GlcNAc)2 (Fuc)1 to (GlcNAc)6(Man)3(GlcNAc)2 were obtained from human 1-acid glycoprotein and fibrinogen, hen ovomucoid and ovalbumin, and bovine fetuin, fibrin and thyroglobulin by hydrazinolysis, mild acid hydrolysis and glycosidase treatment. The oligosaccharides hadN-acetylglucosamine at the reducing termini and mannose andN-acetylglucosamine residues at the non-reducing termini and were prepared for use asN-acetylglucosaminyltransferase substrates. Purification of the oligosaccharides involved gel filtration and high performance liquid chromatography on reverse phase and amine-bonded silica columns. Structures were determined by 360 MHz and 500 MHz proton nuclear magnetic resonance spectroscopy, fast atom bombardment-mass spectrometry and methylation analysis. Several of these oligosaccharides have not previously been well characterized.Abbreviations bis
bisecting GlcNAc
- DMSO
dimethylsulfoxide
- FAB
fast atom bombardment
- Fuc
l-fucose
- Gal
d-galactose
- GLC
gas-liquid chromatography
- GlcNAc or Gn
N-acetyl-d-glucosamine
- HPLC
high performance liquid chromatography
- Man or M
d-mannose
- MES
2-(N-morpholino)ethanesulfonate
- MS
mass spectrometry
- NMR
nuclear magnetic resonance
- PIPES
piperazine-N,N-bis(2-ethane sulfonic acid)
the nomenclature of the oligosaccharides is shown in Table 1. 相似文献
45.
Prediction and demonstration of a novel Epstein-Barr virus nuclear antigen. 总被引:5,自引:0,他引:5 下载免费PDF全文
The protein sequence predicted by the Epstein Barr virus (EBV) BERF4 open reading frame includes a tetrapeptide, Lys-Arg-Pro-Arg (KRPR), shown for other proteins to be a component of a signal for rapid nuclear localization. A subgenomic fragment of EBV DNA containing BERF4 has been incorporated into an expression vector, transfected onto primate cells and the nuclear distribution of the resulting protein established by immunofluorescence using EBV positive human sera. These sera contained high titres of antibodies to a fusion protein, produced in E. coli, consisting of beta-galactosidase and the C-terminal 167 amino acids of BERF4. Immunoaffinity purified antibodies reactive with the EBV component of the fusion show the molecular weight of this antigen in EBV immortalized B-cell lines to be about 160 kD. The demonstration that BERF4 contains an exon encoding a nuclear protein identifies a new EBNA gene (EBNA-6) and suggests that KRPR is a signal sequence common to a number of viral and cellular nuclear polypeptides which bind to nucleic acids and may therefore be of predictive value in identifying karyophilic proteins. 相似文献
46.
Novel biotransformations of 4-chlorobiphenyl by a Pseudomonas sp 总被引:7,自引:0,他引:7
A bacterium, tentatively identified as a representative of the genus Pseudomonas (strain MB86), was isolated from soil contaminated by wood-preservation chemicals by using 4-chlorobenzoate as an enrichment substrate. The pseudomonad was able to grow on 4-chlorobenzoic acid and 4-chlorobiphenyl as sole carbon and energy sources. Spent culture medium from 4-chlorobiphenyl-grown cells contained 4-chlorobenzoic acid, 4'-chloroacetophenone, 2-hydroxy,2-[4'-chlorophenyl] ethane, and 2-oxo,2-[4'-chlorophenyl] ethanol as metabolites. 4'-Chloroacetophenone was produced in large amounts, possibly as a dead-end metabolite. 相似文献
47.
The cytogenetics of domestic geese 总被引:5,自引:0,他引:5
Hybrids were produced between an African male and several Pilgrim female domestic geese. Partial karyotypes revealed a difference in the fourth largest pair of autosomal chromosomes. This chromosome pair was metacentric in the African, submetacentric in the Pilgrim, and heteromorphic in the hybrids. A similar difference between the putative wild ancestors of the African and Pilgrim breeds has been reported by others. These findings provide cytological evidence to support the traditional opinion that the African breed was derived from the Asiatic swan goose (Anser cygnoides) and the Pilgrim breed was derived from the European greylag goose (Anser anser). 相似文献
48.
Rod W. Fry Alan R. Morton Peter Garcia-Webb G P M Crawford David Keast 《European journal of applied physiology and occupational physiology》1992,64(4):335-344
Five subjects undertook 10 days of twice daily interval training sessions on a treadmill followed by 5 days of active recovery. On days 1, 6, 11, and 16 the subjects were required to undertake a test of submaximal and maximal work capacity on a treadmill combined with a performance test consisting of a run to exhaustion with the treadmill set at 18 km.h-1 and 1% gradient. Also on these days a pre-exercise blood sample was collected and analysed for a range of haematological, biochemical and immunological parameters. The subjects experienced a significant fall in performance on day 11 which had returned to pretraining levels on day 16. Serum ferritin concentrations were depressed significantly from pretraining concentrations at the conclusion of the recovery period while the expression of lymphocyte activation antigens (CD25+ and HLA-DR+) was increased both after the training phase and the recovery phase. The number of CD56+ cells in the peripheral circulation was depressed at the conclusion of the recovery period. Several parameters previously reported to change in association with overload training failing to reflect the decrease in performance experienced by subjects in this study, suggesting that overtraining may best be diagnosed through a multifactorial approach to the recognition of symptoms. The most important factor to consider may be a decrease in the level of performance following a regeneration period. The magnitude of this decreased performance necessary for the diagnosis of overtraining and the nature of an "appropriate" regeneration period are, however, difficult to define and may vary depending upon the training background of the subjects and the nature of the preceding training. It may or may not be associated with biochemical, haematological, physiological and immunological indicators. Individual cases may present a different range of symptoms and diagnosis of overtraining should not be excluded based on the failure of blood parameters to demonstrate variation. However, blood parameters may be useful to identify possible aetiology in each separate case report of over-training. An outstanding factor to emerge from this study was the difficulty associated with an objective diagnosis of overtraining and this is a possible reason why there have been new accounts of overtraining research in the literature. 相似文献
49.
C. E. Vallejos C. D. Chase 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1991,81(3):413-419
Summary Backcross and F2 progenies were produced between two bean genotypes, XR-235 and Calima, which differ in seed weight by a factor of two. The small-seeded XR-235 was used as the pistillate and recurrent parent. These genotypes showed polymorphisms at nine isozyme loci and at the phaseolin locus. Seed size parameters (weight, length, width, and thickness) were determined for each BC1 and F2 individual, i.e., for seeds harvested from XR-235 after pollination with F1 and from the F1 after selfing, respectively. A combination of starch gel electrophoresis and enzyme activity staining was used to determine the genotype of each BC1 and F2 individual at the segregating loci. SDS-PAGE and Coomassie blue staining were used to determine geno-type at the phaseolin locus. Tests for independent assortment using two-way contingency and maximum likelihood tables revealed three linkage pairs: Aco-1 — 20 cM — Dia-1; Adh-1 — 2 cM — Got-2; and Est-2 — 11 cM — Pha. Statistical comparisons were made between the means of genotype classes at each segregating locus for all seed size parameters. The results from two independently obtained BC1s and the F2 consistently indicated that the Adh-1-Got-2 segment was linked to a locus that affected seed size and overcame maternal control over seed size. This locus has been designated Ssz-1. This gene exhibited additive gene action and accounted for 30–50% of the seed size difference between the parents.Florida Agricultural Experiment Station, Journal Series No. R00696 相似文献
50.
C. E. Vallejos C. D. Chase 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1991,82(3):353-357
Summary The linkage relationship of 11 bean (Phaseolus vulgaris) seed proteins (including phaseolin), 9 enzyme loci, and theP locus were analyzed in backcross and F2 progenies by use of the software package Mapmaker. The progenies were obtained by crossing the breeding line XR-235-1 and the cultivar Calima. Allelic differences for seed protein loci were detected with SDS-PAGE and those for enzyme loci with starch gel electrophoresis and activity stains. The seed coat color of Calima is a red/beige mottled pattern and that of XR-235-1 is white. Segregation at theP locus was followed by recording the phenotype of the BC1S1 and F3 seed. A linkage group comprising ca. 90 cM was detected with the following gene order:Est-2 — 11 —Pha — 8 — (Spe/Spg) — 24 — P — 9 — (Spa/Spv) — 16 —Spba — 22 —Mdh-1. In addition, another linkage group was detected: (Spd/Spf/Sph) — 5 -Spca. Therefore, the seed proteins appear to be organized in clusters in the bean genome.Florida Agricultural Experiment Station, Journal Series No. R-01131 相似文献