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101.
Radioiodination of highly purified human follicle-stimulating hormone (hFSH) (4000 IU/mg) was performed every other week for 23 weeks using 2 mCi carrier free Na 125I (Amersham Corp., 15 mCi/μg I2) in the presence of lactoperoxidase. Incorporation of 125I into hFSH was determined by the method of [7.]Biochem. J. 89, 114). Hormone binding was studied in vitro under steady-state conditions (16 h, 20°C) using different calf testis membrane preparations having similar receptor characteristics. Each 125I-hFSH preparation was characterized for maximum bindability, specific activity of bindable radioligand as determined by self-displacement analysis, and by determination of Ka and Rt. Incorporation of 125I into FSH was relatively constant over the large number of experiments (62.4 ± 6.4 μCi/μg; n = 23). By comparison, however, specific radioactivity of the receptor bindable fraction of 125I-hFSH was related to the lot of 125I utilized, and was significantly (P ≤ 0.01) lower and more variable (28.7 ± 10.5 μCi/μg). Maximum bindability of 125I-hFSH was not correlated to specific activity (r = 0.06) but was negatively correlated to hFSH 125I incorporation (r = −0.47; P ≤ 0.05). These observations demonstrate the need to assess the quality of each batch of radioligand before undertaking radioligand-receptor assays and suggest that differences in Na125I lots affect specific radioactivity of the radioligand and its receptor binding characteristics. 相似文献
102.
George A. Antonelis Jr. Mark S. Lowry Douglas P. DeMaster Clifford H. Fiscus 《Marine Mammal Science》1987,3(4):308-322
Stomach lavaging was used to study the feeding habits of northern elephant seals ( Mirounga angustirostris ) found on San Miguel Island, California, during the spring of 1984. Fifty-nine elephant seals were chemically immobilized with an intramuscular injection of ketamine hydrochloride. Once immobilized, an animal's stomach was intubated, filled with 3–4 liters of water to create a slurry of the undigested food items, and evacuated into a collection device. The stomachs of 57 (96.6%) of the animals lavaged contained identifiable parts of prey. Twenty-nine different food items were identified, 12 of which have not been previously reported as prey of the northern elephant seal: two teleost fish, Coryphaenoides acrolepis (Pacific rattail) and another unidentified macrourid; two crustaceans, Pasiphaea pacifica (glass shrimp) and Euphausia sp.; six squid, Abraliopsis felis, Gonatus berryi, Histioteuthis dofleini, Cranchia scabra, Taonius pavo, and Galiteuthis sp. and two octopi, Octopus dofleini and Octopus rubescens. 相似文献
103.
Robert L. Brownell Jr. Lloyd T. Findley Omar Vidal Alejandro Robles Silvia Manzanilla N 《Marine Mammal Science》1987,3(1):22-30
The vaquita, Phocoena sinus , is a porpoise in the family Phocoenidae that lives only in the Gulf of California. The external appearance of P. sinus was unknown until 13 fresh specimens were recently examined. The most obvious morphological feature distinguishing P. sinus from its two congeners is the proportionately higher dorsal fin. The most striking features of the pigmentation pattern are the large black eye patches and the black upper and lower lip patches. In both areas, the pigmentation contrasts sharply with the surrounding light gray coloration. The total lengths of the specimens ranged from 70.3 cm (a neonate) to 143.5 cm (an adult female). 相似文献
104.
105.
K D Bradshaw M R Waterman R T Couch E R Simpson M X Zuber 《Molecular endocrinology (Baltimore, Md.)》1987,1(5):348-354
To provide a basis for investigation of the molecular mechanisms underlying the hormonal regulation of steroid 17 alpha-hydroxylase (P-450 17 alpha) activity in adrenal, ovary, and testis as well as human 17 alpha-hydroxylase deficiency, we have isolated from a human fetal adrenal cDNA library a cDNA sequence complementary to the mRNA that encodes the human P-450 17 alpha enzyme. Of 75,000 colonies from the library that were screened by use of a nick-translated 5'-specific bovine P-450 17 alpha cDNA probe, 10 positive colonies were isolated and the clone with the longest insert (pcD-17 alpha H) was selected for further characterization. pcD-17 alpha H encodes the complete human P-450 17 alpha protein having approximately 78% homology at the nucleotide level and 71% homology at the amino acid level when the sequence of pcD-17 alpha H is compared to the bovine P-450 17 alpha cDNA sequence. By transient expression of the human P-450 17 alpha cDNA clone in COS 1 cells, we have demonstrated that the 17 alpha-hydroxylase and 17,20 lyase activities reside within the same human P-450 17 alpha polypeptide chain. The insert was also used as a probe to investigate, by means of Southern blot analysis, possible alterations in the P-450 17 alpha gene sequence in DNA isolated from skin fibroblasts from three patients with clinically characterized 17 alpha-hydroxylase deficiencies. No changes were detected in the DNA of any of the patients by this analysis. 相似文献
106.
Extensive degradation of Aroclors and environmentally transformed polychlorinated biphenyls by Alcaligenes eutrophus H850. 总被引:14,自引:8,他引:6
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D L Bedard R E Wagner M J Brennan M L Haberl J F Brown Jr 《Applied microbiology》1987,53(5):1094-1102
We have isolated and characterized a strain of Alcaligenes eurtrophus, designated H850, that rapidly degrades a broad and unusual spectrum of polychlorinated biphenyls (PCBs) including many tetra- and pentachlorobiphenyls and several hexachlorobiphenyls. This strain, which was isolated from PCB-containing dredge spoils by enrichment on biphenyl, grows well on biphenyl and 2-chlorobiphenyl but poorly on 3- and 4-chlorobiphenyl. Capillary gas-chromatographic analysis showed that biphenyl-grown resting cells of H850 degraded the components of 38 of the 41 largest peaks of Aroclor 1242 and 15 of the 44 largest peaks of Aroclor 1254, resulting in an overall reduction of PCBs by 81% for Aroclor 1242 (10 ppm) and 35% for Aroclor 1254 (10 ppm) in 2 days. Furthermore, H850 metabolized the predominantly ortho-substituted PCB congeners that resulted from the environmental transformation of the more highly chlorinated congeners of Aroclor 1242 by the upper Hudson River anaerobic meta-, para-dechlorination agent system C (J. F. Brown, R. E. Wagner, Jr., D. L. Bedard, M. J. Brennan, J. C. Carnahan, R. J. May, and J. J. Tofflemire, Northeast Environ. Sci. 3:167-179, 1984). The congener selectivity patterns indicate that a two-step process consisting of anaerobic dechlorination followed by oxidation by H850 can effectively degrade all of the congeners in Aroclor 1242 and possibly all those in Aroclor 1254. 相似文献
107.
Design, construction, and use of an electroporator for plant protoplasts and animal cells 总被引:2,自引:0,他引:2
H D Bradshaw W W Parson M Sheffer P J Lioubin E R Mulvihill M P Gordon 《Analytical biochemistry》1987,166(2):342-348
We have designed and constructed an electroporation device capable of efficient transfer of DNA into both plant cell protoplasts and cultured murine lymphocytes. The electroporator design allows various combinations of voltage and capacitance to be used to optimize the electric pulse. Switching of large voltages and currents is accomplished with a silicon-controlled rectifier, yielding excellent reproducibility and long component life. A safety switch is provided to permit complete discharge of the device. Conditions suitable for high levels of transient expression and high frequencies of stable transformation for both plant and animal cell systems have been found. 相似文献
108.
Structure of porcine heart cytoplasmic malate dehydrogenase: combining X-ray diffraction and chemical sequence data in structural studies 总被引:1,自引:0,他引:1
The amino acid sequence of cytoplasmic malate dehydrogenase (sMDH) has been determined by a combination of X-ray crystallographic and chemical sequencing methods. The initial molecular model incorporated an "X-ray amino acid sequence" that was derived primarily from an evaluation of a multiple isomorphous replacement phased electron density map calculated at 2.5-A resolution. Following restrained least-squares crystallographic refinement, difference electron density maps were calculated from model phases, and attempts were made to upgrade the X-ray amino acid sequence. The method used to find the positions of peptides in the X-ray structure was similar to those used for studying protein homology and was shown to be successful for large fragments. For sMDH, X-ray methods by themselves were insufficient to derive a complete amino acid sequence, even with partial chemical sequence data. However, for this relatively large molecule at medium resolution, the electron density maps were of considerable help in determining the linear position of peptide fragments. The N-acetylated polypeptide chain of sMDH has 331 amino acids and has been crystallographically refined to an R factor of 19% for 2.5-A resolution diffraction data. 相似文献
109.
Amino acid sequence of guinea pig prostate kallikrein 总被引:1,自引:0,他引:1
The primary structure of the major arginine esteropeptidase from guinea pig prostate has been deduced from automated Edman degradation of peptides generated by clostripain, cyanogen bromide, endoproteinase Lys-C, and Staphylococcus aureus V8 protease digestion of the protein. The esteropeptidase is a single polypeptide chain comprised of 239 amino acids and contains 2 apparent sites of carbohydrate attachment, Asn-78 and Asn-169. Both occur in consensus sequences for N-linked glycosylation sites. The esteropeptidase exhibits approximately 35% homology with trypsin including conservation of the catalytic residues and the aspartic acid which confers specificity toward basic amino acids. The sequence identity, however, extends to greater than 60% with the kallikrein family of serine proteases. In addition to the overall homology, the guinea pig enzyme displays a number of features characteristic of kallikreins including 10 conserved half-cystine residues, a C-terminal proline, and the "kallikrein loop". On the basis of this structural relatedness, the enzyme has been designed as guinea pig prostate kallikrein. In contrast to many of the kallikreins of other species and tissues, this enzyme does not contain any sites within the kallikrein loop sensitive to proteases that result in internal breaks in the polypeptide chain. 相似文献
110.
Cardiac tissue can be considered macroscopically as a bidomain, anisotropic conductor in which simple depolarization wavefronts produce complex current distributions. Since such distributions may be difficult to measure using electrical techniques, we have developed a mathematical model to determine the feasibility of magnetic localization of these currents. By applying the finite element method to an idealized two-dimensional bisyncytium with anisotropic conductivities, we have calculated the intracellular and extracellular potentials, the current distributions, and the magnetic fields for a circular depolarization wavefront. The calculated magnetic field 1 mm from the tissue is well within the sensitivity of a SQUID magnetometer. Our results show that complex bisyncytial current patterns can be studied magnetically, and these studies should provide valuable insight regarding the electrical anisotropy of cardiac tissue. 相似文献