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31.
The kinetics of NADH oxidation by the outer membrane electron transport system of intact beetroot (Beta vulgaris L.) mitochondria were investigated. Very different values for Vmax and the Km for NADH were obtained when either antimycin A-insensitive NADH-cytochrome c activity (Vmax= 31 ± 2.5 nmol cytochrome c (mg protein)?1 min?1; Km= 3.1 ± 0.8 μM) or antimycin A-insensitive NADH-ferricyanide activity (Vmax= 1.7 ± 0.7 μmol ferricyanide (mg protein)?1 min?1; Km= 83 ± 20 μM) were measured. As ferricyanide is believed to accept electrons closer to the NADH binding site than cytochrome c, it was concluded that 83 ± 20 μM NADH represented a more accurate estimate of the binding affinity of the outer membrane dehydrogenase for NADH. The low Km determined with NADH-cytochrome c activity may be due to a limitation in electron flow through the components of the outer membrane electron transport chain. The Km for NADH of the externally-facing inner membrane NADH dehydrogenase of pea leaf (Pisum sativum L. cv. Massey Gem) mitochondria was 26.7 ± 4.3 μM when oxygen was the electron acceptor. At an NADH concentration at which the inner membrane dehydrogenase should predominate, the Ca2+ chelator, ethyleneglycol-(β-aminoethylether)-N,N,-tetraacetic acid (EGTA), inhibited the oxidation of NADH through to oxygen and to the ubiquinone-10 analogues, duroquinone and ubiquinone-1, but had no effect on the antimycin A-insensitive ferricyanide reduction. It is concluded that the site of action of Ca2+ involves the interaction of the enzyme with ubiquinone and not with NADH. 相似文献
32.
Kathleen W. Miller 《Applied microbiology》1992,58(7):2176-2179
Dibenzyldisulfide was reductively degraded by a methanogenic mixed culture derived from a sewage digestor. Toluene was produced with benzyl mercaptan as an intermediate in sulfur-limited medium. Toluene production was strictly associated with biological activity; however, the reducing agent for the culture medium, Ti(III), was partially responsible for production of benzyl mercaptan. Sulfide was not detected. Additions of sodium sulfide did not inhibit toluene production. Additions of 2-bromoethane sulfonic acid prevented methanogenesis but did not adversely affect toluene yields. 相似文献
33.
Hillary A. Hahm Margot M. Ip Kathleen Darcy Jennifer D. Black Wendy K. Shea Suzanne Forczek Masami Yoshimura Takami Oka 《In vitro cellular & developmental biology. Plant》1990,26(8):803-814
Summary A serum-free primary culture system is described which allows normal rat mammary epithelial cells (RMECs) embedded within
a reconstituted basement membrane to undergo extensive growth and functional differentiation as detected by synthesis and
secretion of the milk products casein and lipid. RMECs isolated from mammary glands of immature virgin rats were seeded within
an extracellular matrix preparation derived from the Engelbreth-Holm-Swarm sarcoma and cultured in a serum-free medium consisting
of Dulbecco's modified Eagle's medium-F12 containing insulin, prolactin, progesterone, hydrocortisone, epidermal growth factor,
bovine serum albumin, transferrin, and ascorbic acid. Casein synthesis and secretion were documented at the electron microscopic
level as well as by an enzyme-linked immunosorbent assay (ELISA) assay using a polyclonal antibody against total rat caseins.
Numerous secretory vesicles with casein micelles were noted near the apical surface of the RMECs, and secreted casein was
observed in the lumen. These ultrastructural data were confirmed by the ELISA assay which showed that microgram amounts of
casein per well were synthesized by the RMECs and that the amount of casein increased with time in culture. Using immunoblot
analysis it was demonstrated that the full complement of casein proteins was synthesized. In addition to casein protein, β-casein
mRNA levels were shown to increase with time. Synthesized lipid was detected at both the light and electron microscopic levels.
Phase contrast photomicrographs demonstrated extensive intracellular lipid accumulation within the ductal and lobuloalveolarlike
colonies, and at the electron micrograph level, lipid droplets were predominantly localized near the apical surface of the
RMECs. The lipid nature of these droplets was verified by oil red O staining. Results from this study demonstrate that RMECs
from immature virgin rats proliferate extensively and rapidly develop the capacity to synthesize and secrete casein and lipid
when grown within a reconstituted basement membrane under defined serum-free conditions. This unique system should thus serve
as an excellent model in which the regulation of mammary development and gene expression can be investigated.
This work was supported by grants CA 33240 and CA 35641 and by core grant CA 24538 from the National Institutes of Health,
Bethesda, MD. 相似文献
34.
Kathleen D. Murray Eleanor Groden Francis A. Drummond A. Randall Alford Richard H. Storch Michael D. Bentley 《Entomologia Experimentalis et Applicata》1996,80(3):503-510
The effects of citrus limonoids, applied topically to potato (Solanum tuberosum L. var. Katahdin) foliage, on Colorado potato beetle (Leptinotarsa decemlineata) (Say) (Chrysomelidae) larval development, growth, and survival were quantified in laboratory assays and a small-plot field test. In laboratory assays, survival, development rate, and body weight decreased with increasing limonoid concentration, however these measures of larval response did not significantly differ among varying periods of limonoid exposure (three, six, or nine days). Significant limonoid application concentration and frequency effects on survival, development rate, and defoliation were observed in the field test. These results indicate the potential utility of lethal and non-lethal effects of citrus limonoids for management of the Colorado potato beetle. 相似文献
35.
Kathleen L. Summers Dr. Patrick Lippiello Ezio Giacobini 《Neurochemical research》1996,21(10):1181-1186
Transcortical dialysis was employed to investigate the effects of subcutaneous (s.c.) injections of RJR-2403 (1.2–7.2 μmol/kg)
on extracellular levels of acetylcholine (ACh), norepinephrine (NE), dopamine (DA), and serotonin (5-HT) in rat. Systemic
administration of RJR-2403 produced a 90% increase of cortical extracellular ACh levels that persisted for up to 90 minutes
after injection. Norepinephrine and DA release were increased 124% and 131% above basal values, respectively. Serotonin (5-HT)
levels in the dialysate were also significantly elevated by RJR-2403 (3.6 μmol/kg, s.c.) 70% above baseline at 90 minutes
post-injection. Comparison of these responses to those of (−)nicotine from a previous study reveals little difference between
the two compounds in their ability to influence cortical neurotransmitter release following systemic administration. 相似文献
36.
Charlene Chang Ben A. Moll Kathleen B. Evenson Mark J. Guiltinan 《Plant Cell, Tissue and Organ Culture》1996,45(1):61-66
In vitro plantlet regeneration systems for the seed geranium (Pelargonium x hortorum Bailey) using cotyledon, hypocotyl and root explants were optimized by studying the influence of seedling age, growth regulators and excision orientation on organogenesis. Indole-3-acetic acid combined with zeatin yielded the highest rate of shoot production on cotyledon explants (0.2–2 shoots per explant). More shoots were produced on explants cut from the most basal region of cotyledons from 2 to 4-day-old seedlings than from older seedlings or more distal cut sites. Hypocotyl explants produced the highest number of shoots, up to 40 shoots per explant, on indole-3-acetic acid (2.8–5.6 mM) + zeatin (4.6 mM) or thidiazuron (4.5 mM). Maximum shoot formation (0.3–1.4 shoots per explant) on root explants occurred when they were cultured on medium containing zeatin. Regenerated shoots rooted best on a basal medium containing no growth regulators. There were substantial differences among cultivars in shoot formation from each of the explant systems.Abbreviations BA
6-benzylaminopurine
- 2,4-d
2,4-dichlorophenoxyacetic acid
- IAA
indole-3-acetic acid
- NAA
naphthaleneacetic acid
- TDZ
thidiazuron 相似文献
37.
Kathleen Miller 《Biology & philosophy》1994,9(1):99-103
In Can Abstractions be Causes, David Johnson defends the view that abstractions can have causal force. He offers as his own example of natural kinds ecological niches, arguing that the causal force of these niches in nature is akin to the force of Aristotelian final causes. He concludes that, rooted as it is in seventeenth century mechanism, the currently-accepted model of causality which recognises only efficient causes is inadequate to the needs of contemporary science. In Natural Kinds and Ecological Niches — Response to Johnson's Paper, Melinda Hogan offers a critique of Johnson's paper which, by begging the question in favor of the very sort of causality Johnson seeks to supplement, misses the epistemological implications of his idea. In this paper I will attempt to clarify and defend Johnson's position, pointing out some of its implications for the epistemology of science in general. 相似文献
38.
Simon C. Allen K. Ravi Acharya Kathleen A. Palmer Robert Shapiro Bert L. Vallee Harold A. Scheraga 《Journal of Protein Chemistry》1994,13(7):649-658
The three-dimensional structure of human angiogenin has been determined by X-ray crystallography and is compared here with an earlier model which predicted its structure, based on the homology of angiogenin with bovine pancreatic ribonuclease A. Comparison of the predicted model and crystal structure shows that the active-site histidine residues and the core of the angiogenin molecule, including most of the-strands and-helices, were predicted reasonably well. However, the structure of the surface loop regions and residues near the truncated C-terminus differs significantly. The C-terminal segment includes the active-site residues Asp-116, Gln-117, and Ser-118; Gln-117 in particular has been shown to be important in affecting the ribonucleolytic activity of angiogenin. Also, the orientation of one helix in the model differed from the orientation observed experimentally by about 20°, resulting in a large displacement of this chain segment. The difficulty encountered in predicting the surface loop regions has led to a new algorithm [Palmer and Scheraga (1991),J. Comput. Chem.,12, 505–526; (1992),J. Comput. Chem.,13, 329–350] for predicting the conformations of surface loops. 相似文献
39.
Locus Heterogeneity for Waardenburg Syndrome is Predictive of Clinical Subtypes 总被引:5,自引:4,他引:1
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Lindsay A. Farrer Kathleen S. Arnos James H. Asher Clinton T. Baldwin Scott R. Diehl Thomas B. Friedman Jacquie Greenberg Kenneth M. Grundfast Christopher Hoth Anil K. Lalwani Barbara Landa Kate Leverton Aubrey Milunsky Robert Morell Walter E. Nance Valerie Newton Rajkumar Ramesar Valluri S. Rao Jennifer E. Reynolds Theresa B. San Agustin Edward R. Wilcox Ingrid Winship Andrew P. Read 《American journal of human genetics》1994,55(4):728-737
Waardenburg syndrome (WS) is a dominantly inherited and clinically variable syndrome of deafness, pigmentary changes, and distinctive facial features. Clinically, WS type I (WS1) is differentiated from WS type II (WS2) by the high frequency of dystopia canthorum in the family. In some families, WS is caused by mutations in the PAX3 gene on chromosome 2q. We have typed microsatellite markers within and flanking PAX3 in 41 WS1 kindreds and 26 WS2 kindreds in order to estimate the proportion of families with probable mutations in PAX3 and to study the relationship between phenotypic and genotypic heterogeneity. Evaluation of heterogeneity in location scores obtained by multilocus analysis indicated that WS is linked to PAX3 in 60% of all WS families and in 100% of WS1 families. None of the WS2 families were linked. In those families in which equivocal lod scores (between −2 and +1) were found, PAX3 mutations have been identified in 5 of the 15 WS1 families but in none of the 4 WS2 families. Although preliminary studies do not suggest any association between the phenotype and the molecular pathology in 20 families with known PAX3 mutations and in four patients with chromosomal abnormalities in the vicinity of PAX3, the presence of dystopia in multiple family members is a reliable indicator for identifying families likely to have a defect in PAX3. 相似文献
40.
Synopsis The early development of the southern mouthbrooder,Pseudocrenilabrus philander, is documented from activation until the early stages of the juvenile period. The duration of the embryonic period is about 14 days at 25°C. Development is direct and there is accelerated exogenous feeding into the embryonic period. The pattern of development and the timing of ontogenetic events and structure formation are a reflection of both internal and external environmental conditions. During mouthbrooding, oxygen uptake is facilitated by embryonic respiratory plexuses and flapping of the pectoral fins. At the time of first release from the buccal cavity, the embryos are in an advanced state of development. The switch-over from the temporary embryonic respiratory system to the adult branchial system has occurred. The yolksac serves as a supplemental source of nutrition as the embryos develop their external food-gathering abilities. The skeletal and sensory systems are sufficiently developed to allow the young to return to the safety of the female's buccal cavity. Pigmentation may provide disruptive colouration. The rate and pattern of development of another mouthbrooding cichlid,Oreochromis mossambicus, is similar to that ofP. philander despite their phylogenetic differences, and may be a consequence of similar life-history styles. 相似文献