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11.
Mary Taub Milton H. Saier Lorraine Chuman Sue Hiller 《Journal of cellular physiology》1983,114(2):153-161
Prostaglandin E1(PGE1), one of the components in the hormone-supplemented, serum-free medium for Madin Darby Canine Kidney (MDCK) cells (Medium K-1), is required for both long-term growth and for dome formation. Variant cells have been isolated from MDCK populations, which lack the PGE1, requirement for long-term growth in Medium K-1. These variants will be useful in identifying the molecular events initiated by PGE1 which are necessary for the growth response to be observed. The growth and functional properties of five independently isolated PGE1 independent clones have been examined. Normal MDCK cells grew at an equivalent rate in Medium K-1 and in serum-supplemented medium; the growth rate was lower in Medium K-1 lacking PGE1. In contrast, PGE1 independent clone 1 grew at an equivalent rate in Medium K-1 minus PGE1, and in serum-supplemented medium. When PGE1 was added to K-1 minus PGE1, less growth of PGE1 independent clone 1 was observed. A similar observation was made with one other PGE1 independent clone which was studied. A hormone deletion study indicated that PGE1 independent clone 1 still retained growth responses to the other four supplements in Medium K-1 (insulin, transferrin, T3, and hydrocortisone). The molecular alterations associated with loss of the PGE1 requirement for long-term growth were examined. At confluency, all of the PGE1 independent clones studied had higher intracellular cyclic AMP levels following PGE1 treatment, as compared with normal MDCK cells. The increased cyclic AMP levels in the variant cells could result from a number of different types of defects, including reduced cyclic adenylic acid (cyclic AMP) efflux, an increased affinity of PGE2 for the PGE1 receptor, or a defect in cyclic AMP metabolism. However, in all of the variant clones studied a decreased rate of cyclic AMP degradation by cyclic AMP phosphodiesterase was observed. Thus, the increased cyclic AMP levels in the PGE1 independent variants may result from alterations which affect cyclic AMP metabolism. The effect of PGE1 on dome formation by the variant cells was also examined. The frequency of dome formation by PGE1 independent clone 1 was enhanced in a dosage-dependent manner, like normal MDCK cells. This observation suggests that PGE1 affects MDCK cell growth and dome formation by different mechanisms. 相似文献
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Jeff Reeve Lorraine H. Kligman Robert Anderson 《Applied microbiology and biotechnology》1990,33(2):161-166
Summary Isolated lipids from Deinococcus radiodurans were reconstituted at final concentrations of 1 mg/ml into dioleoyl phosphatidyl choline (DOPC) vesicles and assayed for the ability to protect cells of Escherichia coli against killing by UV light (254 nm). Values of D37 (UV dose required to reduce the number of surviving cells to 37% of the original number) were calculated from killing curves. E. coli was afforded the greatest protection with an individual lipid, identified as vitamin MK8 (D37=310 J//m2, compared to D37=67 J/m2 for E. coli irradiated in the presence of DOPC alone). Liposome-mediated protection was dependent on UV254 absorbance and not on turbidity-related light-scattering. BOth vitamin MK8 from D. radiodurans and vitamin K1, which is available commercially, showed a similar degree of UV254-protection for E. coli. The UV-protective properties of vitamin K1 were also investigated on mammalian cells in comparison with other natural lipids and known sunscreens. Survival curves were obtained for mouse fibroblast (L) cells irradiated at UV254 in the absence or presence of DOPC liposomes into which were incorporated various natural lipids or standard sunscreen ingredients, all at final concentrations of 1 mg/ml. Experimentally determined values of D37 were as follows: Vitamin K1, 73 J/m2; \-carotene, 44 J/m2; -tocopherol, 20 J/m2; sulisobenzone, 156 J/m2; p-aminobenzoic acid (PABA), 113 J/m2; benzophenone, 80 J/m2; oxybenzone, 61 J/m2 and DOPC alone. 23 J/m2. Vitamin K1, the most protective lipid tested, was also compared with PABA and oxybenzone (all at concentrations of 20 mg/ml; applied topicall) for its ability to protec Skh-hairless mice from UV254-induced erythema, yielding a UV254 protection factor of 3.5. In addition, vitamin K1 (at 100 mg/ml) was able to provide hairless mice with a small degree of UVB protection, as indicated by an experimentally determined Solar Protection Factor of 1.5–2.0. Although it is concluded that vitamin K is not likely to account for the extraordinarily high degree of UV-resistance of D. radiodurans, vitamin K does show characteristics worthy of its consideration as a UV-screening agent.
Offprint requests to: R. Anderson 相似文献
14.
T. C. Hsu Edward J. Shillitoe Lorraine M. Cherry Qi Lin Stimson P. Schantz Cynthia Furlong 《In vitro cellular & developmental biology. Plant》1990,26(1):80-84
Summary Forty lymphoblastoid (lymphoid) lines were established from 42 volunteer blood donors, including healthy individuals and patients
with head and neck carcinomas. Each peripheral blood sample was split into two portions, one for the establishment of a lymphoid
line and the other for short-term culture, which was used to estimate bleomycin sensitivity by cytogenetic procedures. Twenty
lymphoid lines were selected at random to compare bleomycin sensitivity with data obtained from short-term lymphocyte cultures.
In each set, bleomycin sensitivity of lymphoid cells was similar to that of the lymphocytes. The lymphoid lines, which can
be propagated for an unlimited supply of relatively homogeneous cellular material, will be useful for a variety of future
investigations.
This investigation was supported by grants from the John S. Dunn Foundation, Houston, TX, the Esther Knispel Fund administered
by The University of Texas M. D. Anderson Cancer Center, Houston, TX, and Department of Health and Human Services PHS grant
DE 07007. 相似文献
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Changes in maximum oxygen uptake during prolonged training, overtraining, and detraining in horses 总被引:2,自引:0,他引:2
Tyler Catherine M.; Golland Lorraine C.; Evans David L.; Hodgson David R.; Rose Reuben J. 《Journal of applied physiology》1996,81(5):2244-2249
Tyler, Catherine M., Lorraine C. Golland, David L. Evans,David R. Hodgson, and Reuben J. Rose. Changes in maximum oxygenuptake during prolonged training, overtraining, and detraining inhorses. J. Appl. Physiol. 81(5):2244-2249, 1996.Thirteen standardbred horses were trained asfollows: phase 1 (endurance training, 7 wk),phase 2 (high-intensity training, 9 wk),phase 3 (overload training, 18 wk), andphase 4 (detraining, 12 wk). Inphase 3, the horses were divided intotwo groups: overload training (OLT) and control (C). The OLT groupexercised at greater intensities, frequencies, and durations than groupC. Overtraining occurred after 31 wk of training and was defined as asignificant decrease in treadmill run time in response to astandardized exercise test. In the OLT group, there was a significantdecrease in body weight (P < 0.05).From pretraining values of 117 ± 2 (SE)ml · kg1 · min1,maximal O2 uptake(O2 max) increased by15% at the end of phase 1, and when signs of overtraining werefirst seen in the OLT group,O2 max was 29%higher (151 ± 2 ml · kg1 · min1in both C and OLT groups) than pretraining values. There was nosignificant reduction inO2 max until after 6 wk detraining whenO2 max was 137 ± 2 ml · kg1 · min1.By 12 wk detraining, meanO2 max was134 ± 2 ml · kg1 · min1,still 15% above pretraining values. When overtraining developed, O2 max was notdifferent between C and OLT groups, but maximal values forCO2 production (147 vs. 159 ml · kg1 · min1)and respiratory exchange ratio (1.04 vs. 1.11) were lower in the OLTgroup. Overtraining was not associated with a decrease inO2 max and, afterprolonged training, decreases inO2 max occurredslowly during detraining. 相似文献
20.
The mechanism and specificity of amino-acid transport at the plasma membrane of Ricinus communis L. roots was investigated using membrane vesicles isolated by phase partitioning. The transport of glutamine, isoleucine, glutamic acid and aspartic acid was driven by both a pH gradient and a membrane potential (internally alkaline and negative), created artificially across the plasma membrane. This is consistent with transport via a proton symport. In contrast, the transport of the basic amino acids, lysine and arginine, was driven by a negative internal membrane potential but not by a pH gradient, suggesting that these amino acids may be taken up via a voltage-driven uniport. The energized uptake of all of the amino acids tested showed a saturable phase, consistent with carrier-mediated transport. In addition, the membrane-potential-driven transport of all the amino acids was greater at pH 5.5 than at pH 7.5, which suggests that there could be a direct pH effect on the carrier. Several amino-acid carriers could be resolved, based on competition studies: a carrier with a high affinity for a range of neutral amino acids (apart from asparagine) but with a low affinity for basic and acidic amino acids; a carrier which has a high affinity for a range of neutral amino acids except isoleucine and valine, but with a low affinity for basic and acidic amino acids; and a carrier which has a higher affinity for basic and some neutral amino acids but has a lower affinity for acidic amino acids. The existence of a separate carrier for acidic amino acids is discussed.Abbreviations PM
plasma membrane
- TPP+
tetraphenylphosphonium ion
-
pH
pH gradient
-
membrane potential
This work was supported by the Agricultural and Food Research Council and The Royal Society. We would like to thank Mrs. Sue Nelson for help with some of the membrane preparations. 相似文献