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1.
Abstract— Primary cultures of glial cells prepared from brains of newborn rats were grown for 1 week and then exposed to 5 × 10−4 m -pentobarbital (PB) for 4 weeks. Compared with glial cells grown in drug-free medium throughout, exposure to PB significantly increased hexokinase activity, primarily in its mitochondrial form. Furthermore, cellular protein and RNA concentrations were significantly higher in barbiturate-cultivated than in control cells. Pulse labelling with [3H]thymidine after 4 weeks of PB exposure resulted in a significant increase (332%) in 3H incorporation into mitochondrial DNA while 3H incorporation into nuclear DNA was reduced by 58%. In addition, there was a time dependent increase in the size and number of mitochondria as determined in electron micrographs. These results are interpreted to reflect an increased mitochondrial metabolism in glial cells after chronic exposure to the barbiturate and may constitute a compensatory mechanism to the depressant action of this drug.  相似文献   

2.
Cultured guinea pig epidermal cells and dermal fibroblasts were chosen as model systems to study possible growth inhibition by cyclic AMP (cAMP)-elevating drugs. The rate of DNA synthesis was used to assay growth rate in control cultures and those treated with agents which increase intracellular cAMP, including dibutyryl cAMP, the phosphodiesterase inhibitors papaverine and theophylline and agents which stimulate adenylate cyclase, iso-proterenol and prostaglandin E2 methyl ester. Treatment for 24 h with dibutyryl cAMP (10−4 to 10−2 M) inhibited cell growth by 50 to 95%, whereas butyrate(10−4M) showed essentially no effect. This inhibition could not be attributed to decreased precursor transport or to drug toxicity. Papaverine (10−6 to 10−4 M) and theophylline (10−4 to 10−3 M) also gave dose-dependent growth inhibition as did isoproterenol and prostaglandinE2methyl ester. Radioautographic analysis of grain density after dibutyryl cAMP treatment and 3H-thymidine incorporation indicated no S-phase inhibition. Cyclic AMP-elevating drugs appear to inhibit growth of guinea-pig epidermal cells and dermal flbroblasts by blocking the cell cycle in G−2, M1, or G. −1  相似文献   

3.
The virucidal properties of iodophor, chlorine (sodium hypochlorite), formalin, thimerosal (organic mercurial compound), malachite green, and acriflavine were tested on infectious pancreatic necrosis virus (IPNV). Iodine and chlorine showed good activity, but efficacy depended on the concentration of virus, the presence of organic matter (calf serum), and water pH. Water hardness (0-300 mg 1−1 as CaCO3) did not affect virucidal activity. In a 5 min exposure, 4 mg 1−1 available iodine inactivated 103.9 TCID50 m1−1 IPNV but 16 mg 1−1 iodine were needed for inactivation of 106.3TCID50m1−1. The addition of 0-5% calf serum significantly reduced the iodine concentration and the virucidal activity. In comparison, 4 mg 1−1 chlorine were needed to inactivate 1046 TCID50 m1−1 IPNV in 5 min. However, the addition of 0-07 % serum greatly reduced the chlorine concentration and extended the virucidal contact time to 30 min or more. IPNV at 106.3 TCID60 m1−1 was not inactivated by exposures for 60 min to 0-2% formalin, 10 min to 0-2% thimerosal, 60 min to 5 mg 1−1 malachite green, or 20 min to 500 mg 1−1 acriflavine. However, acriflavine at 0-5 mg 1−1 in cell culture media prevented the development of cytopathology caused by IPNV and may be useful in the treatment of the disease.  相似文献   

4.
Abstract: Muscarinic receptors found in the N IE-115 mouse neuroblastoma cell line were tested for their ability to mediate stimulation of phosphatidylinositol (PI) turnover. This study was facilitated by the development of a new solvent system (acetone: butanol: acetic acid: water, 5: 5: 1: 1) for the rapid and consistent separation of PI by one-dimensional thin-layer chromatography. Cholinergic stimulation caused as much as a 680% increase in the incorporation of 32P into PI. Enhanced incorporation of 32P into PI could be measured as early as 4 min after stimulation began. By 20 min, the rate of incorporation by stimulated cells had decreased to that of unstimulated cells, indicating desensitization. The magnitude of the response was dependent on the extent of receptor occupancy and the response elicited by a saturating dose of carbamylcholine was blocked completely by 10−7 M at-ropine, a specific muscarinic antagonist. Chronic stimulation, known to cause a loss of receptor binding sites, led to a 90% decrease in the maximum response even after a 40-min withdrawal period. Replacement of Na+ ions in the medium with choline or K+ severely impaired the ability of the cells to incorporate added 32P into PI (90 and 50%, respectively). Removal of the putative second messenger Ca2+ for short periods of time by the addition of excess EGTA did not alter either basal or muscarinic-stimulated PI turnover.  相似文献   

5.
Treatment of sea urchin embryos with cAMP phosphodiesterase (PDE)-inhibitors such as caffeine (4×10−3 M), theophylline (8×10−3M), or nicotinamide (10−2M), at the morula stage for only a couple of hours, yields vegetalized larvae. Most of the embryos treated with these reagents before the morula stage develop to blastulae filled with mcsenchyme-like cells. Almost all embryos at the blastula stage develop normally even if they are treated with a PDE-inhibitor for a considerable period. The rate of 3H-valine incorporation into protein in the morulae is reduced by caffeine and theophylline, but does not decrease in the presence of nicotinamide. Actinomycin D cancels the vegetalizing effect of PDE-inhibitors on the morulae.  相似文献   

6.
The phenotypic effects of a new recessive mutation mas −1, which in homozygous condition induces testicular development in XX animals of common carp ( Cyprinus carpio L.), are described. Sexual differentiation of XX; mas −+/ mas −1 and XX; mas −1/ mas −1 animals was compared with the gonad development of XX wild type females and XY males. In XX females gonadal differentiation starts with the formation of an ovarian cavity and entry into meiosis of germ cells at around 80 days post hatching (ph). Male gonads remain quiescent until 120 days ph during which period they develop a network of loose connective tissue. Spermatogenesis starts with tubule formation and the differentiation of germ cells into spermatogonia type B. Heterozygous XX; mas −+/ mas −1 animals developed as normal females, but in homozygous XX; mas −1/ mas −1 animals two types of gonad development were observed. In the first type, germ cells did not enter meiosis until 100 days ph when they differentiated as spermatogonia. An ovarian cavity was not formed but male specific connective tissue developed instead. These gonad developed as normal testes. In the second type, germ cells differentiated at 80 days ph as either oocytes or spermatocytes, which resulted in the gonads developing as ovotestes. The formation of an ovarian cavity was in most cases incomplete. The phenotypic effects of mas −1 are interpreted as a timing mismatch between mas activation and female sex differentiation.  相似文献   

7.
Abstract— Synthetic substance P initially increased cyclic AMP levels and subsequently induced neurite extension in cultured neuroblastoma N 18 cells. The magnitude of these effects depended on the concentration of fetal calf serum (FCS) in the culture medium, being more evident in the presence of a lower (0.1%) concentration of FCS.
In Eagle's medium containing 0.1% FCS, low concentrations of substance P (10−7-10−5 M) increased cyclic AMP levels and stimulated neurite extension.
In Eagle's medium containing 5%FCS, both substance P at concentrations of 10−5-10−3M and dibutyryl cyclic AMP at concentrations of 10−4-10−2M increased cyclic AMP levels and stimulated neurite extension. The activities of acetylcholinesterase, (Na++ K+)-, HCO3 and Mg2+ -stimulated-ATPase were also increased. Cell growth was inhibited.
Substance P at concentrations of 10-7-10−5M also stimulated the adenylate cyclase activity of a particulate fraction of N 18 in a concentration-dependent manner.  相似文献   

8.
Epithelial cells are the initial sites of host invasion by group A Streptococcus pyogenes (GAS), and their infection of epithelial cells has been suggested to induce apoptosis. However, the mechanism responsible for bacteria–host interaction and the induction of apoptosis has not been clearly understood. We demonstrate here that human pharyngeal epithelial HEp-2 cells became apoptotic with DNA fragmentation by invasion of GAS strains JRS4 (M6+, F1+) and JRS145 (M6, F1+ mutant of JRS4), whereas apoptotic cellular changes were not observed in SAM1 (M6+, F1 mutant) or SAM2 (M6, F1 mutant) infected HEp-2 cells. Confocal microscopy revealed that Bax translocation to mitochondria and cytochrome c release occurred after 4 h of infection. Western blot analyses showed that the amounts of Bcl-2 and Bcl-xL were decreased in the mitochondria of infected cells. In addition, we demonstrated that the release of nuclear histone from infected cells was prevented by the addition of caspase-9 inhibitor (Ac-LEHD-CHO). We conclude that the internalization of GAS in epithelial cells is necessary and sufficient for the induction of apoptosis, which is initiated by mitochondrial dysfunction, and the mechanism of GAS-induced apoptosis is clearly different from that induced by other intracellular invasive bacteria, e.g. Shigella and Salmonella species.  相似文献   

9.
The complex of bioelectrical paramenters (membrane potential, membrane resistance and capacitance) of internodal cells of Nitellopsis obtusa was measured over a wide range of IAA concentration (10−10 to 10−4 M ) with two intracellular microelectrodes. Primary effects of IAA at a concentration as low as 10−10 M were observed. The optimum range of IAA action was from 10−9 to 10−6 M . The type of IAA-induced electroresponse depended on the initial level of membrane potential, which characterized the energetic state of the plasmalemma. In the energized state (ca −200 mV) N. obtusa cells appeared to have 3 typical reactions: hyperpolarization (membrane potential less than K+-equilibrium potential), depolarization (membrane potential higher than K+-potential) and absence of response at K+-electrochemical equilibrium. Membrane capacitance was found constant at 0.74 ± 0.05 μF cm−2, but membrane resistance increased up to 50% independently of the sign of the electrogenic reaction. Increase of membrance capacitance and decrease of the membrane resistance was a feature of the de-energized state (ca −135 mV) and may be explained by lower viscosity of membrane lipids, which interacted with IAA. The complex of parameter, including cytoplasmic steaming taken as an indicator of energy supply, is discussed as indicating slow IAA penetration combined with a primary action of IAA on the plasmalemma receptor sites.  相似文献   

10.
Abstract: In human astrocyte cultures established from second-trimester fetal brain tissue, ∼5–10% of total astrocyte population in unstimulated cultures were GD3+/glial fibrillary acidic protein (GFAP)+. The GD3+ cells were always GFAP+ and grew as flat, highly spread cells but changed to process-bearing cells after interleukin-1β (IL-1β) stimulation. It is interesting that IL-1β, a known mitogen for rat astrocytes, suppressed human fetal astrocyte proliferation as determined by [3H]thymidine incorporation, bromodeoxyuridine (BrdU) labeling, and cell counting. The GD3+ population, however, consistently increased in absolute number after IL-1β stimulation, in a dose- and time-dependent manner. The IL-1β-mediated increase in number of GD3+ astrocytes was independent of initial cell density or serum concentration. By flow cytometry, IL-1β enhanced both the mean fluorescence intensity and the percentage of GD3+ cells. To investigate whether the increase in GD3+ astrocyte cell number was due to proliferation of preexisting GD3+ astrocytes or due to conversion of GD3 to GD3+ cells, we performed BrdU/GD3 double immunocytochemistry. BrdU/GD3 double-positive cells were extremely rare in both control and IL-1β-stimulated cultures. Moreover, an increase in number of GD3+ astrocytes was still observed in control and IL-1β-stimulated cultures where GD3+ cells had been initially eliminated by cell sorting. These results indicate that GD3+ astrocytes in human fetal culture may represent a postmitotic, differentiated, distinct phenotype.  相似文献   

11.
Killing of wild-type spores of Bacillus subtilis with formaldehyde also caused significant mutagenesis; spores (termed αβ) lacking the two major α/β-type small, acid-soluble spore proteins (SASP) were more sensitive to both formaldehyde killing and mutagenesis. A recA mutation sensitized both wild-type and αβ spores to formaldehyde treatment, which caused significant expression of a recA - lacZ fusion when the treated spores germinated. Formaldehyde also caused protein–DNA cross-linking in both wild-type and αβ spores. These results indicate that: (i) formaldehyde kills B. subtilis spores at least in part by DNA damage and (b) α/β-type SASP protect against spore killing by formaldehyde, presumably by protecting spore DNA.  相似文献   

12.
SYNOPSIS. Experiments were designed to investigate the effects of insect juvenile hormone (JH) on the over-all growth and macromolecular synthesis of Crithidia sp. in vitro. Cells grown in the presence of 10−5M-10−3M JH showed a concentration-dependent inhibition of growth, which appeared to result from both a prolongation of generation time and a delay in the onset of logarithmic growth. Juvenile hormone (10−3M) inhibited the incorporation of [3H]thymidine, [3H]uridine and [3H] leucine into logarithmically growing cells by 50, 70 and 40% respectively. The incorporation of [3H]uridine into acid insoluble material could be stopped within 1 hr of application of the hormone (10−3M). The inhibitory effect was reversible in terms of cell numbers in subcultures of washed cells but an examination of the reversibility of RNA synthesis inhibition suggested that the resumption of RNA synthesis at an optimal level would require a lag period of at least 1–3 hr. It is suggested that JH may act by interfering with RNA synthesis either directly or indirectly by primarily acting at the level of the plasma membrane.  相似文献   

13.
During starch degradation in intact isolated chloroplasts from Chlamydomonas reinhardtii gas exchange was studied with a mass spectrometer. Oxygen uptake by intact chloroplasts in the dark never exceeded 1.5% of the starch degradation rate [maximum 15 nmol O2 (mg Chl)−1 h−1 consumed. 1 000 nmol glucose (mg Chl)−1h−1 degraded]. Evolution of CO2 under aerobic conditions [9.8–28 nmol (mg Chl)−1 h−1] was stimulated by addition of 0.1–0.5 m M oxaloacetate [393–425 nmol CO2 (mg Chl)−1 h−1]. Pyridoxal phosphate (5 m M ) inhibited starch degradation by more than 80%, but had no effect on O2 uptake. Starch degradation rates and CO2 evolution did not differ under acrobic and anaerobic conditions. Increasing Pi in the reaction medium from 0.5 m M to 5.0 m M stimulated starch degradation by 230 and 260% under aerobic and anaerobic conditions, respectively. A rapid autooxidation of reduced ferredoxin was observed in a reconstituted system consisting of purified Chlamydomonas ferredoxin, purified Chlamydomonas NADP-ferredoxin oxidoreductase (EC 1.6.7.1) and NADPH. Addition of isolated thylakoids from C. reinhardtii did not affect the rate of O2 uptake. Our results clearly indicate the absence of any oxygen requirement during starch degradation in isolated chloroplasts.  相似文献   

14.
In vitro shoots of cv. Doyenne ďHiver pear ( Pyrus communis L.) were irradiated under controlled environments for 6 h per day at 5 different levels of biologically effective UV-B radiation (UV-BBE). UV-B exposure caused a progressive increase in apical necrosis above background levels and stimulated leaf abscission. Shoots grown for 2 weeks at 7. 8 mol m−2 day −1 of photosynthetic photon flux (PPF) and treated with 8. 4 or 12. 0 kJ m−2 day −1 UV-BBE produced up to 4 times more ethylene than those given 2. 2 or 5. 1 kJ m−2 day−1 UV-BBE or untreated controls. Exposure of shoots to 12 kJ m−2 day −1 of UV-BBE caused an increase in free putreseine content after 4 to 14 days of irradiation. Shoots showed a decrease in CO2 uptake after 3 days of UV-B: thereafter, they appeared to recover their photosynthetic capacity. Under typical PPF conditions used in micropropagation (90 μmol m−2 S−1). 8. 4 kJ m−2 day −1 of UV-B radiation was injurious to realatively tender tissues of in vitro pear shoots: increasing the level of UV-BBE to 12 kJ m−2 day−1 produced even more adverse effects.  相似文献   

15.
Abstract Estuarine microcosms were used to follow conjugal transfer of a broad host range IncP1 plasmid from a Pseudomonas putida donor to indigenous bacteria. Donor cells were added at a concentration similar to the natural abundance of bacteria in the water column (106 cells ml−1). Transfer was not detected in any of the test microcosms (calculated limit of detection of 10−7 and 10−4 transconjugants donor−1 in water column and sediment, respectively), with the exception of transfer to an isogenic recipient (added at 105 cells ml−1) in sediments of controls that had been inoculated with both donors and recipients. The same plasmid was transferred with high efficiencies (10−1 to 10−3) to a variety of recipients in filter and broth matings. These results suggest that if conjugal gene transfer occurred, it was at efficiencies that were not detectable in estuarine microcosms simulating natural population densities.  相似文献   

16.
Sperm structure and motility of the freshwater teleost Cottus gobio   总被引:1,自引:0,他引:1  
When motility of spermatozoa of Cottos gobio was initiated with distilled water, the motility rate decreased to 0% within 1 min, and significant signs of osmotic alterations were observed at the end of the motility period. By contrast, in 50 mmol 1−1 NaCl solution, the motility rate persisted for 120–140 min. In both distilled water and in 50 mmol 1−1 NaCl solution, the main swimming type of spermatozoa was linear motion during the whole motility period. The initial swimming velocity (50.0 ± 2.1 μm s−1) measured 10 s after motility initiation was similar in both distilled water and in 50 mmol 1−1 NaCl solution. In distilled water, the velocity decreased to <20 μm s−1 (locally motile) during the first minute of the motility phase. In 50 mmol 1−1 NaCl solutions, it remained at a constant level during the first 60 min of the motility period, but then started to decrease to <20 μm s−1 after 120 min. When 5 mmol 1−1 potassium cyanide, antimycin or atractyloside was added to the 50 mmol 1−1 NaCl solution, the motility period was reduced to ≤2min. Ten millimoles per litre 2-deoxy-D-glucose, malonate or a mixture of 5 mmol 1−1 atractyloside and 5 mmol 1−1 carnithine did not effect the duration of the motility period. This indicates that sperm energy metabolism depends mainly on respiration rate and fatty acid metabolism.  相似文献   

17.
In crude extracts of Chlorella kessleri Fott and Novákóva cells grown autotrophically in white light the activity of phosphofructokinase (PFK, EC 2.7.1.11) is 62.9 ± 1.5 nmol (mg protein)−1 min−1 under optimized test conditions. It is greatly increased in red [88.3 ± 1.8 nmol (mg protein)−1 min−1], but somewhat decreased [57.0 ± 0.5 nmol (mg protein)−1 min−1] in blue light of equal productivity. Mixtures of blue and red light yield the low activity as long as blue light represents at least 35% of the total quantum fluence rate. The rough wavelength dependence of the counteracting effect of short wavelength light on the increasing effect of red light exhibits a broad peak at 460 nm, reminiscent of action spectra of the blue/UV photoreceptors(s). Upon transfer of red light-grown cells to blue light, the decrease develops slowly within 72 h; it cannot be prevented by 3-(3,4-dichlorophenyl)-1,1-dimethyl urea (DCMU). Since there is less carbohydrate in blue than in red light-exposed cells, correlations between biosynthesis of PFK and level of carbohydrate are discussed, based on the assumption that red light decreases and/or blue light increases the transport of metabolites across the chloroplast envelope.  相似文献   

18.
Increase in the extracellular Ca2+ concentration from low (≤ 10−7 M) to normal (10−3 M) caused morphological changes of cultured myocardial cells obtained from fetal mouse heart. The extracellular Na+ and K+ concentrations of the normal medium (10−3 M Ca2+) did not significantly affect the genesis of these morphological changes. Like Ca2+, Ba2+ and Sr2+, but not Mg2+, Co2+ or Ni2+, could induce morphological changes. Increase in the extracellular Ca2+ concentration from 10−8 M to 10−3M also caused excess uptake of 45Ca2+ by cultured myocardial cells. B–16CW 1 cells, which did not show these morphological changes, did not take up excess 45Ca2+ on this treatment. Treatments, such as addition of verapamil or incubation at pH 6.3, which reduced the genesis of morphological changes, reduced the rate of 45Ca2+ uptake by myocardial cells. These facts show that the morphological changes of myocardial cells induced by increasing the extracellular Ca2+ concentration from low to normal are due to excess uptake of Ca2+ by the myocardial cells.
The morphological changes of cultured myocardial cells induced by increasing the extracellular Ca2+ concentration from low to normal were reversed on further incubation of the cells in medium with or without Ca2+.  相似文献   

19.
A 5-Bromodeoxyuridine-sensitive Interval during Drosophila Myogenesis   总被引:2,自引:0,他引:2  
Drosophila myogenesis was monitored in vitro and the cells were treated with 5-bromodeoxyuridine (BrdU) or with thymidine at certain intervals. Muscle cells were scored for survival, contractility, and the uptake of thymidine and BrdU. Results indicated that the final S period for myoblasts takes place in vitro between 1.3 and 3.3 h following the initiation of gastrulation in the donor embryos. Treatment with 10-4 M BrdU during this interval inhibited myogenesis, but later treatment did not. Thymidine reversed or prevented the BrdU effect if given before the final myoblast division, but not if given afterwards. All results support the hypothesis that BrdU inhibits Drosophila myogenesis through its incorporation into DNA.  相似文献   

20.
Efficiency of imidacloprid and pymetrozine on population growth parameters of cabbage aphid, Brevicoryne brassicae L. (Homoptera: Aphididae) was determined using demographic toxicology by leaf dip method. At first, bioassay tests were performed. The LC50 value and confidence limit for imidacloprid and pymetrozine were 1.61 × 10^-5 mol/L (0.74 × 10^-5-2.66 × 10^-5) and 2.14 × 10^-4 mol/L (1.24 × 10^-4-3.40 × 10^-4), respectively. To evaluate the sublethal effect of two insecticides on population growth parameters of cabbage aphid, LC30 concentrations of imidacloprid and pymetrozine were used at 5 mol/L and 30 mol/L. The experiments were carried out in a incubator at 20±1℃, 60% ± 5% RH and 16:8 (L:D) photoperiod on canola seedlings, Brassica napus L. var. 'PF'. Net fecundity rate decreased in both insecticide-treated populations. Intrinsic rates of increase (rm) were lower in imidacloprid and pymetrozine treatments than in controls. Intrinsic birth rates also decreased in treated populations. There was a relative increase in intrinsic death rates of treated populations. The mean generation times and doubling time were also lower in populations treated with insecticides than in controls. There was a considerable reduction in the average numbers of nymphs reproduced per female as compared with the control. The average longevity of female adults in the control was significantly different from those treated with imidacloprid and pymetrozine. However, there was no significant differences in aphid life-table parameters between the two insecticide-treated populations (P 〉 0.01).  相似文献   

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