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81.
Margareta Lirvall Pia Ljungqvist-Höddelius Åke Wasteson Karl-Eric Magnùsson 《Bioscience reports》1996,16(3):227-238
Growth factor receptors transmit biological signals for the stimulation of cell growth in vitro and in vivo and their autocrine stimulation may be involved in tumorigenesis. It is therefore, of great value to understand receptor reactions in response to ultraviolet (UV) light which certain normal human cells are invaribly exposed to during their growth cycle. UV irradiation has recently been shown to deplete antioxidant enzymes in human skin. The aims of the present study were a) to compare the lateral mobility of epidermal growth factor receptors (EGF-R) in cultured human keratinocytes and human foreskin fibroblasts, b) to investigate effects of ultraviolet B radiation on the mobility of EGF-R in these cells, and c) study the response of EGF-R on addition of antioxidant enzymes. The epidermal growth factor receptors were labeled with rhodaminated EGF, the lateral diffusion was determined and the fraction of mobile EGF-R assessed with the fluorescence recovery after photobleaching (FRAP). We found that human keratinocytes display a higher basal level of EGF-R mobility than human skin fibroblasts, viz. with diffusion coefficients (D ± standard error of the mean, SEM) of 4.2±0.2 × 10–10 cm2/s, and 1.8±0.2 × 10–10 cm2/s, respectively. UVB-irradiated fibroblasts showed an almost four-fold increase in the diffusion coefficient; D was 6.3±0.3 × 10–10 cm2/s. The keratinocytes, however, displayed no significant increase in receptor diffusion after irradiation; D was 5.1±0.8 × 10–10 cm2/s. In both cell types the percentage of EGF-R fluorescence recovery after photobleaching, i.e. the fraction of mobile receptors, was significantly increased after irradiation. In keratinocytes it increased from 69% before irradiation to 78% after irradiation. Analogous figures for fibroblasts were 61% and 73%. The effect of UVB on fibroblast receptors was abolished by prior addition of superoxide dismutase (SOD) and catalase (CAT). It is concluded that UVB radiation of fibroblasts and keratinocytes can affect their biophysical properties of EGF-R. The finding that addition of antioxidant enzymes prevented the UVB effect in fibroblasts may indicate the involvement of reactive oxygen metabolites.Abbreviations CAT
Catalase
- D
Lateral diffusion coefficient
- EDTA
Ethylenediaminetetraacetic acid
- EGF
Epidermal growth factor
- E-MEM
Eagle's minimum essential medium
- FCS
Fetal calf serum
- FRAP
Fluorescence recovery after photobleaching
- KRG
Krebs-Ringer phosphate buffer
- PBS
Phosphate-buffered saline
- R
Mobile fraction
- ROS
Reactive oxygen species
- SEM
Standard error of the mean
- SOD
Superoxide dismutase
- UVA
Ultraviolet light-A (315-400 nm)
- UVB
Ultraviolet light-B (280-315 nm) 相似文献
82.
Ondrej Prasil Zbigniew Kolber Joseph A. Berry Paul G. Falkowski 《Photosynthesis research》1996,48(3):395-410
The oxygen flash yield (YO2) and photochemical yield of PS II (PS II) were simultaneously detected in intact Chlorella cells on a bare platinum oxygen rate electrode. The two yields were measured as a function of background irradiance in the steady-state and following a transition from light to darkness. During steady-state illumination at moderate irradiance levels, YO2 and PS II followed each other, suggesting a close coupling between the oxidation of water and QA reduction (Falkowski et al. (1988) Biochim. Biophys. Acta 933: 432–443). Following a light-to-dark transition, however, the relationship between QA reduction and the fraction of PS II reaction centers capable of evolving O2 became temporarily uncoupled. PS II recovered to the preillumination levels within 5–10 s, while the YO2 required up to 60 s to recover under aerobic conditions. The recovery of YO2 was independent of the redox state of QA, but was accompanied by a 30% increase in the functional absorption cross-section of PS II (PS II). The hysteresis between YO2 and the reduction of QA during the light-to-dark transition was dependent upon the reduction level of the plastoquinone pool and does not appear to be due to a direct radiative charge back-reaction, but rather is a consequence of a transient cyclic electron flow around PS II. The cycle is engaged in vivo only when the plastoquinone pool is reduced. Hence, the plastoquinone pool can act as a clutch that disconnects the oxygen evolution from photochemical charge separation in PS II.Abbreviations ADRY
acceleration of the deactivation reactions of the water-splitting enzyme (agents)
- Chl
chlorophyll
- cyt
cytochrome
- DCMU
3-(3,4-dichlorophenyl)-1,1-dimethylurea
- FO
minimum fluorescence yield in the dark-adapted state
- FI
minimum fluorescence yield under ambient irradiance or during transition from the light-adapted state
- FM
maximum fluorescence yield in the dark-adapted state
- FM
maximum fluorescence yield under ambient irradiance or during transition from light-adapted state
- FV, FV
variable fluorescence (FV=FM–FO ; FV=FM–FI)
- FRR
fast repetition rate (fluorometer)
- PS II
quantum yield of QA reduction (PS II=(FM – FO)/FM or PS II)=(FM= – FI=)/FM=)
- LHCII
Chl a/b light harvesting complexes of Photosystem II
- OEC
oxygen evolving complex of PS II
- P680
reaction center chlorophyll of PS II
- PQ
plastoquinone
- POH2
plastoquinol
- PS I
Photosystem I
- PS II
Photosystem II
- RC II
reaction centers of Photosystem II
- PS II
the effective absorption cross-section of PHotosystem II
- TL
thermoluminescence
- YO2
oxygen flash yield
The US Government right to retain a non-exclusive, royalty free licence in and to any copyright is acknowledged. 相似文献
83.
Thylakoids and Photosystem II particles prepared from the cyanobacterium Synechococcus PCC 7942 washed with a HEPES/glycerol buffer exhibited low rates of light-induced oxygen evolution. Addition of either Ca2+ or Mg2+ to both thylakoids and Photosystem II particles increased oxygen evolution independently, maximal rates being obtained by addition of both ions. If either preparation was washed with NaCl, light induced O2 evolution was completely inhibited, but re-activated in the same manner by Ca2+ and Mg2+ but to a lower level. In the presence of Mg2+, the reactivation of O2 evolution by Ca2+ allowed sigmoid kinetics, implying co-operative binding. The results are interpreted as indicating that not only Ca2+, but also Mg2+, is essential for light-induced oxygen evolution in thylakoids and Photosystem II particles from Synechococcus PC 7942. The significance of the reactivation kinetics is discussed. Reactivation by Ca2+ was inhibited by antibodies to mammalian calmodulin, indicating that the binding site in Photosystem II may be analogous to that of this protein.Abbreviation HEPES
n-2-Hydroxyethylpiperazine--2-ethane sulphonic acid 相似文献
84.
The role of the N-terminus of the extrinsic 33 kDa protein of Photosystem II has been investigated by means of site-directed mutagenesis and cross-linking. Replacement of Asp-9 resulted in a dramatic increase in proteolytic sensitivity leading to the degradation of the protein forming a 31 kDa fragment with an undefined N-terminus. This fragment was unable to restore oxygen evolution. However, the variants of the 33 kDa protein which remained intact could reconstitute oxygen evolution as effectively as the wild-type protein. Cross-linking experiments with a water-soluble carbodiimide revealed that mutagenesis of residue D9 led to the disruption of an intramolecular salt bridge. Therefore we suggest that the N-terminus of the 33 kDa protein is necessary for maintaining the binding ability of the protein to Photosystem II but might not be involved in binding itself. 相似文献
85.
Flooding-induced membrane damage, lipid oxidation and activated oxygen generation in corn leaves 总被引:19,自引:1,他引:18
Flooding effects on membrane permeability, lipid peroxidation and activated oxygen metabolism in corn (Zea mays L.) leaves were investigated to determine if activated oxygens are involved in corn flooding-injury. Potted corn plants were flooded at the 4-leaf stage in a controlled environment. A 7-day flooding treatment resulted in a significant increase in chlorophyll breakdown, lipid peroxidation (malondialdehye content), membrane permeability, and the production of superoxide (O
2
-
) and hydrogen peroxide (H2O2) in corn leaves. The effects were much greater in older leaves than in younger ones. Spraying leaves with 8-hydroxyquinoline (an O
2
-
scavenger) and sodium benzoate (an .OH scavenger) reduced the oxidative damage and enhanced superoxide dismutase (SOD) activity. A short duration flooding treatment elevated the activities of SOD, catalase, ascorbate peroxidase (AP), and glutathione reductase (GR), while further flooding significantly reduced the enzyme activities but enhanced the concentrations of ascorbic acid and reduced form glutathione (GSH). It was noted that the decline in SOD activity was greater than that in H2O2 scavengers (AP and GR). The results suggested that O
2
-
induced lipid peroxidation and membrane damage, and that excessive accumulation of O
2
-
is due to the reduced activity of SOD under flooding stress. 相似文献
86.
Oxygen-dependent electron transport and protection from photoinhibition in leaves of tropical tree species 总被引:7,自引:0,他引:7
The roles of photorespiration and the Mehlerperoxidase pathway in sustaining electron transport and protection from photoinhibition were studied in outer canopy leaves of two species of tropical trees: the drought-deciduous Pseudobombax septenatum (Jacq.) Dug. and the evergreen Ficus insipida Willd. Ficus had a higher photosynthetic capacity than Pseudobombax and also a greater capacity for light-dependent electron transport under photorespiratory conditions (in the absence of CO2). As a consequence, in the absence of CO2, Ficus was able to maintain a largely oxidized electron-transport chain at higher photon flux densities than Pseudobombax. Under the same light conditions, photoinhibition (reduction in Fv/Fm) was always greater in Pseudobombax than Ficus, was increased when leaves were exposed to 2% O2 in nitrogen compared to 21% O2 in CO2-free air, but was not increased by the absence of CO2. Rates of electron transport due to the Mehler-peroxidase pathway (assessed in 2% O2 in nitrogen) ranged between 16–40 mol · m–2·s–1 in both species. As the dry season approached and Pseudobombax neared leaf senescence there was a decline in the capacity for photorespiratory flux to maintain electron transport in Pseudobombax, but not in Ficus. Ratios of light-dependent electron transport to net CO2 fixation for Pseudobombax, Ficus and two other species in the field, Luehea seemannii Tr. & Planch, and Didymopanax morototoni (Aubl.) Dec. & Planch., ranged from 6.2 (Ficus) to 16.7 (Pseudobombax). High in-situ rates of photorespiration combined with the decreased capacity of Pseudobombax for photorespiratory flux as the dry season approached indicates a decreased capacity to protect against photooxidative damage. This may contribute to the promotion of leaf senescence in Pseudobombax during the transition from wet to dry season.Abbreviations Fv/Fm
ratio of variable to maximum chlorophyll a fluorescence
- NPQ
nonphotochemical fluorescence quenching
- PFD
photon flux density
- QA
primary electron acceptor of PSII
This research was supported by a grant from the Mellon Foundation. We thank Monica Mejia and Juan Posada for assisting with the fluorescence measurements and Aurelio Virgo for assisting with the field CO2-exchange measurements. 相似文献
87.
Robert de Jonge Mario Bedu Nicole Fellmann Stephen Blonc Hilde Spielvogel Jean Coudert 《European journal of applied physiology and occupational physiology》1996,74(4):367-374
We have previously observed that 11-year-old children of low socio-economic status (LSES) showed a delayed physical growth of approximately 2 years and developed lower normalized short-term power output than children of high socio-economic status (HSES) of the same age. In contrast, maximal oxygen uptake
per unit of fat free mass was no different in either group. The aim of this study was to evaluate the effect of anthropometric characteristics between HSES and LSES prepubertal children in aerobic and anaerobic performance. To compare children of the same body dimensions, 11-year-old boys (n = 30) and girls (n = 31) of LSES and 9-year-old boys (n = 21) and girls (n = 27) of HSES were studied. Anthropometric measurements,
(direct test), maximal anaerobic power (P
max, force-velocity test) and mean anaerobic power (
, Wingate test) were determined. In these children having the same body dimensions: mean
were the same in LSES and HSES children [1.2 (SD 0.2)1-min–1];P
max and
were lower in LSES subjects [154.0 (SD 33.2) vs 174.6 (SD 38.4) W and 116.3 (SD 23.3) vs 128.2 (SD 28.0) W, respectively]; the linear relationships between
and fat free mass were the same in LSES and HSES boys but, in the girls, the LSES group had lower values. For anaerobic performance, the relationships were significantly different: the slopes were the same but LSES values for the both sexes were lower. These results would suggest that factors other than differences in body dimensions alone were responsible for the lower performance of LSES girls and boys. Cultural factors and motor learning, structural and functional alterations of muscle induced by marginal malnutrition have been discussed. 相似文献
88.
Myosin molecules contacting an actin filament in the presence of ATP were found to regulate the filamental fluctuations due to ATP hydrolysis in a communicative manner along the filament. As an evidence of the occurrence of the communication, ATP-activated fluctuating displacements of the filament in the direction perpendicular to its longitudinal axis were identified to propagate at a finite velocity not less than about 0.2 μm/s unidirectionally along the filament. 相似文献
89.
Although in vitro studies have shown that oxygen free radicals depress the sarcolemmal Ca2+-pump activity and thereby may cause the occurrence of intracellular Ca2+ overload for the genesis of contractile failure, the exact relationship between changes in sarcolemmal Ca2+-pump activity and cardiac function due to these radicals is not clear. In this study we examined the effects of oxygen radicals on sarcolemmal Ca2+ uptake and Ca2+-stimulated ATPase activities as well as contractile force development by employing isolated rat heart preparations. When hearts were perfused with medium containing xanthine plus xanthine oxidase, the sarcolemmal Ca2+-stimulated ATPase activity and ATP-dependent Ca2+ accumulation were depressed within 1 min whereas the developed contractile force, rate of contraction and rate of relaxation were increased at 1 min and decreased over 3–20 min of perfusion. The resting tension started increasing at 2 min of perfusion with xanthine plus xanthine oxidase. Catalase showed protective effects against these alterations in heart function and sarcolemmal Ca2+-pump activities upon perfusion with xanthine plus xanthine oxidase whereas superoxide dismutase did not exert such effects. The combination of catalase and superoxide dismutase did not produce greater effects in comparison to catalase alone. These results are consistent with the view that the depression of heart sarcolemmal Ca2+ pump activities may result in myocardial dysfunction due to the formation of hydrogen peroxide and/or hydroxyl radicals upon perfusing the hearts with xanthine plus xanthine oxidase. 相似文献
90.
Dissolved oxygen tension and oxygen uptake rate are critical parameters in animal cell culture. However, only scarce information of such variables is available for insect cell culture. In this work, the effect of dissolved oxygen tension (DOT) and the utility of on-line oxygen uptake rate (OUR) measurements in monitoring Spodoptera frugiperda (Sf9) cultures were determined. Sf9 cells were grown at constant dissolved oxygen tensions in the range of 0 to 30%. Sf9 metabolism was affected only at DOT below 10%, as no significant differences on specific growth rate, cell concentration, amino acid consumption/production nor carbohydrates consumption rates were found at DOT between 10 and 30%. The specific growth rate and specific oxygen uptake rate followed typical Monod kinetics with respect to DOT. The calculated max and
max were 0.033 h-1 and 3.82×10-10 mole cell-1h-1, respectively, and the corresponding saturation constants were 1.91 and 1.57%, respectively. In all aerated cultures, lactate was consumed only after glucose and fructose had been exhausted. The yield of lactate increased with decreasing DOT. It is proposed, that an apparent DOT in non-instrumented cultures can be inferred from the lactate yield of bioreactors as a function of DOT. Such a concept, can be a useful and important tool for determining the average dissolved oxygen tension in non-instrumented cultures. It was shown that the dynamic behavior of OUR can be correlated with monosaccharide (fructose and glucose) depletion and viable cell concentration. Accordingly, OUR can have two important applications in insect cell culture: for on-line estimation of viable cells, and as a possible feed-back control variable in automatic strategies of nutrient addition.Abbreviations DOT
Dissolved oxygen tension
- OUR
Oxygen uptake rate
-
specific oxygen uptake rate
-
specific growth rate
- Xv
viable cell concentration
- CL, C*, and
oxygen concentrations in liquid phase, in equilibrium with gas phase, and medium molar concentration, respectively
- H
Henry's constant
- KLa
volumetric oxygen transfer coefficient
- PT
total pressure
-
oxygen partial pressure
-
oxygen molar fraction
- i
discrete element 相似文献