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191.
Ortega A Ferrer P Carretero J Obrador E Asensi M Pellicer JA Estrela JM 《The Journal of biological chemistry》2003,278(41):39591-39599
B16 melanoma (B16M) cells with high GSH content show high metastatic activity. However, the molecular mechanisms linking GSH to metastatic cell survival are unclear. The possible relationship between GSH and the ability of Bcl-2 to prevent cell death was studied in B16M cells with high (F10) and low (F1) metastatic potential. Analysis of a Bcl-2 family of genes revealed that B16M-F10 cells, as compared with B16M-F1 cells, overexpressed preferentially Bcl-2 (approximately 5.7-fold). Hepatic sinusoidal endothelium-induced B16M-F10 cytotoxicity in vitro increased from approximately 19% (controls) to approximately 97% in GSH-depleted B16M-F10 cells treated with an antisense Bcl-2 oligodeoxynucleotide (Bcl-2-AS). l-Buthionine (S,R)-sulfoximine-induced GSH depletion or Bcl-2-AS decreased the metastatic growth of B16M-F10 cells in the liver. However, the combination of l-buthionine (S,R)-sulfoximine and Bcl-2-AS abolished metastatic invasion. Bcl-2-overexpressing B16M-F1/Tet-Bcl-2 and B16M-F10/Tet-Bcl-2 cells, as compared with controls, showed an increase in GSH content, no change in the rate of GSH synthesis, and a decrease in GSH efflux. Thus, Bcl-2 overexpression may increase metastatic cell resistance against oxidative/nitrosative stress by inhibiting release of GSH. In addition, Bcl-2 availability regulates the mitochondrial GSH (mtGSH)-dependent opening of the permeability transition pore complex. Death in B16M-F10 cells was sharply activated at mtGSH levels below 30% of controls values. However, this critical threshold increased to approximately 60% of control values in Bcl-2-AS-treated B16M-F10 cells. GSH ester-induced replenishment of mtGSH levels (even under conditions of cytosolic GSH depletion) prevented cell death. Our results indicate that survival of B16M cells with high metastatic potential can be challenged by inhibiting their GSH and Bcl-2 synthesis. 相似文献
192.
Ruiz-Palacios GM Cervantes LE Ramos P Chavez-Munguia B Newburg DS 《The Journal of biological chemistry》2003,278(16):14112-14120
The most common cause of infant mortality is diarrhea; the most common cause of bacterial diarrhea is Campylobacter jejuni, which is also the primary cause of motor neuron paralysis. The first step in campylobacter pathogenesis is adherence to intestinal mucosa. We found that such binding was inhibited in vitro by human milk and, with high avidity, by alpha1,2-fucosylated carbohydrate moieties containing the H(O) blood group epitope (Fuc alpha 1,2Gal beta 1,4GlcNAc em leader ). In studies on the mechanism of adherence, campylobacter, which normally does not bind to Chinese hamster ovary cells, bound avidly when the cells were transfected with a human alpha1,2-fucosyltransferase gene that caused overexpression of H-2 antigen; binding was specifically inhibited by H-2 ligands (lectins Ulex europaeus and Lotus tetragonolobus and H-2 monoclonal antibody), H-2 mimetics, and human milk oligosaccharides. Human milk oligosaccharides inhibited campylobacter colonization of mice in vivo and human intestinal mucosa ex vivo. Campylobacter colonization of nursing mouse pups was inhibited if their dams had been transfected with a human alpha1,2-fucosyltransferase gene that caused expression of H(O) antigen in milk. We conclude that campylobacter binding to intestinal H-2 antigen is essential for infection. Milk fucosyloligosaccharides and specific fucosyl alpha1,2-linked molecules inhibit this binding and may represent a novel class of antimicrobial agents. 相似文献
193.
FKBP12 binding to RyR1 modulates excitation-contraction coupling in mouse skeletal myotubes 总被引:2,自引:0,他引:2
Avila G Lee EH Perez CF Allen PD Dirksen RT 《The Journal of biological chemistry》2003,278(25):22600-22608
The skeletal muscle sarcoplasmic reticulum (SR) Ca2+ release channel or ryanodine receptor (RyR1) binds four molecules of FKBP12, and the interaction of FKBP12 with RyR1 regulates both unitary and coupled gating of the channel. We have characterized the physiologic effects of previously identified mutations in RyR1 that disrupt FKBP12 binding (V2461G and V2461I) on excitation-contraction (EC) coupling and intracellular Ca2+ homeostasis following their expression in skeletal myotubes derived from RyR1-knockout (dyspedic) mice. Wild-type RyR1-, V246I-, and V2461G-expressing myotubes exhibited similar resting Ca2+ levels and maximal responses to caffeine (10 mm) and cyclopiazonic acid (30 microm). However, maximal voltage-gated Ca2+ release in V2461G-expressing myotubes was reduced by approximately 50% compared with that attributable to wild-type RyR1 (deltaF/Fmax = 1.6 +/- 0.2 and 3.1 +/- 0.4, respectively). Dyspedic myotubes expressing the V2461I mutant protein, that binds FKBP12.6 but not FKBP12, exhibited a comparable reduction in voltage-gated SR Ca2+ release (deltaF/Fmax = 1.0 +/- 0.1). However, voltage-gated Ca2+ release in V2461I-expressing myotubes was restored to a normal level (deltaF/Fmax = 2.9 +/- 0.6) following co-expression of FKBP12.6. None of the mutations that disrupted FKBP binding to RyR1 significantly affected RyR1-mediated enhancement of L-type Ca2+ channel activity (retrograde coupling). These data demonstrate that FKBP12 binding to RyR1 enhances the gain of skeletal muscle EC coupling. 相似文献
194.
195.
The karyotypes of 10 species and one variety of South American Lathyrus were determined and compared with those obtained of five entities from the Northern Hemisphere. Although all the species have a chromosome number of 2n = 14, they could be differentiated by their karyotype formula and quantitative parameters of the karyotypes. Phenetic distance and principal component analysis showed that in spite of the differences observed among entities, they can be grouped in clusters that coincide with the taxonomic sections established by F. K. Kupicha and with the life cycle of the species. South American species form a homogeneous group and can be distinguished by the presence of a subtelocentric pair, which has a macrosatellite in the long arm, and the lack of a short metacentric pair characteristic of most species of the Northern Hemisphere. From an evolutionary point of view, variation in total chromosome length without major changes in the karyotype formula suggests that changes in the amounts of genomic DNA are proportional to the relative length of each chromosome arm and that species of Notolathyrus evolved in a concerted fashion. Variation in genome size, however, is congruent with morphological variation of some reproductive organs as well as with the life cycle and minimum generation time, as predicted by the nucleotype hypothesis. 相似文献
196.
197.
Hernández-Montalvo V Martínez A Hernández-Chavez G Bolivar F Valle F Gosset G 《Biotechnology and bioengineering》2003,83(6):687-694
In Escherichia coli, the uptake and phosphorylation of glucose is carried out mainly by the phosphotransferase system (PTS). Despite the efficiency of glucose transport by PTS, the required consumption of 1 mol of phosphoenolpyruvate (PEP) for each mol of internalized glucose represents a drawback for some biotechnological applications where PEP is a precursor of the desired product. For this reason, there is considerable interest in the generation of strains that can transport glucose efficiently by a non-PTS mechanism. The purpose of this work was to study the effect of different gene expression levels, of galactose permease (GalP) and glucokinase (Glk), on glucose internalization and phosphorylation in a E. coli PTS(-) strain. The W3110 PTS(-), designated VH32, showed limited growth on glucose with a specific growth rate (mu) of 0.03 h(-1). A low copy plasmid family was constructed containing E. coli galP and glk genes, individually or combined, under the control of a trc-derived promoter set. This plasmid family was used to transform the VH32 strain, each plasmid having different levels of expression of galP and glk. Experiments in minimal medium with glucose showed that expression of only galP under the control of a wild-type trc promoter resulted in a mu of 0.55 h(-1), corresponding to 89% of the mu measured for W3110 (0.62 h(-1)). In contrast, no increase in specific growth rate (mu) was observed in VH32 with a plasmid expressing only glk from the same promoter. Strains transformed with part of the plasmid family, containing both galP and glk genes, showed a mu value similar to that of W3110. Fermentor experiments with the VH32 strain harboring plasmids pv1Glk1GalP, pv4Glk5GalP, and pv5Glk5GalP showed that specific acetate productivity was twofold higher than in W3110. Introduction of plasmid pLOI1594, coding for pyruvate decarboxylase and alcohol dehydrogenase from Zymomonas mobilis, to strain VH32 carrying one of the plasmids with galP and glk caused a twofold increase in ethanol productivity over strain W3110, also containing pLOI1594. 相似文献
198.
199.
Ortega AL Carretero J Obrador E Gambini J Asensi M Rodilla V Estrela JM 《The Journal of biological chemistry》2003,278(16):13888-13897
High GSH content associates with high metastatic activity in B16-F10 melanoma cells cultured to low density (LD B16M). GSH homeostasis was investigated in LD B16M cells that survive after adhesion to the hepatic sinusoidal endothelium (HSE). Invasive B16M (iB16M) cells were isolated using anti-Met-72 monoclonal antibodies and flow cytometry-coupled cell sorting. HSE-derived NO and H(2)O(2) caused GSH depletion and a decrease in gamma-glutamylcysteine synthetase activity in iB16M cells. Overexpression of gamma-glutamylcysteine synthetase heavy and light subunits led to a rapid recovery of cytosolic GSH, whereas mitochondrial GSH (mtGSH) further decreased during the first 18 h of culture. NO and H(2)O(2) damaged the mitochondrial system for GSH uptake (rates in iB16M were approximately 75% lower than in LD B16M cells). iB16M cells also showed a decreased activity of mitochondrial complexes II, III, and IV, less O(2) consumption, lower ATP levels, higher O(2) and H(2)O(2) production, and lower mitochondrial membrane potential. In vitro growing iB16M cells maintained high viability (>98%) and repaired HSE-induced mitochondrial damages within 48 h. However, iB16M cells with low mtGSH levels were highly susceptible to TNF-alpha-induced oxidative stress and death. Therefore depletion of mtGSH levels may represent a critical target to challenge survival of invasive cancer cells. 相似文献
200.
García-Barrios L Mayer-Foulkes D Franco M Urquijo-Vásquez G Franco-Pérez J 《Bulletin of mathematical biology》2001,63(3):507-526
Spatial disposition of plants in intercrops, and differences in sowing time between species, can strongly affect their ecological
interactions and, in consequence, the system’s viability and performance. Empirical exploration of a wide range of spatial
and temporal plant arrangements is costly and time-consuming. Modelling the growth of mixed crops is a tool which, combined
with empirical tests, can greatly reduce the time and investment required for this task. Spatially explicit, individual-based
dynamic models seem well suited for this purpose; their exploration and experimental validation for the case of simple, two-species,
artificial plant communities, can also provide further insight as to how the spatial and temporal scales of a plant’s multispecific
neighbourhood affect its growth and performance. The aim of this investigation was to further develop a published spatially
explicit individual-based mixed crop growth model [Vandermeer, J. H. (1989). The Ecology of Intercropping, Cambridge, U.K.: Cambridge University Press, p. 237], and to validate it experimentally. With this purpose in mind: (1)
computer programs to simulate individual plant growth and to perform statistical analysis of both deterministic and stochastic
versions of the model were developed; (2) the model was parametrized using a complex experimental diculture with several cohorts
and spatial arrangements; (3) the predictive capacity of the model was tested using independent spatio-temporal experimental
arrangements; (4) a modified version of the model was written, which abandons the assumption of linearity of the neighbourhood
index at the cost of increasing the number of parameters; (5) The performance of stochastic versions of both Vandermeer’s
and our modified model were compared, employing a non-parametric measure of goodness of fit. We conclude that this approach
to modelling plant growth subject to intra and interspecific competition is a remarkably efficient, general, conceptually
elegant, heuristic tool whose predictive power can be further improved when nonlinear terms are introduced into the neighbourhood
competition index, as done in our modified version of Vandermeer’s model. 相似文献