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Genetic assessment of the importance of galectin-3 in cancer initiation, progression, and dissemination in mice 总被引:1,自引:0,他引:1
Eude-Le Parco I Gendronneau G Dang T Delacour D Thijssen VL Edelmann W Peuchmaur M Poirier F 《Glycobiology》2009,19(1):68-75
The galectin family of β-galactoside binding lectins isinvolved in normal and pathological processes. Altered expressionof galectin-3 has been described in many cancers, and studiesof cancer cell lines have implicated this lectin in variousaspects of the tumorigenic cascade. The goal of this reportwas to directly assess the importance of galectin-3 in tumorbiology by introducing the galectin-3 null mutation (galectin-3–/–)into mouse lines genetically programmed to develop cancers.We used two mouse models of human intestinal cancer, the ApcMinand Apc1638N lines, to study tumor initiation and tumor progression.We also crossed the galectin-3–/– mice with PyMTtransgenic animals, a model in which primary mammary gland tumorsgive rise to lung metastases at high frequency. Unexpectedly,we show that the absence of galectin-3 does not affect the evolutionof the disease in any of these three situations. 相似文献
3.
SEARLE LINDA M.; CHAMBERLAIN KEITH; RAUSCH THOMAS; BUTCHER DENNIS N. 《Journal of experimental botany》1982,33(5):935-942
[Methylene-14C]-3-indolylmethylglucosinolate (14C-IMG) was convertedin vitro to [methylene-14Cl-3-indolylacetonitrile (14C-IAN)by myrosinase over a pH range of 4.0-6.0 and this conversionwas enhanced by ferrous ions. Other products of the reactionincluded 3-indolylmethanol, 3, 3'-diindolylmethane and ascorbigenA. Trace amounts of 14C-IAN were produced non-enzymically from14C-IMG in the presence of ferrous ion over a similar pH range.Furthermore, swede tissues (Brassica napus cv. Danestone) infectedwith Plasmodiophora brassicae Woron. could convert 14C-IMG to14C-IAN. These results were consistent with the hypothesis thatthe overgrowth symptoms of the clubroot disease are caused bythe conversion of IMG to the auxin precursor IAN. 相似文献
4.
Squalamine, a novel cationic steroid, specifically inhibits the brush-border Na+/H+ exchanger isoform NHE3 总被引:1,自引:0,他引:1
Akhter S.; Nath S. K.; Tse C. M.; Williams J.; Zasloff M.; Donowitz M. 《American journal of physiology. Cell physiology》1999,276(1):C136
Squalamine, anendogenous molecule found in the liver and other tissues ofSqualus acanthias, hasantibiotic properties and causes changes in endothelial cell shape. Thelatter suggested that its potential targets might include transportproteins that control cell volume or cell shape. The effect of purifiedsqualamine was examined on clonedNa+/H+exchanger isoforms NHE1, NHE2, and NHE3 stably transfected in PS120fibroblasts. Squalamine (1-h pretreatment) decreased the maximalvelocity of rabbit NHE3 in a concentration-dependent manner (13, 47, and 57% inhibition with 3, 5, and 7 µg/ml, respectively) and alsoincreasedK'[H+]i.Squalamine did not affect rabbit NHE1 or NHE2 function. The inhibitoryeffect of squalamine was 1) timedependent, with no effect of immediate addition and maximum effect with1 h of exposure, and 2) fullyreversible. Squalamine pretreatment of the ileum for 60 min inhibitedbrush-border membrane vesicleNa+/H+activity by 51%. Further investigation into the mechanism of squalamine's effects showed that squalamine required the COOH-terminal 76 amino acids of NHE3. Squalamine had no cytotoxic effect at theconcentrations studied, as indicated by monitoring lactate dehydrogenase release. These results indicate that squalamine 1) is a specific inhibitor of thebrush-border NHE isoform NHE3 and not NHE1 or NHE2,2) acts in a nontoxic and fullyreversible manner, and 3) has adelayed effect, indicating that it may influence brush-borderNa+/H+exchanger function indirectly, through an intracellular signaling pathway or by acting as an intracellular modulator. 相似文献
5.
Total uptake and incorporation of 3H-thymidine, 3H-thymine,and 3H-deoxyuridine into DNA have been investigated in the apical3 cm of the primary root of Vicia faba. Evidence has been obtainedthat endogenous TdR in these roots may be transported eitherapically or basally; apical movement being greater than movementfrom the apex towards the base of the root. The results havebeen discussed with respect to the possible distribution ofendogenous pools of thymidine, thymine, and deoxyuridine inthe primary root of V. faba. 相似文献
6.
A permanent cell line with inducible expression of the humananion exchanger protein 1 (hAE1) was constructed in a derivative ofhuman embryonic kidney cells (HEK-293). In the absence of the inducer,muristerone A, the new cell line had no detectable hAE1 protein byWestern analysis or additional36Cl flux. Increasing dose andincubation time with muristerone A increased the amount of protein(both unglycosylated and glycosylated). The4,4'-dinitrostilbene-2,2'-disulfonate(DNDS)-inhibitable rapid Cl exchange flux was increased up to40-fold in induced cells compared with noninduced cells. There was noDNDS-inhibitable rapid flux component in noninduced cells. This resultdemonstrates inducible expression of a new rapid Cl transport pathwaythat is DNDS sensitive. The additional transport of36Cl and35SO4had the characteristics of hAE1-mediated transport in erythrocytes: 1) inhibition by 250 µM DNDS,2) activation of36Cl efflux by external Cl with aconcentration producing half-maximal effect of 4.8 mM,3) activation of36Cl efflux by external anionsthat was selective in the orderNO3 = Cl > Br > I, and4) activation of35SO4influx by external protons. Under the assumption that the turnovernumbers of hAE1 were the same as in erythrocytes, there was good agreement (±3-fold) between the number of copies ofglycosylated hAE1 and the induced tracer fluxes. This is the firstexpression of hAE1 in a mammalian system to track the kineticcharacteristics of the native protein. 相似文献
7.
Structure, function, and genomic organization of human Na(+)-dependent high-affinity dicarboxylate transporter 总被引:6,自引:0,他引:6
Wang H Fei YJ Kekuda R Yang-Feng TL Devoe LD Leibach FH Prasad PD Ganapathy V 《American journal of physiology. Cell physiology》2000,278(5):C1019-C1030
We have clonedand functionally characterized the human Na+-dependenthigh-affinity dicarboxylate transporter (hNaDC3) from placenta. ThehNaDC3 cDNA codes for a protein of 602 amino acids with 12 transmembrane domains. When expressed in mammalian cells, the clonedtransporter mediates the transport of succinate in the presence ofNa+ [concentration of substrate necessary for half-maximaltransport (Kt) for succinate = 20 ± 1 µM]. Dimethylsuccinate also interacts with hNaDC3. TheNa+-to-succinate stoichiometry is 3:1 and concentration ofNa+ necessary for half-maximal transport(KNa+0.5) is 49 ± 1 mM as determined by uptake studies withradiolabeled succinate. When expressed in Xenopuslaevis oocytes, hNaDC3 induces Na+-dependent inwardcurrents in the presence of succinate and dimethylsuccinate. At amembrane potential of 50 mV,KSuc0.5 is 102 ± 20 µM andKNa+0.5 is 22 ± 4 mM as determined by the electrophysiological approach. Simultaneous measurements of succinate-evoked charge transfer andradiolabeled succinate uptake in hNaDC3-expressing oocytes indicate acharge-to-succinate ratio of 1:1 for the transport process, suggestinga Na+-to-succinate stoichiometry of 3:1. pH titration ofcitrate-induced currents shows that hNaDC3 accepts preferentially thedivalent anionic form of citrate as a substrate. Li+inhibits succinate-induced currents in the presence of Na+.Functional analysis of rat-human and human-rat NaDC3 chimeric transporters indicates that the catalytic domain of the transporter lies in the carboxy-terminal half of the protein. The humanNaDC3 gene is located on chromosome20q12-13.1, as evidenced by fluorescent in situ hybridization. Thegene is >80 kbp long and consists of 13 exons and 12 introns. 相似文献
8.
Lamaze, T., Sentenac, H. and Grignon, C. 1987. Orthophosphaterelations of root: NO3effects on orthophosphate influx,accumulation and secretion into the xylem.J. exp. Bot.38: 923934. Orthophosphate (Pi) accumulation by barley (Hordeum vulgareL.) roots was specifically inhibited by NO3 as comparedto Cl and SO42 , and Pi secretion into the xylemwas stimulated. The inhibition of Pi accumulation by NO3was also observed in roots of intact photosynthesizing horsebean(Vicia faba L.), rice (Oryza sativa L.) and soybean (Glycinemax L.) plants. NO3 effects on Pi transport by rootswere more thoroughly investigated with corn (Zea mays L.). Theywere due to intracellular NO3. Pi secretion was stillstimulated by NO3 after Pi withdrawal from the absorptionsolution. 32Pi influx decreased during Pi accumulation, supportingthe hypothesis that this ion allosterically regulated its owntransport system by feedback control. This control was modulatedby other anions: the decrease was more pronounced in the presenceof nitrate. Chronologically, the depressive effect of NO3on 32Pi influx appeared after the inhibition of Pi accumulation.Furthermore, under conditions where Pi accumulation was notaffected by NO3, 32Pi influx and Pi secretion into thexylem became insensitive to the presence of nitrate. Our hypothesisis that the stimulative effect of NO3 on Pi secretionand the depressive one on 32Pi influx are the repercussionsof an increase in the Pi cytosolic concentration due to an NO3-induced decrease in Pi uptake by the vacuoles. Key words: Root, orthophosphate fluxes, orthophosphate accumulation, nitrate, ionic interaction 相似文献
9.
Phosphate transport by the human renal cotransporter NaPi-3 expressed in HEK-293 cells 总被引:1,自引:0,他引:1
The human renal Na-PO4cotransporter gene NaPi-3 was expressed in human embryonic kidneyHEK-293 cells, and the transport characteristics were measured in cellstransfected with a vector containing NaPi-3 or with the vector alone(sham transfected). The initial rate of32PO4influx had saturation kinetics for external Na andPO4 with K Na1/2 of 128 mM(PO4 = 0.1 mM) andK PO41/2of 0.084 mM (extracellular Na = 143 mM) in sham- and NaPi-3-transfectedcells expressing the transporter. Transfection had no effect on theNa-independent 32PO4influx, but transfection increased Na-dependent32PO4influxes 2.5- to 5-fold. Of the alkali cations, only Na significantly supported PO4 influx. Arsenateinhibited flux with an inhibition constant of 0.4 mM. The phosphatetransport in sham- and NaPi-3-transfected cells has nearly the sametemperature dependence in the absence and presence of extracellularNa. The Na-dependent phosphate flux decreased with pH insham-transfected cells but was pH independent in transfected cells. TheNa-dependent32PO4influx was inhibited byp-chloromercuriphenylsulfonate,phosphonoformate, phloretin, vanadate, and5-(N-methyl-N-isobutyl)-amiloridebut not by amiloride or other amiloride analogs. These functional characteristics are in general agreement with the known behavior ofNaPi-3 homologues in the renal tubule of other species and, thus,demonstrate the fidelity of this transfection system for the study ofthis protein. Commensurate with the increased functional expression,there was an increase in the amount of NaPi-3 protein by Westernanalysis. 相似文献
10.
Nonhcbcl, H. M. 1986. Measurement of the rates of oxindole-3-aceticacid turnover and indole-3-acetic acid oxidation in Zea maysseedlings.J. exp. Bat. 37: 16911697. Oxindole-3-acetic acid is the pnncipal catabolite of indole-3-aceticacid in Zea mays seedlings. In this paper measurements of theturnover of oxindole-3-acetic acid are presented and used tocalculate the rate of indole-3-acetic acid oxidation. [3H]Oxindolc-3-acetic acid was applied to the endosperm of Zeamays seedlings and allowed to equilibrate for 24 h before thestart of the experiment. The subsequent decrease in its specificactivity was used to calculate the turnover rate. The averagehalf-life of oxindole-3-acetic acid in the shoots was foundto be 30 h while that in the kernels had an average half-lifeof 35 h. Using previously published values of the pool sizesof oxindole-3-acetic acid in shoots and kernels from seedlingsof the same age and variety, and grown under the same conditions,the rate of indole-3-acetic acid oxidation was calculated tobe I-I pmol plant1 h1 in the shoots and 7·1pmol plant1 h1 in the kernels. Key words: Oxindole-3-acetic acid, indole-3-acetic acid, turnover, Zea mays 相似文献
11.
SCHMITT ANDREAS K.; LEE HELEN S. J.; LUTTGE ULRICH 《Journal of experimental botany》1988,39(11):1581-1590
Gas exchange measurements were undertaken on 2-year-old plantsof Clusia rosea. The plants were shown to have the ability toswitch from C3-photosynthesis to CAM and vice versa regardlessof leaf age and, under some conditions, CO2 was taken up continuously,throughout the day and night. The light response was saturatedby 120 µmol m2 s1 typical of a shade plant. Gas exchange patterns in response to light, water and VPD wereexamined. All combinations of daytime and night-time CO2 uptakewere observed, with rates of CO2 uptake ranging from 2 to 11µmol m2 s1 depending upon water status andlight. Categorization of this plant asC3, CAM or an intermediateis impossible. Differing VPD affected the magnitude of changesfrom CAM to C3-photosynthesis (0 to 0.5 and 0 to 6.0 µmolm2 s1 CO2, respectively) when plants were watered.Under well-watered conditions, but not under water stress, highPPFD elicited changes from CAM to C3 gas exchange. This is unusualnot only for a shade plant but also for a plant with CAM. Itis of ecological importance for C. rosea, which may spend theearly years of its life as an epiphyte or in the forest understorey,to be able to maximize photosynthesis with minimal water loss. Key words: Clusia rosea, CAM, C3, stress 相似文献
12.
Molecular Characterization of the waxy Locus of Rice (Oryza sativa) 总被引:10,自引:0,他引:10
13.
Delhon Patricia; Gojon Alain; Tillard Pascal; Passama Lucien 《Journal of experimental botany》1995,46(10):1585-1594
The effect of light on NO3 utilization was investigatedin non-nodulated soybean (Clycine max L. Merr., cv. Kingsoy)plants during a 14/10 h light/dark period at a constant temperatureof 26C. A 3050% decrease of net NO3 uptake ratewas observed 26 h after the lights were turned off. Thiswas specifically due to an inhibition of NO3 influx asmeasured by 15N incorporation during 5 min. The absolute valuesof NO3 efflux depended on whether the labelling protocolinvolved manipulation of the plants or not, but were not affectedby illumination of the shoots. Darkness had an even more markedeffect in lowering the reduction of 15NO3 in both rootsand shoots, as well as xylem transport of 15NO3 and reduced15N. Concurrently with this slowing down of transport and metabolicprocesses, accumulations of NO3 and Asn were significantlystimulated in roots during the dark period. These data are discussedin view of the hypothesis that darkness adversely affects NO3uptake through specific feedback control, in response to alterationsin the later steps of N utilization which are more directlydependent on light. Key words: Glycine max, light/dark cycles, nitrate uptake, nitrate reduction 相似文献
14.
Brette F Lacampagne A Sallé L Findlay I Le Guennec JY 《American journal of physiology. Cell physiology》2003,285(2):C310-C318
Inactivation of the L-type Ca2+ current (ICaL) was studied in isolated guinea pig ventricular myocytes with different ionic solutions. Under basal conditions, ICaL of 82% of cells infused with Cs+-based intracellular solutions showed enhanced amplitude with multiphasic decay and diastolic depolarization-induced facilitation. The characteristics of ICaL in this population of cells were not due to contamination by other currents or an artifact. These phenomena were reduced by ryanodine, caffeine, cyclopiazonic acid, the protein kinase A inhibitor H-89, and the cAMP-dependent protein kinase inhibitor. Forskolin and isoproterenol increased ICaL by only 60% in these cells. Cells infused with either N-methyl-D-glucamine or K+-based intracellular solutions did not show multiphasic decay or facilitation under basal conditions. Isoproterenol increased ICaL by 200% in these cells. In conclusion, we show that multiphasic inactivation of ICaL is due to Ca2+-dependent inactivation that is reversible on a time scale of tens of milliseconds. Cs+ seems to activate the cAMP-dependent protein kinase pathway when used as a substitute for K+ in the pipette solution. L-type calcium current; calcium-dependent inactivation; facilitation; phosphorylation; cesium 相似文献
15.
Cultures of water fern Azolla pinnata R. Br. exposed for 1 weekto either 30, 50 or 80 nl l-1 O3 showed significant reductionsin rates of growth and N2 fixation, and had fewer heterocysts.Although the levels of glutamine synthetase (GS) and glutamatedehydrogenase (GDH) activity were decreased by low concentrationsof O3 exposures (30 or 50 nl l-1), significant increases inlevels of the same enzymes were caused by higher concentrationsof O3 (80 nl l-1). Increased levels of total protein, polyamines(putrescine and spermidine), and the xanthophyll-cycle precursorof abscisic acid (ABA), violaxanthin, were also found with higherlevels of O3 (80 nl l-1). Levels of ABA itself were significantlyincreased by low level O3 fumigation (30 nl l-1) but significantlydecreased by exposure to 80 nl l-1 O3. This may indicate thathigher levels of atmospheric O3 inhibit the final stages ofABA biosynthesis from violaxanthin.Copyright 1994, 1999 AcademicPress Abscisic acid, nitrogen assimilation, nitrogen fixation, ozone pollution, polyamines, violaxanthin 相似文献
16.
A Cytochrome P450 Mediated Naringenin 3'-Hydroxylase from Sweet Orange Cell Cultures 总被引:1,自引:0,他引:1
Doostdar Hamed; Shapiro Jeffrey P.; Niedz Randall; Burke M.D.; McCollum T. Gregory; McDonald Roy E.; Mayer Richard T. 《Plant & cell physiology》1995,36(1):69-77
A microsomal flavonoid 3'-hydroxylase (F3'H) catalyzing themetabolism of naringenin to eriodictyol in Citrus sinensis (L.)Osbeck cv. Hamlin cell suspension cultures wasshown to be a cytochrome P450 enzyme. This reaction requiredO2 and NADPH and was inhibited by CO, with partial reversalof CO-inhibition by light at 450 nm. Cytochrome P450 contentranged from 1020 pmol (mg microsomal protein)1.The F3'H reaction was shown to be linear in regard to proteinconcentration between 2.5 and 25 µg of microsomal protein.The optimum pH for the reaction was 7.47.6 and the temperatureoptimum was between 30 and 37°C. The apparent Km and Vmaxfor naringenin were 24 µM±3.2 and 81.4±7.9pmol eriodictyol min1 (mg protein)1, respectively.The microsomal F3'H was also capable of forming dihydroquercetinfrom dihydrokaempferol (40 pmol min1 (mg protein)1)and of quercetin from kaempferol (3.25 pmol min1 (mgprotein1). Cytochrome c and ketoconazole were the bestinhibitors of WH activity followed by piperonyl butoxide anda-naphthoflavone. Light was shown to be an inducer of the F3'Halmost doubling the specific activity and increasing the microsomalcytochrome P450 content by 30% over that of dark grown cells.F3'H activity was also confirmed in microsomal preparationsof young (new flush) leaves from Hamlin treesand flavedo of Hamlin oranges, Marshgrapefruit, and Lisbon lemon. No activity wasobserved in older, hardened leaves and albedo of all the fruittested. Initiation of embryogenesis in the Hamlincell suspension cultures by switching from a sucrose mediumto a glycerol-based medium resulted in the down-regulation ofF3'H.
1Mention of a trademark, warranty, proprietary product, or vendordoes not constitute a guarantee by the U.S. Department of Agricultureand does not imply its approval to the exclusion of other productsor vendors that may also be suitable. 相似文献
17.
Summer coastal zooplankton biomass and copepod community structure near the Italian Terra Nova Base (Terra Nova Bay, Ross Sea, Antarctica) 总被引:7,自引:0,他引:7
Pane Luigi; Feletti Mirvana; Francomacaro Barbara; Mariottini Gian Luigi 《Journal of plankton research》2004,26(12):1479-1488
The structure of the zooplankton biotic community and of copepodpopulation in the coastal area of Terra Nova Bay (Ross Sea,Antarctica) was investigated during the 10th Italian AntarcticExpedition (1994/1995). Zooplankton biotic community consistedmainly of pteropods (Limacina helicina and Clione antarctica),Cyclopoid (Oithona similis), Poecilostomatoid (Oncaea curvata)and Calanoid (Ctenocalanus vanus, Paraeuchaeta antarctica, Metridiagerlachei and Stephos longipes) copepods, ostracods, larvalpolychaetes and larval euphausiids. Zooplankton abundance rangedfrom 48.1 ind m3 to 5968.9 ind m3, and copepodabundance ranged from 45.2 ind m3 to 3965.3 ind m3.The highest peak of zooplankton abundance was observed between25 m and the surface and was mainly due to the contributionof O. similis, O. curvata and C. vanus. Zooplankton biomassranged from 5.28 mg m3 to 13.04 mg m3 dry weight;the maximum value was observed between 25 m and the surface.Total lipid content varied from 216.44 to 460.73 mg g1dry weight. 相似文献
18.
The use of chlorate as an analogue for NO3 during nitrateuptake into Chara corallina cells has been investigated. NO3inhibits 36C1O3 influx into Chara over the concentrationrange 01000 mmol m3. Lineweaver-Burke plots ofthe data are characteristic of competitive inhibition by NO3in the low concentration range (0300 mmol m3 ClO3)and apparent KINO3 is 140 mmol m3 which is of a similarorder of magnitude as apparent KmCIO3- 180 mmol m3. Athigher substrate concentrations the inhibition by NO3was not characteristic of competitive or uncompetitive inhibition. 36C1O3/NO3 influx was dependent on K+ and Ca2+in the external medium and inhibited by FCCP. NO3 pretreatmentor N starvation increased subsequent 36C1O3/NO3influx into Chara. A comparison between rates of net NO3uptake and 36C1O3/NO3 influx supported the previoushypothesis that NO3 efflux is an important componentin the determination of overall uptake rates. Key words: Nitrate, Chara, 36CIO3 相似文献
19.
Total uptake of 3H-thymidine, 3H-thymine, and 3H-deoxyuridineand incorporation of these substances into DNA have been investigatedin excised roots in Vicia faba. Total uptake was found to behigher in excised roots than in intact ones. This was a consequenceof exogenous 3H-DNA precursors entering the excised roots throughthe cut surface. Where the cut surface was not immersed in the3H-thymidine solution 3H uptake was also higher than in intactroots. In this case 3H uptake scemed to be correlated with water-lossfrom the cut surface. Incorporation of 3H-thymidine, 3H-thymine,and 3H-deoxyuridine into DNA was found to be higher in excisedroots than in intact ones, probably as a result of a decreasein the size of the relevant endogenous precursor pools. It issuggested that this decrease resulted from the cessation ofthe supply of DNA precursors to these roots from the cotyledons. 相似文献