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Herman Yeger Diane Forget Jennifer Alami Bryan R. G. Williams 《In vitro cellular & developmental biology. Animal》1996,32(8):496-504
Summary The temporal and spatial expression patterns of the Wilms tumor gene, WT1, were studied during the organogenesis of the mouse
kidneyin vitro. In situ hybridization and immunocytochemistry localized cellular expression of WT1 in whole kidney organ cultures to the induced
metanephric mesenchyme and developing podocytes. Organ cultures were further characterized immunocytochemically with antibodies
that specifically labeled the different tubular epithelial components and supporting mesenchyme of the developing nephrons.
In organ cultures, the WT1 expression pattern could be visualized in induced metanephric mesenchyme and entire cell cohorts
of differentiating podocytes. Expression of WT1 and cell specific markers were retained in short-term monolayer cultures of
dissociated kidneys. The development of the metanephric kidneyin vitro involves a highly restricted temporal and spatial cellular expression pattern of WT1 which closely follows that observed
in tissue sections from gestational kidney isolated during organogenesis in the mouse. 相似文献
44.
Klaus Werner Wolf Sabine Glatzel Frauke Niedereichholz Bryan M. Turner 《Biotechnic & histochemistry》1996,71(3):137-144
We describe a method for isolating chromosomes from testes of the desert locust, Schistocerca gregaria, and their subsequent incubation with antibodies directed against chromosomal proteins. The procedure involves hypotonic pretreatment of the germ cells, centrifugation onto coverslips in a cytocentrifuge and immunolabeling, while still unfixed, using a chromatin-stabilizing buffer. In the present case, an antibody specific for the acetylated isoforms of his tone H4 was tested. After the antibody treatment, the preparations are fixed using formaldehyde, stained with a DNA-specific fluorescent dye and mounted. Analysis of the preparations revealed good preservation of chromosome structure in prophase spermatogonia and late prophase I spermatocytes. Fully condensed chromosomes were not observed and are probably lost during preparation. The bright fluorescence of the autosomes indicates that the reaction between the antibody against acetylated histone H4 and its chromosomal antigen is not impeded. In contrast, the X univalent remained unstained with the exception of a small terminal band. Thus, cytospin preparations of locust germ cells allow high resolution immunolabeling with antibodies against chromosome-associated proteins. 相似文献
45.
Summary A cellular model, where each individual is explicitly defined, is used to describe a population of a mycophagous species ofDrosophila. Patches represent single fungal fruiting bodies which are only available as oviposition sites for a single fly generation. Standard competition equations are used to describe the interaction between larval genotypes at each patch. Dispersal of adults is obligatory and uses a simple model of patch choice to produce aggregated arrivals of adults at fresh patches. The degree to which aggregation of adults and eggs can promote coexistence of genotypes in a one-locus, two-allele system with dominance is explored. When both phenotypes (A- andaa) are aggregated, a polymorphism can be maintained for over 1000 generations even when the selective disadvantage of one phenotype (aa) is great. This model enhances the degree of polymorphism in a population, using aggregation. It does not preclude the operation of other methods which enhance the coexistence of genotypes. Therefore, it is acting to augment the degree of polymorphism maintained in species which exploit patchy and ephemeral habitats, including allDrosophila and a wide range of other organisms. 相似文献
46.
The effects of calystegines isolated from edible fruits and vegetables on mammalian liver glycosidases 总被引:1,自引:0,他引:1
Asano Naoki; Kato Atsushi; Matsui Katsuhiko; Watson Alison A.; Nash Robert J.; Molyneux Russell J.; Hackett Lucy; Topping Joanna; Winchester Bryan 《Glycobiology》1997,7(8):1085-1088
The polyhydroxylated nortropane alkaloids called calyste-ginesoccur in many plants of the Convolvulaceae, Solanaceae, andMoraceae families. Certain of these alkaloids exhibit potentinhibitory activities against glycosidases and the recentlydemonstrated occurrence of calystegines in the leaves, skins,and sprouts of potatoes (Solatium tuberosum), and in the leavesof the eggplant (S.melongena), has raised concerns regardingthe safety of these vegetables in the human diet. We have surveyedthe occurrence of calystegines in edible fruits and vegetablesof the families Convolvulaceae, Solanaceae, and Moraceae byGC-MS. Calystegines A3, B1, B2, and C1 were detected in allthe edible fruits and vegetables tested; sweet and chili peppers,potatoes, eggplants, tomatoes, Physalis fruits, sweet potatoes,and mulberries. Calystegines B1 and C1 were potent competitiveinhibitors of the bovine, human, and rat β-glucosidaseactivities, with K1 values of 150, 10, and 1.9 µM, respectivelyfor B1 and 15,1.5, and 1 µM, respectively, for C1. CalystegineB2 was a strong competitive inhibitor of the -galactosidaseactivity in all the livers. Human β-xylosidase was inhibitedby all four nortropanes, with calystegine C1 having a K1 of0.13 µM. Calystegines A3 and B2 selectively inhibitedthe rat liver β-glucosidase activity. The potent inhibitionof mammalian β-glucosidase and -galactosidase activitiesin vitro raises the possibility of toxicity in humans consuminglarge amounts of plants that contain these compounds. edible plants calystegines glycosidase inhibitors bovine, human, and rat liver 相似文献
47.
R. A. Virginia L. M. Baird J. S. La Favre W. M. Jarrell B. A. Bryan G. Shearer 《Plant and Soil》1984,79(2):273-284
Summary The relative nitrogen fixation efficiencies (RE 1-[H2 evolved÷C2H2 reduced]·100) of four mesquite (Prosopis glandulosa var.torreyana) rhizobia (Strains WR 1001, WR 1002, L5, L9) and a cowpea rhizobia (Strain 176A32) on mesquite were evaluated in a glasshouse experiment. Plant yield, shoot N accumulation, and the natural15N abundance (15N) of nodule tissue were determined. Strain WR 1002 failed to nodulate mesquite and strain L5 produced ineffective nodules. Among the three effective strains (WR 1001, L9, 176A32) the cowpea strain (176A32) and strain L9 had significantly higher RE than strain WR 1001. Differences in RE, however, were not accompanied by significantly higher plant yield and shoot N accumulation. The difference in15N abundance between foliar tissue and nodules (nodules minus leaves) was 0.47 15N for the ineffective L5 nodules, while for the effective WR 1001, L9, and 176A32 nodules, respectively, this difference was 8.35, 7.81, and 8.35 15N. This indicates a similar relationship between N2-fixing effectiveness and natural15N enrichment of nodules that was previously observed in soybeans (Glycine max, L. Merr.). Strains WR 1001 and L9 produced elongate, indeterminate nodules typical for mesquite. The ineffective L5 nodules had few infected cells and an abundance of cortical amyloplasts. Mesquite nodules produced by the cowpea strain were spherical and were somewhat more similar in internal morphology to determinate nodules typical of cowpea than indeterminate nodules normally associated with mesquite. 相似文献
48.
Secretion of melanophore-stimulating hormone (MSH) in long-term cultures of pituitary neurointermediate lobes 总被引:1,自引:0,他引:1
Summary Neurointermediate lobes from pituitaries of the frog, Rana berlandieri forreri (Rana pipiens, sensu lato), were maintained in organ culture in media with and without serum for up to six months. The cultured tissues were examined periodically by light microscopy and transmission electron microscopy and by bioassay of the melanophore-stimulating hormone (MSH) secreted and present in the culture media. Light-microscopic observations revealed a high degree of preservation of the pars intermedia at four weeks with isolated areas of some glands maintaining histological integrity for the entire six months. Similarly, at the ultrastructural level the cells appeared morphologically intact and to be actively synthesizing and secreting hormone. Bioassays showed the glands to be continuously secreting MSH; however, larger yields of hormone were obtained in media lacking serum. No significant ultrastructural differences between cells grown in the presence or absence of serum were detected. The difference in concentration of MSH between the two groups therefore apparently results from enzymatic degradation of the hormone by the serum. Organ culture of the vertebrate neurointermediate lobe may provide a unique method for the production of large quantities of MSH and for the study of other melanotropic and opiate peptides as they may be synthesized and secreted by the pars intermedia. 相似文献
49.
Geoffrey L.R. Gordon V.Bryan Lawlis Bruce A. McFadden 《Archives of biochemistry and biophysics》1980,199(2):400-412
2-Carboxy-d-hexitol 1,6-bisphosphate (CHBP) has been prepared from d-fructose 1,6-bisphosphate and cyanide. DEAE-Sephadex chromatography separated the reaction products into two fractions which were identified as CHBP and CHBP-lactone. CHBP is presumably a mixture of two diastereomers, 2-carboxy-d-glucitol 1,6-bisphosphate and 2-carboxy-d-mannitol 1,6-bisphosphate, but an attempt to separate these compounds was not successful. The material in the CHBP-lactone peak had no effect on d-ribulose 1,5-bisphosphate (RuBP) carboxylase. However, CHBP was a potent reversible inhibitor of RuBP carboxylases. This compound displayed an inhibition constant (Ki at pH 8.0 and 30 °C) of 1–2 μm with the enzymes from spinach and barley, while the Ki was 60–70 μm with bacterial RuBP carboxylases from Pseudomonas oxalaticus and Rhodospirillum rubrum. The mode of inhibition was competitive with respect to RuBP for all the carboxylases, and noncompetitive with respect to CO2 for the enzymes from spinach, P. oxalaticus and R. rubrum. The results indicate that, in the binding of certain organic phosphates by RuBP carboxylases, there may be a fundamental difference between the enzymes isolated from microbial and from higher plant sources. RuBP oxygenase activities from spinach and P. oxalaticus were also inhibited by CHBP, with Ki values which were similar to those obtained with the carboxylase activity of the same enzymes. The mode of inhibition of the oxygenase activities was also competitive with respect to RuBP. Thus, it seems that the binding of CHBP is similar for the carboxylase and oxygenase reactions of the same enzyme. 相似文献
50.
Clofilium is the most promising member in a new series of antifibrillatory agents to selectively prolong cardiac action potential duration (APD) and effective refractory period (ERP). In normal superfused canine Purkinje fibers, clofilium prolonged APD and ERP by a maximum of 35% (ED50=1.3 × 10?8 M). The effect of clofilium reached equilibrium in 61±3 min but APD did not return toward control during several hr of superfusion with drug-free medium. No change in rate of rise, amplitude, resting potential or rate of diastolic depolarization was noted in the presence of clofilium (3 × 10?7 M). Clofilium increased the canine ventricular fibrillation threshold (VFT) measured using gated trains of electrical stimuli. This effect occurred in a dose-related fashion following a 30 min infusion of a total of 0.5 or 1.0 μmole/kg of clofilium. The increase was evident within 30 min after ending the infusion and persisted for at least 4 hr. Following the infusion of clofilium (1.0 μmole/kg) 22% of the episodes of ventricular fibrillation (VF) spontaneously reverted to normal sinus rhythm without the use of direct current countershock; this phenomenon did not occur in dextrose-infused dogs. 相似文献