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181.
The use of bromodeoxyuridine labeling in the human lymphocyte HGPRT somatic mutation assay 总被引:4,自引:0,他引:4
P Ostrosky-Wegman M R Montero C C de Nava R R Tice R Albertini 《Mutation research》1987,191(3-4):211-214
The autoradiographic assay developed by Strauss and Albertini (1979) to quantitate human in vivo somatic mutation at the hypoxanthine guanine phosphoribosyl-transferase locus uses tritiated thymidine to identify mutant cells by their ability to pass through 'S' phase in the presence of 6-thioguanine. An alternative method, based on the incorporation of bromodeoxyuridine (BrdUrd) into the DNA of proliferative cells, followed by differential staining with the fluorescence-plus-Giemsa method, was used to identify 3 classes of lymphocyte nuclei: (a) small darkly stained nuclei, (b) large, reddish-colored nuclei with an apparent nucleolus, and (c) large, bluish-colored nuclei. By double labeling with BrdUrd and tritiated thymidine, it was determined that only the nuclei of the third class had incorporated BrdUrd. These results demonstrate that the technique used for sister-chromatid differentiation can be used to detect putative HGPRT mutants and to determine variant frequencies at the HGPRT locus. 相似文献
182.
Induction of SOS and adaptive responses by alkylating agents in Escherichia coli mutants deficient in 3-methyladenine-DNA glycosylase activities 总被引:3,自引:0,他引:3
The induction of SOS and adaptive responses by alkylating agents was studied in Escherichia coli mutants tagA and alkA deficient in 3-methyladenine-DNA glycosylase activities. The SOS response was measured using an sfiA::lacZ operon fusion. The sfiA operon, in the double mutant tagA alkA, is induced at 5-50-fold lower concentrations of all tested methylating and ethylating compounds, as compared to the wild-type strain. In all cases, the tagA mutation, which inactivates the constitutive and specific 3-alkyladenine-DNA glycosylase I (TagI), sensitizes the strain to the SOS response. The sensitization effect of alkA mutation, which inactivates the inducible 3-alkyladenine-DNA glycosylase II (TagII), is observed under conditions which allow the induction of the adaptive response. We conclude that the persistence of 3-methyladenine and 3-ethyladenine residues in DNA most likely leads to the induction of the SOS functions. In contrast, the adaptive response, evaluated by O6-methylguanine-DNA methyltransferase activity in cell extracts, was not affected by either tagA or alkA mutations. The results suggest that the SOS and adaptive responses use different alkylation products as an inducing "signal". However, adaptation protein TagII inhibits the induction of the SOS response to some extent, due to its action at the level of signal production. Finally, we provide conditions to improve short-term bacterial tests for the detection of genotoxic alkylating agents. 相似文献
183.
Freshly harvested seeds of Agrostemma githago L. do not germinate when they are imbibed at 20°C. The block is located in the embryo and is relased by dry storage at 20°C (after-ripening). Freshly harvested seeds complete only a small part of the processes that occur in after-ripened seeds during the lag phase prior to germination (radicle protrusion). After-ripening removed the block on lag phase processes much faster than the block on germination. This was shown both by direct determinations of the completion of lag phase processes and by measurements of the rate of axial protein synthesis, which approximately doubles when seeds are progressing through the lag phase. It is concluded that the percentage germination does not adequately reflect the extent to which the dormancy mechanism has been overcome. 相似文献
184.
Frank M. Maas Luit J. de Kok Ingrid Hoffmann Pieter J. C. Kuiper 《Physiologia plantarum》1987,70(4):713-721
Exposure of spinach (Spinacia oleracea L. cv. Monosa) to 0.25 μl l?1 H2S reduced the relative growth rate by 26, 47 and 60% at 15, 18 and 25°C, respectively. Shoot to root ratio decreased in plants fumigated at 18 and 25°C. Growth of spinach was not affected by a 2-week exposure to 0.10 or 0.25 μl l?1 SO2. Both H2S and SO2 fumigation increased the content of sulfhydryl compounds and sulfate. A 2-week exposure to 0.25 μl l?1 H2S resulted in an increase in sulfhydryl and sulfate content of 250 to 450% and 63 to 248% in the shoots, respectively, depending on growth temperature. Exposure to 0.15 and 0.30 μl l?1 H2S at 20°C for 2 weeks resulted in a 46% increase in sulfate content of the shoots at 0.30 μl l?1 and no detectable increase at 0.15 μl l?1 H2S; the sulfate content of the roots increased by 195 and 145% at 0.15 and 0.30 μl l?1 H2S, respectively. Fumigation with 0.25 μl l?1 SO2 at 20°C for 2 weeks resulted in an increase in sulfhydryl content and sulfate content in the shoots of 285% and 300 to 1100%. H2S fumigation during the 12 h light period or only during the dark period resulted in identical growth reduction and accumulation of sulfhydryl compounds; they were about 50 and 67% of those observed in continuously exposed plants. H2S- and SO2-exposed plants showed an increased transpiration rate, which was mainly caused by an increased dark-period transpiration. No effect of H2S and SO2 on the water uptake of the plants and the osmotic potential of the leaves was detected. Plants fumigated with 0.25 μl l?1 H2S for 2 weeks were smaller and differed morphologically from the control plants by slightly more abaxially curved leaf margins. Cross sections of the leaves showed smaller cells at the margins and smaller and fewer air spaces. The increased transpiration in the H2S-exposed plants is discussed in relation to the observed morphological changes. 相似文献
185.
Immature seeds of apricot (Prunus armeniaca L.) were fed the native gibberellin A5 (GA5) as 1- and 1,2-[3H]GA5 (5.3 Curies per millimole to 16 milliCuries per millimole) at doses (42 nanograms to 10.6 micrograms per seed) 2 to 530 times the expected endogenous level. After 4 days of incubation, seeds were extracted and free [3H]GA-like metabolites were separated from the highly H2O-soluble [3H]metabolites. For high specific activity feeds the retention times (Rts) of radioactive peaks were compared with Rts of authentic GAs on sequential gradient-eluted → isocratic eluted reversed-phase C18 high performance liquid chromatography (HPLC) -radiocounting (RC). From high substrate feeds (530 and 230 × expected endogenous levels) HPLC-RC peak groupings were subjected to capillary gas chromatography-selected ion monitoring (GC-SIM), usually six characteristic ions. The major free GA metabolites of [3H] GA5 were identified as GA1, GA3, and GA6 by GC-SIM. The major highly water soluble metabolite of [3H]GA5 at all levels of substrate GA5 had chromatographic characteristics similar to authentic GA1-glucosyl ester. Expressed as a percentage of recovered radioactivity, low substrate [3H]GA5 feeds (2 × expected endogenous level) yielded a broad spectrum of metabolites eluting at the Rts where GA1, GA3, GA5 methyl ester, GA6, GA22, GA29 (17, 14, 1.6, 7, 1.1, 0.5%, respectively) and GA glucosyl conjugates of GA1, GA3, GA5, and GA8 (33, 11, 1, 0.1%, respectively) elute. Metabolites were also present at Rts where GA glucosyl conjugates of GA6 and GA29 would be expected to elute (8 and 0.1%, respectively). Only 5% of the radioactivity remained as GA5. Increasing substrate GA5 levels increased the proportion of metabolites with HPLC Rts similar to GA1, GA6, and especially GA1 glucosyl ester, primarily at the expense of metabolites with HPLC Rts similar to GA3, GA3-glucosyl ester, and a postulated conjugate of GA6. There was evidence that high doses of substrate GA5 induced new metabolites which often, but not always, differed from GA1, GA3, and GA6 in HPLC Rt. These same metabolites, when analyzed by GC-SIM yielded m/e ions the same as the M+ and other characteristic m/e ions of the above GAs, albeit at differing GC Rt and relative intensities. 相似文献
186.
Different Characteristics of the Two Glutamate Synthases in the Green Leaves of Lycopersicon esculentum 总被引:4,自引:2,他引:2
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The two glutamate synthases, NAD(P)H- and ferredoxin-dependent, from the green leaves of tomato plants (Lycopersicon esculentum L. cv Hellfrucht frühstamm) differed in their chemical properties and catalytic behavior. Gel filtration of NAD(P)H enzyme gave an apparent molecular size of 158 kilodalton, whereas the ferredoxin enzyme molecular size was 141 kilodalton. Arrhenius plots of the activities of the two enzymes showed that the NAD(P)H enzyme had two activation energies; 109.6 and 70.5 kilojoule per mole; the transition temperature was 22°C. The ferredoxin enzyme however, had only one activation energy; 56.1 kilojoule per mole. The respective catalytic activity pH optima for the NAD(P)H- dependent and the ferredoxin dependent enzymes were around 7.3 and 7.8. In experiments to evaluate the effects of modulators aspartate enhanced the NAD(P)H-linked activity, with a Ka value of 0.25 millimolar, but strongly inhibited that of the ferredoxin-dependent glutamate synthase with a Ki of 0.1 millimolar. 3-Phosphoserine was another inhibitor of the ferredoxin dependent enzyme with a Ki value of 4.9 millimolar. 3-Phosphoglyceric acid was a potent inhibitor of the ferredoxin-dependent form, but hardly affected the NAD(P)H-dependent enzyme. The results are discussed and interpreted to propose different specific functions that these activities may have within the leaf tissue cell. 相似文献
187.
We studied the ability of the hemopoietic organ stroma to recover from damage inflicted by 5 or 7 Gy gamma radiation administered during a period of stromal growth in 4-week-old mice. Irradiation resulted in an immediate depletion of femoral colony-forming fibroblastic progenitors (CFU-F) down to 10-20% of age-matched control values. A full recovery to normal numbers occurred between 120 and 240 days after irradiation and was followed by a secondary decrease 1 year after irradiation. This secondary decrease was accompanied by a decrease in the femoral CFU-S and CFU-C content. Femoral CFU-F attained normal numbers and it was demonstrated to occur from surviving CFU-F and could not be enhanced or prolonged following infusion of unirradiated bone marrow cells after irradiation. During the transient CFU-F recovery the hemopoietic stroma remained severely damaged as judged by the regenerative capacity of spleen and femur stroma after subcutaneous implantation, and the ability of the spleen to accumulate CFU-S in response to lipopolysaccharide injection. We have reported earlier that in similarly irradiated adult mice, no restoration of femoral CFU-F was observed. This difference between 4-week-old and adult mice could not be explained by a difference in in vitro radiosensitivity of CFU-F or in their in vivo regeneration kinetics following irradiation and subsequent lipopolysaccharide injection. We conclude from these observations that the recovery kinetics of the CFU-F population is different in young and adult irradiated mice, infused CFU-F do not contribute to CFU-F regeneration in an irradiated femur, CFU-F are not the sole determinants of stromal regeneration in femur and spleen following irradiation. 相似文献
188.
J Yajeya A Colino J Criado-Gutiérrez A Fernández de Molina 《Revista Espanola de Fisiología》1987,43(2):185-189
The commissural component of the stria terminalis (S. T.) was studied in Equi-Thesin (92.7 mg/kg) anesthetized rats after their exposure in the caudothalamic surface of both hemispheres. Two types of connection between right and left S. T. across the anterior commissure are described: A "direct" connection set up by fibres that run through the S. T. and join the contralateral S. T., and another "indirect" component, formed by cell axons that receive excitatory synaptic contacts from fibres running in the S. T. 相似文献
189.
Glycogen phosphorylase (GP), Hexokinase (HK), Phosphofructokinase (PFK), Pyruvate kinase (PK) and Lactate dehydrogenase (LDH) activities from skeletal muscle and liver were measured in Rana perezi for the four seasons of the year. Skeletal muscle showed a decrease in PFK, PK and LDH activity during winter and summer. Liver displayed an increase in GP activity in spring and in PK and LDH in autumn. 相似文献
190.
L. Rebordinos M. Pérez de la Vega 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1987,74(6):767-772
Summary Further data on the inheritance of seed peroxidases of hexaploid wheat (Triticum aestivum L.) and rye (Secale cereale L.) have been obtained from the genetic analysis of several progenies of both species. Additional data on the inheritance and the chromosomal location and linkage have been obtained for peroxidases of wheat embryo and rye endosperm. The general presence of null alleles in peroxidase loci has been confirmed in both species. In addition to simple monogenic inheritance, epistatic segregations have been observed in both species. These epistatic segregations again suggest the presence of regulatory genes controlling the expression of individual peroxidases in both species and also the existence of several duplicate homoeologous genes in wheat. Known linkage relationships have been confirmed and new ones are indicated. Loci for embryo wheat peroxidases seem to be in chromosomes of the homoeology group 3. The rye endosperm ones should be in chromosome 7R, although it is hypothesized that a duplication of gene EPer1 is located in chromosomes 4R and 7R. 相似文献