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11.
Fructose 1,6-bisphosphatase and glucose 6-phosphatase were induced in organ cultures of liver tissues from 15- and 19-day-old fetal mice, using a culture method that allowed the tissues to be maintained for 7 days in the absence of serum. In cultures from 15-day-old fetal liver, both enzyme activities increased significantly per milligram of DNA after a lag period of 1 to 3 days. In cultures from 19-day-old fetal liver only glucose 6-phosphatase increased in the absence of inducer. N6,O2'-Dibutyryladenosine 3',5'-monophosphate enhanced the rate of increase in fructose 1,6-bisphosphatase and glucose 6-phosphatase activities. The minimum effective concentration of the cyclic nucleotide was approximately 10(-6) M. Dexamethazone inhibited the increase in fructose 1,6-bisphosphatase during culture for 7 days. Glucose 6-phosphatase activity was enhanced by dexamethazone in cultures from 19-day-old fetal liver, but was without effect on glucose 6-phosphatase in cultures from 15-day-old fetal liver. The minimum inhibitory concentration of dexamethazone was less than 10(-8) M. The results suggest a complicated effect of the cyclic nucleotide on the two enzymes in fetal mouse liver as well as different mechanisms of action of dexamethazone on the induction of two enzymes. 相似文献
12.
Sodium-dependence of glycylglycine (Gly-Gly) influx and stimulation of Na+ transport by Gly-Gly were studied in everted sacs, sheet preparations and brush-border membrane vesicles isolated from guinea-pig ileum. Gly-Gly influx was found to be independent of the presence of Na+, while Na+ transport was stimulated by Gly-Gly as evidenced by increases in transmural potential difference (PDt), short-circuit current (Isc) and Na+ influx. The change in PDt (ΔPDt) induced by Gly-Gly was a saturable function of Gly-Gly concentration, showing a Michaelis-Menten type relationship. The half-saturation concentration for Gly-Gly estimated from the electrical data was nearly identical with that estimated from influx data. At a constant Gly-Gly concentration the relationship between Isc and Na+ concentration was sigmoid, and the Hill coefficient was 1.5. Kinetic analysis according to Garay Garrahan indicates that each Gly-Gly carrier has two equivalent non-interacting binding sites for Na+, and that translocation of Na+ occurs when the two Na+ sites on the carrier loaded with Gly-Gly are occupied by Na+. However, our results indicate that the resultant Na+ flow is not capable of stimulating Gly-Gly translocation. 相似文献
13.
Establishment of five human myeloma cell lines 总被引:3,自引:0,他引:3
Masayoshi Namba Takemi Ohtsuki Masaharu Mori Atsushi Togawa Hideho Wada Takashi Sugihara Yoshihito Yawata Tetsuo Kimoto 《In vitro cellular & developmental biology. Plant》1989,25(8):723-729
Summary Five human myeloma cell lines, KMM-1, KMS-5, KMS-11, KMS-12- PE, and KMS-12-BM, have been established at Kawasaki Medical
School since 1980. As the KMS-12-PE and KMS-12-BM lines were obtained from the same patient, these five cell lines have been
derived from four patients with multiple myeloma. The five myeloma cell lines are stably growing at present in RPMI 1640 medium
supplemented with 10% fetal bovine serum. They can also grow in a defined culture medium without serum. That these cell lines
were, human myeloma cells was confirmed by the following findings. Ultranstructually, all five cell lines showed features
characteristic of plasma cells. KMM-1 and KMS-11 cells secreted lambda and kappa chains into the culture medium, respectively,
but the other cell lines produced no immunoglobulins. KMM-1 expressed cytoplasmic lambda antigen, KMS-5 showed cytoplasmic
delta, and KMS-11 expressed surface kappa, whereas KMS-12-PE and KMS-12-BM cells showed no surface or cytoplasmic immunoglobulins.
Regarding reaction with a monoclonal plasma cell antibody (PCA-1), four of the five lines were positive, the exception being
KMS-5. Another monoclonal antibody (CD38), which also recognizes plasma cells, reponded to KMM-1, KMS-12-PE, and KSM-12-BM.
KMS-5 cells expressed acute lymphoblastic leukemia antigens (CALLA). These data suggest that such lines as KMM-1, KMS-11,
KMS-12-PE, and KMS-12-BM represent later stages of B-cell differentiation, and that KMS-5 represents a relatively early stage
of B-cell differentiation. All the cell lines lacked Epstein-Barr virus nuclear antigen, showed abnormal karyotypes of human
origin, and differed from each other in the isozyme patterns examined. Only KMS-5 was tumorigenic when transplanted subcutaneously
into nude mice. 相似文献
14.
Masayoshi Takahashi Hiroshi Yokota Dai Ayusawa Michio Oishi Tetsuo Kunieda 《Biochemical genetics》1992,30(9-10):537-544
A novel restriction fragment length polymorphism (RFLP) in inbred rats was revealed by Southern blot analysis with a clone arbitrarily chosen from a rat genomic library as a probe. A clone named alpha 403 showed interstrain variations in the length of the EcoRI and HindIII fragments. The EcoRI fragments were either 0.7 or 3 kb, those of HindIII were either 4.5 or 5 kb, and three types were identified as combinations of those fragments in 20 inbred rat strains. These types segregated in backcross progeny as codominant alleles. The locus for the RFLP was thus named A403. Analysis of linkage between the RFLP locus and 13 other loci reveal that the A403 locus was closely linked to the Cs-1 locus (15 +/- 5.2%), which belongs to rat linkage group XIII. 相似文献
15.
Kawaguchi Masayoshi; Imaizumi-Anraku Haruko; Fukai Shungo; Syono Kunihiko 《Plant & cell physiology》1996,37(4):461-470
We studied the effects of several plant-growth regulators onthe induction of nodule-like structures on roots of Lotus japonicus,which has been proposed as a candidate for a leguminous plantfor molecular genetic analysis. Contrary to our expectations,the addition of gibberellin A3 (GA3) at concentrations of 10-4M to 10-4 M resulted in the formation of nodule-like structureson roots when seedlings were plated on nitrogen-free Fahraeusagar medium. GA4 also induced such outgrowths but was less activethan GA3. Application of an inhibitor of auxin transport, N-(1-naphthyl)-phthalamicacid (NPA) and of kinetin, which have been reported to inducepseudonodules in other legumes, had no effect on L. japonicus.Microscopic observations showed that GA3-induced nodule-likestructures were caused by cell divisions within the pericycleon the roots. In addition, the outgrowths elicited by GA3 couldbe completely suppressed by the addition of 15 mM potassiumnitrate or ammonium nitrate. These results show that the pericyclecells of the roots of L. japonicus are specifically sensitiveto gibberellins and that potential for cell division might bemodulated by nitrogen compounds. We also examined the effectsof ancymidol and uniconazole [S-3307; (E)-1-(4-chIorophenyl)-4,4-dimethyl-2-(1,2,4-triazol-1-yl)-1-penten-3-ol],two synthetic plant-growth retardants. Both compounds at 3 x10-5 M significantly increased the number of stunted lateralroots. The unusual branching could not be counteracted by theexogenous addition of GA3 but by 10-6 M brassinolide. We discussthe physiological role of brassinolide in the initiation oflateral roots. (Received August 4, 1995; Accepted March 11, 1996) 相似文献
16.
Because of the importance of indole-3-acetic acid (IAA) in thegrowth and development of plants, extensive studies of the biosynthesisof IAA have been performed during the four decades since thediscovery of IAA as a plant hormone. The pathway for the biosynthesisof IAA in plants remains, however, to be unelucidated, eventhough studies within the past decade have revealed unexpectedaspects of such biosynthesis. By contrast, two pathways to IAAhave been characterized in bacteria at the molecular level:the indole-3-acetamide (IAM) pathway (L-tryptophan 相似文献
17.
Masayoshi Yamaguchi Reiko Makino Noriaki Shimokawa 《Molecular and cellular biochemistry》1996,165(2):145-150
18.
The effect of -alany-L-histidinato zinc (AHZ) on bone cell function was investigated in osteoblastic MC3T3-E1 cells. Cells were cultured for 3 days at 37°C in a CO2 incubator in plastic dishes containing -modified minimum essential medium supplemented with 10% fetal bovine serum. After the cultures, the medium was exchanged for that containing 0.1% bovine serum albumin plus AHZ (10–7–10–5 M) or other reagents, and the cells were cultured further for appropriate periods of time. The presence of AHZ (10–7–10–5 M) produced a remarkable increase of alkaline phosphatase activity and protein concentration in osteoblastic cells. Thus increases were seen with the prolonged cultivation (12–21 days). With the culture of 1, 3 and 12 days, the effect of AHZ (10–6 M) to increase alkaline phosphatase activity and protein concentration was more intensive than the effect of zinc sulfate, (10–6 M). The AHZ effects were completely abolished by the presence of cycloheximide (10–6 M), indicating that AHZ stimulates protein synthesis in the cells. The present study suggests that AHZ has a stimulatory effect on cell differentiation, and that this effect is partly involved on protein synthesis in osteoblastic cells. 相似文献
19.
Distribution of Vacuolar H+-Pyrophosphatase and a Membrane Integral Protein in a Variety of Green Plants 总被引:2,自引:0,他引:2
Vacuolar membranes isolated from several species including fernand moss exhibited pyro-phosphate-dependent H+ transport activity.On immunoblot analysis, H+ -pyrophosphatase was detected inthe vacuolar membranes. A membrane integral protein of 23,000daltons was not found in the membranes of Chara, Conocephalum,or Kalanchoë. Thus, H+-pyrophosphatase may be a universalenzyme among green plants, but the 23-kDa protein is not a commonprotein of central vacuoles. (Received September 10, 1993; Accepted November 29, 1993) 相似文献
20.
The effect of -alanyl-L-histidinato zinc (AHZ) on bone metabolism was investigated in osteoblastic MC3T3-El cells. Cells were cultured for 3 days at 37°C in a CO2 incubator in plastic dishes containing -modified minimum essential medium supplemented with 10% fetal bovine serum. After the cultures, the medium was exchanged for that containing 0.1% bovine serum albumin plus various concentrations of AHZ or other reagents, and the cells were cultured further for appropriate periods of time. The presence of AHZ (10–7–10–5M) stimulated the proliferation of cells. AHZ (10–6 and 10–5M) increased deoxyribonucleic acid (DNA) content in the cells with 48hr-culture. This increase was completely blocked by the presence of cycloheximide (10–6M) or hydroxyurea (10–3M). Also, the presence of cycloheximide (10–6M) completely inhibited the AHZ (10–5M)-induced increase in the proliferation of cells. Meanwhile, parathyroid hormone (10–7M), estrogen (10–9M) and insulin (10–M) significantly increased cellular DNA content. However, these hormonal effects clearly lowered in comparison with that of AHZ (10–5M). Dibutyryl cyclic AMP (10–4M) and zinc sulfate (10–5M) did not cause a significant increase in cellular DNA content. The present results support the view that AHZ has a direct specific proliferative effect on osteoblastic cellsin vitro and that this effect is dependent on protein synthesis. 相似文献