DNA and protein synthesis in leukemic leukocytes in patients with leukemia in vitro

2015 ◽  
pp. 583-585
Author(s):  
V. A. Almasow ◽  
V. A. Lapotnikow ◽  
A. L. Shishkow
Keyword(s):  
Protein Synthesis ◽  
Dna And Protein Synthesis

L-methionine uptake, incorporation and effects on proliferative activity and protein synthesis in bovine claw tissue explants in vitro

2007 ◽  
Vol 146 (1) ◽  
pp. 103-115 ◽  
Author(s):  
N. L. HEPBURN ◽  
C. H. KNIGHT ◽  
C. J. WILDE ◽  
K. A. K. HENDRY ◽  
H. GALBRAITH
Keyword(s):  
Protein Synthesis ◽  
Culture Medium ◽  
Culture Media ◽  
Tissue Level ◽  
Normal Function ◽  
Regulation Of Proliferation ◽  
Methionine Uptake ◽  
Dna And Protein Synthesis

SUMMARYL-methionine is a sulphur-containing nutritionally essential amino acid. It has a number of important roles in epidermal and dermal tissues of the integument of animals. Failure of normal function of these tissues in the hoof (claw) is a cause of lameness in cattle. Little is known about quantitative relationships between post-absorptive concentrations of nutrients including sulphur-containing amino acids and uptake and utilization by epidermis and dermis of the bovine claw. These parameters were studied at the tissue level by use of an established in vitro claw explant system using tissue from cattle of beef or dairy origin and L-[35S]-labelled methionine as tracer. The results showed that uptake of L-methionine by freshly prepared solear explants in Dulbecco's Modified Eagle Medium/F-12 Nutrient Mix (DMEM/F12) (1:1) medium containing 1·0 mmol L-methionine/litre was concentrative after 5–8 min, essentially linear for up to 10 min and became curvilinear thereafter. Maximum uptake and steady state conditions were obtained at approximately 30 min. Further measurements were made following 21 h incubation in culture medium. Under conditions of varying concentrations of L-methionine and measurement of uptake after 30 min, the presence of a saturable curve, that obeyed Michaelis–Menten kinetics, was demonstrated. Values of 3·61 mmol/litre and 5·84 mmol/kg intracellular water/30 min were obtained for KM and Vmax, respectively. Uptake was not influenced by L-cysteine and L-cystine concentrations in the culture media.Similar culture and incubation conditions were used in subsequent studies of DNA and protein synthesis. These showed that rates of incorporation of L-methionine into protein fractions and stimulation of DNA synthesis measured by methyl-thymidine incorporation were dependent on L-methionine concentrations in the medium. Maximal rates occurred at approximately 50 μmol/litre, which is in the normal physiological range, and at 1% of maximum uptake capacity. Examination of histological sections by autoradiography showed localization of L-[35S]-labelled methionine in basal and suprabasal epidermal cells with limited retention in dermis. Measurement, by a range of histological, immunohistochemical, electrophoretic, western blotting and autoradiographic techniques, provided further evidence of L-methionine-dependent regulation of proliferation, differentiation and synthesis of proteins under physiological concentrations, by epidermal horn-forming cells.A key role for L-methionine is suggested in the production of horn in bovine claw. The extrapolation of these in vitro data provides guidance for strategies to optimize methionine supply to claw tissues in vivo. Such extrapolation suggests the appropriateness of delivery of systemic concentrations of 50 μmol L-methionine/litre to maximize proliferative and protein depositional activity in solear epidermis and dermis in vivo.

In Vitro Evaluation of Plasticizer Activity on the Growth and Metabolism of Chick Embryo Fibroblasts and on the Development of Chick Embryo Lungs

1992 ◽  
Vol 20 (6) ◽  
pp. 475-482 ◽  
Author(s):  
G Stabellini ◽  
O Fiocchi ◽  
A Pellati ◽  
A Caruso
Keyword(s):  
Cell Proliferation ◽  
Protein Synthesis ◽  
Chick Embryo ◽  
Primary Cultures ◽  
Embryonic Cells ◽  
Cytostatic Effect ◽  
Dna And Protein Synthesis ◽  
Embryo Fibroblasts ◽  
Ethylhexyl Phthalate

Administration of di(2-ethylhexyl) phthalate (DEHP) to primary cultures of chick embryo fibroblasts brought about a decrease in cell proliferation rate after 48 h and an inhibition of both DNA and protein synthesis measured by [3H]thymidine and [3H]leucine, respectively, after 48h. The growth of chick embryo lung rudiments in vitro was also depressed by DEHP treatment. Lung rudiment were smaller in DEHP-treated embryos after 6 days' treatment. These results indicate that DEHP has a cytostatic effect on embryonic cells and tissues. L'administration de di(2-éthylhexyl) phthalate (DEHP) à des cultures primaires de fibroblastes d'embryon de poulet a provoqué une diminution du taux de prolifération cellulaire après 48 heures et une inhibition de la synthèse à la fois d'ADN et de protéines mesurée, respectivement, par [3H]thymidine et [3H]leucine après 48 h. La croissance des ébauches pulmonaires de l'embryon de poulet in vitro a également été abaissée par le traitement DEHP. Les ébauches pulmonaires étaient plus petites dans les embryons traités au DEHP après un traitement de 6 jours. Ces résultats indiquent que le DEHP a un effet cytostatique sur les cellules et tissus embryonnaires.

scholarly journals Evaluation of the Antineoplastic Activity of L-rhamnose in vitro. A Comparison with 2-deoxyglucose

2008 ◽  
Vol 51 (2) ◽  
pp. 113-119 ◽  
Author(s):  
Pavel Tomšík ◽  
Alena Stoklasová ◽  
Stanislav Mičuda ◽  
Mohamed Niang ◽  
Petr Šuba ◽  
...  
Keyword(s):  
Protein Synthesis ◽  
Tumor Cells ◽  
Animal Cells ◽  
Short Term ◽  
Ehrlich Ascites ◽  
Proliferation And Apoptosis ◽  
Dna And Protein Synthesis ◽  
Eat Cells ◽  
Inhibition Of Dna Synthesis

The effect of unsubstituted deoxyhexoses, 2-deoxy-D-glucose (2-DG) and L-fucose, on tumor cells has been reported in several papers throughout the last decades. That of a similar deoxysugar, L-rhamnose, which is synthesized in bacteria and plants but not in animal cells, has until today not been explored. In the present study, we examined the effect of L-rhamnose on DNA and protein synthesis, growth and the potential induction of apoptosis of tumor cells in vitro. Using 2-DG for comparison, we studied the effect of L-rhamnose in concentrations up to 20 (32 resp.) mmol/l on the initial velocity of the incorporation of labeled precursors of DNA and proteins in short term cultures of both mouse Ehrlich ascites tumor (EAT) and human HL-60 cells in vitro, and further, on cell proliferation and apoptosis induction in HL-60 cells. Neither cytotoxic nor cytostatic effects of L-rhamnose were observed with the exception of slightly pronounced inhibition of DNA synthesis in EAT cells. From the lacking inhibition of the protein synthesis it can be considered that L-rhamnose does not interfere with energy metabolism, at least not in a similar manner as 2-DG.

The Use of the Epithelial Kidney OK Cell Line for Studying the Nephrotoxicity of Anticancer Platinum Coordination Complexes

1993 ◽  
Vol 21 (1) ◽  
pp. 30-37
Author(s):  
Hervé Toutain ◽  
Françoise Courjault
Keyword(s):  
Protein Synthesis ◽  
Glucose Uptake ◽  
Succinate Dehydrogenase ◽  
Platinum Complexes ◽  
Marker Enzyme ◽  
Ok Cells ◽  
Dna And Protein Synthesis ◽  
Release Method

Nephrotoxicity is one of the most important dose-limiting side-effects of cis-diaminedichloroplatinum (II) (cDDP) in humans. Quiescent OK cells grown in hormonally-defined, serum-free medium in the total absence of antibiotics were used to study the in vitro nephrotoxicity of three platinum complexes which produce different renal toxicity in vivo: cDDP, its stereoisomer trans-diaminedichloroplatinum (II) (tDDP), and cis-diamine-1,1-cyclobutane-dicarboxylateplatinUm (II) (CBDCA). The uptake and cytotoxicity of these compounds at concentrations of 3-l,600μM and their impact on DNA and protein synthesis, glucose uptake, Na+-K+-ATPase and succinate dehydrogenase activities, as well as intracellular total glutathione level, were measured. The results showed that the cytotoxicity ranking of these three compounds, assessed by the LDH release method, was not in agreement with their in vivo nephrotoxic potentials (tDDP > cDDP > CBDCA00 in OK cells versus cDDP > CBDCA > tDDP in vivo). cDDP and tDDP showed similar uptakes at all the concentrations studied, which demonstrated that their cytotoxic potential was not directly related to intracellular levels of platinum. At non-cytotoxic concentrations, both cDDP and CBDCA decreased DNA and protein synthesis and, to a lesser extent, Na+-K+-ATPase activity, whereas no effect on glucose uptake and succinate dehydrogenase activity (a mitochondrial marker enzyme) was observed. Nevertheless, 5–10 times greater concentrations of CBDCA were required to induce effects similar to those induced by cDDP. Our results did not show a rapid and early depletion of intracellular glutathione after exposure to cDDP or CBDCA. tDDP exhibited the characteristics of a non-specific cytotoxic chemical which was unable to markedly inhibit the biochemical and functional parameters studied at non-cytotoxic concentrations. These results underline the key role of the inhibition of synthetic activities in the pathogenesis of cDDP-induced and CBDCA-induced nephrotoxicity in OK cells.

Paraquat Alters Growth, DNA and Protein Synthesis in Lung Epithelial Cell and Fibroblast Cultures

1992 ◽  
Vol 20 (2) ◽  
pp. 251-257
Author(s):  
Frank A. Barile ◽  
Seeta Arjun ◽  
Jean-Jacques Senechal
Keyword(s):  
Protein Synthesis ◽  
Fibroblast Cell ◽  
Exposure Period ◽  
Lung Fibroblasts ◽  
Type I ◽  
Lung Epithelial ◽  
Dna And Protein Synthesis ◽  
Dose Dependent

In vivo exposure to paraquat, a bipyridyl herbicide, is associated with selective necrosis of type I and type II alveolar pneumocytes. We examined the influence of paraquat on cell growth, DNA and protein synthesis in human and rat lung epithelial and fibroblast cell lines, to determine the effects and affinity of the chemical in vitro for cells of various origins. Cells were incubated with [3H]-thymidine or [3H]-proline in the absence or presence of 0.05–10mM paraquat for periods of 6 or 24 hours. Cell cultures grown in the presence of 50–200μM paraquat exhibited significantly lower cell numbers than comparable controls after 3 days. The suppression of DNA synthesis by paraquat was dose-dependent, but did not rely on the length of exposure. Paraquat inhibited protein synthesis and selectively decreased net production of low molecular weight proteins. NADPH levels were the same in both control and treated cultures. [14C]-Paraquat was accumulated during a 4-hour exposure period via an active non-competitive process. The results suggest that lung fibroblasts are as sensitive to the herbicide as epithelial cells. In addition, paraquat appears to interfere with transcription and/or translation by its ability to generate superoxide radicals, rather than by depletion of reducing equivalents.

Effect of mouse epidermal growth factor on DNA and protein synthesis, progesterone and inhibin production by bovine granulosa cells in culture

1986 ◽  
Vol 111 (1) ◽  
pp. 122-127 ◽  
Author(s):  
P. Franchimont ◽  
M. T. Hazee-Hagelstein ◽  
Ch. Charlet-Renard ◽  
J. M. Jaspar
Keyword(s):  
Protein Synthesis ◽  
Growth Factor ◽  
Epidermal Growth Factor ◽  
Granulosa Cells ◽  
Inhibitory Action ◽  
Inhibitory Effect ◽  
Stimulatory Action ◽  
Epidermal Growth ◽  
Dna And Protein Synthesis

Abstract. The effect of mouse epidermal growth factor (EGF) was investigated on DNA and protein synthesis, progesterone and inhibin production by bovine antral granulosa cells. When incubated for the whole period of culture, EGF inhibited inhibin production the second day of culture, progesterone the third and the fourth days whereas it stimulated DNA and protein synthesis only the fourth day of culture. Inhibition of progesterone and stimulation of DNA and protein were dose-dependent when treatment with EGF (pre-incubation) is followed by 24 h without EGF, a stimulatory effect on DNA and protein synthesis was observed after 48 and 72-h preincubation. Progesterone was reduced after 3 day preincubation and inhibin only after 2-day pre-incubation. Effects observed after 3-day pre-incubation were dosedependent. These experiments demonstrated the stimulatory effect of EGF on growth of granulosa cells and its inhibitory action on hormonal production by these cells in vitro. The inhibitory effect on progesterone and inhibin production is more precocious than stimulatory effect on DNA and protein synthesis. The inhibitory action of EGF on granulosa cell production of progesterone and inhibin could thus be not directly dependent on its stimulatory action on DNA synthesis.

Feedback inhibition of erythropoiesis induced in anaemic Xenopus

Development ◽  
1978 ◽  
Vol 43 (1) ◽  
pp. 221-231
Author(s):  
Vassiliki Aleporou ◽  
Norman Maclean
Keyword(s):  
Molecular Weight ◽  
Protein Synthesis ◽  
Blood Cells ◽  
Feedback Inhibition ◽  
Normal Serum ◽  
Low Molecular Weight ◽  
Erythroid Cells ◽  
Inhibitory Substance ◽  
Dna And Protein Synthesis

Serum from normal Xenopus, when injected into anaemic Xenopus, causes reduction in both DNA and protein synthesis in erythroid cells as indicated by in vitro culture of the blood cells. Experiments with erythrocyte-conditioned medium, reveal that this inhibitory substance can be recovered from mature erythrocytes. Sephadex G-25 fractionation of normal serum and haemolysate demonstrates that the inhibitory factor consists of molecules of low molecular weight, and experiments with cells of Xenopus kidney reveal that the feedback inhibition may be tissue specific to erythroid cells.

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