Cellular control of the tick-borne virus antigen production in persistently infected cell culture.

The influence of inhibition or stimulation of cellular DNA synthesis on tick-borne virus antigen production in persistently infected cell culture was studied. Either mitomycin C or cytosine-arabinoside caused cessation of antigen-containing cell number increase. Stimulation of cellular DNA synthesis by growth medium change increased the level of antigen-containing cells. When HEp-2-Sof culture was synchronized, a correlation was observed between the entrance of cells into DNA synthesis phase and the increase of proportion of antigen-containing cells.     

Cellular control of the tick-borne virus antigen production in persistently infected cell culture

Summary The influence of inhibition or stimulation of cellular DNA synthesis on tick-borne virus antigen production in persistently infected cell culture was studied. Either mitomycin C or cytosine-arabinoside caused cessation of antigen-containing cell number increase. Stimulation of cellular DNA synthesis by growth medium change increased the level of antigen-containing cells. When HEp-2-Sof culture was synchronized, a correlation was observed between the entrance of cells into DNA synthesis phase and the increase of proportion of antigen-containing cells.     

alpha-Mercaptopropionyl-glycine influence on the in vitro proliferative response of human lymphocytes.

The effect of alpha-mercaptopropinyl-glycine on the in vitro proliferative response of healthy subjects' lymphocytes was studied. Low doses of the drug enhanced spontaneous lymphocyte blastigenesis but had no effect on the PHA-induced blastic response. With increasing concentrations of alpha-mercaptopropionyl-glycine inhibition of 3H-thymidine incorporation was found in unstimulated as well as in PHA stimulated lymphocytes.     

Paradoxical effect of 1-β-D-arabinofuranosyl cytosine triphosphate on bleomycin-induced unscheduled DNA synthesis in permeable sarcoma cells

Summary 1--D-Arabinofuranosyl cytosine-5-triphosphate (araCTP), an inhibitor of DNA synthesis, paradoxically enhanced unscheduled DNA synthesis (USD) induced by bleomycin in permeable mouse sarcoma cells. A greater enhancing effect of araCTP on bleomycin-induced USD was observed with lower concentrations of dCTP in the assay mixture. USD measured without bleomycin in nuclei isolated from mouse sarcoma cells was not enhanced, but inhibited by araCTP.Acknowledgments. The authors wish to thank Nippon Kayaku Co. (Tokyo, Japan) for providing copper-free bleomycin A₂. This research was supported in part by a grant from the Japan Ministry of Education, Science and Culture.     

Effects of butyrate and insulin and their interaction on the DNA synthesis of rumen epithelial cells in culture

Rumen epithelial cells (REC) were incubated in the presence of various concentrations of butyrate or insulin or with both of them, to obtain information on their effect on the DNA synthesis of cultured cells. The 24-h values of 3H-thymidine incorporation into cellular DNA were measured in the presence of butyrate, insulin or butyrate plus insulin. While butyrate reduced DNA synthesis, insulin produced an increase over the control. Combined butyrate plus insulin treatment influenced the incorporation of label in accordance with the relative proportion of these two substances.     

Effects of butyrate and insulin and their interaction on the DNA synthesis of rumen epithelial cells in culture

Summary Rumen epithelial cells (REC) were incubated in the presence of various concentrations of butyrate or insulin or with both of them, to obtain information on their effect on the DNA synthesis of cultured cells. The 24-h values of³H-thymidine incorporation into cellular DNA were measured in the presence of butyrate, insulin or butyrate plus insulin. While butyrate reduced DNA synthesis, insulin produced an increase over the control. Combined butyrate plus insulin treatment influenced the incorporation of label in accordance with the relative proportion of these two substances.     

Evidence against prostaglandin E having a physiological role in acetylcholine liberation from Auerbach's plexus of guinea-pig ileum.

The effect of INDO, a PGE synthesis inhibitor, on ACh output from Auerbach's plexus of guinea-pig ileum was investigated. INDO (15-45 mum/ml) failed to alter significantly either spontaneous ACh output or ACh output induced by field stimulation. It is concluded that PGE plays no physiological role in ACh liberation from this tissue.     

Species-specific differences in the mitogenic activity of heparin-binding growth factors in the sera of various mammals

Sera from different mammalian species displayed great differences in mitogenic activity, as measured by stimulation of DNA synthesis in BALB/c 3T3 cells (3T3 cells). Among the sera examined, fetal bovine serum was least active, and increasing activity was detected in calf serum, human serum, rat serum and mouse serum, in that order. Rat and mouse sera exhibited extremely high mitogenic activity with 3T3 cells, but when TIG-1 human fetal lung fibroblasts were used for the DNA assay instead, the activity levels of all of the sera were lower, and the differences between them were smaller. To determine the reasons for these differences, the heparin-binding growth factors in each serum were separated on a heparin affinity column. Five peaks of DNA-stimulating activity were obtained. Three of these were found in all sera examined, with both 3T3 cells and TIG-1 cells. Two other peaks were found only with 3T3 cells; one was peculiar to rat and mouse sera, with extremely high activity in the rat, and the other was specific to fetal serum. The dependence of the activity of these peaks on the cells used for the test was confirmed using normal rat lung fibroblasts and immortalized rat kidney cells. These findings adequately explain the species-specific differences in mitogenic activity of whole sera, and the variation in activity depending on the cells used for assay of DNA synthesis.     

Species-specific differences in the mitogenic activity of heparin-binding growth factors in the sera of various mammals.

Sera from different mammalian species displayed great differences in mitogenic activity, as measured by stimulation of DNA synthesis in BALB/c 3T3 cells (3T3 cells). Among the sera examined, fetal bovine serum was least active, and increasing activity was detected in calf serum, human serum, rat serum and mouse serum, in that order. Rat and mouse sera exhibited extremely high mitogenic activity with 3T3 cells, but when TIG-1 human fetal lung fibroblasts were used for the DNA assay instead, the activity levels of all of the sera were lower, and the differences between them were smaller. To determine the reasons for these differences, the heparin-binding growth factors in each serum were separated on a heparin affinity column. Five peaks of DNA-stimulating activity were obtained. Three of these were found in all sera examined, with both 3T3 cells and TIG-1 cells. Two other peaks were found only with 3T3 cells; one was peculiar to rat and mouse sera, with extremely high activity in the rat, and the other was specific to fetal serum. The dependence of the activity of these peaks on the cells used for the test was confirmed using normal rat lung fibroblasts and immortalized rat kidney cells. These findings adequately explain the species-specific differences in mitogenic activity of whole sera, and the variation in activity depending on the cells used for assay of DNA synthesis.     

Abhängigkeit der Leukozytenwanderung und ihrer Steigerung durch Proteuslipo-polysaccharide von der Glucosekonzentration des Milieus

Summary The migration of leucocytes from the buffy coat is influenced by the glucose concentration in the medium, stimulation of migration results from concentrations of glucose up to 2%.. Higher concentrations of glucose up to 8%. have no effect. The stimulating effect of a constant concentration PLPS increases considerably over the whole range of glucose concentrations up to 8%.. Optimal stimulation by PLPS depends therefore on the glucose concentration.     

DNA synthesis in exocrine and endocrine pancreas after partial hepatectomy in Syrian golden hamsters

3H-thymidine autoradiography showed an enhanced DNA synthesis in acinar and islet cells of pancreas after partial hepatectomy in syrian golden hamsters. A significant nuclear labeling index of acinar cells was observed between 48 and 84 h and reached control levels by 120 h. An increased labeling index of islet cells was also observed, however, this increase was not statistically significant. These results indicate growth factor(s) produced after partial hepatectomy is capable of inducing DNA synthesis in pancreas.     

Antibody-dependent adherence of peritoneal cells of the rat to Trichinella spiralis larvae]

Washed peritoneal exudate cells from normal Rats firmly adhere to Trichinella spiralis larvae in the presence of serum containing anti-Trichinella antibodies. This effect is observed when muscle larvae, cells and dilutions of anti-sera are incubated for 1 hr. at 37 degrees C. No adherence takes place at 4 degrees C. Whole serum or its gammaglobulin fraction are active and effect is inhibited by the addition of Trichinella antigens. Complement is not essential since antiserum heated for 2 hrs. at 56 degrees C is active. Washed cells from infested animals do not adhere to the larvae in the absence of antiserum.     

Differential sensitivity to ethidium bromide of replicative DNA synthesis and bleomycin-induced unscheduled DNA synthesis in permeable mouse sarcoma cells

Replicative DNA synthesis in permeable mouse sarcoma cells was more sensitive to ethidium bromide (EtBr) than bleomycin-induced unscheduled DNA synthesis (UDS). A similar difference in sensitivity to EtBr was observed between DNA polymerases alpha and beta. The difference in sensitivity to EtBr of replicative DNA synthesis and UDS in the present system seems to reflect mainly the sensitivity difference between DNA polymerases alpha and beta.     

Effects of isoxazolyl-naphthoquinoneimines on growth and oxygen radical production in Trypanosoma cruzi and Crithidia fasciculata

Several 4-(aminomethylisoxazolyl)-1,2-naphthoquinones inhibited growth and DNA synthesis in Trypanosoma cruzi and stimulated O2 uptake and O2-. generation by the parasite epimastigotes and their mitochondrial and microsomal membranes; these results support the idea that oxygen radicals play a role in quinone toxicity. Maximal effects on respiration and O2-. generation were observed with antimycin-inhibited cells. Similar results as well as stimulation of H2O2 production were obtained with Crithidia fasciculata despite the presence of catalase in this organism.     

Relationship between DNA and cholesterol synthesis in kidney after refeeding

Summary DNA and cholesterol synthesis were investigated in the kidneys of fasted-refed rats. Refeeding resulted in an increase in kidney DNA synthesis, as measured by³H-thymidine incorporation, starting at 72 h. The increase in DNA synthesis was accompanied by a stimulation of cholesterol synthesis, as measured by¹⁴C-acetate incorporation into cholesterol.     

DNA synthesis in exocrine and endocrine pancreas after partial hepatectomy in Syrian golden hamsters

Summary ³H-thymidine autoradiography showed an enhanced DNA synthesis, in acinar and islet cells of pancreas after partial hepatectomy in syrian golden hamsters. A significant nuclear labeling index of acinar cells was observed between 48 and 84 h and reached control levels by 120 h. An increased labeling index of islet cells was also observed, however, this increase was not statistically significant. These results indicate growth factor(s) produced after partial hepatectomy is capable of inducing DNA synthesis in pancreas.This work is supported by NIH grant CA 36043.     

The effect of temperature and protein synthesis on the renaturation of firefly luciferase in intact H9c2 cells

A mild increase in temperature that does not exert an effect on tolerance development or synthesis of heat shock proteins (Hsps) in control cells can stimulate these processes when applied to cells that have previously been heat shocked. To study the underlying mechanism of this effect, H9c2 cells were stably transfected with the gene encoding firefly luciferase (Luc). Heat-shock-induced inactivation of Luc and its subsequent reactivation is frequently used as a model for cellular protein denaturation and renaturation. Luc reactivation was determined following a damaging heat shock (43 or 44 degrees C for 30 min) in cells that were subsequently exposed to either control temperatures (37 degrees C) or various mild hyperthermic conditions (from 38.5 to 41.5 degrees C for 1 h). To prevent changes in Luc activity consequent to new synthesis of Luc, Luc reactivation was monitored in the presence of cycloheximide, an inhibitor of protein synthesis. The results showed that reactivation of Luc was inhibited when heat-treated cells were post-treated under mild hyperthermic conditions. The observed increase in Hsp synthesis under mild hyperthermic post-heat shock conditions therefore appears to be the result of an increase in the period during which denatured proteins are present. In addition, we studied Luc reactivation in the absence of protein synthesis inhibitors. This condition led to much higher Luc activity. By estimating half-life times of Luc, the contribution of new Luc synthesis in this recovery could be determined, and only partially explained the observed increase in Luc reactivation after heat shock. Thus the synthesis of other proteins must be important for the renaturation of heat-damaged proteins.     

Sister chromatid exchanges in human lymphocytes exposed to 8-methoxypsoralen and long wave UV radiation prior to incorporation of bromodeoxyuridine

Summary Human lymphocytes exposed to the effects of long wave UV radiation in the presence of 8-methoxypsoralen prior to stimulation by PHA show dose related sister chromatid exchanges after 2 replication cycles in vitro. This has implications for interpreting the repair processes involved and for monitoring DNA damaging agents in vivo.     

Fetal calf serum alters cyclic adenosine 3',5'-monophosphate's effect on heme synthesis in vitro.

An investigation of the effect of cAMP on heme synthesis of rat bone marrow cells revealed that at 10(-2) M this cyclic nucleotide inhibits heme synthesis and that optimum stimulation occurs at 10(-4) M. Some unidentified constituent of fetal calf serum in the culture medium modifies the direction and degree of cAMP's effect.     

Hymenolepis nana: Transfer of acquired immunity in mice through sensitized peritoneal exudate cells

Summary Sensitized peritoneal exudate cells from syngeneic donor Swiss albino mice infected with single and repeated doses of viable eggs ofHymenolepis nana produced a significant adoptive immunity in test mice when compared with animals which received non-sensitized (normal) cells. No significant difference was observed among the 2 recipient groups which received singly or repeatedly sensitized peritoneal exudate cells.Acknowledgment. We thank Professor B. M. Sinha for providing facilities and to the University Grants Commission, New Delhi for financial assistance.