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  • Life and Medical Sciences  (1,842)
  • Aircraft Design, Testing and Performance
  • 1970-1974  (1,194)
  • 1950-1954  (721)
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  • 101
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Journal of Cellular Physiology 83 (1974), S. 43-51 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: This paper reports the isolation of a phenotypically stable line of Chinese hamster ovary cells which exhibits a temperature dependent alteration in the transport of some purines. The alteration manifests itself for the uptake of guanine, hypoxanthine, azaguanine and guanosine but not for adenine, adenosine or thymidine. Studies with crude cell extracts suggest that, at the temperature the alteration is being expressed, the HGPRT activity is within the normal range. In cell-cell hybridization studies the alteration behaves as a recessive genetic trait.
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  • 102
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    New York, NY [u.a.] : Wiley-Blackwell
    Journal of Cellular Physiology 83 (1974), S. 53-57 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Enzymatic activities involved in the transformations of uridine nucleotides by intact erythrocytes and their subfractions have been studied and the following enzymatic activities have been identified: UTPase, UDPase, UMPase and uridylate kinase.UTPase activity was present exclusively in the stromal fraction while UMPase and uridylate kinase activities were specific in the soluble fraction. UDPase was present in both the stromal and soluble fractions.This compartmentation in erythrocytes differs from that reported for the enzymes involved in adenine nucleotides transformations. Due to its sensitivity to Zn2+, uridylate kinase could be differentiated from adenylate kinase.
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  • 103
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    New York, NY [u.a.] : Wiley-Blackwell
    Journal of Cellular Physiology 83 (1974), S. 35-41 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Mouse bone marrow forms colonies of granulocytes and monocytic phagocytes when cultured in the presence of human plasma, urine or “feeder layers” prepared from human leukocytes. By contrast, human marrow produces colonies in the presence of leukocyte feeder layers but not in the presence of plasma or urine. It has been tacitly assumed that the response of mouse marrow to human blood leukocyte feeder layers is a measure of physiological substances released by those leukocytes which might control human granulopoiesis. This assumption however, has never been put to the test by comparing the response of mouse and human marrow to stimulation by leukocytes from the same individual. This has been done in the present study by using leukocytes from normal and leukemic subjects. Different human marrows responded similarly to stimulation by the same normal feeder layers, but there was no quantitative or qualitative correlation between the response of human and mouse marrows. Feeder layers from patients with acute granulocytic leukemia did not stimulate colony growth in normal human marrow but were as potent in stimulating mouse marrow colony growth as were feeder layers of normal leukocytes.We conclude that different factors may stimulate human and mouse marrows and that assays of granulopoietic factors of human origin should in future be carried out in human rather than mouse marrows.
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  • 104
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The pattern of changing activities of three lysosomal enzymes, N-acetyl-β-D-glucosaminidase, aryl sulfatase, and DNase II, and that of DNA polymerase was followed in homogenates of 3T3 cells during the logarithmic phase of growth and in stationary cultures. The change in activities of the polymerase and the lysosomal enzymes is antiparallel. DNA polymerase exhibits highest activity in growing cultures, and shows a three-fold decline of the specific activity in stationary cultures. The lysosomal enzymes show a very marked increase in their specific activity after density saturation is reached, which can be prevented by the addition of cycloheximide. Colcemid added to logarithmically growing cultures also causes an increase in the specific activities of lysosomal enzymes.
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  • 105
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    New York, NY [u.a.] : Wiley-Blackwell
    Journal of Cellular Physiology 83 (1974), S. 69-74 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Mammalian cell populations may be enriched for temperature-sensitive (ts) mutants by the tritiated thymidine (3H-TdR) suicide procedure (Thompson et al., '71). Such procedures were carried out on the near-diploid Chinese hamster ovary (CHO) cell line and on a line made “tetraploid” by Colcemid treatment. Clones of ts mutants were obtained from the diploid line, but in spite of repeated attempts, no ts mutants were isolated from the tetroploid line. The phenotypes of the new diploid ts mutants recovered were consistent with the particular selection regime employed.
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  • 106
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The effect of cAMP on a monkey kidney cell line (CV-1) noninfected and infected by SV40 was studied. No effect was found on either growth rate or cell morphology when concentrations up to 1 mM of 3′5′ cAMP were used. However, cAMP was found to increase the incorporation of 3H-thymidine into both cellular and viral DNA without a net increase in DNA synthesis. This increased incorporation was found to be related to an enhanced uptake of thymidine into the nucleotide pool which is reflected in an increase of phosphorylated nucleotides. This, coupled with a lack of effect of cAMP on endogenous deoxyribonucleotide production, produces an increased specific activity of the deoxyribonucleotide triphosphates, with a resultant increase in specific activity of DNA.
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  • 107
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    New York, NY [u.a.] : Wiley-Blackwell
    Journal of Cellular Physiology 83 (1974), S. 187-191 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Leukemic myeloblasts induced by avian myeloblastosis virus in the chicken formed small compact (type II) colonies in semi-solid agar medium. Normal yolk sac cells from 12-day old embryos formed large diffuse (type I) colonies under the same conditions. Type I colony formation (but not type II) was strictly dependent upon the presence in the medium of a colony stimulating factor (CSF) present in fresh chicken serum or conditioned medium. Serum CSF levels were determined for normal, leukemic, and birds which had spontaneously regressed from myeloblastic leukemia. When type I colony formation was used as the assay, serum CSF levels of leukemic birds were found to be significantly lower than levels in either normal or regressed birds. When the same sera were tested for their ability to induce type II colonies, leukemic birds demonstrated a significantly higher CSF level than either normal or regressed sera. Regressed chickens had serum CSF levels similar to normal birds.
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  • 108
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Sera from different strains of mice injected with endotoxin induced clones (D+) from a cultured line of myeloid leukemic cells to undergo normal differentiation to mature granulocytes and macrophages. Other clones (D-) derived from the same cell line were not inducible by these sera to undergo normal cell differentiation. Sera from the same strains of mice that had not been injected with endotoxin, increased the cloning efficiency of D+ and D - clones but did not induce differentiation. Endotoxin serum induced differentiation in D+ cells at dilutions up to 1:64, but increased the cloning efficiency of these cells at dilutions up to 1:2048. The end point of the dilution of endotoxin serum that induced differentiation in D+ cells, was also the end point that induced the formation of colonies with differentiation from normal bone marrow cells. The results indicate that serum from endotoxin treated animals can serve as a good in vivo source to induce normal differentiation in D+ myeloid leukemic cells; that the progeny of a single leukemic cell was induced to undergo differentiation to both macrophages and granulocytes; that endotoxin serum contained two activities, one that increased cloning efficiency and the other that induced cell differentiation; and that the same material in endotoxin serum induced cell differentiation in normal and leukemic cells.
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  • 109
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    New York, NY [u.a.] : Wiley-Blackwell
    Journal of Cellular Physiology 83 (1974), S. 193-201 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Balb/3T3 cells transformed in culture by chemical carcinogens were shown to multiply in a medium supplemented with 2% calf serum or with 10% agamma new-born calf serum. The cell lines that multiply well in medium supplemented with 10% agamma serum produced a higher incidence of tumors in X-irradiated weanling mice than the lines that multiply poorly. The difference in 2-deoxy-D-glucose uptake into exponentially growing transformed and un-transformed cells was 50-100%. In crowded cultures untransformed Balb/3T3 cells ceased taking up the sugar, while chemically transformed cells continued at the same rate even at high cell densities; thus, the difference became greater in crowded cultures. When the serum concentration in the media was reduced from 10% to 2%, untransformed Balb/3T3 cells took up the sugar at a reduced rate, while chemically transformed cells were only slightly affected; agamma new born calf serum supplemented medium had no effect on sugar uptake in any of the cells. When the serum concentration was changed from 2% to 10%, untransformed cells increased sugar uptake followed by cell division. The immediacy (within 15 min) of the response in the sugar uptake to 10% serum concentration suggested that the increased uptake rate and the consequent higher concentration of the sugar (D-glucose in normal situation) within Balb/3T3 cells triggered the cell cycle. Chemical carcinogens appear to alter permanently the uptake mechanism for a key nutrient.
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  • 110
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    New York, NY [u.a.] : Wiley-Blackwell
    Journal of Cellular Physiology 83 (1974), S. 203-210 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The effect of cell density on morphological transformation of chick embryo cells by Rous Sarcoma Virus (RSV) was examined in this study, and a cell density optimum for transformation was found. Less than 10% of the transformed foci appearing at the optimum density (2.5 × 104 cells per cm2) developed at high cell densities, and the diameters of the foci (an indication of the number of cells per focus) decreased with increasing cell density. No correlation was found between the decrease in transformation at high cell densities and the effect of cell density on the initial rate of cell proliferation, although dissociation of transformation from incorporation of radioactive precursors into nucleic acids could not be established. Redistribution of cells infected at high density showed that only a small proportion of successfully infected cells developed into foci. The results indicate that transformation of cells containing the RSV genome can be suppressed by physiological factors accompanying high cell density.
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  • 111
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    New York, NY [u.a.] : Wiley-Blackwell
    Journal of Cellular Physiology 83 (1974), S. 219-230 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: A series of Chinese hamster cell lines were tested and found to be able to proliferate in the absence of added bicarbonate and carbondioxide if hypoxanthine and uridine were present in the medium. Conversely, cells incapable of salvaging one of these precursors, such as hypoxanthine-guanine phosphoribosyltransferase (HGPRT-) deficient cells did not multiply under these conditions.We describe another variant capable of utilizing hypoxanthine and uridine which has an absolute requirement for exogenous CO2/NaHCO3 for growth. These cells appear to be defective in the complete oxidation of pyruvate to carbondioxide, and indications are that the entry of pyruvate into the Krebs cycle is affected.
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  • 112
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    Journal of Cellular Physiology 83 (1974), S. 211-218 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Viral infection by vesicular stomatitis virus (VSV) as well as cellular protection against it were studied in cultured human diploid fibroblast cells (WI38 strain) of varying senescence (passage) level. Full protection against viral infection can be achieved by pretreatment of cells with an interferon inducer (complex of polycytidylate and polyinosinate) or by pretreatment with concentrates of interferon itself. The late passages (over 45) require higher concentration of both agents than the medium passages (25-45); a complete failure of protective mechanisms occurs only close to cellular death. Furthermore, effects of confluency and of duration of induction impulse were studied. WI38 cells are sensitive to VSV infection through their entire life span; however, during the last passage before their death by senescence, VSV replication is significantly decreased. It is concluded that even in relatively senescent cells (a) protection mechanisms which were not used previously can be activated, (b) synthesizing machinery necessary for efficient support of viral replication is sustained, and (c) release of lysosomal enzymes ending the viral replication does not begin sooner than with cells of earlier passage level.
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  • 113
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    New York, NY [u.a.] : Wiley-Blackwell
    Journal of Cellular Physiology 83 (1974), S. 243-250 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Net fluxes of sodium and potassium were studied in Ehrlich mouse ascites tumor cells during contact with the agglutinating protein, concanavalin A. This lectin altered cation transport markedly at concentrations of 20-105 μg/ml (6-47 μg/mg cell protein). Whereas control cells extruded sodium and maintained or accumulated potassium against electrochemical gradients, in the presence of concanavalin A there was rapid net sodium entry and potassium loss. After 10-20 minutes in concanavalin A, sodium extrusion began and potassium loss diminished but these events were prevented by ouabain. The alterations in cation content induced by concanavalin A are unlikely to be the result only of agglutination since soybean agglutinin caused much smaller changes although it agglutinated the cells equally well.
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  • 114
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    New York, NY [u.a.] : Wiley-Blackwell
    Journal of Cellular Physiology 83 (1974), S. 259-261 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Exposure of early sea urchin embryos to 5-bromodeoxyuridine (at concentrations up to 100 μg per ml) severely decreases the uptake of exogenous 3H-uridine into RNA. However, the actual gross rate of DNA or RNA synthesis in these embryos appears not to be affected by the presence of 5-bromodeoxyuridine.
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  • 115
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    New York, NY [u.a.] : Wiley-Blackwell
    Journal of Cellular Physiology 83 (1974), S. 251-257 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The activities of the constant proportion enzymes of the Embden-Meyerhof chain (triose phosphate isomerase (TIM), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), phosphoglycerate kinase (PGK), phosphoglycerate mutase (PGM) and enolase (ENOL)), and the activity of lactic dehydrogenase (LDH) were studied in developing red (trapezius) and white (longissimus) muscles of the pig from a fetal stage to 24 weeks postnatal. Both muscles were differentiated by two weeks postnatal in the sense that they had reached the adult level of enzyme activity. Enzyme activities were two- to three-fold greater in the longissimus than in the trapezius. Enzyme activity ratios based on GAPDH were not consistent in the fetal and day 1 samples but were consistent during later stages of growth. Ratios of enzyme activity based on activity at 105 days gestation revealed that TIM, PGK and PGM are grouped and follow the same pattern, but GAPDH and ENOL are quite different from each other and from the pattern shown by TIM, PGK and PGM. The constant proportion concept in developing muscle is therefore questioned.
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  • 116
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Conditions have been established for the separation of viable mouse lymphoid cells by continuous free-buffer film preparative electrophoresis. The detailed electrophoretic distribution profiles of T and B lymphocytes from mouse spleen and thoracic duct have been determined. Cell surface θ-antigen was used as a marker for T cells, and high surface-density of immunoglobulin as a marker for B cells. Spleen cells from athymic “nude” mice were also studied.In the unselected normal spleen cell populations B lymphocytes are heterogeneous, about 60% being of low mobility with the remainder distributing broadly, and extending into the highest mobility fractions. T lymphocytes are predominantly of high mobility. Lymphoid cells lacking markers of either the B or T lineage are of intermediate mobility. There is only partial separation of T and B cells because of the extensive overlap between the populations.The high mobility B cells, which separate along with T cells, include a substantial proportion of large cells, and include cells with high surface density of immunoglobulin. The majority of these large B cells can be selectively eliminated by their adherence on passage through a glass-bead column.By pre-selecting the 50% non-adherent lymphocytes from spleen as the starting material, a very sharp and more extensive separation of B and T cells can be achieved, with 100% pure B cells and 90% pure T cells in many fractions. However these samples are not representative of the total T and B cell populations of spleen.In thoracic duct lymph high mobility B-cells are absent, there is little overlap between T and B cell mobility. 100% pure T and B cells can be isolated.
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  • 117
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: In cultured fibroblasts, a mutation resulting in deficiency of a pyrimidine salvage enzyme leads to excretion of related pyrimidines. For example, absence of thymidine kinase led to loss of thymidine and deoxyuridine, and absence of deoxycytidine kinase to loss of deoxyuridine. Both wild type and mutant cells excreted uracil; if established lines are representative in this respect, a fully adequate salvage system for uracil does not seem to be present in the fibroblast.
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  • 118
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    Journal of Cellular Physiology 83 (1974), S. 267-273 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Alcohols inhibit the exchange transport of glucose in human erythrocytes. Comparing the inhibition by monohydroxy-alcohols, which have different distribution coefficients between medium and membrane, shows that the degree of inhibition depends mainly upon the concentration of the alcohol in the membrane.1-butanol exerts a mixed-type inhibition; Vmax decreases and Km increases. Since also the Km of the equilibrium transport increases upon the addition of the alcohol, the changes in the Km of exchange transport cannot be attributed solely to the differently affected mobilities of the loaded and free carrier, but the affinity of glucose to the transport system is reduced.The transport system can bind two alcohol molecules. With one alcohol molecule bound the affinity of the transport system for the second alcohol molecule already increases.The nature of the bond of the alcohols to the transport system is discussed and possible explanations for the cooperative effect upon the binding of the second alcohol molecule are offered.
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  • 119
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    Journal of Cellular Physiology 83 (1974), S. 275-286 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Tolbutamide partially inhibited the growth but increased the glycogen content of Tetrahymena pyriformis in logarithmically growing cultures. Tolbutamide slightly increased 14CO2 production from [1-14C] and [6-14HC] glucose and [2-14C] pyruvate, but had little effect on the oxidation of [1-14C] acetate when any of these substrates were added to the proteose-peptone medium in which the cells had been grown. Measurement of 14CO2 production from [1-14C] and [2-I4C]-glyoxylate showed that this substrate was primarily oxidized via the glyoxylate cycle, with little if any oxidation occurring via the peroxisomal glyoxylate oxidase. Addition of tolbutamide inhibited the glyoxylate cycle as indicated by a marked reduction in label appearing in CO2 and in glycogen from labeled acetate. In control cells, addition of acetate strongly inhibited the oxidation of [2-14C]-pyruvate whereas addition of pyruvate had little effect on the oxidation of [1-14C]-acetate. Acetate was more effective than pyruvate in preventing the growth inhibitory and glycogen-increasing effects of tolbutamide. The data suggest that one effect of tolbutamide may be to interfere with the transfer of isocitrate and acetyl CoA across mitochondrial membranes.
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  • 120
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: cAMP, dbcAMP, cCMP, cGMP, theophylline and caffeine caused reversible melanosome dispersion within 5 minutes at 10 mM in the dermal melanophores of the black goldfish, Carassius auratus L. cTMP, cUMP, 5′-AMP, 5′-CMP, 5′-GMP, 5′-TMP, and 5′-UMP did not produce melanosome dispersion or aggregation in this melanophore system. cAMP was the most effective nucleotide in the induction of melanosome dispersion; at 10 mM, cGMP and at 5 mM, dbcAMP were the least effective of those nucleotides inducing melanosome dispersion. At the 10 mM level dbcAMP required 30 minutes to evoke the same degree of melanosome dispersion as 5 minutes cAMP treatment. Theophylline was more effective than caffeine in eliciting melanosome dispersion. At 1 mM, theophylline and caffeine first induced melanosome dispersion which was followed by aggregation in the course of the 30 minute test period. These reactions suggest both a high melanophore phosphodiesterase activity and competitive inhibition of phosphodiesterase by theophylline and caffeine. Induction of melanosome dispersion by several cyclic 3′,5′-nucleotides suggest multi-nucleotide control of melanosome dispersion. These findings also support a proposed mechanism of prostaglandin induced melanosome dispersion as well as the “second messenger” hypothesis.
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  • 121
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    Journal of Cellular Physiology 83 (1974), S. 353-357 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: HTC cells incubated in the absence of serum for more than 14 hours have very low levels of ornithine decarboxylase, the first enzyme on the pathway of polyamine synthesis. Readdition of serum causes an increase in the activity of ODC, reaching a maximum on average 17 times above the basal level after five hours. This increase is due in part to a decrease in the apparent rate of degradation of ODC, and also to a stimulation of its synthesis. Within the first two hours the serum induction of ODC is resistant to Actinomycin D. Insulin at 5 μm/ml alsocauses an increase in ODC activity but only after a delay of two hours, in contrast to its more rapid stimulation of tyrosine transaminase activity.
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  • 122
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    Journal of Cellular Physiology 83 (1974), S. 359-368 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The effect of increasing calcium concentrations on the electrophoretic mobility of cells obtained from early chick blastoderms at successive developmental stages was studied. Cells had zero mobilities at calcium concentrations in the range of 1 to 2 × 10-2 M CaCl2, and became positively charged when calcium concentrations between 2 and 5 × 10-2 M CaCl2 were used. Results using trypsin suggest that while some calcium binding sites disappear from the surface ionogenic layer, the majority of them are not removed by this enzyme. The calcium binding capacity did not vary appreciably in the stages studied. Charge reversal induced by calcium is reversible.
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  • 123
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    Journal of Cellular Physiology 83 (1974), S. 369-378 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Mouse peritoneal exudate cells induced by thioglycollate medium can form colonies in soft agar with a plating efficiency of about 5% (0.6%-10%). Cells from an unstimulated peritoneal cavity form no colonies or have a plating efficiency of less than 0.001 %. These colony-forming cells from the peritoneal exudate are similar to bone marrow colony-forming cells in vitro in that they both require a substance(s) present in conditioned medium from L-cells or mouse embryo fibroblasts or the serum from endotoxin-treated mice for the initiation and the continuation of their growth. However, peritoneal exudate colony-forming cells have a much longer initial lag period (10-14 days) and can survive longer in the absence of L-cell conditioned medium than bone marrow colony-forming cells. Only mononuclear cells, presumably macrophages, are observed in peritoneal exudate colonies, whereas bone marrow cell colonies contain both polymorphonuclear cells and macrophages.
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  • 124
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    Journal of Cellular Physiology 83 (1974), S. 379-388 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The behavior of two established cell lines was found to vary when subcultivated on protein polymers covered with either negatively or positively charged substances. Results indicate that the influence on cell behavior is conditioned by the charge rather than by structural differences or degree of attachment of the substances to the polymer. Furthermore, the substratum seems to have just a physical effect at the cell membrane without any direct influence on cell metabolism.Cells were also seeded on a calf serum polymer and it was found that serum loses its properties on cultivated cells when used as substratum.
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  • 125
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    Journal of Cellular Physiology 83 (1974), S. 401-407 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Nine variant cell lines isolated from cloned 7,12-dimethylbenz(a) -ahthracene transformed Balb/3T3 mouse cells by treatment with FUdR had growth parameters closely resembling nontransformed cells. Chromosome analysis of the variant lines demonstrated that six variants had a diminished number and three variants had an increased number of chromosomes compared to the parental transformed cell line. All variants had unique marker chromosomes not present in the parental transformed Balb/3T3 cells. The distribution of marker chromosomes and heterochromatin suggested that the initial event in variant formation was a reduction in chromosome number with a subsequent polyploidization of the reduced chromosome complement.
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  • 126
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    Journal of Cellular Physiology 84 (1974), S. 429-444 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Cell membrane antigens serve as recognition codes for normal cell functions (substrate transport, cell-cell interaction, etc.). Changes in antigen-function activity are associated with ontogeny and speciation. Some prenatal antigenic configurations are postulated to provide host protection during early development.
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  • 127
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    Journal of Cellular Physiology 84 (1974), S. 409-421 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Nicotinic acid mononucleotide (NaMN) and nicotinamide mononucleotide (NMN) have been proposed as intermediates in the biosynthesis of nicotinamide adenine dinucleotide (NAD) from nicotinic acid and nicotinamide respectively. Although NaMN and NMN were not detected in acid extracts from whole D98/AH2 cells following pulses with 3H-nicotinic acid and 3H-nicotinamide, they were observed following pulses of enucleated cells. Several experiments indicate that enucleated cells are incapable of synthesizing NAD and that biosynthesis stops with mononucleotide formation. These results provide support for the idea, based upon fractionation studies, that NAD pyrophosphorylase (which catalyzes the reaction NMN or NaMN + ATP → NAD or desamido-NAD) is localized exclusively in the nucleus.Acid extraction and chromatography were also used to compare the stability of 3H-NAD in enucleated and whole cells. In the presence of the glutamine analog, azaserine, the intracellular concentration of 3H-NAD decreased exponentially with a half-life of 4 hours in whole cells. In enucleated cells the intracellular concentration of 3H-NAD decreased with a half-life greater than 14 hours. The increased stability of NAD in enucleated cells was confirmed by autoradiography. These results demonstrate the key role of the nucleus in both the biosynthesis and turnover of NAD.
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  • 128
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
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    Topics: Biology , Medicine
    Notes: Three series of neuroblastoma X fibroblast hybrid clones were isolated from crosses between mouse or human fibroblasts and mouse or human neuroblastoma cell lines by virus-mediated cell fusion. The expression of 14-3-2 protein (an acidic protein specific to neurons) and steroid sulfatase activity was studied in parental and hybrid cell lines. Steroid sulfatase was extinguished in hybrids when only one parent expressed the enzyme, but was expressed in one hybrid combination in which both parents expressed the enzyme. The neuron-specific 14-3-2 protein, on the other hand, continued to be expressed in all three series of neuroblastoma x fibroblast hybrids. In most cases where these pheno-types were expressed, they also exhibited temporal modulation; that is, specific activity is low during logarithmic growth and increases markedly during stationary phase. The glial-specific protein S-100 is absent from all parents and hybrids. The results are discussed in terms of mechanisms of regulation of differentiated phenotypes in mammalian cells.
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  • 129
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    Journal of Cellular Physiology 83 (1974), S. 389-400 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: More than 80% of the intracellular pyridine metabolite pool of human culture cells is trapped by OsO4 fixation. The fixed pyridine metabolites fully exchange with nicotinamide and nicotinic acid but not with nicotinamide adenine dinucleotide. Yet, chromatography of the exchanged compounds reveals that NAD and NADP constitute more than 95%. of the fixed material. Although the mechanism of OsO4 fixation is not fully understood, such fixation has permitted the autoradiographic detection of intracellular pyridine metabolites. Cells of the human cell line, D98/AH2, synthesize pyridine nucleotides during all phases of the cell cycle at rates which do not vary by more than six-fold. There is no difference in the apparent concentration of pyridine metabolites between nucleus and cytoplasm after ten minute or three day pulses with 3H-nicotinic acid. The 3H-labeled pyridine ring is lost from D98/AH2 cells upon transfer to unlabeled medium. In general, the rate of loss is uniform among cells in the population. However, in a small proportion of cells there is little or no loss. Non-dividing cells lose the pyridine ring at approximately the same rate as dividing cells, yet the intracellular concentration of pyridine metabolites is 50% greater in non-dividing cells.
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  • 130
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
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    Topics: Biology , Medicine
    Notes: Calcium is required for ACTH stimulated steroidogenesis in adrenal tumor cells in tissue culture. In the absence of calcium, the dose of ACTH required to induce half maximum steroidogenesis was increased 30 fold. In contrast to intact adrenal glands or isolated adrenal cells, high doses of ACTH (50 mU/ml) maximally stimulated steroidogenesis in the absence of calcium. Growth for up to six days in medium with low calcium did not affect basal or ACTH induced steroidogenesis. The addition of calcium to cells incubated with ACTH produced a maximum steroidogenic response in 15 minutes. In contrast to intact adrenal glands, calcium is not required for adenosine-3′,5′-cyclic monophosphate (cyclic AMP) stimulated steroidogenesis in adrenal tumor cells. These experiments support the concept that calcium is important at the level of ACTH-membrane receptor site interaction or activation of adenyl cyclase in adrenal tumor cells.
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  • 131
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
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    Topics: Biology , Medicine
    Notes: Thymocytes incubated as cell suspensions in vitro are known to be markedly dependent upon added glucose for maintenance of maximal rates of incorporation of radiolabelled amino acids into protein. This requirement is only partially satisfied by other added substrates, such as pyruvate. Evidence is presented that incorporation of amino acids into protein associated with the nuclear fraction isolated from these cells is more dependent upon added glucose than is labelling of protein found in the rest of the cell.The dependence of the labelling of nuclear protein upon glucose is shown by comparing the ability of glucose and pyruvate to stimulate the incorporation of [14C-L] valine into the protein of nuclear and cytoplasmic fractions of thymus cells. The fractions are isolated on sucrose gradients after incubating suspensions of cells in substrate-free medium for two hours, adding carbohydrates and labelled L-valine for 30 min and then stopping the incubation by breaking the cells with hypotonic shock. When the protein-synthetic stimulatory effects of glucose and pyruvate are compared, glucose is almost equally capable (90%) at stimulating rates of protein synthesis in nuclear compared to cytoplasmic fractions. Pyruvate is much less effective in nuclear than in cytoplasmic fractions (30%).Evidence is also presented from pulse-chase experiments that the glucosedependent labelling of protein associated with the nuclear fraction occurs within that fraction, as opposed to migration to the nuclear fraction after being synthesized elsewhere.It is suggested from these and other data that a unique ability of glucose to provide non-mitochondrial ATP to the nucleus may be central to the dependence of the labelling of nuclear protein on this substrate.
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  • 132
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    Journal of Cellular Physiology 83 (1974), S. 437-439 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Hydroxyurea and guanazole were used as selective agents in tissue culture to obtain independent Chinese hamster ovary cell lines resistant to the cytotoxic effects of hydroxyurea or guanazole. In all cases tested a cell line selected for resistance to one of the antitumor agents exhibited resistance to both drugs. This result supports the view that these two drugs act at a common site.
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  • 133
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    Journal of Cellular Physiology 83 (1974), S. 425-435 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Respiration as an index of oxidative energy production was investigated in a L-cell suspension culture system previously shown to exhibit density-dependent inhibition of growth. It was found that as cultures progressed from exponential growth to high density nongrowing populations (6-10 × 106 cells/ml) over a 2-week period, the respiratory rate determined from the total amount of oxygen consumed during the daily medium renewal cycle, declined from 5.4 to 1.8 fmoles O2/cell/min. There are two components in this decrement. The first consists of a daily recurrent decline of oxygen uptake resulting from decreased availability of medium oxygen and glutamine and is readily reversed by medium supplementation. The second component which is refractory to medium supplementation and accounts for approximately 50% of the total respiratory decline, is considered to indicate an adaptive change of the respiratory capacity of the cells. This change is reversed during the lag period which precedes resumption of exponential growth upon subculture to low cell densities. The significance of these results is discussed in relation to recent reports indicating a marked depression of respiratory activity in nongrowing dense attached cultures as well.
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  • 134
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    Notes: Redistribution of surface immunoglobulins (Ig), H-2b, Thy-1.2 and TL. 1,2,3 alloantigens, and concanavalin A (Con A) receptors on mouse thymus, lymph node and spleen cells into “caps” induced by bivalent antibodies or ligands was compared by immunofluorescence. Surface Ig was capped rapidly following attachment of anti-Ig antibody at 37°. Capping of alloantigens and Con A receptors occurred very slowly following attachment of alloantibody or Con A, but much more rapidly after addition of a secondary bivalent antibody. An inverse relationship between the number of surface component sites per cell and the extent of capping of that component was observed. Capping of alloantigens sparsely represented on the cell surface was not inhibited by high concentrations of alloantibody, in contrast to capping of alloantigens present in greater quantities. These results suggest that factors in addition to molecular cross-linking may be involved in ligand-induced redistribution of cell surface components.
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  • 135
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    Notes: Hemopoietic colonies were counted macroscopically and microscopically in spleens of hybrid mice seven or eight days after they had been irradiated and given parental bone marrow in donor-host combinations exhibiting poor growth. Colonies counted microscopically were classified as to differentiation pathway. Lymphocytes from the thymus or lymph nodes were injected into some recipients at several different dosages and lymphocyte: bone marrow (L:B) ratios. In confirmation of earlier work it was found that thymocytes increased the number and size of colonies in recipients of marrow. A shift of differentiation toward granulopoiesis was also seen when thymocytes were given, although erythropoietic colonies were still the most frequently seen type except at very high L:B ratios. Lymph node lymphocytes shifted the pattern more markedly toward granulopoiesis, even at low L:B ratios. When lymphocytes from either source were given without marrow, only a few colonies could be found in recipients, and if differentiated they were almost exclusively granulopoietic. Irradiation (900 R) of lymphocyte donors reversed the shift so that a normal pattern of differentiation, like that resulting from marrow alone, was seen; irradiated lymphocytes were nonetheless capable of augmenting the size and total number of hemopoietic colonies.
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  • 136
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    Journal of Cellular Physiology 84 (1974), S. 57-68 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Regulation of the proliferation of transplanted colony forming units (CFUs) was investigated in lethally irradiated mice, pretreated by methods known to accelerate hemopoietic recovery after sublethal irradiation. Prospective recipients were exposed to either hypoxia, vinblastine or priming irradiation and at different intervals thereafter lethally irradiated and transplanted with bone marrow. Repopulation of CFUs was determined by counting the number of splenic colonies in primary recipients or by retransplantation.Regeneration of grafted CFUs was greatly accelerated and their self-renewal capacity increased in mice grafted within two days after hypoxia. Also the number of splenic colonies formed by grafted syngeneic CFUs as well as by C57BL parent CFUs growing in BC3F1 hosts was significantly increased. The effect was not dependent on the seeding efficiency of CFUs and apparently resulted from hypoxia induced changes in the hosts physiological environment. Proliferative capacity of grafted CFUs increased remarkably in hosts receiving vinblastine two or four days prior to irradiation. Priming irradiation given six days before main irradiation accelerated, given two days before impaired regeneration of CFUs. The increased rate of regeneration was not related to the cellularity of hemopoietic organs at the time of transplantation. The growth of CFUs in diffusion chambers implanted into posthypoxic mice was only slightly improved which does indicate that the accelerated regeneration of CFUs in posthypoxic mice is mainly due to the changes in the hemopoietic microenvironment. A short conditioning of transplanted CFUs by host factor(s) was sufficient to improve regeneration. The results might suggest that the speed of hemopoietic regeneration depends on the number of CFUs being induced to proliferate shordy after irradiation, rather than on the absolute numbers of CFUs available to the organism.
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  • 137
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    Notes: Bone cells were isolated by collagenase-hyaluronidase digestion of 19-20 day-old fetal rat calvaria. It is shown that contamination from calcified matrix can give erroneously high values for total cell calcium and can mask intracellular calcium exchange.Filtration of the cell suspension through 35 μ mesh nylon net and a five minute treatment with a cold pH 6.0 isotonic salt solution removed the contamination. Total calcium for cells prepared in this manner was 16.8 mmoles/kg wet weight. 45Ca uptake studies revealed an active component of calcium exchange.
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  • 138
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    Journal of Cellular Physiology 84 (1974), S. 97-100 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Explant cultures were prepared from the slow anterior latissimus dorsi muscle and the fast posterior latissimus dorsi muscle of 15 day chick embryos. The morphology and growth pattern of myotubes from the two types of muscle were very similar. Intracellular microelectrode studies did not reveal consistent differences between the myotube types in regard to resting potential, input resistance, input time constant, or ability to produce active electrogenic responses. It is suggested that specific differentiation of the two muscles is determined by their innervation.
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  • 139
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    Notes: Calcium transport was studied in bone cells isolated from fetal rat calvaria. 45Ca uptake experiments revealed an active component of calcium exchange. Calcium uptake was inhibited by iodoacetamide, DNP, CCCP and oligomycin and appeared to be dependent on medium phosphate concentration. Initial influx values exhibited saturation kinetics from 0.6 mM to 1.5 mM extracellular calcium.Efflux of 45Ca from loaded cells increased in the presence of iodoacetamide, DNP and CCCP. Incubation of the cells af 4° C inhibited both influx and efflux of calcium.Parathyroid hormone had no consistent effect on calcium uptake although characteristic increases in cyclic AMP levels were seen with the hormone. Calcitonin appeared to cause a transient increase in calcium uptake.
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  • 140
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    Notes: During the early developmental stages of the toad, Bufo arenarum, Hensel. up to the stage of gill circulation (150 hr of development at 20-25°C) the total phospholipids content as well as that of phosphoglycerides of choline and of ethanolamine were found unchanged. The subfraction of both phosphoglycerides were separated according to the number of double bonds on silver-ion chromatography and were also found to be unchanged up to the tail bud stage. The distribution of non-polar side chains in the subfractions varied in both phosphoglycerides showing a structural heterogeneity. In the phosphatidylethanolamines predominate the polyenoic containing subfractions.In contrast with the constant concentration of polar lipids, during early embryogenesis a steady increase in 32P incorporation into phospholipids takes place when oocytes labeled during oogenesis are used. These changes were also correlated with the DNA content up to gill circulation stage.It is proposed that most of the nascent membrane polar lipids during early embryogenesis may be derived from a storage site through an active and specific intracellular redistribution process. At the arrival of the polar lipid to the nascent membrane a change in their covalent structure by introduction of a phosphorylbase from a highly labeled pool may explain the raise in specific activity. This change may be necessary to make possible the assembly of the lipid into the membrane structure.
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  • 141
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    Journal of Cellular Physiology 84 (1974), S. 115-125 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Ehrlich cells subjected to anisoosmolar media show very rapid volume changes. In hypertonic media they shrink. In hypotonic media they swell but the rapid initial swelling is followed by a regulatory shrinkage lasting ca. 30 minutes.Cells suspended in media with identical ionic concentrations but different total osmolarity (adjusted by sucrose) were compared. These studies revealed that swollen cells adjust their volume by decreasing the amount of intracellular K+ and ninhydrin positive substances. Intracellular Na+ and ATP concentrations were unchanged. Accordingly 42K+ flux analysis showed that the (passive) cell membrane permeability for K+ is increased to a minor degree and the Na+ permeability unaffected. The increased K+ permeability could not be correlated to an increase in 45Ca2+ influx.
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  • 142
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    Journal of Cellular Physiology 84 (1974), S. 135-140 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: We extracted a pure protein from Tetrahymena pyriformis as the chemotactic substance in Amoeba proteus. The protein was heat-stable, negatively charged at neutral pH (since its isoelectric point was near pH 4.5) and composed of three subunits. Its molecular weight was 3 × 8,000.
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  • 143
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    Notes: Colony-stimulating factor (CSF) is necessary for the clonal growth of human bone marrow in vitro. Human blood monocytes and macrophages produce CSF. Endotoxin was found to increase the level of CSF generated by macrophages, but had no stimulatory effect on monocytes. Several other substances known to influence the pinocytic or phagocytic activity of mononuclear phagocytes failed to enhance cellular CSF generation.
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  • 144
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    Journal of Cellular Physiology 83 (1974) 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
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  • 145
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    Journal of Cellular Physiology 83 (1974), S. 297-308 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: These experiments demonstrate that serum, like insulin, can initiate DNA synthesis in mouse mammary gland epithelium, resulting in a three- to four-fold increase in the rate of DNA synthesis and number of cells synthesizing DNA. Both serum and insulin also increase the tritiated thymidine counts in the acid soluble material of these cells, suggesting that each alters thymidine transport. When combined at any concentration, these agents produce an additive effect on DNA synthesis, number of cells synthesizing DNA and thymidine transport. The factor(s) in serum responsible for these effects is associated with high molecular weight material. These experiments suggest that DNA synthesis and thymidine transport are affected independently by serum and insulin in mammary gland epithelium.
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  • 146
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    Notes: Assay of red cell progenitors by colony formation in culture is expected to allow study of early events in erythroid differentiation in animal models and in man. In this communication, a simplification of the culture method for mouse bone marrow cells originally reported by Stephenson et al. ('71) is described. In the modified procedure, plasma clot is replaced by methyl cellulose, and scoring of erythroid colonies is done directly in the plates without staining. With additional modification the system proved applicable to the culture of erythroid colonies from human bone marrow as well. The development of both mouse and human erythroid colonies was dependent on “erythroid colony stimulating activity” (E-CSA) supplied by extracts of anemic sheep plasma, or by extracts of urine from anemic patients. Over a certain range, the number of colonies was a function of dose of E-CSA.In addition to E-CSA, these extracts also possessed erythropoietin activity as measured in plethoric mice. The possible chemical equivalence of urinary E-CSA and erythropoietin was suggested by their similar behavior on both gel filtration and affinity chromatography on agarose-concanavalin A. The finding further suggests that the culture method might prove useful for the bioassay of erythropoietin.Granulocyte colony stimulating activity (G-CSA) was also present in the urine extracts, as detected in cultures of mouse bone marrow. Virtually all of this activity was bound on agarose-con A, whereas only a small fraction of E-CSA was retained on this material. Agarose-con A may thus be useful for the purification of erythropoietin.
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  • 147
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    Journal of Cellular Physiology 83 (1974), S. 11-18 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: A single hematocytoblast in the yolk sac of the chick embryo has been shown previously to give rise on the average to a clone of 128 erythrocytes. Furthermore, in any given generation the erythroid cell synthesizes a characteristic amount of hemoglobin (Hb). In these experiments day 4 embryos were treated with FUdR for 12 hours, and then reversed with thymidine. We have monitored both the passage of these erythroblasts through the cell cycle, and the effect of this perturbation on the Hb content of single cells. As a result of this disruption the amount of Hb synthesized in a given generation can be varied, but the final amount of Hb/cell in the mature erythrocyte is the same as in the untreated controls. Apparently the total amount of the Hb/cell does not in itself influence the passage of the cell through the cycle. The coefficients of variation of the Hb values in the mature erythrocytes from both normal an perturbed embryos are similar.
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  • 148
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  • 149
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    Journal of Cellular Physiology 84 (1974), S. 343-348 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Agglutinability with Concanavalin was studied as function of cell cycle transition in normal and SV40 virus transformed 3T3 cells. In synchronized cultures of normal cells, agglutinbility was high during mitosis and disappeared rapidly. Agglutinability of transformed cells remained high in G1 phase but diminished gradually upon entering S phase and reached minimum in G1 phase. Decreased agglutinability a the end of the cell cycle was also observed in synchronous SV3T3 cultures by a combined technique of haemadsorption and density gradient centrifugation. In normal 3T3 cells, similar variations in agglutin ability during interphase could not be observed.
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  • 150
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    Journal of Cellular Physiology 84 (1974), S. 333-342 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: We have shown previously that adenosine 3′5′ cyclic monophosphate (cAMP) increased the transport of thymidine into monkey cells (CV-1) without affecting DNA synthesis (Roller et al., '74). Using techniques of cell synchronization, cell separation and DNA synthesis inhibitors in CV-1 and other cell lines, we now report that thymidine transport at low thymidine concentrations increases during the S phase of the cell cycle and that initiation of DNA synthesis is necessary for thymidine transport, although continuation of DNA synthesis is not necessary. Cyclic AMP alters nucleoside transport by increasing the Vmax for the transport reaction, but has no effect on the Km. Cyclic AMP is found not to affect the time in the cell cycle when thymidine transport occurs or the quality of the transport reaction, and only affects the amount of thymidine transported.
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  • 151
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    Journal of Cellular Physiology 84 (1974), S. 365-371 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The effects of elevated temperatures upon protein biosynthesis were determined in L5178Y murine leukemic lymphoblasts. The rate of protein synthesis was inhibited proportionately to the increase in temperature. Efforts were made to determine the mechanism of heat inactivation of protein synthesis by studying the requirements for recovery of activity after the cells were returned to 37°C. The ability of actinomycin to block the recovery process suggests that elevated temperatures destroy or inactivate a species of RNA required for protein synthesis. Loss of RNA during heating of the cells is apparently at least partially dependent on protein synthesis, since the presence of cycloheximide during heat shock, is capable of ameliorating the effects of short duration heat treatment.
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  • 152
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    Notes: An “overlay” method for rapidly and synchronously inducing contact inhibition in normal cultured cells has been developed. Using this method, disaggregation of cytoplasmic polyribosomes has been observed to occur within a matter of hours after overlay, followed by a decrease in cellular ribosomal RNA. Polysome disaggregation was influenced by the extent of cell-cell interaction and was inhibited by pretreatment of overlay cells with cycloheximide. Treatment of underlay cells with cytosine arabinoside also induced polysome disaggregation, but only after an appreciable lag as compared to that observed in overlaid cultures. Disaggregation could be induced by this method in cultured cells derived from normal tissue but not in cells derived from cancerous tissue. Polysome synthesis in growing “normal” cells (as measured by incorporation of tracer uridine into RNA) was markedly decreased when a cell surface membrane preparation was added to cultures.
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  • 153
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    Journal of Cellular Physiology 84 (1974), S. 373-382 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The binding of tritium-labelled cytochalasin B (3H-CB) to a variety of mammalian cells was investigated. Binding studies revealed near-equilibrium binding of 3H-CB within 5 to 10 minutes, but the equilibrium level was influenced by 3H-CB concentration. Binding kinetics revealed strong temperature dependence. Rapid release of up to 70% of cell-bound 3H-CB molecules occurred when cells were washed and returned to fresh medium without CB. The remaining 30% of cell-bound 3H-CB molecules dissociated more slowly. Equilibrium binding studies on a variety of diploid, heteroploid and transformed cells treated with 1 μg/ml 3H-CB revealed between 1.7 X 107 to 5.3 X 107 3H-CB binding sites per cell. Cellular binding of 3H-CB was not affected by inhibition of cellular energy metabolism, RNA or protein synthesis. Modification of the cell surface by proteases, neuraminidase, hyaluronidase, ribonuclease, or occupation of cell surface saccharide residues by a variety of plant lectins did not significantly alter the pattern of 3H-CB binding. Surface pressure measurements on CB-treated lipid monolayers indicated that CB can interact with lipid molecules. The partition of CB in hydrophobic lipid regions of cell membrane systems as a possible mechanism of cellular binding of CB is discussed. Fractionation of 3H-CB-treated cells revealed binding of 3H-CB to both the plasma membrane and by intracellular membranes.
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  • 154
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Molecules of low molecular weight, able to stimulate colony formation by human granulopoietic cells, were prepared from media conditioned by normal and leukemic human peripheral blood leukocytes. When molecules from these sources were studied, heterogeneity was observed in their ability, after radioiodination, to bind and suicide granulopoietic progenitors, in their radiolabelled tryptic digestion products and in their biological activity as stimulators of granulocyte colonies.
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  • 155
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    Journal of Cellular Physiology 84 (1974), S. 397-407 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: A temperature-sensitive growth mutant derived from the BHK 21 cell Line, ts AF8, was found to have greatly reduced DNA synthesis at the nonpermissive temperature. This reduction is mainly due to a decrease in the frequency of cells synthesizing DNA. Upon shift up, ts AF8 becomes blocked in the G1 phase of the cell cycle. The cells acquire elevated cAMP levels and a unimodal distribution of DNA content, equivalent to that of G1 cells at the permissive temperature, Ts AF8 cells blocked at the G1/S boundary with hydroxyurea will enter S when shifted to the nonpermissive temperature. On the other hand, ts AF8 cells arrested m G1 by serum deprivation and shifted to the nonpermissive temperature at the moment of serum addition do not enter S, while those synchronized by isoleucine deprivation and shifted at the time of isoleucine addition will enter S. These data suggest that the cycle arrest point of the ts AF8 mutation is located in G1 between the blocks induced by serum starvation and isoleucine deprivation.The reduction in DNA synthesis caused by the ts AF8 mutation is not reversed by infection or transformation with Polyoma virus. Mitochondrial DNA continues to be synthesized at wild-type levels at the nonpermissive temperature.
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  • 156
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: L1210 murine leukemia cells after treatment with Cl. perfringens neuraminidase at pH 7.0 incorporated six times more N-acetylneuraminic acid-[C14] than control cells when incubated for 30 minutes with cytidine 5′-monophosphate N-acetylneuraminic-[C14] acid and three times more galactose-[C14] when incubated with uridine diphosphate galactose-[C14]. These sugars were incorporated in a 10% trichloracetic acid insoluble fraction and more than 75% of the incorporated N-acetylneuraminic acid- [C14] could be removed by further treatment of these cells with neuraminidase. The incorporation of N-acetylneuraminic acid- [C14] as a function of time was divided into two rates: a rapid one, active during the first 30 minutes followed by a slower one, similar to the rate observed with untreated cells. The addition of Ba++ and Ca++ ions at 8.3 mM increased the incorporation of N-acetylneuraminic acid- [C14] by 25% while 8.3 mM EDTA decreased activity by 58% . The addition of Zn++ or Hg++ at similar concentrations abolished the incorporation almost completely. The optimal pH for the incorporation of N-acetylneuraminic acid- [C14] by these neuraminidase treated cells was 6.5. These data suggest that ectoglycosyltransferases are present on the outer surface of the plasma membrane of L1210 cells and are able to catalyze the addition of radiolabeled nucleotide sugars onto macromolecular acceptors (cell surface glycoproteins and glycolipids) prepared by prior incubation of the cells with neuraminidase. Use of these procedures for labeling outer cell surfaces may also prove to be valuable for the study of plasma membrane glycoprotein and glycolipid structure, synthesis, and turnover.
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  • 157
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    Journal of Cellular Physiology 84 (1974), S. 29-36 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The potassium content of single human red cells was measured with an electron probe. Cells were placed on beryllium discs and coated with a thin layer of dibutyl pthalate to prevent loss of cellular contents. Samples were stable under the electron beam during analysis for more than 15 minutes and could be stored for long periods of time. Primary standards were prepared by loading red cells with varying known amounts of potassium in order to circumvent the corrections for absorption. The X-ray intensity was found to be directly proportional to the potassium content of the cells.
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  • 158
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Monolayer cultures of a mouse teratocarcinoma were established in vitro. These cultures contained embryonal carcinoma, the malignant stem cell, and its differentiated progeny: parietal yolk sac, neuroepithelial, and mesenchymal cells. Tissues such as squamous epithelium, cartilage, striated muscle, neuroepithelium, and glands were produced from embryonal carcinoma that was maintained under conditions of long term culture. Frequent subcultivation with pancreatin allowed the establishment of cell lines of embryonal carcinoma which have been maintained for more than 18 months in vitro and continue to produce differentiated cells under specific culture conditions. Chromosomally these lines of embryonal carcinoma have a stem line of 39 chromosomes. Two lines of parietal yolk sac cells have been established which produce basement membrane, are not tumorigenic, and chromosomally are hypotetraploid. This system may yield information concerning neoplastic differentiation and its possible use in therapy for cancer.
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  • 159
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Changes in the number and erythropoietin sensitivity of erythropoietic progenitor cells (CFU-E or E) capable of erythropoietin-dependent colony formation in culture have been measured in mice subjected to a variety of manipulations known to affect erythropoiesis. Injection of erythropoietin into mice whose E levels had been reduced by transfusion-induced plethora stimulated a striking increase in this population within 24 hours in both spleen and marrow. Following irradiation and marrow transplantation E were found to regenerate in the spleen with a population doubling time of 6 hours. Evidence of erythropoietin-dependent stimulation of E in vivo under conditions of hemopoietic regeneration was also obtained, although substantial numbers of E were detected in both regenerating spleen and marrow of plethoric mice even without erythropoietin administration, In the same experiments comparable effects of erythropoietin on pluripotent stem cells or the progenitors of granulopoietic colonies in culture were not observed. Detailed studies of the dependence of colony formation on erythropoietin concentration in culture showed variations in erythropoietin sensitivity of E from normal, regenerating and plethoric, erythropoietin-stim-ulated spleen and marrow. This finding provides the basis for an extremely fine mechanism regulating the flow of erythropoietic differentiation at the level of the production of E.The number of E present in the marrow of untreated W/WV mice was normal, but their sensitivity to erythropoietin in culture was decreased to a similar extent as E from normal mice exposed to a heightened erythropoietic demand. Plethora had a more marked effect in reducing E numbers in W/WV mice than in +/ + controls and stimulation of E following subsequent erythropoietin administration was highly defective. The results of these studies provide further evidence that cells identified as E represent a stage of differentiation along the erythropoietic pathway that is several steps removed from pluripotent stem cells and support the view that erythropoiesis, granulopoiesis and stem cell self-renewal are regulated to a major degree by independent mechanisms.
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  • 160
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    Journal of Cellular Physiology 84 (1974), S. 127-133 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The Chinese hamster cell line K12 is temperature sensitive for the initiation of DNA synthesis and for the programming of rounding up to enter mitosis. Viability is lost expontentially after twelve hours at the non-permissive temperature. This temperature sensitivity is reversed following exposure of K12 cells to SV40 virus.
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  • 161
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    Journal of Cellular Physiology 84 (1974), S. 141-145 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The membrane potentials of human embryonic lung fibroblasts have been measured in different cellular environments. Sparse cells on plastic have a mean membrane potential of -8.5 mV. As the cells progress to dense culture, the mean membrane potential rises to -14.7 mV. The mean membrane potential of fibroblasts in human embryonic lung fragments by comparison was found to be -16.5 mV. Sparse cells on collagen, at the same density as the sparse cells on plastic, have mean membrane potentials of -10.8 mV.Sparse cells on plastic migrating from dense cellular areas, following a cut being made in a thick sheet of cells, have mean membrane potentials of -5.9 mV.The significance of these results in relation to cellular environments has been discussed.
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  • 162
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    Journal of Cellular Physiology 84 (1974), S. 159-163 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: A transient increase in the number of peritoneal colony-forming cells in agar (PCFC) was noted in the peritoneal cavity of mice given one intraperitoneal injection of thioglycollate medium. This increase of PCFC was not accompanied by a similar degree of increase in the number of either pluripotent hemopoietic stem cells or committed hemopoietic stem cells for granulocytes and/or macrophages that are normally present in the peritoneal cavity in small numbers. Other substances, such as glycogen, starch, Freund's adjuvant, sheep red blood cells and lectins, were also capable of increasing PCFC, but to different degrees. The ability of thioglycollate medium as well as other stimulatory agents to increase PCFC appeared to be dose-dependent.
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  • 163
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    Journal of Cellular Physiology 84 (1974) 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
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  • 164
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
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    Topics: Biology , Medicine
    Notes: The CSF's extracted from 6-hour “in vivo” post-endotoxin mouse lung tissue and 48-hour “in vitro” lung-conditioned medium appear to be glyco-proteins, for like human urinary CSF they were inactivated by proteases and altered in charge but not inactivated by sialidase.Neither endotoxin injection nor length of in vitro incubation significantly affected the apparent S20 W (approximately 2.OS) of the bulk of the CSF in mouse lung-conditioned medium. The lower apparent molecular weight of CSF in mouse lung-conditioned medium contrasted with that of monkey lung-conditioned medium (S20 W approximately 3.3S). CSF's in both monkey and mouse lung-conditioned media exhibited a much higher apparent MW on gel nitration than one would expect from their sedimentation behavior, a characteristic generally observed with most unpurified CSF preparations.Mouse lung-conditioned medium CSF was antigenically distinguishable from other CSF's. Failure to demonstrate this form of CSF in the tissue serum or urine of normal or endotoxin-injected mice suggests that this type of CSF might normally lack in vivo significance.
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  • 165
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    Notes: Using a combination of DNA-cytophotometry and tritiated thymidine-autoradiography, we have shown that the majority of nondividing cells in serially propagated human diploid cell populations have the 2C DNA content consistent with their being arrested in the G1 phase of the diploid cell cycle. Unlabeled 4C cells appear increasingly with time in culture. These may be arrested G2 diploids or they may be G1 tetraploids, since there is an associated increase in polyploidy in older cultures as evidenced by the appearance of labeled 8C cells.
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  • 166
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    Notes: A recently described pituitary chondrocyte growth factor (CGF) is a contaminant of several related glycoprotein hormones (TSH, LH and HCG). Chondrocytes were cultured from rabbits two to three months old. Bovine TSH (NIH) 69.5 μg/ml, used as the source of CGF, reduced the generation time from 16 to 10 hours through a virtual effacement of the G1 period. Incorporation of 3H-thymidine declined rapidly after 48 hours from maximal values (control 300 cpm/μg DNA; CGF, 679). Total DNA accumulated thereafter until 116 hours when the figures were 36 and 98 μg/flask, respectively. Little growth response occurred in spinner cultures. CGF lowered plating efficiency from 4.5 to 2.3%. The stimulatory effect diminished when CGF was removed from the medium. The treated cells were smaller and contained less protein and RNA than controls. They synthesized smaller quantitites of sulfated mucopolysaccharides (chondroitin sulfates 4,6 and doubly sulfated chondroitin as well as some dermatan sulfate) but hyaluronate production was not diminished.
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  • 167
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    Journal of Cellular Physiology 84 (1974), S. 181-192 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Multiplication-stimulating activity (MSA) for chicken embryo fibroblasts was purified from serum-free medium conditioned by the growth of a line of rat liver cells (CRL), The biological activities of purified CRL MSA for chicken embryo fibroblasts were compared with those of calf serum to determine which activities are important for the stimulation of DNA synthesis and mitosis. In a balanced salt solution, only glucose and amino acids were needed in addition to purified CRL MSA to stimulate DNA synthesis maximally. Purified CRL MSA stimulated the rates of uptake of glucose and α-aminoisobutyric acid. Only the stimulation of the rate of glucose uptake appeared to be a primary response to purified CRL MSA since the stimulation was not inhibited by actinomycin D or cycloheximide. The stimulation of the rate of uptake of α-aminoisobutyric acid was inhibited by actinomycin D.CRL MSA differed from calf serum in its inability to commit cells irreversibly to synthesize DNA after the removal of CRL MSA and in its lack of the ability to stimulate the migration or prolong the survival of chicken embryo fibroblasts.Comparative studies indicated that purified CRL MSA had functional similarities to insulin and somatomedin. CRL MSA may be representative of a family of small polypeptide hormones having insulin-like activity which are involved in the control of cell multiplication.
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  • 168
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: A flow-system multiparameter cell analyzer that simultaneously measures and processes fluorescence and cell volume signals from single cells was used to study the binding of fluorescein-conjugated Concanavalin A (Con A-F) to the cell surface. Cells reacted with Con A-F were passed through a flow chamber where sensors measured both cell volume and fluorescence of each individual cell. Sensor signals were electronically processed by first converting the cell volume signals to two-thirds power (proportional to surface area) and then forming the fluorescence-to-surface area ratio. These ratios, which were considered as estimates of the surface density of binding sites, were displayed as frequency distribution histograms using a multichannel pulse-height analyzer for various cell populations differing in cell size. Comparisons between cell lines showed characteristic differences in binding site density. Cell cycle dependent changes were not found for CHO cells synchronized by mitotic selection. An important benefit of this analysis method was the ability to quantitate very weak cell surface fluorescence.
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  • 169
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    Journal of Cellular Physiology 84 (1974), S. 225-235 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: A microphotometric method is introduced that allows measurement of the contraction-relaxation kinetics of Spirostomum in response to electrical stimulation. The time course of contraction includes a rapidly contracting phase of some 4-5 mS during which cells shorten at a rate in excess of 100 cell lengths sec-1. While a stimulus strength-duration curve determines the threshold of the response, the response to above threshold stimuli of different strengths and to trains of stimuli suggest that contraction of Spirostomum may not be an all-or-none event. The kinetics of relaxation following high stimulating voltages and repetitive after contractions also induced by high voltages are explained by excitation-contraction coupling through a stimulus-dependent intermediate effector, possibly the release of calcium ions. Changes in resting membrane potential detected by intracellular recording do not influence the initiation of contraction, while microinjection of calcium buffers above 10-5 M Ca2+ invariably induces contraction.
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  • 170
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    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The factors involved during the pumping of a particle suspended in an electrolyte through a small cylindrical orifice across which exists an electric field are reviewed, and their practical application to cell volume measurement in terms of orifice geometry and particle properties is considered. Procedures are described to determine whether a particular cell type satisfies the operational criteria of a rigid, non-conducting sphere, for which the theoretical expressions can be reduced to an especially simple form under appropriate conditions.Experimental results are presented for the case of chick, mouse and human lymphocytes, all of which are shown to satisfy the above requirements. Volume distributions are provided for chick blood lymphocytes and compared with those from various lymphatic organs; representative data are also reported for different types of mouse lymphocytes. Human blood lymphocytes are found to have normally-distributed diameters (p 〉 98%), a property not shared by their volumes or their surface areas, nor by blood lymphocytes from the chick or from patients with chronic lymphatic leukemia. A possible implication of this finding is mentioned briefly.
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  • 171
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Liver mitochondria from normal and alloxan diabetic rats, isolated in 0.25 M sucrose, were assayed with an oxygen electrode for ADP/O and Ca+2/O ratios, respiratory ratio, and respiratory control index. Mitochondria were incubated with two substrates, succinate and β-hydroxybutyrate; two types of ionic media, Na+ medium (Na+ the major monovalent cation) and K+ medium (K+ the major monovalent cation); and two respiratory stimulants, ADP (352 μM) and Ca+2 (187 μM). Significant differences between respiratory rates and ADP/O ratios were dependent upon the substrate and ionic medium employed. The results confirm previous studies which showed no alteration in ADP/O ratio but decreased State 3 respiratory rates under similar conditions of K+ medium with ADP stimulation in the diabetic. Furthermore, the State 3 respiration was prolonged compared to normal. Ca+2 stimulation was the same in normal and diabetic mitochondria in K+ medium. Studies in Na+ media revealed more significant differences in RCI's, respiratory rates, and ADP/O ratios that were substrate dependent as well as ion dependent. The results from these various studies can be accounted for by an hypothesis linking mitochondrial K+ interaction with alterations in the diabetic mitochondria.
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  • 172
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    Journal of Cellular Physiology 84 (1974), S. 291-299 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: In phototrophic culture of Euglena gracilis, good synchrony was found only under rather restricted programs of light-dark cycles, and rather narrow ranges of temperature and light intensity, when cultures were flushed with air fortified with adequate amounts of CO2. When flushed with air alone, CO2 was found to be limiting, and while cell divisions were rhythmic, less than a doubling of cell number occurred in division bursts. With air as gas phase, rhythmic division activity was maintained over wide ranges of temperature, light intensity, and the ratio of light:dark in a given program; all these factors affected the amplitude of the division burst, however.
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  • 173
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    Topics: Biology , Medicine
    Notes: Formation of granulocytic and macrophage colonies in agar cultures of mouse marrow or spleen cells was stimulated by the addition of medium from pokeweed mitogen-stimulated cultures of mouse spleen cells (PKW-CM). Approximately 5% of the colonies developing were large, dispersed granulocytic colonies (DG-colonies) composed of cells with eosinophilic cytoplasmic granules. The capacity to stimulate DG-colonies was shown by media conditioned by PKW-treated lymphoid and peritoneal cells but not by other cells or organ fragments.Velocity sedimentation studies indicated that cells generating DG-colonies were separable from cells generating regular granulocytic or macrophage colonies. DG-colonies did not survive if transfered to cultures containing other forms of CSF. The active colony stimulating factor in pokeweed mitogen-conditioned medium which stimulates DG-colony formation was antigenically distinct from the factor stimulating granulocytic and macrophage colony formation, was separable electrophoretically from the latter factor and on gel filtration had an apparent molecular weight of 50,000.Although the cells in DG-colonies have not been established to be eosinophils, DG-colonies represent an interesting new system for analysing further aspects of the control of growth and differentiation in hemopoietic populations.
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  • 174
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    Topics: Biology , Medicine
    Notes: When embryoid bodies are grown in suspension culture in vitro, they undergo only a limited amount of morphological development. When these same embryoid bodies are permitted to attach to the surface of a culture dish, a wide variety of new morphological cell types appear.Suspension cultures of embryoid bodies do not contain significant detectable levels of acetylcholine esterase or creatine phosphokinase. These same enzymes however are produced in cell cultures derived from embryoid bodies attached to the culture dish surface. Polyacrylamide gel electrophoresis has been employed to demonstrate that the electrophoretic form of creatine phosphokinase produced by teratoma cells in culture is the brain form of the enzyme.Solid transplantable tumors containing only embryonal carcinoma cells (stem cells) do not contain either of these enzymatic activities. Well differentiated transplantable teratomas contain both enzymes.
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  • 175
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    Journal of Cellular Physiology 83 (1974), S. 75-83 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Differential centrifugation of homogenates of Harding-Passey melanoma demonstrated that aryl sulfatase A and β-glucuronidase sediment with particles (i.e., lysosomes) distinct from those particles bearing tyrosinase (i.e., melanosomes). The sedimentation curves for the lysosomal enzymes and tyrosinase, however, demonstrated that an adequate separation of these particle types could not be obtained by differential centrifugation. Isopycnic density gradient centrifugation was used to obtain the necessary resolution. The results of the density gradient studies demonstrated that lysosomes and melanosomes could be separated by this technique, as judged by enzyme distribution among the fractions recovered from the gradients and from electron microscopic examination of the melanosome fractions. It was further evident that the purified and washed melanosomes contained significant amounts of both acid hydrolase activities. Indeed 24% to 27% of the total acid hydrolase activities recovered from the density gradients were associated with the melanosome fractions. The acid hydrolases associated with the melanosomes could not be solubilized by treatment with 0.1% (v/v) Triton X-100, nor by exposure to hypo-osmotic shock. The melanoma lysosomes, however, did release most of both their hydrolase activities into soluble form after treatment with the same percentage of detergent. The lysosomes were, however, very resistant to rupture by exposure to hypo-osmotic conditions.
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  • 176
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    Journal of Cellular Physiology 83 (1974), S. 85-90 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Skin fibroblast cultures from six patients with Down's syndrome (Trisomy 21) were compared with four in vitro age-matched normal fibroblast cultures. Growth rates were calculated from increases in cell number and total protein during exponential growth, early in culture lifetime (less than 20 doublings). The Down's syndrome (D.S.) cultures had an average population doubling time of 35.6 ± 1.1 hours and average mass doubling time of 38.6 ± 3.2 hours, significantly lower (p〈0.005) than the corresponding normal culture values of 23.0 ± 0.7 hours, and 23.3 ± 1.9 hours. D. S. Cells also contained 4.46 ± 0.19 ± 10-4 μg protein/cell as compared to 3.06 ± 0.13 × 10-4 μg/cell (p〈0.001) for normal fibroblasts.Similar in vitro observations of increased doubling time and protein content have been reported in normal fibroblasts from older donors, and from individuals with premature aging syndromes, as well as in normal fibroblasts near the end of their in vitro lifetime. The present results, obtained from cultures young in vitro, may therefore suggest that D.S. fibroblast cultures age prematurely. This hypothesis is consistent with clinical manifestations of premature aging in D.S. patients and points to a defect in growth regulation, both in vivo and in vitro, resulting from an extra copy of chromosome 21.
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  • 177
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    Journal of Cellular Physiology 83 (1974), S. 91-101 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Strong evidence is given that a nucleoside diphosphate kinase is present to some extent on the surface of intact neoplastic cells in culture. Experiments could be performed with cells cultured in a few plates to which an incubation medium was added. The cells were firmly attached to the supporting medium and remained viable during the incubation procedure.Determinations of lactate dehydrogenase were carried out to rule out any possible contamination from the culturing medium as well as from the cell interior. From these analyses, a procedure was developed which easily removed the last traces of the culturing medium and which showed that there was no leakage of intracellular lactate dehydrogenase during the incubation procedure. There was a rather insignificant diffusion of nucleoside diphosphate kinase into the incubation medium. In common with other nucleoside diphosphate kinases, the glioma cell surface enzyme seemed to be nonspecific with regard to nucleotide substrates.
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  • 178
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    Journal of Cellular Physiology 83 (1974), S. 103-116 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Colchicine resistant (CHR) lines of stable phenotype have been isolated from cultured Chinese hamster (CHO) cells. Successive single-step selections for increasing resistance were performed by isolating resistant colonies at each step. Two complementary assays involving [3H] colchicine uptake by whole cells and binding of [3H] colchicine by cytoplasmic extracts were developed to test for altered permeability and altered intracellular target protein, respectively. All clones isolated appeared to have decreased permeability to the drug while their colchicine-binding ability was not reduced. The amount of reduction in colchicine uptake correlated strongly with cellular resistance. The CHR lines were also cross resistant to other drugs such as actinomycin D, vinblastine and Colcemid; furthermore, the degree of cross resistance was positively correlated with the degree of colchicine resistance. The non-ionic detergent Tween 80 potentiated the cytotoxic action of colchicine on mutant cells as well as its rate of uptake into whole cells.
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  • 179
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    Journal of Cellular Physiology 83 (1974), S. 131-139 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: High density L cell suspension cultures were previously shown to remain viable for indefinite periods of time and to exhibit marked inhibition of DNA synthesis and mitosis while the fraction of total protein synthesis represented by collagen is increased. The present study demonstrates that regulation in this system extends to the activity of acetylcholinesterase found to be approximately 100-fold greater in the high density populations than in low density exponentially growing cultures. Kinetic studies of the increase of the activity, its fluctuation over an extended period of time and its decrease upon resumption of exponential growth after dilution of the cultures were performed. The data obtained indicate that the enzyme does not accumulate in high density populations merely as a result of the absence of net protein synthesis and cell division but that changes of its rates of synthesis and possibly degradation are involved. The expression of regulated acetylcholinesterase activity in a cell line of connective tissue origin is considered in relation to phenotype reprogramming and to cell membrane associated growth control mechanisms.
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  • 180
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    Notes: A perfusion technique is described for the study of melanosome response in ventral tailfin melanophores of Xenopus laevis tadpoles. The melanosomes remain aggregated (punctate melanophores) in Ringer's. Theophylline (15 mM) and caffeine (30 mM) cause a reversible dispersion (stellate melanophores) of melanosomes which is partly blocked by cytochalasin B (10 μg/ml). When added with theophylline or caffeine to stellate cells, cytochalasin B causes a disrupted distribution of pigment granules, characterized by a melanosome free central region. C-AMP (20 mM) and dibutyryl c-AMP (1 mM) cause a reversible dispersion of melanosomes which is partly inhibited by cytochalasin. When cytochalasin plus a nucleotide are added to stellate cells, some show the disrupted distribution of melanosomes. Colchicine (5 mM) causes irreversible, while griseofulvin (0.2 mM) causes a slight, but reversible dispersion of melanosomes, and cytochalasin has little effect on these reactions. Perfused tailfin melanophores remain capable of responding to reversible reagents for at least 12 hours and are unresponsive to changes in illumination.
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  • 181
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    Journal of Cellular Physiology 83 (1974), S. 141-149 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: A clonal strain of epithelial cells has been established from the transplantable Morris hepatoma 7800 and is designated 7800C1. The cells grow with a population doubling time of about three days in serum-supplemented synthetic medium. Cells of the 7800C1 strain have maintained measurable activities of all the enzymes of the urea cycle during 17 months in continuous culture. The activity of argininosuccinate lyase is approximately that found in normal rat liver, while argininosuccinate synthetase, carbamoyl phosphate synthetase, arginase and ornithine carbamoyl transferase activities are, respectively, 40%, 28%, 6%. and 1% of normal values. Treatment of 7800C1 cells with glucagon, dibutyryl 3′,5′-cyclic adenosine monophosphate or hydrocortisone did not increase the activity of any of the five enzymes.
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  • 182
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    Journal of Cellular Physiology 83 (1974), S. 151-158 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The attraction of fern spermatozoids by secretions from the female reproductive structures, and by salts of malic acid, has long been known as a classic example of precise chemotactic orientation by motile cells. Spermatozoids of the bracken fern are attracted by the partially ionized form of malic acid, bimalate ion, and also by calcium ions. Both calcium and bimalate ions must be present for chemotactic response and for response to voltage gradients, Spermatozoids swimming up a bimalate concentration gradient swim in helical paths of abnormally small radius; if they accidentally swim down the concentration gradient the radii of their path helices become abnormally large.These observations suggest that changes in the direction of flagellar beating in response to the rate of change of bimalate ion concentration with time may be the basis for chemotactic orientation. A “coupled diffusion” hypothesis for chemo-reception is presented, which postulates a membrane carrier which can only circulate freely in the membrane if it binds both bimalate and calcium ions. This hypothesis could explain time-differentiation of the stimulus, the coupling of a specific stimulus  -  bimalate ions  -  to a general mediator of intracellular response  -  calcium ions  -  and the quantitative relationship between response of the spermatozoids and the chemical potential of “calcium bimalate.”
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  • 183
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    Journal of Cellular Physiology 83 (1974), S. 159-161 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
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  • 184
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    Journal of Cellular Physiology 83 (1974) 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
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  • 185
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    Journal of Cellular Physiology 83 (1974), S. 287-296 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Elevated calcium and magnesium concentrations promoted mitotic activity in rat thymic lymphocyte cultures. Oestradiol inhibited calcium- but not magnesium-induced mitogenesis. One prerequisite for the mitogenic action of calcium is a raised intracellular concentration of cyclic adenosine 3′5′ monophosphate (cyclic AMP) but cyclic AMP-induced mitogenesis was insensitive to oestradiol. This suggests that the steroid blocks the mitogenic process at a stage preceding the endogenous cyclic AMP elevation. Furthermore the mitogenic actions of adrenaline, which stimulates adenylate cyclase (the enzyme responsible for cyclic AMP biosynthesis), and caffeine, which inhibits phosphodiesterase (the enzyme which degrades cyclic AMP) were also insensitive to oestradiol inhibition. This precludes a direct effect of the steroid on these enzymes. However, oestradiol did inhibit the mitogenic action of parathyroid hormone (PTH). Since the mitogenic action of PTH probably involves increased calcium entry to the cell, oestradiol may block this ion influx.The inhibition of calcium- and PTH-induced mitogenesis must be attributable to some structurally specific action of oestradiol. The steroids cholesterol, progesterone and testosterone all failed to reduce calcium-induced mitogenesis, whereas both α and β oestradiol were effective.In addition to its insensitivity to oestradiol inhibition, magnesium-stimulated mitosis was unaffected by both imidazole and calcitonin at concentrations which significantly reduced calcium-stimulated proliferation. These findings are compatible with the thesis that magnesium-induced mitogenesis does not involve the elevation of cyclic AMP concentrations.
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  • 186
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    Journal of Cellular Physiology 83 (1974) 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
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  • 187
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    Journal of Cellular Physiology 83 (1974), S. 19-25 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Intranuclear Na, K and Mg concentrations were determined in cells of salivary glands incubated for 1h in selected NaCl/KCl/MgCl2 media. By variation of the external milieu beyond “physiological” limits the intranuclear electrolytes can be shifted between ca 100 and 280 mM [K]i, between ca 8 and 100 mM [Na]i and between ca 5 and 75 mM [Mg]i. No significant competition or interactions of the 3 ionic species are apparent. The relationships [K]e : [K]i and [Na]e : [Na]i can best be described by a positive and linear, that between [Mg]e : [Mg]i by a negative and exponential function. Regression parameters are given which permit a computation of intranuclear [Na], [K] and [Mg] as induced by NaCl/KCl/MgCl2 in any binary or triple combination that is tolerated by the explanted gland without visible damage.
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  • 188
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    Journal of Cellular Physiology 83 (1974), S. 321-336 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The incorporation of 3H-labeled deoxyadenosine and deoxyguanosine into nucleic acids by cultured Novikoff rat hepatoma cells is about 80% into RNA and 20% into DNA. The pathways of incorporation have been elucidated in studies with whole cells and cell-free extracts. Deoxyadenosine is very rapidly deaminated to deoxyinosine. Most of the deoxyinosine formed by whole cells is transported out of the cells and accumulates in the medium. A portion of the deoxyinosine, and deoxyguanosine are phosphorolyzed by purine nucleoside phosphorylase to hypoxanthine and guanine, respectively. The latter are subsequently converted by hypoxanthine-guanine phosphoribosyl transferase to IMP and GMP, respectively. Incorporation of the purine deoxyribonucleosides into DNA is mainly via this pathway and the subsequent reduction of ADP and GDP by ribonucleoside reductase, although a small proportion of the deoxyadenosine and deoxyguanosine taken up by the cells seems to be directly phosphorylated to dAMP and dGMP, respectively. Deoxyguanosine is incorporated only into guanine residues of RNA and DNA. Deoxyadenosine is also mainly incorporated into guanine residues of RNA and DNA, although the radioactivity of deoxyadenosine in the acid-soluble pool is almost exclusively associated with ATP. A similar labeling pattern is observed with labeled deoxyinosine, inosine or hypoxanthine. The pyrimidine deoxyribonucleosides, on the other hand, are specific precursors for their respective bases in DNA.Hydroxyurea inhibits the incorporation of all deoxyribonucleosides into DNA. Results from pulse-chase experiments indicate that the inhibition of DNA synthesis is prevented by the presence of high concentrations of deoxyadenosine plus deoxyguanosine in the medium. Either purine deoxyribonucleoside alone or deoxycytidine, hypoxanthine or inosine alone or in combination with deoxyadenosine or deoxyguanosine are ineffective. The results are consistent with the conclusion that the inhibition of DNA synthesis is due to a depletion of the dATP and dGTP pools as a result of the hydroxyurea treatment. On the other hand, hydroxyurea causes an increased incorporation of thymidine and deoxycytidine into the dTTP and dCTP pools, respectively. Evidence is presented to indicate that this effect of hydroxyurea is due to an increased synthesis of dTTP and dCTP rather than to an inhibition of their turnover.
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  • 189
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    Journal of Cellular Physiology 83 (1974), S. 337-343 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The transport of various deoxyribonucleosides by cultured Novikoff rat hepatoma cells (subline N1S1-67) follows normal Michaelis-Menten kinetics. The transport reactions are competitively inhibited by most heterologous deoxy- and ribonucleosides and by Persantin and Cytochalasin B. Comparisons of the transport kinetics of the various deoxyribonucleosides (Km and Vmax ) and of the Km/Ki ratios for the inhibitions indicate that deoxythymidine, deoxyuridine and 5-fluordeoxyuridine are transported by a single system, whereas deoxycytidine and the purine deoxyribonucleosides are transported by other systems. The data suggest that deoxyadenosine, deoxyguanosine and deoxyinosine, are not transported by a single system, but the number of transport systems involved could not be established unequivocally. Similar comparisons also suggest that the deoxyribonucleosides are transported by different systems than the ribonucleosides. All deoxyribonucleoside transport systems are inhibited to about the same extent by Persantin (Ki = 1-2 μM) and Cytochalasin B (Ki = 4-12 μM). The inhibitions of deoxynucleoside transport resulted in corresponding apparent competitive inhibitions of their incorporation into nucleic acids.
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  • 190
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    Journal of Cellular Physiology 83 (1974), S. 345-351 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Ornithine decarboxylase activity in high density, stationary phase rat hepatoma (HTC) cells in suspension culture has an extremely short half-life of between 5 and 15 minutes, as measured after inhibiting protein synthesis. Following dilution of these cells into fresh medium there is a large increase in ornithine decarboxylase activity, reaching a peak often several hundred times the initial level at about four hours. At least part of this stimulation is due to an increase in the apparent half-life of the enzyme, to between 30 and 90 minutes. Evidence is presented that the supply of amino acids can control the turnover of ODC under some conditions. For example supplementing high density cells with glutamine, asparagine, serine, glycine and proline, either singly or together, increases ODC activity and decreases its apparent turnover. The stimulation by amino acids is enhanced by serum.
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  • 191
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    Journal of Cellular Physiology 84 (1974), S. 319-332 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Embryoid bodies are produced when a transplantable testicular teratoma from strain 129 mice is serially passaged in the peritoneal cavity of these mice. These bodies are roughly spherical containing two morphologically distinct cell types. Scanning and transmission electron microscopy have been employed to show that the endodermal cells of embryoid bodies, like those of the mouse embryo, are on the outer surface and have highly convoluted surfaces containing numerous microvilli-like projections. The inner, embryonal carcinoma cells, are the pluripotent stem cells of this tumor.Intracisternal A-type particles have been observed in electron micrographs and are almost exclusively located in the endodermal cells of the embryoid bodies. The A-type complement-fixing antigen has been identified in extracts prepared from this tumor.When embryoid bodies are placed in culture and allowed to attach to the surface of a petri dish, a large number of new morphologically distinct cell types appear. Attachment to the petri dish surface is required for the generation of these new cell types. Cells of similar morphology in culture, display a distinctly “clonal” distribution on the petri dish surface.
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  • 192
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    Notes: Mouse embryo fibroblasts growing asynchronously in vitro stained with Feulgen method and their nuclear chromatin was analysed by means of the image analysing computer Quantimet 720D. Cells with 2C, 3C and 4C content of DNA were considered as being in G1, middle S and G2 phase of cell cycle, respectively. It was found that the projected area of nuclei increases during the cell cycle and that the mean optical density of chromatin increases from G1 through S to G2 phase. The curves showing the areas of chromatin at different optical density thresholds are different for cells in G1, S and G2 phase. The results demonstrate cyclic changes in chromatin morphology in the interphase nuclei during the cell cycle.
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  • 193
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    Notes: Exposure of a thymic lymphocyte population (suspended in serum-free synthetic medium) to the phytomitogen concanavalin A (Con A) causes brief (within the first 8 to 12 minutes) rises in the cellular contents of cyclic AMP and cyclic GMP. However, the rise in the cyclic GMP level is calcium (extracellular)-dependent, but the cyclic AMP rise is not. These changes are followed during the next hour by the initiation of DNA synthesis by a large fraction of the lymphoblast subpopulation which, like the preceding cyclic GMP rise, is calcium-dependent. The stimulated lymphoblasts eventually progress into mitosis. Additional observations indicate that Con A operates by sensitizing lymphoblasts to calcium ions which, in turn, cause the initiation of DNA synthesis by a process mediated by cyclic GMP, but not cyclic AMP.
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  • 194
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    Journal of Cellular Physiology 84 (1974), S. 459-462 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: When chick embryo fibroblasts are grown for two days in the presence of low doses (18 μg/ml) of 5-bromodeoxyuridine (BrdU), their agglu-tinability by Concanavalin A is increased to about the same degree as following viral transformation of the cells by Rous sarcoma virus. Simultaneous addition of a large excess, but not of a low dose of thymidine prevents this effect of BrdU.Treatment with BrdU also enhances the agglutinability of fibroblasts infected with a conditional It mutant of Rcus sarcoma virus at the temperature of 41° which restricts morphological transformation.
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    Journal of Cellular Physiology 84 (1974), S. 463-471 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The discharge of Aplysia pacemaker neurons varies with temperature over the range 10 to 22°C. Three types of frequency-temperature plots are found, with maximal discharge at lowest, intermediate or highest temperatures. In the presence of ouabain, however, all cells show maximal discharge at the highest temperature, suggesting that the steady state activity of an electrogenic sodium pump is an important determinant of membrane excitability. The average magnitude of pump current, as indicated by the applied current necessary to restore discharge to control values after ouabain application, was about 4 namps at 20°C but near zero at 10°C. These neurons may be excellent models of mammalian thermoreceotprs.
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    Journal of Cellular Physiology 84 (1974), S. 481-485 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Following enucleation of a portion of human culture cells containing 3H-pyridine nucleotides, autoradiography revealed no difference in the grain density over enucleated and whole cells. These results provide evidence that the concentration of pydine nucleotides does not differ by more than threefold between nucleus and cytoplasm and probably does not vary at all.
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    Journal of Cellular Physiology 83 (1974), S. 449-456 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: P-chloromercuriphenylsulfonate, PCMBS, and 5, 5′ dithiobis-(2-nitrobenzoic acid), DTNB at a concentration of 1 mM are found to inhibit the rate of water transport across human red cell membrane. In addition PCMBS inhibits the rates of transport of small hydrophilic but not hydrophobic nonelectrolytes. Other sulfhydryl reagents such as N-ethylmaleimide and iodoacetamide have no significant effect on the rate of water transfer in these cells. The results suggest that there are at least two populations of membrane bound SH-groups which differ in their topical location which participate in the control of water transfer. One is located closer to the outer surface of the membrane, and thus is readily accessible to PCMBS while the other component is probably located in the membrane interior. These two populations can be dissociated by pH. The effect of PCMBS on water transfer can be greatly influenced by pH and temperature. The main effect of temperature and pH is on the permeability of the membrane to the drug. The same concentration of PCMBS is also found to inhibit to a lesser degree water transfer across other biological membranes.
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  • 199
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    Journal of Cellular Physiology 84 (1974), S. 49-55 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Variants or “mutants” temperature-sensitive (ts) for growth have been isolated by selection from a near-diploid mouse cell line. Thus far. 10 ts mutants which grow normally at 33° C, but not at 39° C, have been isolated. These ts mutants were then studied to determine if any manifested their defect at a unique point or stage in the cell cycle. This type of ts mutant is termed a “cell cycle” mutant.The first screen involves observing individual cells of an asynchronous culture for residual division after a shift from 33° C (permissive temperature) to 39° (nonpermissive temperature). A cell cycle mutant should show some fraction of the cells dividing only once at a normal rate after the shift. The ts variant B54 met this first criterion for a cell cycle mutant (i.e., 50% residual division) and was further analyzed.The second screening technique monitors (1) the rate of entry into S, (2) the length of G2, and (3) the rate and duration of cells entering mitosis after a shift of an asynchronous culture to 39°. This experiment with B54 revealed that cells in G1 at the time of the shift to 39° failed to enter S while cells already into S completed the cycle at 39°. These results suggest that B54 is defective in a G1 function which is required for entry into S, but which is no longer needed once cells have entered S. Other results are presented which also support this hypothesis.In addition the ts function of B54 is apparently required for recovery from a “high density” G1 arrest.
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    Journal of Cellular Physiology 84 (1974), S. 69-73 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The cAMP levels of the 3T3 cell line and its transformed derivatives were determined. The level was found to be lower in transformed cell lines than in parental 3T3 and to decrease at confluence in all cell lines tested. SV40 transformed 3T3 cell lines which are temperature sensitive in growth control were found to have lower cAMP levels than 3T3, but these levels were not temperature dependent. Retransformation of these cell lines by murine sarcoma virus did not markedly affect their cAMP levels.
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