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  • General Chemistry  (890)
  • ENERGY PRODUCTION AND CONVERSION  (833)
  • GEOPHYSICS  (467)
  • Cell & Developmental Biology  (372)
  • 1980-1984  (2,562)
  • 1970-1974
  • 1915-1919
  • 1980  (2,562)
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  • 1980-1984  (2,562)
  • 1970-1974
  • 1915-1919
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  • 101
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    New York, NY : Wiley-Blackwell
    Journal of Morphology 165 (1980), S. 187-204 
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Common snapping turtles (Chelydra serpentina) lay nearly spherical, flexible-shelled eggs having an outer mineral layer composed of calcium carbonate in the aragonite form. The mineral layer is arranged into loosely organized groups of nodular shell units, with numerous spaces (or pores) between adjacent shell units. Shell units are structurally complex, consisting of an inner tip that is morphologically distinct from the main body of the shell unit. Contained within an intact shell unit at the interface of the tip and the main part of the shell unit is the central plaque, an apparent modification of the shell membrane that may serve to nucleate calcification of shell units during shell formation. The tips of shell units are firmly attached to a single, multilayered shell membrane throughout much of incubation. The calcareous layer begins to detach from the shell membrane about half-way through incubation, and changes in shell morphology attending this detachment indicate that snapping turtles may use the shell as a source of calcium during embryogenesis. The arrangement of the mineral layer into groups of shell units, the large number of spaces between shell units, and little or no interlocking of crystallites of adjacent shell units apparently are factors contributing to the ability of these eggs to swell as they absorb water.
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  • 102
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    New York, NY : Wiley-Blackwell
    Journal of Morphology 165 (1980), S. 255-260 
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Three unusual highly ordered configurations of yolk protein in yolk precursor bodies are described. These differ from the crystalline structure of the main body of mature yolk platelets. One of these is an aggregation of paired membranes with a spacing of about 100 Å between the members of a pair. The paired membranes of such an aggregation may be straight, parallel, and very close together; they may appear as a tight whorl; or they may display an intermediate random arrangement with varying distances between pairs. Another configuration is a tubule with a diameter of about 450 Å, whose wall appears in cross section to consist of particles measuring 50 × 100 Å. A third configuration is a crystalline array of rows of angular-shaped particles with a spacing of about 160 Å. It is suggested that these may represent intermediates in the transition of vitellogenin to lipovitellin and phosvitin.
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  • 103
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    New York, NY : Wiley-Blackwell
    Journal of Morphology 165 (1980), S. 237-254 
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The oral apparatus of neonatal and juvenile golden hamsters was investigated by clearing and staining of whole crania, videotaping of behavior, and electromyography of several jaw muscles. Chewing developed during the first postnatal week and matured in the second; however, suckling was still the primary mode of feeding. Micromovements of the jaws occurred early when the osseous skeleton and joints developed. Macromovements correlated well with EMG records and were limited to jaw opening at birth. Muscles of the oral floor generated large bursts of activity during jaw opening and tongue protrusion from 0 days postnatal (dpn), when simple and stereotyped gaping was induced, until 14 dpn, when movements were spontaneous and not stereotyped nor inducible. However, adductor muscle activity was brief, low in amplitude, and primarily involved with jaw stabilization until 4 dpn, when these muscles became active during closing the jaws; closing activity increased in frequency and amplitude until the end of the second week. Development of frequent, coordinated macromovements of chewing was associated with the refinement of joint structure and dental occlusion and with the growth of the craniofacial skeleton. Jaw movements and associated EMG's correlated better with available data on development of neural circuitry than with that for musculoskeletal development.
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  • 104
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    New York, NY : Wiley-Blackwell
    Journal of Morphology 166 (1980) 
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
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  • 105
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    New York, NY : Wiley-Blackwell
    Journal of Morphology 165 (1980), S. 301-317 
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Light, transmission, and scanning electron microscopy of both adult and third-stage Echinocephalus sinensis shows that the head is characterized by a circular muscle layer in the headbulb wall, a highly unusual arrangement in nematodes. The basal region of cephalic spines is enclosed by all three cuticular zones but the distal region of its shaft is lined only by the cortical layer. The so-called “ballonet-cervical sac” system is actually formed by four modified cervical muscle cells of the circomyarian type, each with a liquid cytoplasmic matrix serving as a hydrostatic chamber. The headbulb is deflated by contraction of the circular wall muscle and six pairs of specialized longitudinal and oblique muscles. Two pairs of oblique somato-oesophageal muscles also serve to shorten the oesophagus. Relaxation of the muscles and an anterior flow of fluid into the cephalic region of the cervical muscle cells inflate the headbulb. In adult worms, the trilobed pseudolabia are lined internally mostly by the oesophageal cuticle. Four radially arranged muscles help to dilate the buccal cavity and the pseudolabia can be retracted into the headbulb by two pairs of oblique muscles inserted at their base. The radial musculature at the anteriormost oesophagus has more abundant and tightly packed myofilaments than other regions. Four pitlike structures of unknown function are located near the base of the pseudolabium. In the third-stage worm the pyriform pseudolabium is internally lined mostly by the body cuticle. Two rows of bulbous structures each with a central process are located on the headbulb a short distance from the pseudolabium. Two pairs of oblique buccal dilatory muscles help to dilate the oral opening and draw the pseudolabia towards the headbulb. Two bands of oblique myofilaments are present within the anterior-most region of the oesophagus. The functional adaptation of the cephalic system in relation to the biology of the parasite is discussed.
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  • 106
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    New York, NY : Wiley-Blackwell
    Journal of Morphology 166 (1980), S. 37-50 
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Intercellular bridge development was compared in Ceropia by transmission electron microscopy in the germ cells of males and females. Bridge formation begins in the fourth instar, and the spindle remnants in newly formed bridges are replaced at this time by an amorphous material known as the fusome. In the four- and eight-celled cystocyte clusters of the female, the newly formed intercellular bridges migrate centripetally, forming a central complex of bridges surrounded by a rosette of germ cells. The fusomes become continuous and occupy all of the bridges in the complex. At each division each mitotic spindle orients itself with one pole toward the continuous fusome. In the female, mitosis stops at the eight-cell stage, and the cystocytes all enter first meiotic prophase. At the end of the fifth instar, when nurse cell differentiation commences in seven of the cells, the continuous fusome is replaced by a continuous mass of mitochondria and microtubules. In the male, bridge migration and rosette formation are abandoned during the later mitotic and meiotic divisions; during these stages the fusome is no longer continuous. The fusome of males disappears during spermatid differentiation and is not replaced by mitochondria and microtubules. The ability of the centrally located continuous fusome to orient the mototic spindles of succeeding mitoses could account for the earlier observations that all pre-existing bridges remain in only one of the daughter cells at each successive cystocyte division in the female.
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  • 107
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    Journal of Morphology 166 (1980), S. 109-126 
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The testicular wall and the process of spermatogenesis in the crinoid, Florometra serratissima, has been studied at the fine structural level. The testicular wall is composed of three layers: a perivisceral layer consisting of nerve processes, muscle fibers, and epithelial cells; a haemal sinus containing haemal fluid, collagen-like fibers, and haemocytes; and a germinal layer consisting of germinal and interstitial cells.The germinal layer is elaborated into numerous folds that project into the lumen of the testis and a branch of the haemal channel extends through the core of each fold. Evidence suggesting that nutrients are carried to the testis and germinal cells via the haemal system is presented. Spermatogonia are concentrated around the base of each fold and spermatocytes line the more distal regions. Spermatids occur at the luminal surface of the germinal layer and spermatozoa fill the testicular lumen. Interstitial cells phagocytize spermatozoa and may also transfer nutrients to spermatids.The nucleus of spermatogonia is large and contains one or two nucleoli. The cytoplasm contains numerous organelles, lipid granules, and a distal and proximal centriole, each with a satellite complex. A striated rootlet extends from the distal centriole. During first meiotic prophase, the distal centriole loses its striated rootlet and produces a flagellum, the proximal centriole loses its satellite complex, the nucleolus disappears, and proacrosomal vesicles are synthesized by the Golgi complex. During spermiogenesis, most of the mitochondria appear to fuse to form a single, large mitochondrion, the nuclear chromatin condenses, and superfluous cytoplasm is lost by autophagocytosis. The formation and definitive positioning of the acrosomal vesicle and periacrosomal material at the apex of the nucleus is described in detail.
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  • 108
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    Journal of Morphology 166 (1980), S. 197-202 
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The scutate scales are entirely missing in chick embryos homozygous for the gene, “scaleless.” Reticulate scales of this mutant are present; however, they have undergone abnormal morphogenesis into irregular mounds and crevices. The pattern of keratinization seen along the anterior metatarsus of normal embryos differs dramatically from that seen along the anterior metatarsus of scaleless embryos. In contrast, we find that the unique pattern of keratinization seen in the epidermal cells of normal reticulate scales is retained in mutant reticulate scales, even though these scales are morphologically abnormal. We believe that differences in the initial tissue interactions (which establish the inductive ability of the dermis) of these two types of scales are responsible for the differences seen in their responses to the scaleless gene. The pleiotropic nature of the scaleless gene is discussed.
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  • 109
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    Journal of Morphology 166 (1980), S. 258-258 
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
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  • 110
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    Journal of Morphology 166 (1980), S. 231-257 
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Electron microscopy was used to follow the transformation of the endostyle to a thyroid gland in the anadromous sea lamprey, Petromyzon marinus L., throughout metamorphosis (stages 1-7). Transformation of the larval (ammocoete) endostyle begins at the first signs of external change (stages 1-2), and the adult form of the gland is reached by stage 5. Only slight modifications of the gland accompany further development to the end of metamorphosis.Development of the thyroid gland involves degeneration, proliferation, and reorganization of the cells in the endostyle, and changes in their fine structure. Ultrastructural changes during early stages are most obvious in the type 1 cells that make up the shrinking glandular tracts, and involves the accumulation of cytoplasmic microfilaments and a variety of cytoplasmic inclusions. The glandular tracts and their cells gradually disappear through autolysis and, apparently, through phagocytosis by neighboring epithelial cells and macrophages. Although the fine structure of the type 2, 3, 4, and 5 cells is not altered in the early stages, by stage 3, many of these cells become either vacuolated, undergo autolysis, or are extruded. Phagocytosis of some of each of these cell types likely occurs.Thyroid follicles are first observed during stage 4. Some of their lumina seem to arise from the accumulation of material in intercellular spaces and from vacuoles among cell clusters. Other lumina may represent a portion of the original lumen of the endostyle. Many follicles appear to be comprised of cells with cytological characteristics similar to those of larval cell types 3 and 2c. Some of the other larval cell types, such as type 5, may also be involved. In young adult lampreys follicles are composed of cuboidal to columnar cells that lack the dilated cisternae of rough endoplasmic reticulum seen in follicular cells of higher vertebrates. Dense collagenous connective tissue surrounding the follicles contains relatively few blood vessels.The transformation process described may have some relevance to our understanding of the development and evolution of the vertebrate thyroid gland.
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  • 111
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    New York, NY : Wiley-Blackwell
    Journal of Morphology 166 (1980), S. 275-288 
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Serial sections ranging from very young embryos to hatched juveniles and whole embryos of Scyliorhinus show that dentition and dermal skeleton belong to two independent secondary developmental fields that differ both developmentally and structurally. The development of the dentition starts very early, with a thickening of the ectoderm in the region of the mouth (stage 04), the invagination of the dental lamina (stage 18), and the formation of the germs of the first generation (stage 20). Tooth replacement movements start only near the end of embryogenesis (stage 35). Scale germs, on the other hand, first begin to form at stage 24. Scales erupt shortly before the animal hatches (stage 43). Only one scale generation is formed during embryogenesis. The forces which erupt the scales may come from fluid pressures in vacuoles of the fibrous layer of the dermis. Those which erupt the teeth probably also result from similar fluid pressures. The crown and upper part of the base of scales and teeth are formed by cells of the inner dental epithelium which are differentiated from the ectoderm. They are also formed by odontoblasts which are derived from the vascular layer of the dermis. However, the basal plates of scales and teeth containing the anchoring fibers are formed by osteoblasts, which are derived from the fibrous layer of the dermis.
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  • 112
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    Journal of Morphology 166 (1980), S. 217-230 
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: A broad survey of muscle unit properties in 14 muscles of the cat hind limb is presented which emphasizes some general features of unit properties in mammalian muscles. A more detailed analysis of muscle unit properties in three muscles of the posterior compartment of the lower leg is then presented using Burke's tetrapartite (FF, FI or F (Int.), FR, and S) unit classification scheme. Our data on the properties of motor units in cat tibialis posterior (TP) have been compared to those generated by Burke and colleagues on units in flexor digitorum longus (FDL) and medial gastrocnemius (MG). In all three muscles, twitch contraction time was distinctly slower for type S units and specific tension outputs were substantially greater for type FF units than for type S units. The innervation ratios of type FR units were slightly lower than for type S units but the specific tension of the FR units was closer to FF units than to type S units. The FF units controlled 70-74% of the cumulative force output of each muscles, indicating a substantial capacity for powerful rapid contractions of all three of these muscles despite their differences in “size,” action, and force generation. Distinctive features of the three muscles included differences in the unit types' force producing capabilities and in the relative representation of “nonfatigable” type FR and S units in each muscle. In particular, TP is endowed with some unusually powerful type FF units and a high percentage (42%) of type S units. In contrast, FDL has units that develop relatively little force and an unusually high representation (56%) of type FR units. The possible relationships between these muscle features and their presumed role in posture and locomotion is discussed.
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  • 113
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    Journal of Morphology 166 (1980), S. 127-127 
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
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  • 114
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    Journal of Cellular Physiology 102 (1980), S. 63-70 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Colchicine resistant (CchR) mutants have been isolated from Friend erythroeleukemic cells by successive single-step selections. Measurements of the rate of uptake of [3H]-colchicine into whole cells, and the binding of [3H]-colchicine to cytoplasmic extracts, suggest that these mutants are colchicine-resistant due to a reduced membrane permeability to colchicine, rather than an altered intracellular colchicine-binding target. Consistent with this conclusion is the observation that non-toxic concentrations of Tween-80, a non-ionic detergent, potentiated colchicine uptake into mutant cells. In addition, these Friend cell mutants, like CchR mutants of other cell types, are cross-resistant to a variety of unrelated drugs, including daunomycin, puromycin, emetine, and actinomycin D.A comparison of the dose-response curves for the induction of Friend cell differentiation by actinomycin D of both wild-type and two CchR cells suggests that actinomycin D permeation is required for its effects on Friend cell differentiation. Potentiation of actinomycin D uptake by Tween-80 significantly lowered the concentration of drug required to induce hemoglobin synthesis in the CchR cells, but had no significant effect on either actinomycin D induction of CchS cells or DMSO induction of both CchS and CchR cells. In common with other chemical inducers of Friend cell differentiation, the addition of actinomycin D results in an early decrease in 86 RbCl uptake, although this effect on transport occurred 14 hours later than that observed with DMSO.
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  • 115
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    Journal of Cellular Physiology 102 (1980), S. 91-98 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The characteristics of glucose and amino acid metabolism over a 98-hour incubation period were studied in a primary culture of neonatal rat skeletal muscle cells. The cells formed large myotubes in culture, were spontaneously highly contractile, and had cell phosphocreatine levels exceeding ATP concentrations. Medium glucose fell from 7.2±0.2 to 1.5±0.1 mM between 0 and 98 hours; intracellular glucose was readily detectable, indicating glycolysis was limited by phosphorylation, not glucose transport. Alanine levels in the medium increased from 0.06±0.01 to 0.82±0.04 mM between 0 and 48 hours and decreased to 0.72±0.04 mM by 98 hours. The period of net alanine production correlated with the rise in the cell mass action ratio of the alanine aminotransferase reaction. Cell aspartate, glutamate, and calculated oxalacetate levels were inversely related to the cell NADH/NAD+ ratio, as represented by the intracellular lactate/pyruvate ratio (r=0.78-0.88). The branched chain amino acids (leucine, isoleucine, valine) were actively utilized, e.g., medium leucine fell from 0.70±0.01 to 0.30±0.06 mM between 0 and 98 hours. In addition, arginine and serine consumption was observed in conjunction with ornithine, proline, and glycine production. Conclusions: (1) A major driving force for the high rates of alanine production by skeletal muscle cells in tissue culture is the active utilization of branched chain amino acids. (2) Intracellular aspartate and glutamate pools are linked, probably via the malate-aspartate shuttle, to the cell NADH/NAD+ redox state. (3) Muscle cells in tissue culture metabolize significant amounts of arginine and serine in association with the production of ornithine and proline, and these pathways may possibly be related to creatine production.
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  • 116
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    Journal of Cellular Physiology 102 (1980), S. 81-89 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The synthesis of ribosomes in HeLa cells was studied during recovery from a 20-hour deprivation for valine. The rates of incorporation of labeled precursors into ribosomal pre-RNA, processed rRNA, total cellular proteins, and proteins of the 60S ribosomal subunit returned to normal or nearly normal levels immediately after restoration of valine to the medium. Specific proteins of the 60S ribosomal subunit, whose apparent net synthesis is reduced more than that of the other proteins of the 60S ribosomal subunit during valine deprivation, were no longer undersynthesized after valine was restored. This rapid recovery suggests that the apparent decrease in the net rate of synthesis of these ribosomal proteins during valine deprivation is effected at the translational or post-translational level. No evidence of significant synchrony in any particular stage of the cell cycle was observed after a 20-hr valine deprivation. Key words: 60S ribosomal subunit; HeLa, cells; valine deprivation.
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  • 117
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    Journal of Cellular Physiology 102 (1980), S. 305-316 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Stable mutants resistant to pactamycin (PacR), a polypeptide chain initiation inhibitor, have been selected in a single step in Chinese hamster ovary (CHO) cells. The sensitivity of protein synthesis in mutant cell extracts to pactamycin indicates that resistance involves an alteration in the permeability of this drug. The failure of PacR mutants to show cross-resistance to other compounds provides further indication that the lesion is presumably specific for pactamycin. Cell hybrids formed between PacR × PacS lines show intermediate sensitivity towards pactamycin, suggesting that the PacR lesion behaves codominantly under these conditions. In the presence of subinhibitory concentrations of pactamycin, CHO cells, which are normally short, polygonal and disoriented, became greatly elongated and aligned themselves in parallel fashion to produce highly oriented colony morphologies, reminiscent of normal diploid fibroblasts. This effect of pactamycin on cellular morphology was seen much more clearly with the PacR mutants, although somewhat higher concentrations of the drug were required to produce this change.
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  • 118
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    Journal of Cellular Physiology 102 (1980), S. 323-331 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Leukemic cells in the myeloblastic stage from a murine myeloid leukemia cell line (M1) were induced to differentiate to macrophages by lipopolysaccharide (LPS) from Gram-negative bacteria. A granulocyte-macrophage colony-stimulating factor (CSF) was produced only during differentiation. After induction of differentiation, the continued presence of LPS was necessary to stimulate the macrophages to release CSF.In contrast, a macrophage cell line (Mm-1) derived from the M1 line produced CSF without LPS-stimulation, but CSF release was stimulated by the presence of LPS.
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  • 119
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    Journal of Cellular Physiology 102 (1980), S. 333-341 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Secretion of a granulocyte-macrophage colony-stimulating factor (GM-CSF) was accomplished by L-P3 cells in culture with a serum-free medium. Cell proliferation per se was not requisite for the production of GM-CSF; the cells continued secreting GM-CSF even after their growth had been suspended. The amount of GM-CSF accumulated in the conditioned medium was reasonably accounted by the daily rate of production, and the addition of a proteinase inhibitor such as leupeptin and pepstatin did not result in greater accumulation of GM-CSF in the culture. It is thus postulated that there is no significant proteolytic inactivation of the secreted GM-CSF in the culture. However, when partially purified GM-CSF preparation was chromatographed on a gel-filtration column in the presence of 0.1% Triton X-100, a derivative of the GM-CSF was yielded which had been diminished in the molecular weight and altered in the isoelectric point. On the other hand, when leupeptin was included in the solution during production and isolation of the factor, the yielded GM-CSF did not manifest such a detergent-induced transformation and maintained its isoelectric point at pH 3.5. It is thus assumed that, in the presence of the detergent, GM-CSF suffers deterioration by an endogenous proteinase and releases a sialoglycopeptide fragment without loosing its colony-stimulating activity.
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  • 120
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    Journal of Cellular Physiology 103 (1980), S. 179-191 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: UV damage to CHO cell DNA, measured by formation of thymine-containing dimers, increases from mitosis to early S phase. Computer simulation of UV absorption by the DNA of an idealized CHO cell at different stages in the cell cycle resembles the cycle dependence of UV damage.Incision at UV damage sites, measured by the accumulation of breaks in preexisting DNA during 30 minutes' post-irradiation incubation with the DNA synthesis inhibitors 1-β-D arabinofuranosylcytosine and hydroxyurea, increases from mitosis to interphase. Analysis of the dose dependence of DNA break accumulation indicates that both the affinity of the endonuclease for dimer sites and the maximum enzyme activity at saturating levels of dimers are significantly lower in mitosis than in interphase.The killing of CHO cells by UV is enhanced if repair is temporarily inhibited by ara C. The DNA gyrase inhibitor novobiocin prevents UV-induced incision.
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  • 121
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    Journal of Cellular Physiology 103 (1980), S. 173-178 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: When primary cultures of hepatocytes are maintained for 2 weeks from the time of perfusion, the activity of the enzyme glucokinase decreases rapidly, so that the activity can no longer be detected after the fourth day in culture. Concomitantly, there occurs an increase in the activity of hexokinases, the low-KM isozymes, which predominate in fetal liver. We have made several modifications of the culture medium in an attempt to prevent the decrease in glucokinase activity. When the medium was supplemented with a mixture of insulin, thyroxine, glucagon, dexamethasone, testosterone, and estradiol, the activity of the enzyme in the hepatocytes was present at approximately 15% of in vivo levels after 2 weeks in culture. When this hormone mixture was present during the first 4 hrs of culture and when the hepatocytes were allowed to attach to the collagen support and were maintained thereafter in medium supplemented with fetal bovine serum, insulin, and dexamethasone, the activity of glucokinase increased after an initial decrease for 3 days and was maintained thereafter at levels comparable to those observed in vivo. This effect of the hormone mixture was found to be the result of the presence of glucagon in the mixture, since the presence of glucagon with no other hormones added, except insulin, during the attachment period produced the same pattern of increased glucokinase activity. Immunoprecipitation of glucokinase from the hepatocytes, using monospecific antibody, indicated that the increase in enzyme activity was the result of increased glucokinase enzyme protein and not an increased synthesis of the other hexokinase isozymes. These studies demonstrate the specific hormonal requirements for the maintenance of glucokinase levels in primary hepatocyte culture at those seen in vivo and lends support to the hypothesis that fetal gene expression in primary hepatocyte cultures is selectively regulated rather than being a general effect with a common regulatory mechanism.
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  • 122
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    Journal of Cellular Physiology 103 (1980), S. 201-208 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Treatment of murine bone marrow cultures with the cholinergic agonist carbamylcholine enhanced megakaryocytic colony growth by as much as 65%. In contrast, adrenergic agonists had no such effect. Addition to cultures of dibutyryl cyclic GMP (db-cGMP) also enhanced megakaryocytic colonies up to 50%, whereas dibutyryl cyclic AMP (db-cAMP) had no effect. Sodium nitroprus-side and sodium nitrite, putative guanyl cyclase activators, also enhanced colony numbers, as did imidazole, a postulated cGMP phosphodiesterase inhibitor. Preincubation of marrow for two hours with carbamylcholine resulted in both an increase in colony numbers (58%) and percent of progenitors in DNA synthesis (48%, compared to 14% for controls) as determined by tritiated thymidine suicide studies. Treatment of mice with the acetylcholinesterase inhibitor neostigmine resulted in an increase in CFU-M/humerus (62%) and percent in DNA synthesis (45%). These data indicate that (1) cholinergic, but not adrenergic, agonists modulate megakaryocytopoiesis in culture; (2) this effect may be mediated by cyclic GMP; and (3) only a brief period of exposure of marrow cells to agonist results in enhancement of megakaryocytic colonies.
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  • 123
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    Journal of Cellular Physiology 103 (1980), S. 209-216 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Two predictions of the error/mutation hypothesis of cellular sensecence (Orgel, '73) namely, (a) exponential accumulation of somatic mutations during the replicative lifespan and (b) shortening of culture lifespan upon treatment with mutagens have been examined experimentally in a strain of cultured human diploid fibroblasts. Our studies show that as cells traverse the replicative lifespan (from 10 to 75 mean population doublings (MPD); total lifespan ⋍ 95 MPD), no rapid and exponential increase occurs in the accumulation of mutations measured by the frequencies of Thgr (thioguanine resistance) and Dipr (diphtheria toxin resistance) mutants. Furthermore, repeated cycles of treatment (from 1- to 14-times) of human fibroblasts with two mutagens, ethyl methane sulfonate (EMS) and N-methyl-N′nitro-nitrosoguanidine, which led to a marked increase in the mutation frequency for the Dipr marker (⋍ 100-fold), failed to shorten the lifespan of cultured fibroblasts. On the contrary, repeated mutagen treatment (12 times with EMS) prolonged the lifespan of one replicative culture (110 MPD versus 94-98 MPD). These results strongly indicate that mutations are unlikely to be the primary event in cellular senescence and suggest instead that senescence is probably controlled by one or more (specific) gene(s) whose expression can be modified by mutations.
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  • 124
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Molecular changes occur at the surface of hemopoietic cells during differentiation from progenitor cells to mature granulocytes and macrophages. The differential expression of surface carbohydrate residues has been probed using lectins and the results used to purify normal mouse granulocyte-macrophage progenitor cells. Ten different lectins were screened for selective interaction with mouse hemopoietic colony-forming cells (CFCs), using agglutination or a quantitative analysis of the number of fluoresceinated lectin molecules bound per cell using a fluorescence activated cell sorter (FACS). Pokeweed mitogen (PWM), Helix pomatia agglutinin (HPA), soybean agglutinin (SBA), and peanut agglutinin (PNA) preferentially bound to CFCs so that it was possible to enrich 4 to 10-fold for these progenitor cells by sorting for the highly fluorescent cells. Further analysis of the low and high angle light scattering characteristics of the CFCs indicated that these cells were polydisperse, but could be enriched ten-fold by selecting for cells with high intensity low angle (90°) scatter and low intensity high angle (90°) scatter. PWM gave the best enrichment (10 to 15-fold) for adult bone marrow CFCs, for CFCs from fetal sources (fetal liver, fetal blood), and for CFCs from the spleens of mice injected previously with outer membrane lipoprotein from E. coli. Three parameter sorting for CFC using the FACS (low angle scatter, high angle scatter, and PWM-fluorescence) resulted in large enrichment factors (16 to 50-fold) for CFCs from all the above sources. Over 7% of the cells sorted from bone marrow, 10% of the cells sorted from post-lipoprotein spleen, and 28% of the cells sorted from fetal peripheral blood were hemopoietic CFCs. Ninety percent of the cells in these fractions had the morphology of blast cells or myelocytes. Thus, it was possible to identify the morphological characteristics of the hemopoietic progenitor cells. Screening of other developmental systems using quantitation of fluorescence with lectins should prove of general value for the purification of selected differentiation states.
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  • 125
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    Journal of Cellular Physiology 103 (1980), S. 239-246 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Addition of prostaglandin E2 (PGE2) significantly altered the cellular composition of murine long-term bone marrow cultures. After 4-5 weeks of culture, increased cellularity in the suspension phase was observed in all cultures containing prostaglandin. These suspension cells contained markedly higher proportions of differentiated neutrophils than did cells cultured in the absence of PGE2. Granulocyte-macrophage progenitor cell levels in the suspension layer were increased 3-20 fold after five weeks in prostaglandin-containing cultures compared with control cultures. Fewer cells comprised the adherent layer in cultures containing prostaglandin. The number of macrophages in this layer was reduced 3-8 fold in these cultures compared with control cultures, while the number of granulocytes was increased 2-3 fold. The progenitor cells biased toward macrophage development were selectively inhibited in the cultures with PGE2. There was no significant effect of PGE2 on pluripotent stem cell levels or on the longevity of the cultures. It is concluded that excessive monopoiesis in bone marrow may be limited by PGE2 without influencing either stem cell maintenance or the development of other marrow-derived cell types.
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  • 126
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
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    Topics: Biology , Medicine
    Notes: Bovine vascular endothelial cells continuously maintained and grown in the presence of FGF adopt at confluence the configuration of a cell monolayer composed of contact-inhibited cells which do not overgrow each other and which are highly flattened and closely apposed. Such cultures exhibit structural and morphological characteristics similar to those observed with their in vivo counterparts. These include the production of an extracellular matrix consisting mostly of basement membrane collagen and fibronectin localized exclusively beneath the cell monolayer, but not on top of it, as well as a nonthrombogenic, blood-compatible apical cell surface. Removal of fibroblast growth factor (FGF) from adult bovine aortic endothelial cell (ABAE) cultures results within three passages in the loss by the cells of their characteristic contact-inhibited morphology. The cells, which during their logarithmic growth phase divide with a greatly increased doubling time, become larger and more elongated. Confluent cultures, instead of adopting the morphology of a contact inhibited cell monolayer, are now composed of overgrowing cells. Parallel with the morphological alterations taking place within the culture, the cells also lose the polarity of cell surfaces characteristics of the vascular endothelium. Formation of an extracellular matrix composed primarily of fibronectin and collagen types I, III, and IV is observed on both the apical and basal cell surfaces. Platelets which previously did not bind to the apical cell surface now become capable of binding to it. CSP-60, a major cell surface protein present in highly confluent and contact-inhibited vascular endothelial cell cultures, can no longer be detected. Exposure of confluent endothelial cell cultures, maintained in the absence of FGF to medium conditioned by cells which had been grown in the presence of FGF, but maintained in its absence upon reaching confluence led, within four to eight days, to a reversion of the altered phenotype. This medium has little or no mitogenic activity and retains a full activity in the absence of serum or after depletion of its fibronectin content by affinity chromatography on a gelatin-Sepharose column. Cultures which were previously composed of cells growing in multiple layers reorganized into a single cell monolayer composed of closely apposed and highly flattened cells. The cultures thereby regained the contact-inhibited morphology characteristic of the vascular endothelium. Concomitant with this cellular reorganization, the extracellular matrix disappeared from the apical cell surface, the cells regained their nonthrombogenic properties, and CSP-60 reappeared as one of the major cell surface proteins. These results suggest that vascular endothelial cells secrete a soluble factor(s) which can restore the normal morphology and function lost following removal of FGF from the medium. Such a factor(s) may be involved in maintaining the differentiated state of the vascular endothelium.
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  • 127
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    Journal of Cellular Physiology 103 (1980), S. 355-362 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The numbers of CFU-S which developed in spleen colonies were measured 11 days after injection of irradiated mice with marrow from normal mice or mice which had been treated in one of a variety of ways. The broad spread of CFU-S numbers, seen by other authors, in colonies derived from normal marrow was confirmed. However, the range and distribution of CFU-S per colony was generally different in colonies derived from the marrow of mice which were recovering or had recovered from some form of depopulation. From the data obtained, the mean CFU-S/colony, M1, and the probability of self-renewal, p, of the CFU-S were calculated. These values are used to calculate the number of cell cycles undergone during development of the colony and, by making certain assumptions, the cell cycle time of the CFU-S. The plot of p against log M for the various samples measured should be linear if all CFU-S proliferate at the same rate in a growing colony. It is not linear, however, so that CFU-S obtained under different experimental conditions do not all undergo the same number of cycles. In general, treatments given to the mice result in a lowering of the capacity for self-renewal of their CFU-S and also to a shortening of their cell cycle time. Some of the possible implications of these findings are discussed.
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  • 128
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    Journal of Cellular Physiology 103 (1980) 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
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  • 129
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    Journal of Cellular Physiology 103 (1980), S. 385-392 
    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 culture conditions on calf dorsal aorta endothelial cells was studied. Population doubling time varied as a function of the cell seeding density, growth medium, serum supplement, and concentration of fibroblast growth factor (FGF). The shortest population doubling time was found for cells (population doubling level 0-30) grown in Eagle's Minimal Essential Medium (MEM) supplemented with 10% fetal bovine serum (FBS) and 100 ng/ml FGF. The stimulatory effect of FGF on bovine endothelial cell proliferation was dependent on cell inoculation density. FGF significantly increased cell division rate at cell inocula less than 1 ± 104 cells/cm2 but not at higher densities. The population doubling time and cell size increased as the mass culture population doubling level increased. The replicative lifespans of bovine endothelial cells grown in medium supplemented with 20% FBS were 10-15% greater than parallel cultures supplemented with 10% FBS. Cultures grown in medium supplemented with 10% FBS and 50 ng/ml FGF showed a 50% increase in replicative lifespan compared to cultures grown in medium supplemented with 10% FBS alone. When FGF was used the increase in the number of doublings was a function of the length of time the cells were grown in the presence of FGF. This report extends comparable observations on the in vitro aging of human diploid fibroblasts to bovine endothelial cells.
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  • 130
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    Journal of Cellular Physiology 103 (1980), S. 371-383 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Erythroid colony formation in agar cultures of CBA bone marrow cells was stimulated by the addition of pokeweed mitogen-stimulated spleen conditioned medium (SCM). Optimal colony numbers were obtained when cultures contained 20% fetal calf serum and concentrated spleen conditioned medium. By 7 days of incubation, large burst or unicentric erythroid colonies occurred at a maximum frequency of 40-50 per 105 bone marrow cells. In CBA mice the cells forming erythroid colonies were also present in the spleen, peripheral blood, and within individual spleen colonies. A marked strain variation was noted with CBA mice having the highest levels of erythroid colony-forming cells.In CBA mice erythroid colony-forming cells were mainly non-cycling (12.5% reduction in colony numbers after incubation with hydroxyurea or 3H-thymidine). Erythroid colony-forming cells sedimented with a peak of 4.5 mm/hr, compared with CFU-S, which sedimented at 4.25 mm/hr. The addition of erythropoietin (up to 4 units) to cultures containing SCM did not alter the number or degree of hemoglobinisation of erythroid colonies.Analysis of the total number of erythroid colony-forming cells and CFU-S in 90 individual spleen colonies gave a correlation coefficient of r = 0.93 for these two cell types.In addition to benzidine-positive erythroid cells, up to 40% of the colonies contained, in addition, varying proportions of neutrophils, macrophages, eosinophils, and megakaryocytes. Taken together with the close correlation between the numbers of CFU-S in different adult hemopoietic tissues, including individual spleen colonies, the data indicate that the erythroid colony-forming cells expressing multiple hemopoietic differentiation are members of the hemopoietic multipotential stem cell compartment.
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  • 131
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    Journal of Cellular Physiology 103 (1980), S. 393-398 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Epithelial and fibroblast cells from adult rat liver were found to differ markedly in their metabolism of the purine hypoxanthine. Both cell types took up hypoxanthine and possessed hypoxanthine-guanine phosphoribosyl transferase for phosphoribosylating the purine. However, in the transferase assay, lysates from epithelial cells converted hypoxanthine predominantly to inosine monophosphate, with small amounts of the nucleoside inosine as product, whereas fibroblast cell lysates converted hypoxanthine predominantly to inosine. The inosine appeared not to be produced by direct ribosylation of the base, since fibroblast cell lysates had less purine nucleoside phosphorylase activity than epithelial cell lysates. Rather, the inosine produced by fibroblast lysates appeared to be derived from inosine monophosphate through catabolism of the mononucleotide by 5′ nucleotidase. An inhibitor of 5′ nucleotidase, thymidine triphosphate, reduced the amount of inosine formed.
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  • 132
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    Journal of Cellular Physiology 103 (1980), S. 407-416 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The water permeability of V-79 Chinese hamster lung fibroblasts was determined by measuring the rate of cell shrinkage in hypertonic medium using a cell sizer. The water permeability appears to follow Arrhenius kinetics as a function of temperature with a sharp discontinuity at 21°C. An activation energy of 7.0±1.6 kcal/mole was found below 21°C and 22.8±3.1 kcal/mole above 21°C. The correlation time of rotation of the spin label 2,2-dimethyl-5-dodecyl-5-methyloxazolidine-N-oxide was measured as a function of temperature in the cellular membranes, and shows a break at 20°C. A discontinuity was also found in the membrane to water partitioning of the spin label 2,2-dimethyl-5-pentyl-5-butyloxazolidine-N-oxide near 20°C. These breaks may correspond to a membrane lipid phase transition. Dimethylsulfoxide, in the concentration range of 0.2-0.5 M, decreases the water permeability by a factor of two.
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  • 133
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    Journal of Cellular Physiology 103 (1980), S. 447-453 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Mouse myeloma cell lines synthesize large amounts of immunoglobulin on their membrane-associated polyribosomes. Variants which no longer synthesize immunoglobulins have been studied and shown to have the same number of tightly bound membrane-associated polyribosomes as the parental cell lines. These polyribosomes are still active in the synthesis of nonimmunoglobulin proteins.
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  • 134
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    Journal of Cellular Physiology 104 (1980) 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
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  • 135
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
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    Topics: Biology , Medicine
    Notes: The transport of selected neutral and cationic amino acids has been studied in Balb/c 3T3, SV3T3, and SV3T3 revertant cell lines. After properly timed preincubations to control the size of internal amino acid pools, the activity of systems A, ASC, L, and Ly+ has been discriminated by measurements of amino acid uptake (initial entry rate) in the presence and absence of sodium and of transportspecific model substrates. L-Proline, 2-aminoisobutyric acid, and glycine were primarily taken up by system A; L-alanine and L-serine by system ASC; L-phenylalanine by system L; and L-lysine by system Ly+ in SV3T3 cells. L-Proline and L-serine were also preferential substrates of systems A and ASC, respectively, in 3T3 and SV3T3 revertant cells. Transport activity of the Na+-dependent systems A and ASC decreased markedly with the increase of cell density, whereas the activity of the Na+-independent systems L and Ly+remained substantially unchanged. The density-dependent change in activity of system A occurred through a mechanism affecting transport maximum (Vmax) rather than substrate concentration for half-maximal velocity (Km). Transport activity of systems A and ASC was severalfold higher in transformed SV3T3 cells than in 3T3 parental cells at all the culture densities that could be compared. In SV3T3 revertant cells, transport activity by these systems remained substantially similar to that observed in transformed SV3T3 cells. The results presented here add cell density as a regulatory factor of the activity of systems A and ASC, and show that this control mechanism of amino acid transport is maintained in SV40 virus-transformed 3T3 cells that have lost density-dependent inhibition of growth, as well as in SV3T3 revertant cells that have resumed it.
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  • 136
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    Journal of Cellular Physiology 105 (1980), S. 105-127 
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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 cellular differentiation on the osmotic properties of a proliferating line of bone marrow cells has been investigated. This population proliferated and differentiated into distinct maturation phases which have been separated quantitatively by velocity sedimentation at 1g in a Ficoll gradient.Certain osmotic characteristics were evaluated. The volume of osmotically active water increased linearly with maturation. The osmotically inactive volume was variable during proliferation and maturation. Several correlations that concerned the exosmotic movement of water were noted, as follows: (1) membrane permeability to water increased with an increase in the diameter of the nucleus, (2) the ratio of osmotically active water to mean corpuscular volume increased with an increase in the diameter of the nucleus, (3) membrane permeability to water increased with an increase in the osmotically active water normalized to the mean cell volume, and (4) the heat of activation associated with the permeability of the membrane to water increased during maturation and varied inversely with the ratio b/MCV.These results were used to assess the effects of cellular maturation on membrane function and the state of water in a maturing cell population. The permeability data suggest that charged groups on membrane proteins and phospholipids can vary the state of water in the membrane from a thermodynamically mobile state to an “ice-like” state. These unusual properties may be a direct result of a dynamic and functional relationship between cellular water and active biological macromolecules.
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  • 137
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    Journal of Cellular Physiology 105 (1980), S. 137-141 
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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 artificial cryptorchidism and its surgical reversal on spermatogenesis were examined in germ cell mutant, S1/+ and wild type, +/+, mice. In cryptorchid testes no difference was found between S1/+ and +/+ mice in the number of undifferentiated type A spermatogonia. The activity of type A spermatogonia in mutant mice appeared normal as judged by its mitotic cell number and DNA synthesis. The surgical reversal of cryptorchidism resulted in regenerative differentiation of mature germ cells in both types of mice, but the pattern of cellular differentiation in the mutant testes was completely different from that of the wild type testes. At two steps of cellular differentiation, intermediate or type B spermatogonia and spermatid, the numbers of cells were much smaller in the S1/+ testes than those in the +/+ testes. The steel gene was therefore suggested to exert its effects on the differentiation of type A spermatogonia to intermediate or type B spermatogonia, on meotic division and/or the survival rate of these cells, but not on the undifferentiated type A spermatogonia or stem cells.
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  • 138
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    Journal of Cellular Physiology 105 (1980), S. 163-180 
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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 examined culture fluids from a variety of Kirsten murine sarcoma virus (KiMSV) transformed rat and mouse cells for the presence of factors which induce normal Rat-1 cells to assume the transformed phenotype. All KiMSV transformants produced transforming factor (TF). Revertants of KiMSV transformed rat or mouse cells failed to relase TF as did normal rat or mouse cells. Cells transformed by a temperature sensitive mutant of KiMSV produced TF at the permissive temperature but not at the nonpermissive temperature. Further, cells from a spontaneous transformant of Rat-1 cells also produced TF. TF is a small polypeptide which competes for the epidermal growth factor receptor. Its effect upon normal cells is reversible and requires de novo RNA and protein synthesis. Cells treated with TF lose the actin fibers observed in normal fibroblasts, assume a transformed cell morphology, become anchorage independent for growth, grow in low concentrations of serum, grow to a high cell density, and have an increased rate of hexose uptake.
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  • 139
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    Journal of Cellular Physiology 105 (1980), S. 185-185 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
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  • 140
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    Journal of Cellular Physiology 105 (1980), S. 191-196 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Macrophages, when maintained in vitro, take up glucose from the medium and oxidize it to CO2. The rate of oxidation of glucose varies considerably, depending on the physical state of the cell preparation. Cells in suspension oxidize glucose at a level six-fold that of cells in monolayers. The differences cannot be attributed to change in the rates of transport of glucose. On the other hand, an increse in intracellular glycogen (about three-fold) and free glucose plus glucose-6-P (many-fold) was found in the cells prepared as monolayers. During subsequent incubation with glucose-14C, this could be the cause of an isotope dilution effect and could explain the lower production of 14CO2 by the adherent cells. Since oxidation of glucose-1-14C to 14CO2 is used by many investigators to indicate the functional state of macrophages, we suggest close attention be paid to the system used, i.e., monolayers vs. suspensions.
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  • 141
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    Journal of Cellular Physiology 105 (1980), S. 221-225 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Media concentration of total soluble CO2 increases with culture age of Tetrahymena pyriformis. CO2 is a weak acid and is capable of acidifying intracellular pH (pHi). Changes in pHi have been demonstrated to affect cell metabolism and growth in many systems. For these reasons, we investigated whether the concentrations of CO2 produced in vitro were sufficient to affect cell proliferation and pHi in Tetrahymena. In this study, we used DMO to mimic the weak acid properties of CO2. DMO is freely permeable to membranes in its uncharged form and has a pKa similar to that of CO2/HCO3-. In addition, it has the advantages of being metabolically inert and non-volatile. At concentrations similar to endogenously produced CO2, DMO acidifies pHi and arrests culture growth. In addition, procedures are described which decrease the media CO2 concentrations in both growing and non-growing cultures. These conditions lead to increased maximum culture density at stationary phase. The data indicate that, under our conditions, accumulation of CO2 in the culture leads to cessation of growth, probably through elimination of transmembrane pH gradients, which are necessary for regulation of metabolism and growth.
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  • 142
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    Journal of Cellular Physiology 105 (1980), S. 247-258 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The involvement of altered protein metabolism in the expression of the temperature-sensitive (ts) pleiotropic phenotype of ts A1S9 cells was investigated. Cells are ts in growth and DNA replication. They undergo decondensation of their heterochromatin, interruptions of chromatin synthesis, and changes in cell size and morphology at the non-permissive temperature (npt) of 38.5°C. Whereas the rates of incorporation of 3H-leucine, 35S-methionine, and 3H-fucose into proteins were unaffected at 38.5°C, net protein accumulation was greatly reduced. This imbalance resulted from a rapid increase in the rate of protein degradation at the npt. Enhancement of protein degradation was detected within 2-4 hours after temperature upshift and constitutes the earliest metabolic alteration thus far observed during expression of the temperaturesensitive phenotype. The average half-life of proteins preformed in ts A1S9 cells at 34°C was decreased four-fold at the npt, and all major cytoplasmic proteins were affected equally. Enhanced protein degradation at the npt was shown to be sensitive to cycloheximide, ammonia, chloroquine, and vinblastine at concentrations that did not affect the basal protein degradation of normally cycling cells. Increased protein degradation at 38.5°C did not involve an equivalent increase in total cellular protease activity. The data obtained are compatible with a model that suggests that temperature inactivation of the ts A1S9 gene product results in activation of a lysosome-mediated mechanism for the rapid degradation of cytoplasmic proteins.
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  • 143
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    Journal of Cellular Physiology 105 (1980), S. 301-311 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: In rat pancreatic fragments, stimulation of amylase and labeled protein release by carbachol, caerulein, and ionophore A 23187 results within minutes in a short rise in cyclic GMP levels. Cyclic AMP levels do not change significantly. The secretory response elicited by each secretagogue is not modified when combined in pairs. Under intracellular calcium depleting conditions, both the cyclic GMP and the secretory responses to secretagogues are inhibited in parallel, suggesting a good correlation between both processes. Furthermore, 8-Bromocyclic GMP induces pancreatic secretion, but to a lesser extent, and fails to alter the increase in secretion caused by the various secretagogues. However, other agents such as imidazole, ascorbic acid, phenylhydrazine, and sodium azide also increase cyclic GMP levels but fail to stimulate pancreatic secretion. On the other hand, dibutyryl cyclic AMP also stimulates amylase and labeled protein discharge and potentiates the increase caused by cabachol, caerulein, and ionophore A 23187. These results do not permit conclusions regarding a cause and effect relationship between cyclic GMP and secretion. A role for calcium seems to be the most likely.
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  • 144
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    Journal of Cellular Physiology 105 (1980), S. 327-334 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The viability of neutrophils in the condition under which they kill neoplastic cells was studied. In the presence of phorbol myristate acetate (PMA) the 51Cr-release by human neutrophils was markedly stimulated. The PMA-induced 51Cr-release by neutrophils correlated well with the number of nonviable neutrophils as determined by the uptake of trypan blue. Phorbol myristate acetate had no effect on the 51Cr-release by lymphocytes, LPC-1 myeloma cells, ovarian ascites tumor cells, or neutrophils from a patient with chronic granulomatous disease. This suggests that the effect of PMA is not due to its nonspecific toxic effect; instead, it is dependent on the reactive oxygen species produced by the normal neutrophils. Catalase, cytochrome C, histidine, and methionine inhibited the PMA-induced 51Cr-release by human neutrophils, whereas superoxide dismutase, myeloperoxidase inhibitors, and some hydroxyl radical scavengers or singlet oxygen quenchers had no effect. The clumping of neutrophils induced by PMA was also important in the PMA-induced 51Cr-release by human neutrophils.
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  • 145
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    Journal of Cellular Physiology 105 (1980), S. 347-354 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: tsAF8, ts13, tsHJ-4, and TK-ts13 cells are G1-specific temperature-sensitive (ts) mutants of BHK cells that do not enter S phase when serumstimulated from quiescence at nonpermissive temperature (39.6°-40.6°). TK-ts13 are, in addition, defective in thymidine kinase. Different G1 functions must be involved in these cells, since the first three cell lines complement each other when forming heterokaryons.We have used these cells to study the role of the nucleus in the cytoplasmic expression of these G1 functions during the transition of cells from the non-proliferating to the proliferating state. We fused cytoplasts from either serumstarved (G0) or serum-stimulated (S) tsAF8 cells with G0-ts13, G0-tsHJ-4, and G0-TK-ts13 recipient cells and determined, after serum stimulation of the fusion products, which type of cytoplasts could complement the defective G1 functions. Cytoplasts from S-tsAF8 cells complemented all three functions, i.e., cybridoids between S phase cytoplasts and ts13 or tsHJ-4 recipient cells entered S at the nonpermissive temperature, and TK-ts13 recipient cells incorporated exogenous thymidine. Cytoplasts isolated from G0-tsAF8 cells (3 days of serum starvation) complemented ts13 cells but not tsHJ-4 and TK-ts13 cells. Cytoplasts from 6-day starved tsAF8 cells lost the complementing capacity for ts13 cells. However, when the 6-day starved tsAF8 cells were fused with G0-ts13 cells, the heterokaryons entered S phase at the nonpermissive temperature. Also, cytoplasts isolated from the 6-day starved cells that were serum stimulated for 40 hr before enucleation regained the capacity to complement ts13 cells.These results demonstrate that three functions required in G1 cannot be detected in the cytoplasm of serum-starved cells, although they are present in the cytoplasm of S-phase cells. These results suggest that a functional nucleus is required for the cytoplasmic appearance of certain G1 functions in serumstimulated cells.
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  • 146
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    Journal of Cellular Physiology 105 (1980), S. 287-300 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Regulation of A system amino acid transport was studied in primary cultures of the R3230AC mammary adenocarcinoma. Higher rates of carrier-mediated Na+-dependent proline transport, vc, was decreased and was attributed to a two-fold decrease in Vmax and a two-fold increase in Km. When compared to cells grown in standard media (Eagle's minimal essential medium, MEM), cells grown in media supplemented with A system substrates (alanine, serine, glycine, and proline) demonstrated adaptive decreases in proline transport; the decrease was due to two-fold reduction in Vmax, with no change in Km for proline. Even in the presence of preferred substrates for the A system, a density-dependent decrease in proline transport was manifested. Both fast- and slow-growing cultures maintained in MEM exhibited rapid increases in proline transport when switched to buffers devoid of amino acids; two-fold increases in Vmax were seen within 4 hr, but Km was unchanged. This starvation-induced adaptation was completely prevented by inclusion in the buffer of 10 mM proline, 0.1 mM -(methylamino)-isobutyric acid (MetAIB) or 10 mM serine, whereas inclusion of the poorer A system substrate, phenylalanine (10 mM), had no effect. The effects of MetAIB to prevent starvation-induced increases in proline transport were dose-related, rapid, and reversible. Amino acid starvation-induced increases in proline transport were partially blocked by cycloheximide or actinomycin D. Data were obtained demonstrating a temporal relationship between increasing intracellular [proline] and decreasing vc for proline uptake. In addition, efflux of proline from preloaded cells preceded the increase in initial rates of proline entry. Taken together, we concluded that: (1) A system transport in primary cultures of this mammary adenocarcinoma is regulated by cell density as well as by availability of A system substrates, but these two types of regulation are kinetically distinct; and (2) starvation-induced enhancement of proline transport appears to be due to release from transinhibition, but may also involve a derepression-repression type of mechanism.
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  • 147
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    Journal of Cellular Physiology 102 (1980) 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
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  • 148
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    Journal of Cellular Physiology 102 (1980), S. 51-54 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Transcriptional activity of nuclei from human diploid fibroblast (WI-38) cells at different passage levels was studied with endogenous RNA polymerase. The rate and extent of the RNA synthesis decreased when the nuclei were were prepared from senescent cells. The decreased activity of RNA synthesis in senescent cell nuclei was demonstrated from both the transcriptional assay and the estimation of the processing activity, i.e., polyadenylation of the transcripts.
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  • 149
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    Journal of Cellular Physiology 102 (1980) 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
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  • 150
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    Journal of Cellular Physiology 102 (1980), S. 259-266 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Cultured rat embryo fibroblasts, when placed in media with 10% serum containing 20 mM NH4Cl, show an inhibition of protein degradation and, concurrently, an accumulation of numerous, large vacuoles, partially filled with cellular debris. Cells placed in a serum-free media exhibit an enhanced degradation of cell protein, which is also inhibited by NH4Cl. When these cells are removed from media containing NH4Cl and placed in fresh media, the material accumulated in these vacuoles is rapidly and quantitatively released to the media in both an acid-soluble and acid-insoluble form. NH4Cl inhibits rapidly and specifically the lysosomal proteolytic mechanism, and is without effect on the basal turnover mechanism. The lysosomal proteolytic mechanism accounts for approximately 25% of protein turnover, and, at least in low density cultures, can be stimulated to levels which account for more than half of the protein turnover in the cell. The major pathway for the degradation of fast turnover proteins appears to be separate from lysosomal mechanism.
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  • 151
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    Journal of Cellular Physiology 102 (1980), S. 279-286 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The effects of L-cell conditioned medium which contains granulocyte/macrophage colony stimulating factor (CSF); of highly purified L-cell CSF; and the antiserum directed against L-cell CSF, have been investigated in long-term murine bone marrow cultures. Treatment of cultures with CSF containing conditioned medium led to a rapid decline in haemopoiesis. However, this inhibition of in vitro haemopoiesis is probably caused by materials other than CSF, since the addition of highly purified L-cell CSF had no appreciable effect upon long-term haemopoietic cell proliferation or differentiation. Furthermore, the inhibitory activity of L-cell conditioned medium was not abrogated following neutralization of the CSF activity by CSF antiserum. The direct addition of CSF antiserum did not inhibit granulocyte or macrophage formation. These results suggest that long-term cultures of murine marrow cells may show extensive interactions with stromal cells which are not influenced by exogenous stimulatory or inhibitory factors.
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  • 152
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    Notes: BFU-E from the blood of 14 normal adults have been grown by the plasma clot technique. The hemoglobins synthesized in burst colonies were purified from other proteins by affinity chromatography on Sepharose-haptoglobin. The radioactivity incorporated in the globin chains was estimated by CM-cellulose chromatography in urea. The number of bursts scored at the 14th day of culture fluctuated between 50-130 (average 86, s: 29) for 106 mononuclear plated cells. A constant reactivation of fetal hemoglobin was found (from 1.4% to 11%, mean value 5.8%, s: 3.07), but was lower than previously described, mainly because of the highly selective purification of Hb. This reactivation of fetal hemoglobin was not dependent upon the concentration of erythropoietin (from 1 U/ml to 6 U/ml) nor on the purity of the erythropoietin preparations (from 6 U/mg of protein to 70 000U/mg of protein). In addition, the same subject exhibited a constant proportion of Hb F synthesized in culture over a period of time up to 6 months. A positive correlation exists between the proportion of Hb F in culture and that of F cells present in the blood, with the exception of two subjects. Such findings suggest that Hb F in culture is a characteristic of each individual and that this reactivation often represents an amplification of the Hb F synthesis in vivo.
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  • 153
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    Journal of Cellular Physiology 102 (1980), S. 317-321 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The growth of human mammary epithelial cells is stimulated by cholera toxin and analogues of cyclic AMP, while the growth of breast derived fibroblasts is inhibited. These compounds have little effect on DNA synthesis in the absence of other mitogens but show a synergistic effect with serum and/or EGF. The results suggest that high intracellular levels of cyclic AMP in human mammary epithelial cells increase the growth response of the cell to mitogens.
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  • 154
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    Journal of Cellular Physiology 102 (1980), S. 343-349 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Lectin-dependent neutrophil cytotoxicity against autologous human red cells was studied using an 111In(indium)-release assay. Human red cells were not readily killed by neutrophils in the presence of phytohemagglutinin (PHA). However, removal of red cell membrane sialic acids (desialylation) markedly enhanced their susceptibility to PHA-dependent neutrophil cytotoxicity. This neutrophil cytotoxicity was dependent on the energy supplied by anaerobic glycolysis, but it was independent of erythrophagocytosis. Catalase, superoxide dismutase, KCN, and Na azide did not inhibit PHA-dependent neutrophil cytotoxicity. Neutrophils from a patient with chronic granulomatous disease, in the presence of PHA, also killed desialylated red cells normally. On the other hand, desialylation of neutrophils had no effect on the expression of their cytotoxic effect. The results suggest that desialylated red cells are much more susceptible to lectin-dependent neutrophil cytotoxicity than normal red cells, and that lectin-dependent neutrophil cytotoxicity is independent of reactive oxygen species.
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  • 155
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    Journal of Cellular Physiology 102 (1980), S. 351-365 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Granulocytic and erythrocytic colonies developed on the chick embryo chorioallantoic membrane (CAM) following the inoculation of chick embryo spleen cells. Dose response and kinetic experiments showed that the colonies were derived from cell aggregates present in the inoculum. Dissociation and reaggregation studies of the CAM colony-inducing cells (CAM-CIC) indicated that these cells must be present as aggregates in order to form colonies. Results from the morphology and cell marker experiments suggested that the colony-inducing aggregates (CAM-CIA) attract and support the differentiation of primitive host hemopoietic cells. The physical characteristics of the CAM-CIC, which are different from those of the hemopoietic progenitor cells, indicated that they represent a stromal cell population of the chick embryo spleen. Further evidence supporting this notion was provided by the radiation studies which showed that the colony-inducing ability of the CAM-CIC is radioresistant. The above characteristics of the CAM-CIC strongly suggest that they represent the stromal cells of the chick embryo spleen which influence hemopoiesis.
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  • 156
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    Notes: Nontransformed mouse embryo derived AKR-2B cells stop growing in the G1 phase of the cell cycle at saturation density due to depletion of serum growth factors, whereas a chemically transformed derivative line (AKR-MCA) arrests in G1 at a higher saturation density due to depletion of amino acids and glucose. Stimulation of DNA synthesis is inhibited in the AKR-2B cells, but not in the AKR-MCA cells, by two inhibitors of RNA metabolism, α-amanitin and 5-fluorouridine (5-FU). To determine whether the AKR-MCA cells growth arrest at a uniue point in G1 or whether they arrest in a physiologic state which can also be achieved by the nontransformed cells, AKR-2B cells were maintained in medium with 10% serum containing the mitogens, epidermal growth factor (EGF) or 12-O-tetradecanoylphorbol-13-acetate (TPA), until they reached saturation density or were arrested at subconfluence by artificial deletion of amino acids from the medium. The AKR-2B cells maintained in EGF or TPA stopped growing in G1 at a higher saturation density, due to depletion of amino acids. Cells arrested in EGF or TPA or in amino acid deficient medium had a shortened interval between stimulation and the onset of DNA synthesis, and the stimulation of DNA synthesis was not inhibited by α-amanitin or 5-FU. The data show that the nontransformed AKR-2B cells have two different arrest states which may represent two separate and distinct G1 arrest points - a growth factor deficiency arrest point and a nutrient deficiency arrest point. The nutrient deficiency arrested cells were very similar to the G1 arrested transformed AKR-MCA cells.
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  • 157
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    Journal of Cellular Physiology 102 (1980), S. 379-384 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The rate of NAD turnover in microplasmodia of Physarum polycephalum was investigated using a double labeling technique with (14C)-adenine or adenosine and (3H)-nicotinamide.The half-life of an NAD molecule in Physarum was estimated to be 25 min, which is shorter than in either E. coli or human cell lines. The half-life of NAD in the presence of an inhibitor of NADase and poly ADPR synthase, 5-methyl-nicotinamide, was also investigated, but found to be indistinguishable from controls. The possible reasons for this and for the rapid turnover is discussed in the light of the known functions for NAD in prokaryotes and eukaryotes.
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  • 158
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    Journal of Cellular Physiology 102 (1980), S. 385-393 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Rat peritoneal macrophages were studied for their ability to undergo metabolic cooperation. Macrophages were unable to cooperate with human fibroblasts. This was true for macrophages which had been activated in vivo as well as for macrophages treated with various agents in vitro. Macrophages were also unable to undergo metabolic cooperation with rat fibroblasts or with other macrophages. In contrast, rat reticular cells, mesothelial cells, and fibroblasts were able to cooperate with human fibroblasts.
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  • 159
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    Notes: Phenylalanine in high concentrations inhibits the growth of mouse A9 cells. Protein synthesis is inhibited earlier and more severely than RNA or DNA synthesis. Phenylalanine inhibits the uptake and decreases the intracellular pool of several amino acids. Certain amino acids added in excess reverse the phenylalanine inhibition. The strongest reversing amino acids appear to function by excluding phenylalanine. The phenylalanine inhibition does not appear to be due to a deficiency of any amino acid, but to the high intracellular phenylalanine concentration and/or an amino acid imbalance resulting from the large ratio of phenylalanine to other amino acids.
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  • 160
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    Journal of Cellular Physiology 103 (1980) 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
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  • 161
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    Journal of Cellular Physiology 103 (1980), S. 1-9 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The specific activity of lactate dehydrogenase (LDH; EC 1.1.1.27) is induced two-fold by l-norepinephrine (NE) in C6TK- rat glioma cells, but not in NA mouse neuroblastoma cells or various other nonglial cells. Previous reports have shown that the induction is mediated by cyclic AMP (cAMP) and possibly protein phosphorylation, and that it requires RNA and protein synthesis. To study the block to LDH induction in nonglial cells, we hybridized C6TK- cells with NA cells and isolated a hybrid clone in which LDH is inducible by NE. Mouse and rat LDH from hybrid cells were separated by electrophoresis and quantitated by two independent methods, and it was found that mouse and rat LDH were induced equally when cells were exposed to NE. The results suggest that inducibility of LDH is not determined by a cis-acting control at the gene level, but rather by the presence or absence of an earlier component in the cAMP-mediated induction system, and that the induction system acts indiscriminately on all active LDH gene copies in the cell.
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  • 162
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    Journal of Cellular Physiology 103 (1980) 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
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  • 163
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    Journal of Cellular Physiology 103 (1980), S. 169-172 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: High pressure liquid chromatography was used to determine the base, nucleoside, and nucleotide levels in wild type and a series of respiration-deficient Chinese hamster cell mutants. From this analysis the size of the total adenylate pool and the energy charge could be calculated for each cell line. We find a constant energy charge, as expected, but the adenylate pool seems to be considerably lower in the respiration-deficient mutants.
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  • 164
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    Journal of Cellular Physiology 103 (1980), S. 159-168 
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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 insulin, glucagon or Dexamethasone (DEX) and of glucagon with insulin or DEX were examined on the uptake of 2-amino [1-14C]isobutyric acid (AIB) and N-Methyl-2-amino [1-14C]isobutyric acid (NMe AIB) in monolayer cultures of rat hepatocytes. Insulin and glucagon stimulated the uptake of both the amino acids and DEX inhibited it, showing that all three of these hormones regulate the A system (the sodium-dependent system that permits the transport of NMe AIB) for amino acid transport in these cultures. Experiments investigating the transport of aminocyclopentane-1-carboxylic acid, 1- [carboxyl-14C] in the presence of excess AIB or in the absence of sodium showed that insulin had no effect on the activity of the L system (the sodium-independent system that prefers leucine). Experiments on the uptake of AIB in the presence of excess NMe AIB showed insulin had no effect on the transport activity of the ASC system (the sodium-dependent system that does not transport NEe AIB). Insulin concentrations ranging from 0.1 nM to 100 nM did not antagonize the stimulatory effect of optimum or suboptimum concentrations of glucagon on the uptake of either AIB or NMe AIB. Similarly, glucagon did not antagonize the stimulatory effect of optimum or suboptimum concentrations of insulin on the uptake of both the amino acids. The combined effect of insulin and glucagon was additive on the rate as well as the cumulative uptake of both AIB and NMe AIB. DEX alone inhibited the transport of both AIB and NMe AIB by about 25%, while glucagon caused a 2-3-fold increase; however, the addition of glucagon to cultures containing DEX caused a 7-8-fold increase in the uptake of both AIB and NMe AIB when compared to cultures containing DEX alone. The effect of insulin on the levels of cAMP was also investigated. Insulin had no effect on the cAMP levels in cultures treated or untreated with optimum or suboptimum concentrations of glucagon.
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  • 165
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    Notes: In this study, the intracellular concentrations of six elements (mmole/kg dry weight) were directly measured in the muscle fibers of pectoralis major muscles of eight week old, genetically dystrophic and normal chickens by the X-ray microanalysis technique. The extent of muscle degeneration was evaluated by morphometric measurements of muscle fiber diameter and other histological changes. A significant increase in the concentration of intracellular sodium and chlorine was evident in dystrophic muscles. The concentration of intracellular sodium was 127.0 ± 35.0 in the muscle fibers of dystrophic chicks compared to 65.7 ± 16.5 in normal controls. The concentration of chlorine was 90.5 ± 27.5 and 54.1 ± 5.5 in the muscle fibers of dystrophic and normal chicks respectively. The intracellular concentrations of potassium, magnesium, phosphorous, and sulfur remained unchanged in the dystrophic condition. Morphometric studies revealed that the dystrophic pectoralis muscles contain fewer but thicker fibers per unit area compared to normal pectoralis muscles. The importance of these findings are discussed in relation to the results of earlier investigations.
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  • 166
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    Journal of Cellular Physiology 103 (1980), S. 255-262 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Mitochondrial protein synthesis was measured in line CHO cells after phases of the cell cycle were synchronized by isoleucine deprivation or mitotic selection. Maximum incorporation of [3H] leucine into mitochondrial polypeptides occurred within 2 hours after isoleucine was added to initiate G1 traverse. In cells synchronized in G1 by mitotic selection, the rate of mitochondrial protein synthesis was fairly constant throughout the cell cycle. SDS-polyacrylamide gel electrophoretic profiles of labeled mitochondrial polypeptides were similar in cells synchronized by either isoleucine deprivation or mitotic selection. Obvious changes in the distribution of polypeptides were not detected during various phases of the cell cycle. The increased rate of incorporation of [3H] leucine into mitochondrial polypeptides after reversal of G1-arrest may indicate that mitochondrial protein synthesis and possibly mitochondrial biogenesis are synchronized in CHO cells deprived of isoleucine.
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  • 167
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    Journal of Cellular Physiology 103 (1980), S. 247-254 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Suspending anchorage-depent fibroblasts in methocel results in marked reduction in a number of macromolecular metabolic processes. These are restored when cells make contact with a solid substrate and are allowed to spread. The response of 3T6 cells to suspension culture and their recovery upon reattachment has been extensively studied. (Benecke et al., '78; Farmer et al., '78). In these cells, message production and its turnover rate both drop abruptly when cells are suspended, while protein synthesis declines very slowly but extensively. Recovery of protein synthesis in 3T6 cells only requires surface contact and not cell spreading and proceeds rapidly, reaching control values within a few hours. The recovery of messenger RNA production in spread cells is much slower, and commences about 18 hours after cells are replated.The present report extends the studies on RNA regulation in suspended and reattached 3T6 fibroblasts. All RNA synthesis systems respond to cell suspension. Production of small nuclear RNA species is shown to behave very similarly to the production of messenger RNA, suggesting these may be coordinately regulated. Ribosomal precursor RNA synthesis ceases promptly upon cell suspension and recovers only slowly after reattachment. The control of ribosome formation appears independent of the regulation of protein synthesis in this system. The products of polymerase III including transfer RNA, 5S RNA, and species K and L are all regulated together and show a distinct pattern of behavior unlike that of any other RNA species. Their formation is rapidly inhibited at the beginning of suspension, but then recovers to the level of control cells, even during continued suspension culture. This unusual property of polymerase III RNA species is quite different from the behavior of the small nuclear (snRNA) RNA species, which strengthens the previous suggestion that these two classes of RNA molecules are formed by different mechanisms and regulated independently.
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  • 168
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    Journal of Cellular Physiology 103 (1980), S. 305-311 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: This study suggests that membrane perturbations can affect oral morphogenesis in Stentor, possibly by a mechanism involving calcium ions. Exposure of regenerating Stentor to micromolar concentrations of the membrane active local anesthetics dibucaine, tetracaine, or procaine greatly delayed the progress of oral regeneration. In the case of tetracaine and dibucaine the greatest delays were observed in the early stages of regeneration prior to stage 4, when the majority of essential synthetic activity is occurring. The effects of dibucaine were generally readily reversible upon removal of the cells from the drug, with some residual effects occurring at higher dibucaine concentrations. Regenerating cells in the presence of dibucaine and excess extracellular calcium were not delayed, suggesting that the effects of dibucaine were reversible by calcium ions. The effects of tetracaine were not reversible by calcium ions, however. Exposure of regenerating cells to medium either lacking in, or containing an excess of, extracellular calcium had no effect on the time required to complete oral regeneration. The plant lectin, phytohemagglutinin, can also delay oral regeneration. The possible implications of these findings on the control of oral regeneration are discussed.
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  • 169
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    Journal of Cellular Physiology 103 (1980), S. 323-331 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The growth requirements of SV40 transformed Balb/c-3T3 cells have been studied in the absence of serum. For growth in serum-free medium, the cells require (i) insulin, (ii) transferrin, and (iii) cis-unsaturated fatty acids added in combination with fatty acid free bovine serum albumin. The growth rate, saturation density, and morphology of cells grown in this serum-free medium are the same as those of cells grown in serum supplemented medium. This mixture also supports the growth of SV40 transformed Swiss-3T3 cells and SV40 transformed primary mouse embryo cells, but does not support the growth of untransformed Balb/c-3T3 cells.The addition of fibronectin to this mixture allows routine subculture, repeated passage, and indefinite propagation of SV40 transformed Balb/c-3T3 cells. Cells grown in this medium for a period of two months retain their ability to induce tumors when injected into athymic nude mice.
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  • 170
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    Journal of Cellular Physiology 103 (1980), S. 363-370 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Growth-contingent alterations in potassium and sodium fluxes, ouabain binding, and potassium ion content were examined following serum stimulation of quiescent, density-inhibited chicken embryo fibroblasts. Serum stimulation resulted in very rapid 1.5- to 1.8-fold increases in ouabain-sensitive potassium influx and lesser 1.4- to 1.5-fold increases in potassium efflux and sodium influx. Potassium influx stimulation was maximal after addition of 5-20% calf serum and was unaffected by cycloheximide inhibition of protein synthesis. Reflecting the slightly greater stimulation of potassium influx versus potassium efflux, potassium ion levels were 10-15% higher in serum-stimulated compared to unstimulated cells. Specific ouabain binding levels in stimulated and unstimulated control cells were initially similar, however, by four hours after stimulation a 40-50% increase in specific ouabain binding was observed. Incubation with ouabain was found also to inhibit later serum-stimulated hexose uptake and thymidine incorporation; this blockage may be a consequence of subnormal potassium levels rather than ouabain inhibition of the serum-stimulated potassium influx.
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  • 171
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
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    Topics: Biology , Medicine
    Notes: The incubation of linoleic acid with cells causes profound effects on membrane associated phenomenon. Using the fluorescent probe diphenyl hexatriene (DPH) to monitor lipid changes in the microenvironment of the cell surface, we find that linoleic acid reduces the polarization values (P) in mouse lymphocytes and BHK cells. Measurements on lipids extracted from the cells grown in linoleic acid produce similar results. We also find in the mouse lymphocyte that capping of Ig is inhibited and con A stimulated mitogenesis is unaffected. In contrast to the latter effect, LPS and PHA stimulated mitogenesis is inhibited and in the rat lymph node, con A stimulated mitogenesis, greatly enhanced. We also show that linoleic acid alters the binding of antibodies to the cell surface of EL-4 lymphoma cells. These observations suggest that linoleic acid alters cellular function by interfering with protein/lipid interactions within the surface membrane.
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  • 172
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    Journal of Cellular Physiology 103 (1980), S. 467-474 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The cell line B16/C3 will undergo melanogenesis at a specific time after plating. We have found that this time can be modulated by varying the pH of the culture medium. At high pH levels (8.2-8.6) the onset of melanogenesis occurs in 3 or 4 days, while at lower pH (6.7-7.2) it occurs in 7 or 8 days. Furthermore, the time of onset is also sensitive to the extracellular ionic strength. The addition of sodium lactate, sodium chloride, or any other salt tested delays or blocks completely the onset of melanogenesis. These effects are not simply consequence of growth inhibition, nor can they be correlated with patterns of lactate accumulation.These cells are sensitive to pH or ionic strength after entering the stationary phase just prior to the time of onset of melanogenesis. The existence of a specific pH-and ionic-strength-sensitive phase may provide an important clue to the events responsbile for differentiation in this system.
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  • 173
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
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    Topics: Biology , Medicine
    Notes: Quiescent 3T3 cells can be stimulated to enter S by defined factors. When used in combination, three polypeptide hormones (EGF, vasopressin, and insulin), or a tumor promotor and insulin, are very effective in stimulating DNA synthesis. Like serum, the defined factors also stimulate deoxyglucose uptake and induce the synthesis of ornithine decarboxylase during G1. The second stage of deoxyglucose uptake and the induction of ornithine decarboxylase are protein synthesis-dependent events. When added with the growth factors, mouse interferon inhibits the synthesis of DNA and the induction of ornithine decarboxylase but has no effect on the uptake of deoxyglucose. Kinetic experiments comparing the effect of inhibitors of translation or transcription on induction of ornithine decarboxylase with the effect of interferon suggest that interferon may affect the synthesis of enzyme by inhibiting both transcription and translation of message. The findings provide further support for the proposition that interferon exerts a differential effect on mitogen-stimulated events which are dependent on continuous protein synthesis.
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  • 174
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    Journal of Cellular Physiology 104 (1980), S. 27-33 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Two clone of mouse C1300 neuroblastoma cells (clones NB1R and NB6R) bind mouse 2.5S Nerve Growth Factor (NGF) in vitro. The ligand is then capped and internalized by the cells. This step requires active cell processing. In serum-free or low serum conditions, clear effects of NGF are seen with both clones. Cultured NB1R cells are stimulated, after a lag interval of a few hours, to synthesize DNA and to proliferate, whereas NB6R cells are stimulated to cell differentiation and maintain viability under these conditions much longer than similar cultures in the absence of NGF. Stimulation of clone NB1R occurs within an optimal dose concentration of the same order as that used in the Levi-Montalcini bioassay with 8-day-old chick embryo-sensitive ganglia; the effects of clone NB6R, however, need higher NGF concentrations. Both effects are protein-specific since they are inhibited in the presence of added anti-NGF antibodies. This system could provide a convenient means to study the control of cell division in susceptible malignant neuroblastoma clones and to correlate NGF binding to receptors and biological activity.
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  • 175
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    Journal of Cellular Physiology 104 (1980), S. 47-52 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Growth of human neuroblastoma strain SK-N-SH in a plasminogen-deficient medium results in about a 40% increase in the number of differentiated cells (cells with a neurite-like process at least 50 μm in length) and about a five-fold increase in the amount of plasminogen activator liberated per cell. Plasminogen deficiency has no effect on the growth rate of SK-N-SH cells. These results are consistent with the hypothesis that plasminogen activator is involved in neuroblast development.
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  • 176
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    Journal of Cellular Physiology 104 (1980), S. 83-96 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The effects of bromodeoxyuridine (BrdUrd) on the growth, morphology, and tumorigenicity of the spontaneously transformed rat liver cell line R72/3 were studied. These cells grow either in suspension or in a monolayer and are tumorigenic. In monolayer cultures, cells treated with low concentrations (2.5 μg/ml) of BrdUrd were larger, more spread out, and more firmly attached to the substratum than were untreated controls. Treated cells failed to grow in suspension or on confluent monolayers of 3T3 cells and did not form colonies in soft agar. Scanning electron microscopy revealed extensive flattening of treated cells and a dramatic reduction in the number of microvilli on the cell surface. Transmission electron microscopy showed an increase in polyribosomes and rough endoplasmic reticulum, as well as an enlargement of endoplasmic reticulum cisternae and a complete absence of the bundles of intermediate size filaments that were conspicuous in untreated cells. The persistence of these changes required the continuous presence of BrdUrd in the medium. The effects of BrdUrd were readily reversed by withdrawal of BrdUrd and were not expressed in the presence of excess thymidine.
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  • 177
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    Journal of Cellular Physiology 104 (1980) 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
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  • 178
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    Journal of Cellular Physiology 104 (1980), S. 121-125 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Electron probe x-ray microanalysis was used to measure cytoplasmic elemental content (in mmoles/kg dry weight) of the basal layer of cells of the vaginal epithelium of ovariectomized rats. Measurements were made both before estradiol injection and at 2 hr, 17 hr, and 24 hr after estradiol administration. Mitotic figures first appeared in the basal cell layer at 24 hr. During the course of the study significant time-dependent differences were seen in the content of all elements measured. A pattern of change in cytoplasmic content was seen for Na, P, S, and Cl; all of which decrease significantly by 17 hr and then return to approximately the nonstimulated concentration by 24 hr. On the other hand K, and to a lesser extent Mg, show an early and continued increase in cytoplasmic content after estradiol injection. Thus, the marked increase in the intracytoplasmic content of K in the estradiol treated cells suggests that K, or the ratio of Na to K, may be directly or indirectly involved in growth stimulation.
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  • 179
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    Journal of Cellular Physiology 104 (1980), S. 177-186 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: A morphologic and growth control variant of bovine aortal endothelial cells has been isolated and shown to synthesise factor VIII antigen (McAuslan and Reilly '79). The variant also possesses the endothelial surface markers angiotensin converting enzyme and α2-macroglobulin. The normal cell synthesises fibronectin and deposits it underneath the cells; the variant also synthesises fibronectin. At least three times more fibronectin is distributed over the upper cell surface of variants. This correlates with the three-fold increased binding of the replication inhibitor Con A and suggests a role of fibronectin in endothelial cell growth control. When stimulated to migrate by CuII ions, the variant leaves deposits of fibronectin in its trail; in contrast, migrating normal cells do not, but they do redistribute their surface fibronectin. As revealed by scanning electron microscopy, variant cells are unusual in that they grow over or under cultured normal endothelial cells. It is proposed that during the process of neovascularisation, variant cells have a special function as lead cells that lay down fibronectin on which an endothelium can become established.
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  • 180
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    Journal of Cellular Physiology 104 (1980), S. 209-214 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Using a stopped-flow device, the osmotic water permeability of human red cells to D2O and H2O was studied as a function of temperature and under the influence of the sulfhydryl reagent paracholoromercuribenzene sulfonic acid (PCMBS), an inhibitor of water transport. The ratio, permeability (D2O)/permeability (H2O) at each temperature can be predicted simply by assuming that permeability varies inversely with macroscopic viscosity. When water permeability is inhibited with PCMBS, this dependency on viscosity vanishes; the inhibited permeabilities in D2O and H2O are indistinguishable.
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  • 181
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    Journal of Cellular Physiology 104 (1980), S. 199-207 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The production of plasminogen activator (PA) by two cloned cell lines, one from an ethylnitrosourea-induced glioma (Al5A5) and the other from normal adult rat brain (ARBO C9), has been investigated. Three assays were used to detect and measure PA in harvest fluids, cells and cell lysates. Similar levels were detected in harvest fluids from both cell lines. However, the cell and lysate assays indicated much higher levels in the tumor line. When actively growing cells were compared Al5A5 cells had approximately 16 × more fibrinolytic activity than the control cells with a limit of detection in the order of 103 cells or 1 μg protein (cell lysate). In contrast for the control cells PA could only be detected when upwards of 104 cells or 5 - 10 μg protein were assayed. Plaminogen activator in as few as 103 tumor cells could be detected in the presence of 104 non-tumor cells. Plasmingoen activator in 26 μg protein of Al5A5 cell lysate could also be detected in the presence of 44 μg protein from ARBO C9 lysates indicating no inhibitory activity in the control cell lysates. Levels of PA in both harvest fluids and cell lysates wre determined as cultures progressed through the growth cycle. For cell lysates this showed a build-up of PA in the normal cell line as the cells approached and attained confluence. A much higher level was measured in the tumor cells soon after seeding and maintained to confluence. No differences in growth cycle-associated changes in secreted PA could be determined in harvest fluids: both cell lines showing similar levels at confluence.
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  • 182
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    Journal of Cellular Physiology 104 (1980), S. 225-232 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Indian muntjac diploid cells that have only three pairs of easily discernible large chromosomes were fused with hamster cells deficient in hypoxanthine guanine phosphoribosyltransferase (HGPRT) using polyethylene glycol. Cells that survived in hypoxanthine-aminopterine-thymidine (HAT)-oubaine medium were analyzed.Hybrid cells containing both muntjac and hamster chromosomes in a given cell were not found. Instead, the cells had the same chromosomal sets as those of either parental muntjac or hamster cells.A clonal isolate that had the same chromosomal sets as those of parental hamster cells was analyzed in detail and showed the following characteristics: (1) portions of the survival curve in various concentrations of HAT medium were intermediate between those of parental cells; (2) expressions of both muntjac and hamster antigen(s) were detected by immunofluorescence staining; (3) the mobility of the enzyme HGPRT in gel electrophoresis differed from that of parental hamster or muntjac cells. These results indicate that the clonal isolate (AD202h) is a somatic cell hybrid of hamster and muntjac that contains chromosomal sets of hamster with an inserted segment of the muntjac genome, including HGPRT.The formation of such an unusal hybrid and a possible explanation of transfer of some gene segments in the hybrid cell in this system are discussed.
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  • 183
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    Journal of Cellular Physiology 104 (1980), S. 233-240 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: A rat ovarian tumorous cell line (OV1N) has been isolated and established in tissue culture whose growth can be controlled by the addition of epidermal growth factor (EGF) and transferrin to serum-free medium. EGF at 1 ng/ml (1.5 × 10-10M) stimulates growth about 300% in a six-day incubation assay. A combination of 1 ng/ml EGF plus 1 μg/ml (1.25 × 10-8M) transferrin stimulates growth 1700% above controls in six days. EGF and transferrin at these concentrations completely replaced fetal calf serum (FCS), which maximally stimulates the cells at 2%. At maximal stimulation (either by FCS or EGF and transferrin) a doubling time of from 16-20 hours is obtained. The growth of OV1N cells was also studied in vivo and compared to another rat ovarian cell line, 31A-F2. Cells from both cell lines were injected intrasplenically into normal, ovariectomized and hypophysectomized female Fischer 344 rats. Animal data have indicated that 31A-F2 cells behave more like “normal” ovarian cells in that they grew only in ovariectomized hosts. OV1N cells, on the other hand, grew metastatically in all hosts and killed 36% of those individuals.The binding of 125I-EGF was also studied in OV1N and 31A-F2 cells. OV1N cells bound about eightfold more 125I-EGF than did 31A-F2 cells at 37°C. The dose of 125I-EGF required to specifically saturate 31A-F2 sites was 10 ng/ml (1.5 × 10-9M) or half-maximally at 4 ng/ml (6 × 10-10M). Scatchard plots indicate that 31A-F2 cells contain roughly fourfold less EGF-specific sites than OV1N cells. Incubation of either OV1N or 31A-F2 cells with a 100-fold excess addition of various proteins plus a maximal or a half-maximal addition of 125I-EGF demonstrated that unlabeled EGF was most effective in reducing 125I-EGF binding to both OV1N and 31A-F2 cell sites. Insulin, ovarian growth factor, transferrin, or their combinations, had minimal, if any effect on 125I-EGF binding.
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  • 184
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  • 185
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
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    Topics: Biology , Medicine
    Notes: Derepression of hexose transport in a line of Syrian hamster fibroblasts (Nil) and polyoma-transformed (PyNil) hamster fibroblasts is obtained when cells are either starved for glucose or fed with fructose as the only hexose source. D-glucosamine feeding of these cells does not alter the repressed state with regard to hexose transport. High, derepressed rates of galactose transport were changed to low, repressed rates, within 18 hours of refeeding glucose-starved cells with D-glucosamine as the only hexose source. Nil and PyNil cells, when cultured in the presence of D-glucosamine, undergo rapid reductions in total cellular uridine 5′-triphosphate (UTP) pool sizes. By contrast, the total cellular pools of adenosine 5′-triphosphate, guanosine 5′-triphosphate, and cytosine 5′-triphosphate (ATP, GTP, and CTP) were only moderately affected by the treatment of the cells with glucosamine. The metabolic drain of the UTP pools in PyNil cells was much more pronounced than in the untransformed cells. The larger and more rapid metabolic lability of UTP pools in the transformed cells may be the primary reason for the selective toxicity of glucosamine on tumor cells. A comparison of the effects of glucosamine on hexose-starved Nil and PyNil cells demonstrated that only the untransformed cells were able to utilize glucosamine to increase the hexose starvation-depleted pools of all nucleoside triphosphates. Accumulation of UDP-glucosamine and UDP-N-acetylglucosamine followed the reduction in the UTP pools. Inhibition of protein synthesis by cycloheximide during glucosamine feeding led to higher levels of UDP-glucosamine and UDP-N-acetylglucosamine accumulation. It is suggested that the drain of UTP pools during glucosamine treatment proceeds through the formation of the UDP-aminosugars which turn over due to the action of intracellular UDP-aminosugar pyrophosphatase activities.
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  • 186
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    Journal of Cellular Physiology 102 (1980), S. 19-26 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: An intraperitoneal injection of the β-adrenergic agonist dl-isoproterenol hydrochloride (100 mg/Kg body weight) into a rat caused an early, very large (400-fold) cyclic AMP surge (peaking at 10 minutes) in the parotid gland which was followed by a second, much smaller (two-fold) surge 12 to 16 hours later. DNA synthesis began about 16 to 20 hours after the isoproterenol injection and peaked between 24 and 28 hours. The maximum level of DNA-synthetic activity at 24 hours was correlated positively to the magnitude of the small cyclic AMP surge at 12 hours, but not to the size of the much larger cyclic AMP surge at 10 minutes. An intraperitoneal injection of dl-propranolol hydrochloride (59 mg/Kg body weight) at 8 hours after isoproterenol injection abolished the second cyclic AMP surge at 12 hours and markedly (65-75%) reduced the incorporation of [3H]-thymidine into DNA. Injection of dibutyryl cyclic AMP (6.3 mg/Kg body weight) and theophylline (25 mg/Kg body weight) at 8 hours prevented propranolol from inhibiting DNA synthesis. Propranolol appeared specifically to affect the cyclic AMP-dependent pre-DNA-synthetic step because it did not reduce [3H]-thymidine incorporation when injected after the second cyclic AMP surge had passed and DNA synthesis had just begun. Thus, the initial, large cyclic AMP surge following β-adrenergic stimulation may not be necessary for the initiation of prereplicative development, while the much smaller second surge may be needed for the initiation of DNA synthesis.
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  • 187
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    Journal of Cellular Physiology 103 (1980), S. 429-433 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Using a colony formation assay, the cytotoxic effects of steroids and an anit-steroid on an established human breast tumor line and two human diploid fibroblast strains were studied. Experiments involving 17 a-estradiol, 17 β-estradiol, dexamethasone, cortisone, dihydrotestosterone, and the anti-estrogen Tamoxifen showed no killing at concentrations below 10-7M following a 24-hour exposure to these agents. A maximum of 80% killing was observed at 10-5M with dexamethasone in one fibroblast strain and at the same dose of 17 β-estradiol in the breast tumor line. The extent of killing observed is insufficient to account for many of the clinical remissions observed with steroid therapy. The data also suggest that at therapeutic doses, there is no selective killing of malignant cells by these agents.
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  • 188
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    Journal of Cellular Physiology 103 (1980), S. 417-428 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Five to six percent (by mass) of AKR-2B mouse embryo cell polysomal RNA consists of messenger RNA sequences which may exist in polyadenylated form. In the steady state, however, only 30-40% of these molecules are retained by extensive passage over oligo(dT)-cellulose, the remainder being present in the form of poly(A)-deficient analogues. Within experimental limits, these poly(A)-deficient analogues contain representatives of all poly(A)-containing mRNA sequences in these cells. An analysis of the kinetics of hybridization of cDNA probes enriched for either abundant or rare poly(A)-containing mRNA sequences suggests that the frequency distributions of poly(A)-containing and poly(A)-deficient analogues are dissimilar, and that a relationship exists between the intracellular frequency of a given mRNA sequence and the number of poly(A)-deficient analogues of that sequence. High frequency sequences appear to be enriched in the poly(A)-containing fraction, while low frequency sequences are predominately associated with the poly(A)-deficient fraction, thus, poly(A) may play a role in the regulation of mRNA frequency in the cytoplasm.
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  • 189
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    Journal of Cellular Physiology 104 (1980), S. 433-441 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Human diploid fibroblasts metabolize up to 13% of the glutamine in tissue culture medium to lactate. Four μCi of glutamine-U-14C were added to media containing 5 mM or 65 μM glucose or medium containing no added glucose, but supplemented with purine and pyrimidine nucleosides (HGTU). Aliquots of the media were taken at daily intervals and were assayed for glucose, lactate, pyruvate, malate, citrate, aspartate, glutamine, and glutamate. The label incorporation into these compounds was determined, except for glutamine and glucose. The distribution of label from glutamine-U14C in 5 mM glucose medium by day 4 was lactate (10.2%), glutamate (15.2%), citrate (1.9%), pyruvate (2.0%), malate (1.1%), and aspartate (〈 0.1%). The accumulation of label in lactate and glutamate occurred continuously during the growth cycle. Malate, citrate, and aspartate accumulation occurred primarily in confluent cultures. The label in aspartate was seen only in stationary phase cells or when the glucose concentration was decreased to 65 μM or less; net aspartate accumulation was increased twofold in low glucose media. These data demonstrate an actively functioning pathway for the conversion of 4-carbon TCA-cycle intermediates to 3-carbon glycolytic intermediates in human diploid fibroblasts.
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  • 190
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    Journal of Cellular Physiology 105 (1980), S. 25-32 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The serum supplement used in the culture of a variety of mammalian cells can be replaced by known growth factors. Diploid Chinese hamster fibroblasts (CHEF/18) will grow for several days in a medium (4F) supplemented with four growth factors: epidermal growth factor (EGF), insulin, fibroblast growth factor (FGF), and transferrin. The growth rate is only about 50% as fast as when fetal calf serum is added. This difference is eliminated by thrombin (10-100 ng/ml; 0.3-3 nM). The CHEF/18 cell line is unique in that no other cell line responds to thrombin in this concentration range.Thrombin acts synergistically with other growth factors to stimulate CHEF/18 cell growth. By itself, thrombin is only mitogenic at elevated concentrations. Thrombin can largely compensate for the absence of EGF and partly for the absence of insulin in serum-free media. Chemically and “spontaneously” transformed cell lines related to CHEF/18 have lost requirements for both EGF and thrombin, and have retained requirements for insulin and transferrin expressed by CHEF/18. No CHEF cells in this work required FGF. These results suggest that the mechanisms by which EGF and thrombin stimulate cells to grow are related.
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  • 191
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    Journal of Cellular Physiology 105 (1980), S. 33-38 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Immunostimulants from various microorganisms were tested on a myeloid leukemia cell line (Ml) for the ability to induce production of CSF and to cause differentiation of these cells. Based on their activities, the compounds were divided into two general classes: those inducing extensive cellular differentiation and those devoid of this effect. The stimulants which were active in this regard always produced large quantities of CSF, whereas those devoid of a differentiating effect did not cause CSF production. Even the potent stimulants had no effect on the D- subline, in which the differentiation was not inducible.
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  • 192
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    Journal of Cellular Physiology 104 (1980), S. 443-459 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Human lymphocytes contain a large, saturable fraction of K+ that exchanges slowly with K+ in the external medium, and a small non-saturable fraction that exchanges rapidly. We determined whether or not Na+ exchanges in a similar manner with external Na+. Cells were pre-equilibrated to ensure absence of net ion movements. Efflux was studied by loading with 22Na and transferring without washing to a non-labeled medium. Influx was studied by transferring to labeled medium and separating large samples of cells at 6,000g. There are fast, intermediate, and slow fractions of Na+ exchange, with half-times of 2, 14, and 120 minutes. At normal external K+, most cell Na+ exchanges rapidly, while at lower external K+ the Na+ that replaces cell K+ exchanges slowly. Parallel sources of fast and slow fractions, such as extracellular ones and subpopulations of cells, were ruled out by simultaneous 42K and 22Na fluxes and by a quantitative analysis of the combined K+ and Na+ content and flux data over a range of external K+ and Na+ levels. Five possible models of ion fluxes occurring in series were considered. Surface matrix, surface binding sites, and cytoplasmic channels with rapid nuclea exchange were eliminated as sources of the fast fractions. Therefore, the fast fractions of K+ and Na+ must reflect the permeability of the surface membrane. This left only two possible sources of the slow fractions. One, a subcellular compartment (e.g., nucleus), was eliminated by the combined content and flux data. We conclude that the slow fractions of ion flux are rate-limited by adsorption onto and desorption from cellular macromolecules. The data support the association-induction hypothesis and are understood by reference to two fundamental concepts: that of rapid solute exclusion from cell water existing in a polarized state; and that of solute accumulation limited by adsorption onto fixed anionic sites within the cell.
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  • 193
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    Journal of Cellular Physiology 105 (1980), S. 93-103 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Viable interspecies cytoplasmic-nuclear hybrid cells were constructed by fusion of karyoplasts prepared from the highly tumorigenic A9 mouse fibroblast cell line and cytoplasts prepared from the Detroit 532 normal human diploid cell strain. The identity of the hybrid cells was ascertained using a variety of morphological, immunological, and genetic criteria, including: nuclear pattern of staining with the fluorochrome Hoechst 33258, appearance of the actin-myosin containing cytoskeleton, presence of fibronectin, and resistance to azaguanine and diphtheria toxin. About 90% of the hybrid cells were viable, that is, capable of division. Changes in the morphology of the hybrid cells, apparently nuclear directed, were observed before cell division occurred. Using the techniques described here, large numbers of interspecies hybrid cells suitable for many types of biochemical analyses can be routinely produced.
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  • 194
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    Journal of Cellular Physiology 105 (1980), S. 143-152 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The influence upon differnt cellular and humoral parameters of hemopoiesis of three structurally unrelated, highly purified bacterial cell-wall components (BCWC) was investigated. The spleens of C57BL/6 mice assayed 6 days after the injection of either lipid A or outer-membrane lipoportein, but not murein, showed a marked increase in granulocyte-macrophage, eosinophil, and megakaryocyte progenitor cell levels. The number of pluripotent hemopoietic stem cells (CFU-S) also increased in the spleens of mice treated with either lipid A or lipoprotein. Similar results were obtained following the injection of lipoprotein or lipid A into CBA or C57BL/6.nu mice. Genetically anemic Wf/Wf mice were found to have spontaneously elevated numbers of splenic progenitor cells, which increased further after the injection of lipid A. The proportions of the different splenic progenitor cell types were similar in both untreated and lipid A treated Wf/Wf mice, and in normal littermate controls.When tested in vitro, unfractionated or partially purified post-lipid A serum was found to stimulate the growth of granulocyte-macrophage progenitor cells (GM-CFC), but no detectable stimulation of eosinohphil, megakryocyte, or erythroid progenitor cells was observed. The data suggest that the rise in splenic levels of the different progenitor cells is not mediated by the corresponding types of CSF, but more likely by proliferation and differentiation of CFU-S.
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  • 195
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    Journal of Cellular Physiology 105 (1980), S. 181-184 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
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  • 196
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    Journal of Cellular Physiology 105 (1980) 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
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  • 197
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    Journal of Cellular Physiology 105 (1980), S. 209-220 
    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 (ts) cell cycle mutant of Chinese hamster fibroblasts with a block in G1 was investigated. Attention was on the expression of the activity of three enzymes: ornithine decarboxylase (ODC), S-adenosylmethionine decarboxylase (SAMDC), and thymidine kinase (TK). ODC and SAMDC activities are normally induced in the middle of, or late in, the G1 phase, while TK activity starts to appear at the G1/S boundary. In the ts mutant released from serum starvation at the nonpermissive temperature (40.8°C), we find no effect on the expression of SAMDC activity, a significantly reduced level of ODC activity compared to the control at the permissive temperature (34°C), and no induction of TK activity. Results presented here and in a previous publication (Landy-Otsuka and Scheffler, '78) suggest that the decrease in ODC activity is due to an effect of the nonpermissive temperature on a post-transcriptional step, possibly a very rapid inactivation of the enzyme. The absence of TK activity, on the other hand, appears to be due to a block in transcription at the nonpermissive temperature.
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  • 198
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    Journal of Cellular Physiology 105 (1980), S. 235-246 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: A human osteosarcoma cell line, U-2 OS, cultured under serumfree conditions, was shown to produce a growth factor (osteosarcoma-derived growth factor, ODGF) for human-cultured glial cells, fibroblasts, and other cells. ODGF, collected from the spent medium of 2 OS cultures, was purified by a sequence involving heparin-Sepharose chromatography, hydrophobic chromatography, gel chromatography, and preparative gel electrophoresis in SDS. Purified ODGF, at a concentration of 3 ng/ml, elicited a mitogenic response in human glial cells equivalent to 50% of that afforded by human serum at a final concentration of 1%. The preparation was estimated to be 〉 50% pure.The biological activity of ODGF resided in a cationic, relatively heat-resistant, reduction-susceptible protein with a molecular weight of 30,000 (by gel chromatography and SDS-gel electrophoresis). The electrophoretic behaviour of radioiodinated ODGF suggested that the protein was composed of two different polypeptide chains (about 13,000-14,000 and 16,000-17,000 daltons, respectively) linked via disulphide bonds. The molecular makeup of ODGF was thus similar to that of platelet-derived growth factor. 125I-ODGF could be precipitated by an antibody to platelet-derived growth factor, indicating that the two factors were immunologically related. Resemblance with platelet-derived growth factor was also indicated by the finding that the latter (but not, e.g., fibroblast growth factor or epidermal growth factor) competed with 125I-ODGF for binding to human-cultured glial cells.
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  • 199
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    Journal of Cellular Physiology 105 (1980), S. 259-266 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: We have analyzed ongoing DNA replication in ts BN-2, a dna- mutant of BHK-21 cells (Nishimoto et al. '78). At the non-permissive temperature of 39.5°C, inhibition of 3H-thymidine into acid-precipitable material begins 1 to 2 h after the cells are released from a block at the start of the S-phase. The fraction of nuclei incorporating 3H-thymidine is similar to that of wild-type cells through the synchronized S-phase of 8 h. Alkaline sucrose gradient analysis shows tht pulse-labeled DNA from mutant cells is incorporated into high molecular weight material after 3 h at eithr the permissive or non-permissive temperature. DNA fiber autoradiograms reveal that, at 39.5°C, the rate of replication fork movement is about 30% increased in the mutant as compared to wild-type cells. In the mutant cells, however, the interval between adjacent initiation sites is increased and the relative frequency of initiation events is decreased at the restrictive temperature. The results indicate that there is a block to ongoing replication in ts BN-2 at the level of initiation of synthesis on individual replication units; elongation of nascent chains is not inhibited.
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  • 200
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    Journal of Cellular Physiology 105 (1980), S. 335-346 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Protein degradation has been measured in confluent monolayers of eleven lines of contact-inhibited cells and ten transformed lines as the rate of release of trichloroacetic acid-soluble radioactivity after prelabeling cell protein with [3H]leucine. Insulin, at concentrations from 10-12 M to 10-6 M, has been added at the beginning of the 4-hour degradation period to detect selective effects of this hormone as an inhibitor of the inducible proteolysis occurring in serumfree medium. In addition insulin binding measurements have been performed on selected cell lines in an attempt to relate receptor properties to insulin action. Substantial effects of insulin are found in most cells with a selective inhibition at low insulin concentrations noted in several of the transformed lines. The difference in insulin sensitivity is not entirely definitive because temperature-sensitive transformation mutants of NRK cells are not more sensitive to insulin at a temperature where they show the transformed phenotype. Although insulin receptors on different cell lines have similar binding properties, two of the hepatomas used, H35 and MH1C1, show inhibition of protein degradation at insulin concentrations where receptor occupancy is extremely low. Calvarial osteoblast-like cells have a high rate of protein degradation which can be reduced by growth factors but not by insulin. The lack of an insulin response is a consequence of poor insulin binding to the cells. Insulin binds to the osteogenic sarcoma cells in substantial amounts. However, its normal action to inhibit the induced proteolysis is restricted because with these cells no increase of proteolysis occurs in serum-free medium. Generally higher rates of protein degradation are observed in the contact-inhibited lines than the transformed cells. We suggest that this difference may provide a selective growth advantage to transformed cells.
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