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  • Cell & Developmental Biology  (372)
  • 1980-1984  (372)
  • 1950-1954
  • 1980  (372)
  • 101
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Journal of Morphology 166 (1980), S. 337-386 
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Three categories of dietary adaptation are recognized - faunivory, frugivory, and folivory - according to the distinctive structural and biochemical features of animal matter, fruit, and leaves respectively, and the predominance of only one in the diets of most species.Mammals subsisting mainly on animal matter have a simple stomach and colon and a long small intestine, whereas folivorous species have a complex stomach and/or an enlarged caecum and colon; mammals eating mostly fruit have an intermediate morphology, according to the nature of the fruit and their tendency to supplement this diet with either animal matter or leaves. The frugivorous group are mostly primates: 50 of the 78 mammalian species, and 117 of the 180 individuals included in this analysis are primates.Coefficients of gut differentiation, the ratio of stomach and large intestine to small intestine (by area, weight, and volume), are low in faunivores and high in folivores; the continuous spread of coefficients reflects the different degrees of adaptation to these two dietary extremes.Interspecific comparisons are developed by allowing for allometric factors. In faunivores, in which fermentation is minimal, the volume of stomach and large intestine is related to actual body size, whereas these chambers are more voluminous in larger frugivores and mid-gut fermenting folivores; fore-gut fermenters show a marked decrease in capacity with increasing body size. Surface areas for absorption are related to metabolic body size, directly so in frugivores; area for absorption is relatively less in larger faunivores and more in larger folivores, especially those with large stomachs.Indices of gut specialization are derived from these regressions by nonlinear transformation, with references to the main functional features of capacity for fermentation and surface area for absorption.These are directly comparable with the dietary index, derived from quantitative feeding data displayed on a three-dimensional graph, with all species within a crescentic path from 100% faunivory through 557ndash;80% frugivory to 100% folivory, perhaps illustrating, at least for primates, the evolutionary path from primitive insectivorous forms through three major ecological grades.
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  • 102
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The ventral surface of the most proximal tarsomere of each mesothoracic leg of the female black fly, Simulium venustum Say, bears approximately 60 bifurcate sensilla. Externally, a sensillum appears as a hair set into an asymmetric socket and with the distal tip flattened into two flared lobes. A single pore opens into a short groove at the base of the lobes. The hair shaft is divided into two lumina, one of which contains the dendrites. Each sensillum is innervated by four neurons, the dendrites of which extend unbranched to the pore. Sensillum liquor bathes the dendritic tips and extends through the pore into the adjacent groove and across part of the lobes. A sieve-like structure exists in the pore region of many if not all sensilla. At least two sheath cells are associated with each sensillum.It is suggested that, although the bifurcate sensilla have the internal structure associated with known contact chemosensilla, they have secondarily acquired an olfactory function which is facilitated by the flattened lobes which increase the adsorptive surface area.Along each side of the bifurcate sensilla is a row of sturdy spines, each innervated by a neuron with a tubular body, a characteristic of cuticular mechanoreceptors. These spines are likely tactile sensilla.
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  • 103
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The morphology and ultrastructure of the blood cells of the freshwater snails Lymnaea stagnalis, Biomphalaria glabrata, and Bulinus truncatus were studied. By performing in vitro experiments and enzyme histochemical studies, special attention was paid to the role of the blood cells in phagocytosis of foreign particles.No fundamental differences were found in the ultrastructure, lysosomal enzyme contents, and phagocytic capacities of the blood cells of these species. It is concluded that only one type of blood cell, the amoebocyte, exists in the freshwater snails. Amoebocytes constitute a morphologically and functionally heterogeneous population of cells, ranging from round (electron-dense) cells with the morphological characteristics of young cells to highly phagocytic spreading cells with a prominent lysosomal system. In addition to acid phosphatase, nonspecific esterase and peroxidase were found within the lysosomes.The presence of enzyme activity in the RER and the Golgi bodies indicates that amoebocytes are able to synthesize lysosomal enzymes continuously.
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  • 104
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    New York, NY : Wiley-Blackwell
    Journal of Morphology 165 (1980), S. 13-29 
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The active motility of the cells of the yolk sac of the living Fundulus embryo was studied by time-lapse cinemicrography with phase contrast optics. In the teleost, the yolk sac lies peripheral to the body of the embryo proper and consists of a fluid-filled space bounded above by a superficial epithelium, the enveloping layer (EVL), and below by the yolk syncytial layer (YSL). The cell types treated in the present study are the enveloping layer epithelial cell, the stellate cell which lies in a layer flattened on the inner surface of the EVL, the epithelioid deep cell, the yolk sac amoebocyte, the yolk sac endothelial cell and the yolk sac melanoblast. The most actively motile cells examined in the present study are the yolk sac amebocyte and the melanoblast, which emigrates from the embryo proper at stages 19-21. The amoebocytes are compact rounded cells that move very rapidly by the extension of lamellipods with scalloped margins. The amoebocytes wander over the yolk sac in an apparently undirected fashion and invade the embryo proper when they happen to encounter it, moving between cells of the lateral mesoderm. The melanoblasts migrate by the gradual extension of elongated branching processes. Cells are sometimes monopodial, with movement being parallel to the long axis of the cell. Alternatively, movement may be perpendicular to the predominant long axis, with processes being extended alternatively from opposite ends of the cell obliquely forward, so the path described is a zig-zag to either side of the overall direction of movement. Although the melanoblasts show irregularity in their movement, the predominant direction of initial movement is away from the embryo proper. The major yolk sac blood vessels form in situ by the collective activities of presumptive endothelial cells that enclose volumes of the yolk sac space with sheet-like processes from the cell body and from the extensions that connect cells into networks.
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  • 105
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    New York, NY : Wiley-Blackwell
    Journal of Morphology 164 (1980), S. 161-166 
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Tactile hairs are present on all three subsegments of the antennal flagellum of the human louse. There is, in addition, a single chemoreceptor (tuft organ) on subsegment 2 and 12 or 13 chemoreceptors (one tuft organ, two pore organs and nine or ten pegs) on subsegment 3. The cuticle surrounding the bases of the pegs at the tip of the antenna is unusual in that parts of it are perforated by many fine pores. This cuticle is underlain by a thin layer of dendrites. This region may also have a chemoreceptor function.
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  • 106
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    New York, NY : Wiley-Blackwell
    Journal of Morphology 164 (1980), S. 139-159 
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Two morphologically distinct structures occur on the surfaces of the oral papillae in several loricariid catfish species; namely, (1) typical vertebrate taste buds composed of receptor and sustentacular cells and (2) brushlike projections, termed epidermal brushes, that represent specialized epidermal cells containing keratin. Both of these structures were studied with the combined use of light microscopy and scanning and transmission electron microscopy. The general body surface, fins, and rostral cutaneous processes of some loricariid catfishes are covered with taste or terminal buds but lack the epidermal brushes. It is suggested that the epidermal brushes found on the oral papillae serve as protective devices for the taste buds and as abrasive surfaces for substrate scraping during feeding. The taste buds on the oral papillae may detect any gustatory stimuli from the resulting substrate disturbance. Comparative studies reveal many differences in the number and spatial arrangement of these two structures on the oral papillae among the several species of the Loricariidae examined. These differences may represent functional adaptations to the various modes of life in the Loricariidae.
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  • 107
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    New York, NY : Wiley-Blackwell
    Journal of Morphology 164 (1980), S. 121-138 
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The heart-body of the marine worm Amphitrite, located within the supraesophageal dorsal vessel, is in the form of a cylinder the thin wall of which is deeply corrugated by luminal projections and folds along its entire length. It is anchored in places to the luminal surface of the dorsal vessel by an extracellular matrix containing collagen fibers. The luminal surfaces of both the heart-body and the dorsal vessel are covered by a basement membrane-like vascular lamina which in turn supports a discontinuous pseudoendothelium of littoral hemocytes.The cells of the heart-body constitute a pseudostratified, high columnar epithelium. They possess extensive rough endoplasmic reticulum (RER), a well developed Golgi zone, ferritin particles and granules, and several types of membrane-bound inclusions. Hemoglobin molecules identical to those in the circulation lie within cytoplasmic, membrane-bound vesicles. Analysis of our electron micrographs suggests the following sequence of hemoglobin production and secretion: Large quantities of a moderately dense flocculent material, probably globin, are synthesized in RER and move to the Golgi zone within partly rough- and partly smooth-surfaced transitional cisternae; small transport vesicles, formed from Golgi cisternae that have fused with transitional cisternae, convey the flocculent material from the convex to the concave face of the Golgi complex; a similar flocculent material and an amorphous, highly dense material are processed in the Golgi complex and are transferred to condensing vacuoles in which clearly identifiable hemoglobin molecules are first observed. Mature secretory vesicles containing only hemoglobin migrate to the cell periphery and discharge their contents by exocytosis. Hemoglobin molecules then cross the vascular lamina to reach the circulation.
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  • 108
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    Journal of Morphology 165 (1980), S. 85-116 
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Lateral cortex is the most laterally placed of the four cortical areas in snakes. Earlier studies suggest that it is composed of several subdivisions but provide no information on their organization. This paper first investigates the structure of lateral cortex in boa constrictors (Constrictor constrictor), garter snakes (Thamnophis sirtalis), and banded water snakes (Natrix sipedon) using Nissl and Golgi preparations; and secondly examines the relation of main olfactory bulb projections to the subdivisions of lateral cortex using Fink-Heimer and electron microscopic preparations.Lateral cortex is divided on cytoarchitectonic grounds into two major parts called rostral and caudal lateral cortex. Each part is further divided into dorsal and ventral subdivisions so that lateral cortex has a total of four subdivisions: dorsal rostral lateral cortex (drL), ventral rostral lateral cortex (vrL), dorsal caudal lateral cortex (dcL) and ventral caudal lateral cortex (vcL). Systematic analyses of Golgi preparations indicate that the rostral and caudal parts each contain distinct populations of neurons. Rostral lateral cortex contains bowl cells whose dendrites arborize widely in the outer cortical layer (layer 1). The axons of some bowl cells can be traced medially into dorsal cortex, dorsomedial cortex and medial cortex. Caudal lateral cortex contains pyramidal cells whose somata occur in layers 2 and 3 and whose dendrites extend radially up to the pial surface. In addition, three populations of neurons occur in both rostral and caudal lateral cortex. Stellate cells occur in all three layers and have dendrites which arborize in all directions. Double pyramidal cells occur primarily in layer 2 and have dendrites which form two conical fields whose long axes are oriented radially. Horizontal cells occur in layer 3 and have dendrites oriented concentric with the ependyma. Fink-Heimer preparations of snakes which underwent lesions of the main olfactory bulb show that the primary olfactory projections to cortex are bilateral and restricted precisely to rostral lateral cortex. Electron microscopic degeneration experiments indicate that the olfactory bulb fibers end as terminals which have clear, spherical vesicles and asymmetric active zones. The majority are presynaptic to dendritic spines in outer layer 1.These studies establish that lateral cortex in snakes is heterogeneous and contains two major parts, each containing two subdivisions. The rostral and caudal parts have characteristic neuronal populations. Primary olfactory input is restricted to rostral lateral cortex and seems to terminate heavily on the distal dendrites of bowl cells. Axons of some of these cells leave lateral cortex, so that the rostral lateral cortex forms a direct route by which olfactory information reaches other cortical areas. The functional role of caudal lateral cortex is not clear.
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  • 109
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    New York, NY : Wiley-Blackwell
    Journal of Morphology 163 (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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  • 110
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    Journal of Morphology 163 (1980), S. 9-12 
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Previously unreported structures found on the head and thorax of several species of microcaddisflies (Trichoptera: Hydroptilidae) are described. Depending on the species, these presumptive pheromone-producing glands are found either (1) on the basal segment of the antenna, (2) on movable and immovable occipital sclerites, (3) as eversible organs from the occipital area of the head, or (4) on structures which are attached near the bases of the front wings.
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  • 111
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    New York, NY : Wiley-Blackwell
    Journal of Morphology 163 (1980), S. 1-8 
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The annular bands were studied by light and scanning electron microscopy in normal and hormonally bursectomized ducks (Anas platyrhynchos). The four annular bands are normal lymphoid structures of 5-10 mm wide and encircle the intestine at regularly spaced position, two on each side of Meckel's diverticulum. The anterior three are well defined, complete rings whereas the posterior-most encompasses about one half of the gut circumference. The bands are characterized by prominent follicles in the tunica muscularis, submucosa, and lamina propria. In addition, large numbers of diffusely organized lymphocytes fill the lamina propria and villus cores. Each nodule possesses germinal centre activity, as revealed by the characteristic macrophage content seen in 1.0 μm sections.The bands were present in rudimentary form at hatching. Lymphoid nodules began to develop at day 3 and were morphologically mature at day 98 posthatching. When viewed in the scanning electron microscope, the mucosa of the lymphoid areas was seen to be arranged in tortuous folds, often with irregular fusions. Following hormonal bursectomy, the bands were present, although difficult to detect, and lacked distinct nodules and germinal centres. The mucosal surface still appeared irregularly folded in the SEM, but the folds were more slender with convoluted surfaces.
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  • 112
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The present investigation describes the ultrastructural changes which occur at the surface and in the cytoplasm of developing oocytes of the lobster, Homarus americanus, during vitellogenesis. The immature oocytes showed no surface specializations of the oolemma and no pinocytotic activity was observed. Horseradish peroxidase (HRP) tracer studies showed penetration of the tracer into the perivitelline space, but no uptake by the oocytes. The surfaces of oocytes examined during vitellogenesis, when yolk protein accumulation was maximal, exhibited numerous microvilli that projected into the perivitelline space, often appearing to be embedded in the follicular cell mass. In addition, the plasma membrane of vitellogenic oocytes contained many pinocytotic pits frequently situated at the bases of microvilli. The perivitelline space was engorged with electrondense material which appeared similar to that contained in pinocytotic structures of the oocytes. Vitellogenic oocytes incubated in HRP showed uptake of tracer reaction product by the coated pits and vesicles of the oolemma. Aggregation and subsequent fusion of these vesicles into large multivesicular bodies of ingested material were also observed in vitellogenic oocytes. Animals artificially induced to undergo vitellogenesis exhibited modulations of oocyte ultrastructure similar to those of normal vitellogenesis, notably, pinocytotic incorporation of extra-oocytic material and hypertrophy of oocyte surface microvilli. This study supports the hypothesis for a dual source of yolk protein in the American lobster.
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  • 113
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    Journal of Morphology 163 (1980), S. 37-44 
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The morphology and organization of chromatophores in the neotropical glass-frog, Centrolenella fleischmanni (family Centrolenidae), were studied with both light and electron microscopes. Four types of pigment cells are described in the dorsal skin. The fine structure of two chromatophores corresponds to the typical amphibian xanthophore and iridophore; one is similar to the unusual melanophore found in phyllomedusine hylids; the fourth cell type is unlike any chromatophore previously described. Pigment granules in the unusual chromatophore are moderately electron-dense and have an irregular shape, suggesting a fluid composition. This pigment appears to be laid down in organelles similar in appearance to pterinosomes. The organization of pigment cells in this species differs from that of other green, leaf-sitting frogs in that there are few discrete groups resembling “dermal chromatophore units.” It is suggested that the unusual new pigment cell contributes significantly to the overall green color of C. fleischmanni.
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  • 114
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    Journal of Morphology 165 (1980), S. 261-284 
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: This study carried out on the posterior caeca of Orchestia in intermolt by means of light and electron microscopy shows that the diverticula of the midgut consist of two segments which are different from an anatomical point of view. The distal segment is in close relationship to the dorsal blood vessel, whereas the proximal segment, twice as long as the distal one, only touches the haemocoel. The cells of the distal segment are characterized by a brush border, some apical extrusions, a great number of ribosomes, rough endoplasmic reticulum, often associated with the mitochondria, the matrix of which is clear, high activity of the Golgi complexes, and a great development of extracellular channels. All these features indicate an activity in synthesizing proteins and transport. In the proximal segment, the cells are characterized by a striated border, reduced intercellular space, and especially by a great development of the smooth endoplasmic reticulum sometimes associated with mitochondria having a dense matrix. These diverse features indicate absorption ion and water transport. From an ultrastructural point of view, the posterior caeca of Orchestia cannot be considered homologous to the Malpighian tubules. Whereas during molting the posterior caeca of Orchestia are sites of calcium storage, during intermolt they are probably involved in the processes of water and mineral regulation and excretion.
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  • 115
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    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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  • 116
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    Journal of Morphology 165 (1980), S. 285-299 
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Neuronal projections from neuroendocrine tracts (nervi corpori cordiaci I and II) in the brains of the locust (Schistocerca vaga), cricket (Acheta domesticus), and cockroach (Periplaneta americana) were studied using reconstructions of silver-intensified cobalt chloride preparations. Collaterals from the NCC I in these species branch extensively in the dorsal protocerebral neuropile, anterior to the stalk of the corpora pedunculata and ventral to its calyces. Other fibers project from the NCC I bilaterally into the medial protocerebral neuropile, anterior to the central body, and posterior to the beta lobes. NCC II collaterals arborize in the medial, dorsal, and lateral protocerebral neuropile, their region of projection partially overlapping with that of the NCC I. Several NCC II fibers terminate in the superior arch of the central body in Acheta but not in the other two species. Tritocerebral cells filled through the NCC I branch in the medial tritocerebral neuropile in all three species, but most extensively in Schistocerca. No NCC fibers were seen to penetrate any part of the corpora pedunculata, protocerebral bridge, olfactory glomeruli, ocellar tracts, or optic lobes.These neuronal projections from the NCC I and II lie anterior to regions of branching of second-order ocellar fibers and thus provide no anatomical basis for direct ocellar input to neurosecretory cells, contrary to previous reports for orthopteroid species (Brousse-Gaury, '71a, b). However, interneurons filled from the optic lobes were found to terminate in the same region of dorsal protocerebral neuropile as NCC I and II fibers in Acheta, thus providing a possible pathway for optic input to the cerebral neuroendocrine system.
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  • 117
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    Journal of Morphology 166 (1980), S. 1-25 
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The paired spermathecae of Rhodnius are simple tubular out-pocketings of the common oviduct. Each consists of a short muscular proximal duct and the distal glandular region with a blind tapering end. The spermathecal wall has a cuticular intima, slender columnar epithelial cells and ensheathing longitudinal striated muscle, connective tissue, tracheoles, and nerves. Glandular epithelial cells possess an elaborate apical secretion-filled tubular inpocketing with an extensively folded plasma membrane. Laterally, cells interact by desmosomes, septate desmosomes, and extensive interdigitations. The cytoplasm is rich in longitudinally oriented microtubules associating with membrane densities along the invagination, lateral, and basal plasmalemmae. Apical concentration of mitochondria suggests their role in secretion or ion transport. The possible role of the spermathecae in maintaining the stored luminal sperm and its role in transmitting the mating stimulus is considered in light of the epithelial ultrastructure. The ultrastructure of the spermathecae of Rhodnius differs significantly from that of other insects.
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  • 118
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    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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  • 119
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    Journal of Morphology 163 (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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  • 120
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    Journal of Morphology 163 (1980), S. 79-93 
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: All lizard species of the subfamily Iguaninae except Amblyrynchus cristatus possess from one to eleven transverse valves in the proximal colon. Valves are of two kinds: circular (sometimes with a sphincter valve) or semilunar. Circular valves (if present) always occur proximally to semilunar valves. Intraspecific variation in the number and type of valves is small, but increase with modal number of valves. No significant ontogenetic change in number of valves could be demonstrated. Colic valves in iguanine lizards apparently evolved as simple infoldings of the medical colic wall.Comparisons are made with colic modifications occurring in other lizard families. Herbivorous species of the Scincidae, Agamidae, and Iguanidae are the only lizards known to exhibit colic partitioning, suggesting that the evolution of these structures is intimately related to the evolution of herbivory in these lizards. The potential taxonomic and phylogenetic importance of lizard colon anatomy is discussed.
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  • 121
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    Journal of Morphology 163 (1980), S. 99-133 
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Study of the visceral anatomy of 41 specimens of amphisbaenians representing 13 genera shows that they share a very distinct structure which differs from that found in either snakes or typical lizards. The left lung is large while the right is rudimentary or absent (unique); the kidneys are freely suspended in the coelom by a mesentery (unique); the spleen is usually embedded in the anterior end of the pancreas (as in snakes); the gall bladder lies in a notch in the liver, and the kidneys lie opposite each other (as in lizards). The distinctness of this pattern supports the recognition of the Amphisbaenia as a separate suborder of the Squamata.
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  • 122
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    Journal of Morphology 163 (1980), S. 135-155 
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Formation of a circular hole 8-10 mm in diameter in the calcified layers of the carapace from crabs in stage C4 of the molt cycle stimulates the tissue under and adjacent to the injury to deposit a unique calcified cuticular material below the intact membranous layer. Deposition was followed for 69 days using light microscopic histology, histochemistry, and scanning electron microscopy. Quantitative analyses of CaCO3 were conducted using atomic absorption spectrophotometry and Gran titration. Spatial distribution of CaCO3 was determined with X-radiography. A scab is formed by day two under the injury. At four days the epithelium changes from squamous to columnar and deposits a PAS-positive layer with an irregular lamellar fine structure, followed by highly organized lamellae structurally similar to normal exocuticle. Histochemically, however, these lamellae resemble normal endocuticle. CaCO3 is evident external to the outermost lamellae by day eleven as a fused mass of aragonite granules. The lamellar region calcifies proximally from the outer surface and is amorphous CaCO3. Repair cuticle is approximately 20%CaCO3 by weight.
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  • 123
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    Journal of Morphology 163 (1980), S. 157-165 
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Electron microscopy of the cerebral ganglionic commissure of the leech Macrobdella decora (Say, 1824) revealed numerous neurosecretory axons terminating in the neural lamella of both the inner and outer capsules, and in the neural lamella deep within the neuropile. The proximal protions of the terminals, with an investment of glial tissue, contain either numerous large homogeneously electron dense granules, or numerous large granules of varying electron density. The distal portions, often devoid of glia, display numerous infoldings, omega profiles, and electron dense focal sites, and contain numerous neurosecretory granules, small lucent vesicles, and, occasionally, acanthosomes. Statistical analysis of the size distribution and morphology of the neurosecretory granules showed that in many individual terminals the granules are not significantly different from those seen within four groups of neurosecretory cells found in the cerebral ganglion. These terminals, because of their diffuse nature, probably represent a neurohemal complex of a primitive nature. The term “intralamellar complexes” is proposed to describe the form and location of these neurosecretory terminals.
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  • 124
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    Journal of Morphology 163 (1980), S. 167-174 
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The paired Y-organs of crustaceans control the molting process. In males of C. antennarius, these glands are opalescent, lobulated, epithelioid structures embedded in brown fatty tissue. Cells in the periphery extend processes to the connective tissue capsule, an arrangement that suggests increased surface area for metabolic exchange. The processes contain mitochondria and are tipped distally with electron dense material. The cytoplasm, scarce relative to nuclear volume, contains vesicles, polymorphic mitochondria with tubular cristae, and numerous free ribosomes, but little in way of smooth or rough endoplasmic reticulum or Golgi complexes. Progressing from intermolt to the premolt stage, mitochondria, as well as vesicles, and electron-dense particles in peripheral processes increase somewhat in number. Also, heterochromatin masses concentrate adjacent to the nuclear envelope. Eyestalk removal, which induces premolt stages in some species, did not produce consistent change in Y-organ substructure in C. antennarius. Although evidence is accumulating that Y-organs secrete a steroid molting hormone during late intermolt-premolt, the substructure of the glands exhibits neither (a) striking changes with the molt cycle, nor (b) all the characteristics typical of vertebrate steroid hormone synthesizing glands. Nevertheless, the structural features, respectively, are consistent with biochemical evidence that Y-organs (a) rapidly take up and convert sterol precursor and secrete a product without its accumulation or change in total sterol pool size, and (b) apparently cannot synthesize the sterol precursor. Y-organ cytology closely resembles that of some vertebrate steroid hormone secreting glands in which this synthetic capacity is minimal.
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  • 125
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    Journal of Morphology 163 (1980), S. 175-190 
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Light and electron microscope studies were made on harvestman oocytes during the course of their origin, differentiation, and vitellogenesis. The germ cells appear to originate from the ovarian epithelium. They subsequently migrate to the outer surface of the epithelium, where they remain attached often by means of stalk cells which suspend them in the hemocoel during oogenesis. The “Balbiani bodies,” “yolk nuclei,” or “nuage” constitute a prominent feature of young, previtellogenic oocytes, and take the form of large, but variable sizes of electron-dense cytoplasmic aggregates with small fibrogranular components. The cytoplasmic aggregates fragment and disperse, and cannot be detected in vitellogenic oocytes. The young oocytes become surrounded by a vitelline envelope that appears to represent a secretory product of the oocyte. The previtellogenic oocytes are impermeable to horseradish peroxidase under both in vivo and in vitro conditions. In addition to mitochondria, dictyosomes, and abundant ribosomes, the ooplasm of the previtellogenic oocyte acquires both vesicular and lamellar forms of the rough-surfaced endoplasmic reticulum. In many areas, a dense homogeneous product appears within the cisternae of the endoplasmic reticulum and represents nascent yolk protein synthesized by the oocyte during early stages of vitellogenesis. Later in vitellogenesis, the oocyte becomes permeable to horseradish peroxidase under both in vivo and in vitro conditions. This change is associated with a massive process of micropinocytosis which is reflected in the presence of large numbers of vesicles of variable form and structure in the cortical ooplasm. Both spherical and tubular vesicles are present, as are coated and uncoated vesicles. Stages in the fusion of the vesicles with each other and with developing yolk platelets are illustrated. In the harvester oocytes, vitellogenesis is a process that involves both autosynthetic and heterosynthetic mechanisms.
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  • 126
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    Journal of Morphology 163 (1980), S. 191-201 
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Colonic organogenesis in rats was studied using light microscopic techniques for the demonstration of mucosubstances, glycogen, and connective tissue fibers.Crypts began as intraepithelial spaces which were in continuity with the colonic lumen. The cells forming the floors of these spaces invaded the nonsulfated acid glycosaminoglycan-rich mesenchyme as the basement membrane became discontinuous. As the diameter of the colon increased, the crypts lengthened and the lamina propria thickened until a layer of collagen and sulfated acid glycosaminoglycans formed at the bases of the crypts and the basement membrane was reestablished. The circular layer of the muscularis externa developed first, then the longitudinal layer, and finally the muscularis mucosae.Three types of mucous cells arose in these newly formed crypts. The initial epithelial cell type contained glycogen and gave rise to cells with apical coats of nonsulfated acid glycoproteins. This cell type was followed by the appearance of cells at the bases of the crypts containing nonsulfated acid glycoproteins. As the crypts lengthened, the goblet cells near the base contained nonsulfated and/or sulfated acid glycoproteins. Closer to and on the surface, the cells contained sulfated acid glycoproteins, a mixture of sulfated acid and neutral glycoproteins, or just neutral glycoproteins. Striated-border cells appeared intermingled with the mucous cells close to the bases of the crypts and continued onto the surface.A comparison was made between regeneration following placement of a surgical lesion in adult rats and events in organogenesis of the colon.
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    Journal of Morphology 163 (1980) 
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  • 128
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    Journal of Morphology 163 (1980), S. 203-215 
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: As part of a study of ulcer formation and healing, regeneration of colonic mucosa in rats was studied following placement of a surgical lesion. Alterations in mucosubstances and connective tissue were examined and their possible significance discussed.The sequence of events in healing was: (1) The mucosa adjacent to the lesion tipped into the lesioned area. The crypts in this mucosa became lined with cells which contained no mucus and had no striated borders. Later in the experimental period, these undifferentiated cells gave rise to cells containing carboxymucins. Cells containing sulfomucin, neutral mucin, or having striated borders arose from the carboxymucin cells. (2) An epithelial ledge of undifferentiated cells migrated onto a sulfated glycosaminoglycan, fibrous interface between necrotic and living tissue in the lesion. (3) Crypt formation began with the appearance of intraepithelial anlagen. (4) Crypts lengthened by a process of epithelial-connective tissue proliferation from the base of the crypt upwards. Following completion of connective tissue regeneration, crypts formed by invading the reestablished lamina propria. (5) The first mucous cells in the ledge contained carboxymucins. As crypt formation occurred, these cells gave rise to typical columnar absorptive cells, to cells containing sulfomucins, and to cells containing neutral mucins. (6) Lengthening of crypts ceased following the appearance of a sulfated acid glycosaminoglycan - collagenous layer deep in the submucosa.
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  • 129
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    Journal of Morphology 163 (1980), S. 219-230 
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Rat submandibular gland was dissociated by enzymatic digestion with collagenase and hyaluronidase, followed by mild mechanical shearing and filtration through a nylon mesh. The dissociated cell populations contained predominantly groups of acinar cells which maintained their acinar arrangement. The morphological and functional viability of the cells was confirmed by electron microscopic examination and a normal secretory response to β-adrenergic or cholinergic stimulation was observed. Both isoproterenol (IPR) and carbachol caused the fusion of secretory granules into large vacuoles which were also continuous with the lumen, and into which the secretory product was released. Secretion was assessed quantitatively from the incorporation of 14C-glucosamine into the acinar cells and its subsequent release into the culture medium as labelled glycoprotein. IPR stimulated secretion to 125% of untreated controls in the concentration range 5 × 10-5 to 5 × 10-7 M, and to 110% of controls at 5 × 10-8 M, after 40 min incubation. Carbachol stimulated secretion to 131% of controls at 5 × 10-5 M and to 115% at 5 × 10-6 M but had no effect at 5 × 10-7 or 5 × 10-8 M. The secretory response was blocked by the respective β-adrenergic and cholinergic antagonists, propranolol and atropine. These findings show that dissociated rat submandibular acinar cells provide a useful in vitro model for the study of mucus synthesis and secretion.
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    Journal of Morphology 163 (1980), S. 231-252 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
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    Notes: The fine structure of the turtle tastebud has been examined by light, transmission, and scanning electron microscopy. It contains five types of cells on the basis of their cytological features, designated types 1,2,3,A, and B. Types 1, 2, and 3 reach the taste pore, whereas types A and B are located basally. The type 2 cell has access to the tongue surface, i.e., the site of gustatory stimuli, and also synapses onto afferent nerves; it probably is a gustatory receptor cell and corresponds to the so-called “light” cell observed in other vertebrate tastebuds. Some cells may be differentiating. In support of this hypothesis, light microscopic autoradiography shows that postmitotic cells occur in the tastebuds within 24 hours after administration of H3-thymidine. The tastebuds of the turtle are similar to those of other vertebrates described electron-microscopically.
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  • 131
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    Journal of Morphology 163 (1980), S. 253-281 
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Mastication has been studied by cinematography with synchronized electromyography (computer quantified and analyzed), while unanesthetized, freely feeding cats (Felis catus) were reducing equivalent-sized chunks of raw and cooked beef and cooked chicken. Cats reduce food on one side at a time, and their chewing cycles show both horizontal and anteroposterior deflections. Food objects are shifted from side to side by lateral jerks of the head and movements of the tongue.During the opening phase, the lower jaw is rotated relatively straight downward, and the digastric muscles are active in bilateral symmetry. Near the end of opening, the head jerks upward, both zygomaticomandibulares start to fire, and opening acceleration of the mandible decreases. Closing starts with horizontal displacement of the mandibular canines toward the working side, accompanied by asymmetrical activities from the working side deep temporalis and the balancing side medial pterygoid, as well as a downward jerk of the head. As closing proceeds, the mandibular canines remain near the working side and the working side zygomaticomandibularis and deep masseter are very active. Near the end of closing, the mandibular canine on the working side moves toward the midline, and adductors, digastrics, and lateral pterygoids of both sides are active. The adductors of the working side are generally more active than those of the balancing side.During a reduction sequence, the number and shape of the masticatory cycles, as well as movements of the head, during a reduction sequence are affected significantly by food type. As reduction proceeds, the duration of bite and the muscular activity (as characterized by number and amplitude of spikes) change significantly among muscles of the working and balancing sides. The adductors of the working side are generally most active when cats chew raw beef, less for cooked beef, and least for cooked chicken. In general, the adductor activity reflects food consistency, whereas that of the digastrics and lateral pterygoids reflects more the vertical and lateral displacements of the mandible. Statistical analysis documents that the methods of electrode insertion and test give repeatable results for particular sites in different animals. Thus, it should be possible to compare these results with those produced while other mammalas are masticating.
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    Journal of Morphology 163 (1980), S. 283-317 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
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    Notes: The comparative functional anatomy of feeding in Polypterus senegalus, Lepisosteus oculatus, and Amia calva, three primitive actinopterygian fishes, was studied by high-speed cinematography (200 frames per second) synchronized with electromyographic recordings of cranial muscle activity. Several characters of the feeding mechanism have been identified as primitive for actinopterygian fishes: (1) Mandibular depression is mediated by the sternohyoideus muscle via the hyoid apparatus and mandibulohyoid ligament. (2) The obliquus inferioris and sternohyoideus muscles exhibit synchronous activity at the onset of the expansive phase of jaw movement. (3) Activity in the adductor operculi occurs in a double burst pattern - an initial burst at the onset of the expansive phase, followed by a burst after the jaws have closed. (4) A median septum divides the sternohyoideus muscle into right and left halves which are asymmetrically active during chewing and manipulation of prey. (5) Peak hyoid depression occurs only after peak gape has been reached and the hyoid apparatus remains depressed after the jaws have closed. (6) The neurocranium is elevated by the epaxial muscles during the expansive phase. (7) The adductor mandibulae complex is divided into three major sections - an anterior (suborbital) division, a medial division, and a posterolateral division.In Polypterus, the initial strike lasts from 60 to 125 msec, and no temporal overlap in muscle activity occurs between muscles active at the onset of the expansive phase (sternohyoideus, obliquus superioris, epaxial muscles) and the jaw adductors of the compressive phase. In Lepisosteus, the strike is extremely rapid, often occuring in as little as 20 msec. All cranial muscles become active within 10 msec of each other, and there is extensive overlap in muscle activity periods.Two biomechanically independent mechanisms mediate mandibular depression in Amia, and this duality in mouth-opening couplings is a shared feature of the halecostome fishes. Mandibular depression by hyoid retraction, and intermandibular musculature, consisting of an intermandibularis posterior and interhyoideus, are hypothesized to be primitive for the Teleostomi.
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    Journal of Morphology 163 (1980), S. 331-348 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
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    Notes: The relationship of the hemipenis to the cloaca in copula and sperm storage and transport in the female oviduct were studied in Anolis carolinensis using light and scanning electron microscopy. During copulation, the hemipenis does not penetrate beyond the cloaca, but the two apical openings of the bifurcate sulcus spermaticus appose the openings of the oviducts from the cloaca. Sperm enter the sperm storage tubules between 2 and 6 hr after insemination and small amounts of sperm reach the infundibulum 6 to 24 hr following mating. Sperm storage tubules are embedded in the wall of the utero-vaginal transition, and are formed by the folding and fusion of the oviducal epithelium. The importance of the hemipenile-cloacal relationship and the role of sperm storage in the life history of A. carolinensis are discussed.
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    Journal of Morphology 164 (1980), S. 213-214 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
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    Notes: The sagitta otolithic membrane of Fundulus heteroclitus consists of two different zones. A structured zone (gelatinous layer), which usually exhibits a reticulated or honeycomb-like architecture, is composed of tightly arranged fibrous material and covers only the sensory region of the macula. The gelatinous layer extends from the otolith surface to the tips of the sensory hairs, and probably functions primarily as a mechanoreceptor. The arrangement of this zone is closely associated with specific overlying structural features of the otolith surface and may also influence the pattern of mineral deposition to some degree. A nonstructured zone (subcupular meshwork) consists of fibers in very loose networks and covers both sensory and nonsensory regions of the macula. Over the sensory region, some of this fibrous material extends from the epithelial surface, through pores in the gelatinous layer, to the surface of the overlying otolith. In the nonsensory region, fibers of the subcupular meshwork are relatively more numerous and extend around the peripheral margin of the otolith. Evidence is presented which suggests that the fibrous material of the subcupular meshwork is incorporated into the otolith as an organic matrix constituent. New aspects on the ultrastructure of the otolith are presented and discussed.
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    Notes: Ultrastructural studies on blood leukocytes of the channel catfish, Ictalurus punctatus, show the presence of heterophils (neutrophils), small lymphocytes, monocytes, and thrombocytes. Monocytes cannot always be distinguished from large lymphocytes. Cells resembling macrophages or transitional forms between monocytes and macrophages are occasionally seen. Blood eosinophils and basophils are not found. Thrombocytes and small lymphocytes are the most abundant leukocytes, while monocytes are the least frequently encountered leukocyte. Glycogen, present in all leukocytes, is most abundant in heterophils and least abundant in monocytes. Although monocytes are similar to heterophils in size and shape, a greater amount of rough endoplasmic reticulum, free ribosomes, and fewer granules are observed in monocytes. Heterophils possess oval or elongate granules, which often contain a crystalline or striated structure; small tubules which resemble smooth endoplasmic reticulum, and cristae which traverse the long axes of the mitochondria are frequently seen. Small lymphocytes are characterized by the presence of pseudopodia, many free ribosomes, numerous large mitochondria, dictyosomes (Golgi), and long profiles of rough endoplasmic reticulum. The dictyosomes are often associated with a large zone of exclusion. Bundles of microtubules are observed near the elongated ends of thrombocytes. Deep indentations of the plasmalemma, which give the appearance of vacuoles, are also seen in thrombocytes.
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  • 139
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    Journal of Morphology 163 (1980), S. 27-35 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The presence of scolopophorous organs in aquatic Heteroptera has been reported in a number of species. This study presents a morphological investigation of these sensory structures of Lethocerus (Belostomatidae) as observed with the scanning electron microscope (SEM). Paired mesothoracic and metathoracic organs are present. Externally, each sensory structure consists of a raised sensory membrane. The distal-most portion consists of thickenings of this sensory membrane (sclerite). The receptor neurons of the mesothoracic organ are of two types - one discolopidial sensillum and 12 monoscolopidial sensilla. The former is attached to the internal wall and distal thickening of the sensory membrane, while the latter are dispersed throughout the interior and attached to the internal wall of the sensory membrane. The structure of the organs suggest that an effective stimulus could be a compression of the membrane. A discussion of possible functions (pressure reception and hearing) is included.
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  • 140
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    Journal of Cellular Physiology 102 (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: It has been previously shown that newly synthesized nuclear low molecular weight RNA species C and D are first detected in the cytoplasm for a few minutes before they are finally found in the nucleus. The following are some of the observations made in the present study, regarding the formation of C and D RNA: (1) The 5′ end cap ribose methylation of the C RNA precursor is complete in its cytoplasmic stage; the internal ribose methylation of the precursor seems to be completed about the time of its apparent transition from cytoplasm to nucleus. (2) The few nucleotides lost from the D RNA precursor during maturation seem to be excised sometime near its apparent cytoplasmic → nuclear transition. Newly synthesized C RNA also appears to lose some of its non-conserved nucleotides about the time of that transition, while the other extra nucleotides are lost later, in the nucleus. (3) The maturation of C and D RNA is inhibited early during suppression of protein synthesis by cycloheximide, while their synthesis is not. (4) The cytoplasmic precursors of C and D RNA are not associated with ribonucleoprotein particles as large as those reported for mature C and D RNA, although they do not appear to be free in the cytoplasm. (5) When the cellular UTP pool is depleted by exposure of the cells to amino sugars, and the synthesis of C, D, and other RNA species decreases, the level of[3H]uridine labeling of C and D RNA increases, while that of 4 S and 5 S RNA does not. These data are compatible with the existence of more than one nuclear UTP pool.
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    Journal of Cellular Physiology 102 (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: Cell cycle studies, using PLM analysis, were carried out on a mouse-Chinese hamster cell hybrid and its derivatives which stably retained all parental chromosomes during the year of study. Parameter estimates were obtained from the PLM curves, using conjugate gradient curve fitting procedures. The hybrid initially grew very slowly, and all phases (especially G1) were longer than those of either parent. During propagation, mean generation time decreased progressively, and the phase times approached those of the mouse parent (which had the longer G1 and S). DNA replication could be scored separately in mouse and hamster chromosome sets; initially termination was highly asynchronous, but during growth asynchrony was progressively reduced as DNA synthesis in the hamster set was prolonged.We conclude that cell hybrids may undergo progressive modifications of the cell cycle, even in the absence of significant chromosome segregation, and suggest that such changes may at least partly account for the great variety of relationships between the growth rates and phase times of parent and hybrid cells which have been reported. Because of the complexity of these changes in the cycles of interspecific cell hybrids, we believe that somatic cell genetic analysis of the regulation of the cell cycle would be more usefully applied to intraspecific hybrids whose parents differ in only one specific cycle characteristic.
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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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  • 143
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    Journal of Cellular Physiology 102 (1980), S. 333-341 
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    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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    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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  • 145
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    Journal of Cellular Physiology 102 (1980), S. 27-36 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Glucocorticoids and insulin were found to act synergistically to promote differentiation in some clones of the L6 rat myoblast cell line. Other hormone effects on these cells were investigated to determine the extent of the synergism. The insulin stimulation of sugar transport was unaffected by glucocorticoids although they did by themselves slightly enhance transport. Glucocorticoids were found to increase the adhesiveness of the cells-an effect not influenced by insulin. Cyclic AMP levels were found to peak just prior to the time of the onset of fusion and insulin broadened this peak, while the combination of both hormones further lengthened the time for which cyclic AMP levels remained elevated.
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  • 146
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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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  • 147
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    Journal of Cellular Physiology 102 (1980), S. 37-43 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Exogenous ATP has been shown to cause a rapid and reversible increase in permeability in transformed 3T3 cells (3T6 and SV3T3) but not in untransformed 3T3 cells. The cells remain viable, but lose intracellular acid-soluble pools. Treatment of transformed cells with ATP greatly reduces incorporation of 14C-leucine into protein, which is restored by the incubation of the cells with Dulbecco's modified Eagle's medium or by the external additions of certain ions and energy sources. tRNA is not required for the restoration of protein synthesis. In the permeabilized cells the energy for protein synthesis can be provided by glycolysis, oxidative phosphorylation, or direct addition of ATP. These studies demonstrate the usefulness of this method for studying the control of metabolism and macromolecular synthesis in monolayer cultures of transformed mammalian cells.
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  • 148
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    Journal of Cellular Physiology 102 (1980), S. 45-50 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: These studies were undertaken to examine the relationship between the inhibition by 5-bromodeoxyuridine (BrdU) of erythroid differentiation in Friend erythroleukemia cells and the incorporation of BrdU into DNA. Experiments were carried out in which the incorporation of BrdU into DNA and the concentration of BrdU to which the cells were exposed were varied independently of each other. In addition, the ability of deoxycytidine (dC) to reverse the effects of BrdU on hemoglobin production and to reduce the amount of BrdU in DNA was analyzed. Under all the conditions tested, the effects of BrdU were correlated with the amount of Brd incorporated into nuclear DNA. These results differ from those of recent studies on the inhibition of pigmentation and the induction of mutations by BrdU in Syrian hamster melanoma cells. The results suggest that BrdU may be producing its biological effects by a variety of different mechanisms.
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  • 149
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    Journal of Cellular Physiology 102 (1980), S. 51-54 
    ISSN: 0021-9541
    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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  • 150
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    Journal of Cellular Physiology 102 (1980), S. 55-62 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Enhanced amino acid transport is observed when quiescent cultures of chicken embryo fibroblasts are stimulated to proliferate by the addition of purified multiplication-stimulating activity (MSA). This increase in amino acid transport is an early event occuring prior to the onset of DNA synthesis in stimulated cells. Results indicate that the changes in transport activity, as measured by α-aminoisobutyric acid (AIB) uptake, are due to stimulation of only the Na+-dependent A transport system. There is little or no change in the activities of transport systems ASC, L, or Ly+ upon exposure to MSA. A kinetic analysis shows this increased activity is due to a change in Vmax while Km remains unaltered. Continuous exposure to the stimulus is required to maintain the increased level of transport activity and the presence of inhibitors of RNA and protein synthesis significantly inhibits the response. Results also indicate that a similar specific increase in the A transport system is initiated when RSV tsNY68 infected cells are shifted to the permissive temperature. It appears that the A system of mediation is emerging as a strategic regulatory site for cell function.
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  • 151
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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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  • 152
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    Journal of Cellular Physiology 102 (1980), S. 71-80 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The internalization of membrane markers during phagocytosis was followed in guinea pig granulocytes as a function of the extent of particle ingestion. The plasmalemma was carefully labeled with diazotized 35S-sulfanilate, or by treatment with periodate and sodium 3H-borohydride. These treatments provided general membrane markers. More specific markers used were 5′-nucleotidase, neuraminidase-releasable membrane sialate, and concanavalin A binding sites. In all cases except the last, internalization of membrane was directly determined on isolated phagosomes; disappearance of binding sites from the cell surface was followed in the last instance. Both phagosomal levels and disappearance from the surface were measured in the case of 5′-nucleotidase, permitting balance studies. Phagocytosis was determined as uptake of paraffin emulsions labeled with Oil-Red 0. “Marker/particle” ratios were determined as the percent external marker internalized per mg paraffin ingested.The “marker/particle” ratios for cells with chemically labeled membranes were considered to reflect random internalization of membrane entities. Colchicine had no effect on those ratios. Internalization of 5′-nucleotidase and neuraminidase-releasable sialate gave “marker/particle” ratios similar to those of the random markers, and these increased in the presence of colchicine. Concanavalin A binding sites did not appear to be removed from the cell surface in the absence of colchicine, but disappearance was observed in its presence. Control experiments indicated that these changes due to colchicine must be very cautiously interpreted. Our results differ from those obtained by others, and the reasons due to species, cell type, experimental design, etc. are discussed. Maximal particle uptake was computed to require internalization of one-fifth to one-third of the total external membrane of the cells - based on the assumption of random internalization of markers.
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  • 153
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    Journal of Cellular Physiology 102 (1980), S. 81-89 
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    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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  • 154
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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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  • 155
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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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  • 156
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    Journal of Cellular Physiology 102 (1980), S. 99-112 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Electrical responses to acetylcholine, noradrenaline, and histamine were recorded from solitary smooth muscle cells. Iontophoresis of each transmitter elicited three fast responses: a hyperpolarization, a depolarization, or a biphasic hyperpolarization-depolarization. Each transmitter activated a specific receptor since responses were specifically blocked by antagonists, two transmitters elicited different responses in solitary cells, and desensitization of response to one transmitter did not cause desensitization of responses to other transmitters. Responses were due to increased ion conductances since input resistance decreased during responses and reversal potentials were measured for deplarizing responses (-5 mV) and hyperpolarizing responses (-60 mV). Regional differences in transmitter sensitivity were mapped on solitary cells. Biphasic responses were due to simultaneous activation of receptors mediating hyperpolarizing responses and receptors mediating depolarizing responses which were segregated in the cell membrane. Noradrenaline enhanced action potential amplitude by regulation of voltage-dependent ion conductances. Finally, noradrenaline and histamine elicited periodic hyperpolarizing potentials, which may be due to increased intracellular Ca++.
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  • 157
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    Journal of Cellular Physiology 102 (1980), S. 113-117 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Weak acid distribution methods demonstrate that mitogenic levels of concanavalin A induce an intravesicular alkalinization of isolated thymocyte membrane vesicles. Experiments with chemical reagents that crosslink the high affinity concanavalin A receptor and extensive correlation with known cellular events suggest that a “membrane Bohr effect” may participate in the initiation of mitogenesis.
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  • 158
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    Journal of Cellular Physiology 102 (1980), S. 119-127 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Rates of fluid pinocytosis by bovine aortic endothelial cells were measured during various manipulations of growth status in vitro. Sparsely seeded cultures grew exponentially until a confluent monolayer was formed, at which time growth slowed. This change in growth rate coincided with a decline in the rate of pinocytosis to about one-third that in the growing cultures. During the subsequent attainment of maximal cell density in the confluent monolayer, the pinocytic rate remained constant. There was close correlation between 3H-thymidine labelling indices, as measured by autoradiography, and the rates of pinocytosis. Mechanical “wounding” of the confluent monolayer resulted in cell migration and proliferation. Twenty-four hours after “wounding,” rates of pinocytosis per mg. cell protein were significantly enhanced. When regeneration of the monolayer was blocked by cytochalasin B, pinocytosis remained at the same rate as in the uninjured, confluent monolayer. These experiments support, and extend to endothelium, earlier observations that in growing cells pinocytosis proceeds at a higher rate than in non-growing, quiescent cells. Furthermore, they raise the possibility that the transendothelial transport of macromolecules such as lipoproteins by receptor-in-dependent fluid pinocytosis in vivo may be altered by the growth status of the endothelium.
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  • 159
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    Journal of Cellular Physiology 102 (1980), S. 141-153 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: An epithelioid clone of Chinese hamster (CHO) cells which is spontaneously transformed was exposed to the mutagen N-methyl-N-Nitro-N-Nitrosoguanidine (MNNG), and a fibroblastic variant, clone CHO-F2, was isolated. This clone is partially reverted in several of the in vitro properties characteristic of transformed cells. When compared to wild type CHO, CHO-F2 has a longer doubling time, a lower saturation density and less piling up at high cell density, and a higher serum requirement. CHO-F2 also elaborates less plasminogen activator and has more abundant microtubules and actin cables. On the other hand, both CHO and CHO-F2 grow in agar suspension (although CHO-F2 grows with a lower efficiency), both lack detectable LETS protein, and both are tumorigenic in nude mice. Thus, expression of the individual properties frequently associated with transformation and tumorigenicity can be dissociated.The most critical biochemical change in CHO-F2 appears to bean increase in the intracellular level of cyclic AMP, when compared to CHO, and several growth and morphological properties of CHO-F2 resemble those induced in wild type CHO exposed to exogenous dibutyryl cyclic AMP. Therole of cyclic AMP in expression of the transformed phenotype and the significance of individual in vitro parameters of transformation with respect to tumorigenicity are discussed.
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  • 160
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
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    Topics: Biology , Medicine
    Notes: The tripeptide H-Gly-His-Lys-OH (GHL) is a human plasma constituent which has been previously shown to modulate the growth and viability of a variety of cell types and organisms. Experimental observations presented herein indicate that GHL is complexed with the transition metal ions Cu++ and Fe++ in vivo and may exert its biological effects as a peptide-metal chelate. At physiological pH in vitro, GHL associates with ionic copper, cobalt, iron, molybdenum, manganese, nickel, and zinc, but has no affinity for calcium, manganese, potassium, and sodium. GHL acts synergistically with copper, iron, cobalt, and zinc to alter patterns of cell growth in monolayer cultures of a tumorigenic hepatoma cell line (HTC4). These transition metals induce cellular flattening and adhesion to support surfaces, and inhibit DNA synthesis and lactic acid production when growth is limited by reduction of serum concentrations in medium. These inhibitory effects are neutralized, and intercellular adhesion and growth are stimulated by GHL in medium at nanomolar concentrations. Cu and Fe are the most active metals when combined with GHL. The results suggest that the inability of HTC4 cultures to replicate without adequate concentrations of serum in medium may reflect deficiency of GHL and transition metals, which appear to form complexes prior to interaction with cells. Chelation of transition metals with GHL and, potentially, with other growth-modulating peptide factors in plasma or medium, may provide a mechanism for expression and regulation of biological activities influenced by transition metals and polypeptide growth factors. The observed effects of GHL-metal complexes, including stimulation of cellular adhesiveness to substratum (flattening) and intercellular attachment (monolayer formation), appear to satisfy requirements for growth of hepatoma cells in monolayer culture.
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  • 161
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    Journal of Cellular Physiology 102 (1980), S. 175-181 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: A procedure has been investigated for sorting viable cells according to their DNA content. Cells are stained with the U.V. activated fluorochromes 4′6-diamidino-2-pheylindole (DAPI), Hoechst 33258 or Hoechst 33342, and sorted with a Fluorescence Activated Cell Sorter. Hoechst 33342 is a suitable vital stain for a varietyof cell types. Hoechst 33258 and DAPI, however, are quantitative vital stains for CHO cells only. Cloning efficiency is unaffected by the sorting procedure, and these stains are not mutagenic at concentrations suitable for vital staining. Potential applications of this procedure to cell biology are discussed.
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  • 162
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    Journal of Cellular Physiology 102 (1980), S. 155-174 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Transport of 3-O-methyl-(1-3H)-D-glucose (3-OMG) was studied in primary cell cultures of the R3230AC rat mammary adenocarcinoma. Fastest rates of carrier-mediated 3-OMG transport (vc) were temporally associated with fastest cell growth, as were the rates of 3H-labeled thymidine, uridine, and leucine incorporation into macromolecules. The decrease in vc for 3-OMG observed as cultured cells approached quiescence was due to a 4-fold decrease in Vmax with the Km remaining relatively constant (4-9mM). Provided adequate time was allowed for cells to adapt, (6-12hr), the vc for 3-OMG transport was found to be inversely related to the concentration of glucose in the medium. Within 6 hours after switching cells from standard growth medium (5mM glucose) to medium containing no glucose (5 mM fructose), a 2-fold increase in vc for 3-OMG transport was observed in both fast and slow growing cells. The glucose-starvation induced increase in 3-OMG transport was due to an increase in Vmax; the Km remained constant, and was not significantly influenced by the presence of serum (10%) or insulin (5 m̈g/ml) or by [5 mM] galactose, mannose, fructose, 2-deoxy-D-glucose, 3-OMG, or mannitol. Readdition of glucose (5 mM) to transport-activated cells (deprived of glucose for 9-11 hr) resulted in a rapid return of 3-OMG transport to basal levels. In the presence of cycloheximide (10 m̈g/ml) or actinomycin D (10 m̈g/ml), this glucose-induced decline in carrier function was largely blocked. In fast-growing cells, the addition of either of these inhibitors, in the presence or absence of glucose, resulted in an initial rise in the rates of 3-OMG transport, followed by a linear decrease. Compared to fast-growing cells, the cycloheximide-induced increase in 3-OMG transport was greater and longer sustained in slow-growing cells. Regardless of their growth rate, cell cultures preincubated in medium containing glucose and cycloheximide exhibited decreases in 3-OMG transport when transferred to medium containing fructose, with or without actinomycin D. Slow-growing cells preincubated in medium containing fructose and cycloheximide exhibited an increase in 3-OMG transport when switched to medium containing fructose, whereas similarly treated fast growing cell cultures displayed a slight decrease. These experiments suggest that the adaptive regulation of glucose transport in these mammary tumor cell cultures occurred at the transcriptional level and was influenced by the rates of cellular growth. A model in which a metabolite of glucose acts to enhance the synthesis or stabilization of a mRNA species specific for a putative carrier inactivator protein is proposed.
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  • 163
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
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    Topics: Biology , Medicine
    Notes: Morphologic and biochemical studies were performed on cultures of bovine aortic endothelial cells which had developed a second growth pattern that has been referred to as “sprouting” (Gospodarowicz and Mecher, '78; Schwartz, '78). These morphologically atypical cells undergrew the intact endothelial cell monolayer and appeared only after the cells had reached confluence. They were ultra-structurally very similar to endothelial cells, but synthesized reduced amounts of fibronectin and a predominance of type I procollagen, rather than the types III and IV procollagens synthesized by monolayer endothelial cells.It is suggested that these cells represent phenotypically altered endothelial cells that differ in biosynthesis of secreted proteins and display a reduced contact-inhibition.
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  • 164
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    Journal of Cellular Physiology 102 (1980), S. 193-197 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Membrane potential was measured in perfused rat liver and was shown to increase from -33 ± 1.0 mV in livers from normal rats to -50 ± 1.1 mV in livers from rats 12 hr after partial hepatectomy. The hyperpolarization of the membrane in regenerating liver was no longer evident after perfusion with 1 mM ouabain for 5 min. Ouabain had a small (4 mV) depolarizing effect on membrane potential in normal liver. The potential measured in normal and regenerating liver decreased as a function of the external potassium concentration above 5 mM; however, the potential was more electronegative in regenerating liver compared to normal liver at all values of external potassium concentration, and the differences in potential between the two kinds of cells did not decrease at higher concentrations of external potassium. Thus, a plot of membrane potential vs external potassium concentration resulted in approximately parallel curves for the two different cell types. We conclude that hyperpolarization of the liver cell membrane is an early event during rat liver regeneration and results from an electrogenic Na-K pump.
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  • 165
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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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  • 166
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    Journal of Cellular Physiology 104 (1980), S. 21-26 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Mitogenesis of human blood lymphocytes in culture is inhibited by concentrations of ouabain that are approximately one order of magnitude lower than those that block Na and K transport. For example, the 50% inhibition (ID50) of Na-K transport, 280 nM, is seven-fold greater than the ID50 for RNA synthesis, DNA synthesis, or blastogenesis, ˜40 nM. Yet, inhibition of transport and consequent reduction in cell K is considered responsible for the effects of ouabain on mitogenesis. Since synthetic processes are assessed at least 24 hours after lymphocyte stimulation, this discrepancy could be explained by either 1) a progressive increase in K leak, or 2) a progressive inhibition of Na-K transport by ouabain during 24 hours of PHA treatment. We found that the lymphocyte membrane leak rate of K increased immediately after PHA treatment but did not increase further from 4 to 24 hours. In contrast, the ouabain sensitivity of 42K uptake was markedly increased with time: ID50 for 42K uptake of 35 nM at 24 hours as compared to 280 nM at 30 minutes. Measurement of ouabain binding revealed a seven-fold increase in the lymphocyte-associated ouabain after 24 hours compared to binding at 1 hour. These data indicate that the dose response of ouabain inhibition of active K transport and lymphocyte proliferation are closely correlated if one considers the slow membrane binding of ouabain at low concentrations.
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  • 167
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    Journal of Cellular Physiology 104 (1980), S. 61-72 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Events that are essential for progression through the G1 period begin immediately or shortly after resting chick embryo cells are given fresh medium with serum. The following observations support the contention that the critical events include the production of non-ribosomal RNAs: (1) Addition to the “shift-up” medium of either of two inhibitors of RNA formation, comptothecin or 5, 6-dichloro-1-β-D-ribofuranosylbenzimidazole, delays the onset of DNA replication by about the length of time the cells are exposed to the drugs. (2) Although entry into the S phase is delayed by the inhibitors, the slopes of the DNA response curves are identical to that of control cultures. (3) Neither drug reduces significantly the rate of overall protein synthesis. Observations (2) and (3) are taken to mean that expansion of the G1 period is not due to cell damage. (4) A third inhibitor of RNA synthesis, cordycepin, also delays passage of stimulated cells throgh the G1 phase, but, in this case, the length of the delay period is greater than that of the exposure period. (5) A low dose of actinomycin D does not impede movement towards the S phase, even though the synthesis of preribosomal RNA is considerably reduced.The possibility is considered that the essential G1 molecules are mRNAs.
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  • 168
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    Journal of Cellular Physiology 104 (1980), S. 41-46 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: A mouse embryo epithelial cell line, MMC-E, was used to study the effects of purified murine sarcoma growth factors (SGFs) on the growth of epithelial cells. Murine SGF was a partially purified preparation from the serum-free culture media of mouse fibroblasts transformed by Moloney murine sarcoma virus. SGFs stimulated DNA-synthesis in resting cells and induced them to grow to higher densities than control cells. With continued exposure to SGFs, MMC-E cells lost their postconfluency inhibition of division and formed small expanding foci. When the SGFs were removed and the cells were subcultured, they regained their normal phenotype, showing that the effects of SGFs are reversible on these epithelial cells. SGFs could also stimulate the MMC-E epithelial cells to grow in soft agar, like the syngeneic fibroblasts (MMC-F) from the same mouse embryo, but slower than the control fibroblastic clone. Microgram quantitites of SGFs were needed to stimulate soft ager growth of MMC-E cells. The results indicate that SGF can bring about a phenotypic change in the growth pattern of epithelial cells as well as fibroblastic cells.
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  • 169
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    Journal of Cellular Physiology 104 (1980), S. 53-60 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Transmembrane potential (Em) and alpha-aminoisobutyric acid (AIB) transport were measured in primary monolayer cultures of rat hepatocytes obtained from unoperated control rats and from rats 12 hr following partial hepatectomy. Measurements were performed 20-24 hr after plating the cells. The capacity of both kinds of cells to concentrate AIB depended upon extracellular sodium; however, the steady-state accumulation in regenerating cells was twice that of control cells. Transmembrane potentials, recorded with glass microelectrodes, were -13 ± 0.6 mV and -27 ± 1.6 mV in control and regenerating cells, respectively. Ouabain (1 mM) depolarlized regenerating cells to -18 ± 1.0 mV, but it had no effect on control cells. The initial rates of 1 mM AIB transport into control and regenerating cells were 1.2 ± 0.1 and 3.1 ± 0.1 nanomoles/mg protein × 4 min, respectively. Ouabain (1 mM) reduced the initial rate of AIB transport into regenerating cells to 2.7 ± 0.1 nanomoles/mg protein × 4 min, but it had no effect on AIB transport into control cells. Glucagon (10-7 M) added to control cells 12 hr before measurements hyperpolarized Em to -31 ± 1.3 mV and increased AIB transport rate to 3.1 nanomoles/mg protein × 4 min. The results suggest a relationship between increases in Em and increases in AIB transport in rat hepatocytes. An electrogenic Na-K pump may be involved in both of these events.
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  • 170
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    Journal of Cellular Physiology 104 (1980), S. 73-81 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Resting cultures of primary chick embryo cells have been labeled with 3H-leucine for the first 2 or 3 hours after feeding basal medium or basal medium supplemented with 10% dialyzed serum. The labeling patterns of the 3H-polypeptides of the soluble cell fraction have been compared by fluorography of two-dimensional gels. Large and consistent differences are seen in only three of the more than 900 spots that can visualized. This report concerns two of the three spots. The 3H contents of the two polypeptides (41,000 daltons, pI 7.1, designated 41-7.1, and 34,000 daltons, pI 6.2, designated 34-6.2) are increased by serum by about ten-fold. The highly selective effect of serum on the labeling of the two spots does not appear to be an artifact related to the extractability, solubility, or state of aggregation of the polypeptides. The radio-intensities of both polypeptides decrease markedly when the cells are labeled later than 3 hours after “shift-up”.Drugs that inhibit RNA synthesis and are known to stop the progression of the chick cells through the G1 period, camptothecin, cordycepin and 5,6-dichloro-β-D-ribofuranosylbenzimidazole, depress, with great specificity, the enhanced labeling of polypeptide 41-7.1 in the stimulated cells, and all but camptothecin have a similar action on polypeptide 34-6.2. A high level of actinomycin D (10 μg/ml), but neither a low level of the drug (0.02 μg/ml) nor 5-fluorouridine prevents the increased labeling of the two polypeptides in serum-fed cells. That 5-fluorouridine enters the chick cells and is converted to its active form is shown by the inhibition of the processing of pre-ribosomal RNA.The observations with the RNA inhibitors are at least consistent with the conclusions that the enhanced labeling of the two sports results from increased rates of synthesis of the polypeptïdes that depend upon mRNA production but not on the formation of ribosomal RNAs, and that the polypeptides play a role in the regulation of DNA replication in the chick cells.
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  • 171
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
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    Notes: We have investigated the effects of the tumor promoter 12-0-tetradecanoyl-phorbol-13-acetate (TPA) on plasminogen activator production, hexose transport and metabolism, and the incorporation of choline into the acid soluble pool and into phosphatidylcholine in suspension cultures of mouse L, mouse P388 leukemia, human HeLa, and Chinese hamster ovary cells, and in monolayer cultures of baby hamster kidney (BHK), mouse 3T3, mouse 3T6, and mouse P388D1 macrophage-like cells. BHK, 3T3, P388D1, and P388 cells produced plasminogen activator constitutively, but no significant production was observed in the other cell lines. Plasminogen activator production was induced or stimulated by TPA only in P388 cells (10- to 20-fold by 100 ng TPA/ml). On the other hand, phosphatidylcholine synthesis was stimulated by TPA only in HeLa cells, and hexose transport, as measured with 3-0-methyl-D-glucose, only in 3T3 and P388D1 cells, as well as in human lymphocytes. The stimulation of hexose transport occurred more rapidly than the induction of plasminogen activator production and seemed to be part of the mitogenic response of cells to TPA treatment. A stimulation of deoxyglucose uptake was similarly limited to 3T3 and P388D1 cells. A significant decarboxylation of carbon 1 of deoxyglucose occurred in P388 and P388D1 cells, but not in Novikoff cells, and any decarboxylation that occurred was not stimulated by TPA. The results indicate that the various investigated responses of cells to TPA are unrelated and occur independent of each other. The time courses of the biochemical responses also differ significantly.
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  • 172
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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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  • 173
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    Journal of Cellular Physiology 104 (1980), S. 187-197 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Bone marrow from barrier-sustained specific pathogen-free (SPF) CBA and C57BL/6 mice gave relatively low numbers of BFU-E colonies in methylcellulose culture, as compared to conventional mice. Addition of thymocytes to the marrow cultures increased the yield of BFU-E colonies more than fourfold in SPF mice but only 1.5-fold in conventional mice. Colony size was also increased.Increased yield of BFU-E colonies was also obtained by co-culture of bone marrow with lymph node cells or with bone marrow or spleen cells from 900R whole-body-irradiated mice. The effect appeared to be cellular rather than humoral. It was not reproduced by conditioned medium from thymus or pokeweed mitogen stimulated spleen cells. The helper effect of thymus cells was eliminated or reduced by freezing and thawing, or by 48 hours of incubation after irradiation. Treatment of bone marrow cells in vitro with anti-theta serum and complement did not decrease the number of BFU-E colonies. The putative helper cells appear not to be T cells, were non-adherent to the plastic culture dish, and were cortisone resistant and radioresistant. The low BFU-E colony yield from SPF mouse marrow is presumed to be largely the result of deficiency of these non-T helper cells in SPF bone marrow, rather than of BFU-E progenitor cells.
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  • 174
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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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  • 175
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    Journal of Cellular Physiology 104 (1980), S. 177-186 
    ISSN: 0021-9541
    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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  • 176
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    Journal of Cellular Physiology 104 (1980), S. 233-240 
    ISSN: 0021-9541
    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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  • 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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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Fibroblast Growth Factor (FGF) stimulates quiescent Swiss 3T3 cells to initiate DNA synthesis and divide. Cells begin to enter the S-phase after a lag of 13-15 hr, and the rate of initiation of DNA synthesis in the population can be quantified by a first order rate constant, k. A subsaturating concentration of FGF may establish the lag phase, while the value of k is dependent on the FGF concentration present during the second half of the lag phase. Insulin and hydrocortisone enhance the effect of FGF by increasing k without changing the lag phase, and they can act when added at any time after FGF. Prostaglandin E1 (PGE1) causes a decrease in k and a lengthening of the lag phase, and acts only when added during the first 8 hr. None of these agents stimulate DNA synthesis in the absence of FGF.These results show that the stimulation of growth by FGF follows the same basic pattern as was previously shown with Prostaglandin F2α (PGF2α). However, since hydrocortisone inhibits stimulation by PGF2α when added during the first 4 hr of the lag phase, there are clearly differences in some events stimulated by the two growth factors.
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  • 179
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    Journal of Cellular Physiology 104 (1980), S. 269-281 
    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 fresh medium and serum on protein synthesis in suspension-cultured HeLa cells after growth to high cell density (〉5 × 105 cells/ml) were studied. Cells which were resuspended in fresh medium plus serum and grown for 24 hours (control) were compared with cells grown for 2 hours after resuspension (stimulated). The spectrum of proteins being synthesized by control and stimulated cells does not appear to be grossly different; that is, the weight and number average molecular weights of newly synthesized whole-cell protein are about the same in both cultures. Also, no significant differences were observed in the number of ribosomes per polysome or in the fraction of total ribosomes in polysomes. However, the transit times (combined elongation and termination times) were found to differ significantly; the average transit time for control cells was 2.24 minutes, while the average transit time for stimulated cells was 1.26 minutes. (An appendex evaluating the methodology involved in measuring the transit time is included.) In agreement with the difference in transit time, the absolute rate of protein synthesis in stimulated cells was approximately 1.8 times the rate measured in control cells. These data are taken as evidence that under certain conditions, the rate of elongtion and/or termination of polypeptide chains limits the overall rate of translation, and that cells can respond to growth conditions by changing the elongation and/or termination rate of protein synthesis.
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  • 180
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    Journal of Cellular Physiology 104 (1980), S. 283-293 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: During the first three months after birth lambs produce sequentially three erthryocyte populations of different mean volume as demonstrated by electric sizing methods (Valet, Franz, and Lauf, J. Cell. Physiol. 94 (1978) 215). We separated by centrifugal elutriation the small volume population (type II) red cells of a genotypically low K+ (LK) lamb from the population containing the larger volume type I and III cells, an admixture of fetal (I) and adult (III) erythrocytes. The cells were separated at various time intervals after birth and analyzed with respect to their volumes, cation contents, and cation flux properties by means of 86Rb uptake. The effect of anti-L on K+ pump and leak fluxes was ascertained in unseparated and separated red cells. It was found that the small red cells of population II, transiently present for several weeks, were fully developed LK cells with K+ pumps responding characteristically to the stimulatory action of anti-L. In constrast, the larger cells of population I and III were of high K+ (HK) nature at early time points, the K+ pump activities approximately ten times higher than adult LK cells. These cells constitute an admixture of type I fetal HK cells, and type III reticulocytes which are precursors for the final type III adult LK cells, since anti-L had a small stimulatory effect. At later times, however, only adult type III LK cells predominated. The data directly support our earlier finding that the HK-LK transition in genotypically LK lambs is primarily governed by cellular replacement.
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  • 181
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    Journal of Cellular Physiology 104 (1980), S. 309-319 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The mode of permeation of uracil, 5-fluorouracil, and orotic acid into cells has been investigated in four established cell lines (Novikoff rat hepatoma, P388 mouse leukemia, mouse L, and Chinese hamster ovary cells) in attempts to assess the rate-determining step(s) in their incorporation into the nucleotide pool and nucleic acids. Uracil and 5-fluorouracil shared a saturable transport system (Km = 5 to 15 mM) capable of rapid equilibration of these substrates across the cell membrane (t1/2 at 25 in first-order range of concentration = 25 to 58 sec). Thus it seems unlikely that transport is limiting the incorporation hypoxanthine. Only the non-ionized form of fluorouracil was a substrate for the transporter; exclusion of charged pyrimidines may explain why orotate was not a substrate at physiological pH. Orotate permeated the cell membrane much more slowly (t1/2 = 2890 to 6930 sec); its permeation was apparently non-mediated and rate-determining in the conversion of extracellular orotate to intracellular nucleotides.
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  • 182
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    Journal of Cellular Physiology 104 (1980), S. 359-366 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Utilizing the high affinity interactions between pure 125I-L cell colony stimulating factor and its receptor(s) on the murine macrophage cell line J774, a murine radioreceptor assay (RRA) has been developed. The murine RRA selectively detects a colony stimulating factor (CSF) subclass (CSF-1) previously defined by murine radioimmunoassay (RIA) (E.R. Stanley, Proc. Nat. Acad. Sci., USA, 76:2969-2973 ('79)). CSF-1 stimulates macrophage production exclusively, and the occurrence of the CSF-1 receptor(s) appears to be restricted to cells of the mononuclear phagocytic system (L.J. Guilbert and E.R. Stanley, J. Cell Biol. 85:153-160 ('80)). The murine CSF-1 RRA failed to detect a variety of other CSF subclasses, growth factors, and hormones. In contrast to data obtained with the murine CSF-1 RIA, human CSF-1 (e.g., human urinary CSF) is detected by the mouse CSF-1 RRA almost as sensitively as murine CSF-1. In addition, there was an absolute correlation between CSF-1 levels determined by murine CSF-1 RRA and those determined by a human CSF-1 RIA for a variety of human CSF-1 sources. The murine CSF-1 RRA is a sensitive (sensitivity 5 units or 1.0 femtomole of CSF-1 protein), rapid, and highly specific assay for CSF-1 in both murine and human sources.
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  • 183
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    Journal of Cellular Physiology 104 (1980), S. 349-357 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Mouse myeloid leukemia cells (Ml) were induced to differentiate into mature macrophages and granulocytes by various inducers. The differentiated Ml cells synthesized and released prostaglandins, whereas untreated Ml cells did not. When the cells were prelabelled with [14C]arachidonate, the major prostaglandins released into the culture media were found to be prostaglandin E2, D2, and F2α in an early stage of differentiation, but the mature cells produced predominantly prostaglandin E2. The synthesis and release of prostaglandins were completely inhibited by indomethacin. Dexamethasone, a potent inducer of differentiation of Ml cells, did not induce production of prostaglandins in resistant Ml cells that could not differentiate even with a high concentration of dexamethasone. These results suggest that production of prostaglandins in Ml cells is closely associated with differentiation of the cells. Homogenates of dexamethasone-treated Ml cells converted arachidonate to prostaglandins, but this conversion was scarcely observed with homogenates of untreated Ml cells. Dexamethasone and the other inducers stimulated the release of arachidonate from phospholipids. Therefore, induction of prostaglandin synthesis during differentiation of Ml cells may result from induction of prostaglandin synthesis activity and stimulation of the release of arachidonate from cellular lipids. Lysozyme activity, which is a typical biochemical marker of macrophages, was induced in Ml cells by prostaglandin E2 or D2 alone, as well as by inducers of differentiation of the cells, but it was not induced by arachidonate or prostaglandin F2α. These results suggest that prostaglandin synthesis is important in differentiation of myeloid leukemia cells.
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  • 184
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    Journal of Cellular Physiology 104 (1980), S. 367-373 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Cyclic AMP (cAMP) levels were measured in 8-hour migrating wound epithelial and non-migrating epithelial cells of the newt. Tissues were collected in vivo and in vitro with and without epidermal-dermal separation by collagenase. Regardless of manner of collection and treatment, cAMP levels were always significantly higher in the migrating cells. Levels were also measured in 28-hour and 36-hour wound epithelia. There was a progressive decline in levels in wound epithelia between 8, 28, and 36 hours, suggesting that levels were in the process of returning to normal. When cells were treated with a dose of cAMP and theo-phylline previously shown to inhibit migration, levels of cAMP were much higher than any migrating epithelium. The fact that cAMP inhibits migration, yet migrating cells have higher cAMP levels, seems contradictory at first, but possible explanations are advanced to account for the apparent discrepancy.
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  • 185
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    Journal of Cellular Physiology 104 (1980), S. 375-389 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Cells of confluent cultures of the established pig renal epithelial line, LLC-PK1, accumulate α-methyl-D-glucoside against a concentration gradient. This transport system is strongly inhibited by phlorizin and 6-deoxy-D-glucose, moderately inhibited by phloretin, and only weakly inhibited by 3-0-methyl-D-glucose, paralleling the situation in mammalian kidney. The time course for the uptake of α-methyl-D-glucoside and for the carrier-mediated but passive uptake of 3-0-methyl-D-glucose are identical to those seen in mammalian kidney. Subconfluent cultures of LLC-PK1 cells are unable to accumulate α-methyl-D-glucoside, and their transport of this glucose analog is less sensitive to phlorizin inhibition than is the transport system in confluent cultures. Transmission electron micrographs show that cells from subconfluent cultures lack the microvillous surface seen in cells from confluent cultures. Cell density is thus a factor in the occurrence of structural and functional differentiated properties related to transport in these cells.
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  • 186
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Release of sulfated proteoglycans into the medium of fetal rat chondrocytes in monolayer culture was studied by contrasting the effects of 10% calf serum, long-acting cyclic nucleotides (8 Br-cAMP or DBcAMP), and lysine vasopressin (LVP). Eight hours after initiation of the experiment, the monolayer was pulsed for 2 hours with Na2[35SO=4], the radioactivity was chased, and the monolayer was reincubated for 6 hours with conditioned medium from replicate cultures. Immediately after labelling, the amount of newly synthesized sulfated proteoglycans was invariably higher in the insoluble matrix than in the medium compartment. Both additives selectively enhanced sulfate incorporation into chondroitin sulfate of the matrix when compared to serum controls, but only LVP stimulation caused increases in the medium. Remodeling (loss of cell layer and release into the medium at 6 hours) was suppressed by cAMP analogues and increased by LVP. This process was more active in cultures of lower cell density. Utilizing calibrated gel columns, no size difference of the glycosaminoglycans was found between the medium and cell layer compartments of the three treatment groups at the two time points. Because the cAMP analogues inhibit, while LVP stimulates cell division, our observations imply that the rate of degradation of the constraining matrix is increased when replication is favored, even when chondroitin sulfate synthesis is selectively stimulated.
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  • 187
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    Journal of Cellular Physiology 103 (1980), S. 149-157 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The methylation of nucleic acids has been investigated during the cell cycle of an asparagine dependent strain of transformed fibroblasts (BHK 21 HS 5). The synchrony was carried out by a partial asparagine starvation of cells for 24 hours. The amino acid supply induced all cells to enter synchronously the G1 phase. Methylation and DNA synthesis were respectively measured by pulsed [methyl-14C] methionine and [methyl-3H] thymidine incorporation. DNA methylation followed a biphasic pattern with maximal methyl incorporations during both S phase and mitosis.A partial desynchronisation induced the S phase of the second cycle to proceed before all the cells have achieved their division. Hydroxyurea was used in order to inhibit the DNA synthesis of cells entering the second cell cycle, which might interfer with the mitosis of the first one. The inhibitor was added either at the first beginning of cell division or during all the G1 phase. In both conditions it suppressed 3H thymidine incorporation of the second cycle. However, mitosis took place and methylations occurred as in previous experiments.The DNA methylation of the mitotic phase in the first cell cycle could thus be dissociated from the classical post-synthetic DNA maturation and did not correspond to any DNA methylation appearing in the course of the second cell cycle.
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  • 188
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    Journal of Cellular Physiology 103 (1980), S. 169-172 
    ISSN: 0021-9541
    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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  • 189
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Epidermal growth factor (EGF) added in a single dose (between 10-16 and 1.7 ± 10-9M) to neonatal rat hepatocytes in primary culture with subsequent incubation for 12 and 24 hours in Eagle's MEM fortified with 10% (v/v) FBS stimulated their entry into S and M phases, as shown by (3H)thymidine labeling and autoradiography and by a 1-hour exposure to colchicine (0.1 mM). Growth stimulation by EGF was detectable after 4 hours, peaking between 12 and 16 hours, and thereafter declining in intensity. Rat hepatocytes exposed for 72 hours (between the fourth and the seventh day in vitro) to no serum or to 10% fresh FBS possessed similar growth rates and absolute numbers in the cultures. A 24-hour exposure to 20 to 50% FBS stimulated hepatocytic DNA synthesis and mitotic activity and resulted (except for the 50% FBS treatment) in increased hepatocytes' numbers, which were relatively greater than the concurrent increases in connective tissue cell numbers. In serum-devoid medium EGF (10-11M) enhanced hepatocytic mitotic, but not DNA-synthetic activity. To be fully effective EGF required a 10% FBS addition to the medium, then eliciting within 24 hours a marked increase in hepatocytes' number with respect to cultures incubated with 10% serum only. When associated with 20 to 30% FBS, EGF stimulated parenchymal cell growth at rates slightly higher, but not significantly different, than those elicited by the same serum concentrations alone. However, when used in conjunction with 10 to 30% FBS, EGF preferentially increased the number of hepatocytes rather than that of non-parenchymal cells. Moreover, comparative proliferation kinetic studies showed that in the presence of 10% FBS, an equimolar (10-14M) mixture of EGF, insulin, and glucagon promoted an early and marked increase in the DNA-synthetic and mitotic activities of hepatocytes, which peaked after 8 hours. Within a 24-hour time lag this growth stimulation was as effective in increasing the final hepatocytes' number as was a 1000-fold higher EGF concentration, and was twice as active as either an equimolar (10-14M) mixture of the two pancreatic hormones or EGF by itself at 10-14M.These results show that the growth-promoting effect of EGF on primary neonatal rat hepatocytes is modulated by serum factor(s) and can be additively amplified by the simultaneous administration of subphysiological doses of glucagon and insulin.
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  • 190
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    Journal of Cellular Physiology 103 (1980), S. 159-168 
    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 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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  • 191
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    Journal of Cellular Physiology 103 (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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  • 192
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    Journal of Cellular Physiology 105 (1980), S. 489-502 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Rabbit reticulocytes were incubated with rabbit transferrin labelled with 59Fe and 125I in media in which the NaCl was replaced by other electrolytes or sucrose. Iron and transferrin uptake by the cells was affected by changes in the pH, ionic strength, ionic composition, and the osmolarity of the medium. Uptake was maximal at pH 7.4. A reduction in ionic strength produced by replacing NaCl with sucrose inhibited the uptake in a concentration-dependant manner, greatest inhibition occurring at lowest salt concentration. Similar results were obtained when KCl, LiCl, RbCl, Na2SO4, or K2SO4 were used instead of NaCl. Low ionic strength was found to inhibit the endocytotic uptake of transferrin labelled with colloidal gold, but had only a small effect on transferrin binding to cell membrane receptors. It was concluded that low ionic strength inhibits iron uptake primarily by blocking the endocytosis of transferrin. Three salts, NH4Cl, CaCl2, and MgCl2, produced different results from the above. NH4Cl inhibited iron uptake at all concentrations used. This action was due to an effect on the release of iron from transferrin, which appeared to be taken up by the cells in a normal manner. When the ionic strength of the sucrose medium was increased by adding low concentrations of CaCl2, iron uptake was greater than with equivalent concentrations of NaCl. However, with CaCl2 concentrations above 10 mM, iron uptake was inhibited, due to inhibition of transferrin uptake, possibly by blocking endocytosis. By contrast, MgCl2 stimulated iron uptake at all concentrations used. The results are discussed in terms of the possible effects of ionic strength, pH, and ionic composition of the extracellular fluid on the three main steps involved in iron uptake by immature erythroid cells: transferrin-receptor interaction, endocytosis, and iron release from transferrin.
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  • 193
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    Journal of Cellular Physiology 105 (1980), S. 1-6 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Uptake of α-methyl-D-glucoside (AMG) by LLC-PK1 cells is inhibited by the uncoupler p-trifluoro-methoxyphenyl-hydrazone (FCCP) and by the absence of extracellular Na+, indicating that the transport system is energy- and Na+ -dependent. We have previously demonstrated that transport of AMG by LLC-PK1 cells proceeds against a concentration gradient and is phlorizin-sensitive (Mullin et al., '80). Uptake of AMG was also inhibited by ouabain (OUA) but not by ortho-vanadate (VAN). Rubidium uptake also was affected by OUA but not by VAN. VAN, however, caused collapse of the three-dimensional domes of confluent LLC-PK1 monolayers much more rapidly and thoroughly than OUA. Since domes are presumably dependent upon the Na+ pump, yet VAN is not acting on transport-related functions of the OUA-sensitive (Na+ + K+)-ATPase, we hypothesize a direct effect of VAN on the water permeability of these cells. We also suggest that OUA does not act on these cells until domes collapse in normal course, and access of the OUA to the extracellular surface of antiluminal membranes is then achieved.
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  • 194
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    Journal of Cellular Physiology 105 (1980), S. 7-15 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: In order to better understand the mechanism of lymphocyte surface receptor redistribution induced by externally added ligands, polycationized ferritin (PCF), a nonconventional ligand, was tested using both fluorescence and electron microscopy for its ability to cause patching and capping of anionic molecules on the surface of both transformed and normal mouse lymphocytes. Binding of PCF at 0°C for 1 hour induces the appearance of patches; subsequent incubation at 37° for 30-60 minutes causes the formation of a cap structure with the lymphoid cells tested (T-lymphoma cells and splenic lymphocytes). Using various experimental treatments (e.g., sodium azide, cytochalasin B and D, colchicine, prefixation, and cold temperatures), PCF-induced capping has been found to be temperature sensitive, and to require metabolic energy and an intact cytoskeletal system. In addition, using double immunofluorescence techniques which involve rhodamine-labeled PCF and fluorescein-conjugated heavy meromyosin, it has been observed that the formation of the PCF-induced cap coincides with an accumulation of intracellular actin directly beneath the cap structure. Furthermore, agents such as dibutyryl cyclic AMP and theophylline, which cause an increase in intracellular cyclic AMP, have been shown to stimulate PCF-associated capping. This study suggests that increasing levels of intracellular cyclic AMP may activate, directly or indirectly, membrane-associated contractile elements required for the aggregation of membrane proteins into patches and caps.
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  • 195
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: We have compared the subcellular sites of H2O2 and presumably also superoxide-(O2-) production, and certain aspects of metabolic responses (O2 consumption, O2- production) of stimulated neutrophils from human blood and those elicited into guinea pig peritonea. Stimulation was accomplished with either opsonized zymosan or phorbol-12-myristate-13-acetate (PMA). Striking quantitative differences were observed between these cell types with regard to the increased respiration and O2- production observed during stimulation. These differences were most apparent when opsonized zymosan served as the stimulating agent. They were minimized when the soluble stimulating agent, PMA, was used. With either stimulus, the subcellular sites of H2O2 production were the same for both types of neutrophils, i.e., the plasmalemma and phagosomal membranes. No H2O2 production could be detected cytochemically in the absence of stimulation.Treatment of both unstimulated human blood and elicited guinea pig peritoneal neutrophils with the nonpenetrating, covalently linking reagent, p-diazobenzenesulfonic acid, failed to diminish O2- production upon subsequent stimulation, in contrast to a previous report. These data are discussed in terms of the possible cytological arrangements of the respiratory enzyme(s), and the different modes of stimulation of neutrophil metabolism by various agents. Ancillary data on elicited mouse peritoneal neutrophils are presented.
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  • 196
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    Journal of Cellular Physiology 105 (1980), S. 565-570 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: For malignant cells cultured from a human astrocytoma, electrophysiological characteristics of the plasma membrane included specific resistivity of 446.82 ± 279.5 ohm·cm2, specific capacitance of 0.758 ± 0.52 microfarads/cm2, time constant 0.318± 0.10 msec. The resting membrane potential averaged-14.07 ± 7.4 mV; the mean input resistance 8.1 ± 4.0 megohms. The average cell area was 1638 ± 585 ±2 for contactual and 1919 ± 989 ±2 for noncontactual cells. Changes in input resistance and resting membrane potential were observed with increasing time in culture, possibly reflecting cell cycling. There did not appear to be electrical coupling in this cell line.
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  • 197
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    Journal of Cellular Physiology 103 (1980), S. 271-278 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Adsorption of Sendai virus to HeLa cells induced in them an increased permeability to K+, Na+, Ca++, deoxyglucose, but not to fluorescein. The stimulation of uptake of 42K was temperature-dependent, did not occur below 15°C, and was not inhibited by ouabain.The virus-induced increase in the uptake and release of 42K and of 3H deoxyglucose could not be mimicked by treatment of cells with linoleic acid, a procedure which increased the fluidity of the cellular membranes. The stimulatory effect of 0.5 mM ATP on the release of deoxyglucose was enhanced several fold in the presence of Sendai virus. These results seem to indicate the possible involvement of membranal enzymes such as e.g. protein kinase in the permeability changes induced by Sendai virus.
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  • 198
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    Journal of Cellular Physiology 103 (1980), S. 299-303 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: To ascertain whether the fidelity of protein synthesis declines during cellular aging in vitro, we have developed a cell-free protein synthesizing system from cultured human fibroblasts which actively incorporates phenylalanine into acid-insoluble material upon addition of poly (U). The accuracy of poly(U)-directed protein synthesis was determined by comparing the ratio of leucine to phenylalanine incorporation in extracts of early- and late-passage fibroblasts derived from normal persons and from subjects with two genetic disorders of premature aging, progeria, and Werner syndrome. The results show no decline in translational fidelity at late passage or in prematurely aging cells, and thus fail to support the error catastrophe theory of cellular aging.
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  • 199
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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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  • 200
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    Journal of Cellular Physiology 103 (1980), S. 289-298 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Cytoplasmic calcium levels are believed to be important in blood platelet activation. Upon activation, the discrete marginal microtubule band, which maintains the discoid shape of non-activated platelets, becomes disrupted. Present studies demonstrate that the extent of assembly of the marginal microtubule band is related to cytoplasmic calcium levels. The divalent cationophore, A23187, causes platelet aggregation, secretion, and contraction by promoting calcium transport from intraplatelet storage sites into the cytoplasm. A23187 caused disassembly of platelet microtubules. Quantitation of electron micrographs revealed that numbers of microtubules were reduced by approximately 80% after A23187 treatment. Secondly, assembled microtubules in homogenates of platelets, in which microtubules were stabilized prior to homogenization, were decreased in favor of free tubulin in A23187-treated platelets. Thirdly, A23187 increased 14C-colchicine binding by intact platelets; this also indicated a shift in the microtubule subunit equilibrium to favor free, colchicine-binding tubulin subunits. In control experiments, A23187 did not affect the stability of platelet tubulin, the colchicine binding reaction, or the total tubulin content of platelets. Stimulation of colchicine binding depended on A23187 concentration (0.05-0.5 μM) and did not require extracellular calcium. A23187-stimulation of colchicine binding was blocked by dibutyryl cyclic AMP (0.80 mM) and/or 3-isobutyl-1-methylxanthine (50 μM) and by indomethacin (10 μM). Cyclic AMP or indomethacin also interferes with A23187-induced platelet activation, but indomethacin is not likely to completely inhibit the perturbation of intraplatelet calcium gradients by A23187. It is suggested that A23187-induced microtubule disassembly may be an indirect effect of calcium on microtubules.
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