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  • Life and Medical Sciences  (2,394)
  • 1
    ISSN: 0886-1544
    Keywords: chondrocytes ; matrix vesicle formation ; actin ; tubulin ; myosin ; vinculin ; alkaline phosphatase ; immunofluorescence ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Matrix vesicles, extracellular microstructures known to eb involved in endochondral calcification, are rich in alkaline phosphatase and have been shown to contain actin. The mechanism of matrix vesicle formation in chondrocytes in not well understood. Chondrocytes from the epiphyseal growth plate, when grown in primary culture, elaborate alkaline phosphatase-rich vesciles. We examined the distribution of the cytoskeletal proteins actin, myosin, tubulin, and vinculin at various time-points during culture using indirect immunofluorescent labeling. Concomitantly, the production of alkaline phosphatase-containing matrix vesicles was also followed. Cell morphology changed noticeably at two distinct stages during the 22-day culture period: Immediately after release from the growth plate the cells were founded, but after 4 days of cultre they began to spread out and acquire irregular shapes with distinct filopodia. By 13 datsm as tge cekks attaubed confluency, they reacquired a rounded, polygonal appearance. At all time-point, tubulin was seen as a dense network of microtubules radiating from the perinuclear region throughout the cytoplasm toward the cell periphery. Initially actin was seen in filamentous from, but displayed a punctate distribution focused at contact points during the cell-spreading stage of culture. After confluency, actin was concentrated at cell-cell junctions. Initially, vinculin was diffusely distributed, but became focused in multiple adhesion plaques and at the termini of filpodia during the cell-spreading stage of culture. Following confluency vinculin became concentrated at cell-cell junctions. Myosin was observed at all time-points in small, intensely localized focal points in the cytoplasmic region of the cells and was consistently absent from the nuclear and peripheral regions. The amount of myosin in the cells increased steadily with time in culture. Elaboration of alkaline phosphatase-rich vesicles, which corresponded closely with the rounded morphology of early and late stages of culture, may be correlated with contact inhibition.
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  • 2
    ISSN: 0197-8462
    Keywords: abnormalities ; chick embryos ; pulsed magnetic fields ; development ; Life and Medical Sciences ; Occupational Health and Environmental Toxicology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Physics
    Notes: Six independent experiments of common design were performed in laboratories in Canada, Spain, Sweden, and the United States of America. Fertilized eggs of domestic chickens were incubated as controls or in a pulsed magnetic field (PMF); embryos were then examined for developmental anomalies. Identical equipment in each laboratory consisted of two incubators, each containing a Helmholtz coil and electronic devices to develop, control, and monitor the pulsed field and to monitor temperature, relative humidity, and vibrations. A unipolar, pulsed, magnetic field (500-μs pulse duration, 100 pulses per s, 1-μT peak density, and 2-μs rise and fall time) was applied to experimental eggs during 48 h of incubation. In each laboratory, ten eggs were simultaneously sham exposed in a control incubator (pulse generator not activated) while the PMF was applied to ten eggs in the other incubator. The procedure was repeated ten times in each laboratory, and incubators were alternately used as a control device or as an active source of the PMF. After a 48-h exposure, the eggs were evaluated for fertility. All embryos were then assayed in the blind for development, morphology, and stage of maturity. In five of six laboratories, more exposed embryos exhibited structural anomalies than did controls, although putatively significant differences were observed in only two laboratories (two-tailed Ps of .03 and 〈.001), and the significance of the difference in a third laboratory was only marginal (two-tailed P = .08). When the data from all six laboratories are pooled, the difference in incidence of abnormalities in PMF-exposed embryos (∼25 percent) and that of controls ( ∼ 19 percent) although small, is highly significant, as is the interaction between incidence of abnormalities and laboratory site (both Ps 〈 .001). The factor or factors responsible for the marked variability of inter-laboratory differences are unknown.
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  • 3
    ISSN: 1040-452X
    Keywords: ES cells ; Pluripotent ; Bovine embryos ; Nuclear transfer ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Inner cell masses (ICM) from in vitro produced day 8 or 9 bovine blastocysts were isolated by immunosurgery and cultured under different conditions in order to establish which of two feeder cell types and culture media were most efficient in supporting attachment and outgrowth of the bovine ICM cells. The efficiency of attachment and outgrowth of the ICM cells could be markedly improved when STO feeder cells were used instead of bovine uterus epithelial cells, and by using charcoal-stripped serum instead of normal serum to supplement the culture medium. More than 20 stable cell lines were obtained. Some of these lines were examined by immunofluorescence for developmentally regulated markers. From these results we conclude that the cell lines resemble epithelial cells, rather than pluripotent ICM cells. The developmental potential of cells of one of the lines was tested in the nuclear transfer assay. The cell line could support the initial development of enucleated oocytes, but none of the reconstructed embryos passed the eight-cell block. © 1995 Wiley-Liss, Inc.
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  • 4
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Molecular Reproduction and Development 35 (1993), S. 293-301 
    ISSN: 1040-452X
    Keywords: Plasma membrane ; Proteolipid ; Epididymis ; Spermatozoa (ram) ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We examine here the biochemical properties and epididymal localization of a maturation dependent ram sperm surface antigen. A monoclonal antibody, ESA152, identifies an antigen that is present on the surface of ejaculated sperm, but is absent from testicular sperm. Crosslinking of the ESA152 antigen with bivalent antibodies induces the acrosome reaction, redistributing the antigen into the anterior region of the sperm head where it associates with the fusion product of the plasma membrane and the outer acrosomal membrane. The ESA152 antigen appears as a polypeptide of 18 kDa on immunoblots of SDS-polyacrylamide gels. The ESA152 epitope includes the sialic acid termini of N-linked oligosaccharides, as shown by its sensitivity to neuraminidase and endoglycosidase F. The ESA152 antigen is a highly hydrophobic integral membrane protein that resists aqueous extraction, partitions into the detergent phase of Triton-X-114, and solubilizes in chloroform-methanol mixtures. The anchoring of ESA152 is unaffected by phosphtidylinositol specific phospholipase C. The antigen is absent from extracts of caput and corpus epididymidis but appears abruptly in the first segment of the cauda. Immunofluorescence reveals that the ESA152 epitope first appears in clusters of cells in the luminal epithelium of the proximal cauda, prior to or concurrent with its appearance on sperm. © 1993 Wiley-Liss, Inc.
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  • 5
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Molecular Reproduction and Development 1 (1989), S. 164-169 
    ISSN: 1040-452X
    Keywords: Growth Hormone-Releasing Factor ; Metallothionein-I ; Lambs ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Pronuclei of fertilized sheep ova were injected with fusion genes consisting of the mouse metallothionein-I promotor/regulator ligated to either the structural gene for bovine growth hormone (mMTbGH) or to a minigene for human growth hormone-releasing factor (mMThGRF). From a total of 842 sheep ova injected with mMTbGH and transferred into recipient ewes, 47 lambs were born. Two of the lambs were transgenic with mMTbGH, and both had bGH mRNA present in liver, kidney, and gut. In one lamb, plasma growth hormone was as high as 700 ng/ml. From a total of 435 sheep ova injected with mMThGRF and transferred to recipients, 54 lambs were born and 9 fetuses were collected. Nine of the 63 had integrated the mMThGRF gene. One of the nine had high concentrations of immunoassayable hGRF in its plasma and high variable plasma concentrations of ovine growth hormone. The lamb that expressed the hGRF gene did not release GH in response to an hGRF challenge. Four of five fetal offspring of a nonexpressing mMThGRF transgenic ram also contained the mMThGRF gene and, like the sire, failed to express the gene as determined by either liver hGRF mRNA or by plasma hGRF. Growth of the single transgenic lamb expressing hGRF was similar to control lambs. These studies demonstrate efficient introduction of genes into the sheep genome and indicate that transgenes are expressed and heritable.
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  • 6
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The length of the oviduct, the thickness of its wall, and the height of its mucosal epithelium and cilia were measured in (a) 0-, 2-, 4- and six-month-old rabbits, (b) rabbits ovariectomized at birth and (c) ovariectomized, estrogen-treated rabbits. The length and external diameter of the oviduct increased progressively until four months of age, after which their rates of increase declined. The thickness of the oviductal wall at the uterotubal junction was twice as large as that of the isthmus at two months of age and six times as large at four and six months of age. The height of the mucosal epithelium in the fimbriae was less than that in other oviductal segments at birth, but exceeded that in other segments at six months of age. Ciliated cells and motile cilia were absent 24 hours after birth; they were first observed two months after birth. The cilia of fimbriae were shorter than cilia elsewhere in the oviduct. Neonatal ovariectomy retarded the development of the oviduct and the mesotubarium and caused pyknosis of ciliated and non-ciliated cells of the oviductal mucosa. Cells with scarcely motile cilia were present five and one-half months after neonatal ovariectomy.
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  • 7
    ISSN: 0730-2312
    Keywords: acute leukemia virus ; transforming gene ; DNA sequencing ; LTRs, nucleotides ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Avian mycloblastosis virus (AMV) is a replication-defective acute leukemia virus, requiring a helper virus to provide the viral proteins essential for synthesis of new infectious virus. The genome of the AMV has undergone a sequence substitution in which a portion of the region normally coding for the “env” protein has been replaced by chicken cellular sequences. These latter sequences are essential for the transforming activity of the virus. We have determined the complete nucleotide sequence of this region. Examination of the AMV oncogenic sequence revealed an open reading frame starting with the initiation codon ATG within the acquired cellular sequences and terminating with the triplet TAG at a point 33 nucleotides into helper viral sequences to the right of helper-viral-cellular junction. The stretch of 795 nucleotides would code for a protein of 265 amino acids with a molecular weight of 30,000 daltons. The eleven amino acids at the carboxy terminus of such a protein would be derived from the env gene of helper virus.
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  • 8
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Medium harvested from cultures of mouse L-cells contains “conditioning factor activity” (CFA) that may be detected by its ability to stimulate colony formation by mouse marrow cells in culture. The active material has been characterized and appears to be a glycoprotein with properties similar to those reported for the colony-stimulating activity in human urine. Characterization of trypsin-digested material indicated that only part of the molecule is essential for biological activity.The CFA has been partially purified from serum-free L-cell conditioned medium, and evidence has been obtained that radioactivity derived from labelled valine or glucosamine may be incorporated into the factor. L-cell conditioned medium appears to be a convenient source of partially-purified, highly active material, suitable for use in studies on its mechanism of action.
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  • 9
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Journal of Cellular Physiology 130 (1987), S. 460-465 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Mouse (MEL) and human (K-562) erythroleukemia cell lines can be induced to undergo erythroid differentiation, including hemoglobin (Hb) synthesis, by extra cellular hemin. In order to study the effect of extracellular hemin on intracellular ferritin and Hb content, we have used Mossabauer spectroscopy to measure the amount of 57Fe incorporated into ferritin or Hb and a fluorescent enzyme-linked immunosorbent assay (ELISA) to measure the ferritin protein content. When K-562 cells were cultured in the presence of a 57Fe source either as transferrin or citrate, in the absence of a differentiation inducer, all the intracellular 57Fe was detected in ferritin. When the cells were cultured in the presence of 57Fe-hemin, 57Fe was found in both ferritin and Hb. 57Fe in ferritin increased rapidly, and after 2 days it reached a plateau at 5 × 10-14g/cell. 57Fe in Hb increased linearly with time and reached the same value after 12 days. Addition of other iron sources such as iron-saturated transferrin, iron citrate, or iron ammonium citrate caused a much lower increase in ferritin protein content as compared to hemin. When K-562 cells were induced by 57Fe-hemin in the presence of 56Fe-transferrin, 57Fe was found to be incorporated in equal amounts into both ferritin and Hb. However, when the cells were induced by 56Fe-hemin in the presence of 57Fe-transferrin, 57Fe was incorporated only into ferritin, but not into Hb, which contained 56Fe iron. These results indicate that in K-562 cells, when hemin is present in the culture medium it is preferentially incorporated into Hb, regardless of the availability of other extra-or intracellular iron sources such as transferrin in ferritin. In MEL cells induced to differentiate by dimethylsulfoxide (DMSO) a different pattern of iron incorporation was observed; 57Fe from both transferrin and hemin was found to incorporate in ferritin as well as in Hb.
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  • 10
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Journal of Cellular Physiology 68 (1966), S. 35-44 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Using paired frog muscles it was shown that fluoride caused no observable change in the rate of oxygen consumption in Ca++-free Ringer's solution, but stimulated the rate in Ca++-containing Ringer's; the obvious explanation for this stimulation is the spontaneous activity of the muscle caused by the rapid lowering of Ca++ ion concentration when fluoride was added. Fluoride had no effect on potassium movement in these experiments. In a Ca++-free solution fluoride (0.03 M) inhibited the insulin + lactate-stimulated oxygen consumption of muscles which had been adapted to the absence of calcium by overnight soaking. In Ca++-containing Ringer's there was no net effect of fluoride, demonstrating the fact that the direct inhibition of oxygen consumption was cancelled by the indirect stimulation caused by the fluoride-lowering of the Ca++ ion concentration. Fluoride depressed the IL-induced K uptake but for this effect some calcium, albeit a very low concentration, was necessary. Fluoride did not alter the muscle glycogen concentrations.The only observable effect of calcium (in the absence of fluoride) was a rise in respiration caused by exposure to a sudden reduction in Ca++ ion concentration. Both the IL-stimulated O2 rate and K uptake were independent of Ca++ ions since they occurred in Ca++-free adapted muscles, in muscles exposed to a rapid reduction of Ca++ ions as well as in muscles in a Ca++-containing Ringer's solution.
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