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  • 1
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: In studying ultrastructural changes in metamorphosis-related degeneration of intersegmental muscles in Antheraea polyphemus, particular attention was directed to the mechanisms and timing of degradation of organelles and myofilaments. At emergence, the muscles are typical slowly contracting insect muscles, with a few dense body lysosomes and occasional autophagic vacuoles containing mitochondria. During the early phases of degradation the number of autophagic vacuoles, dense bodies, and lamellar bodies increases rapidly, along with an expansion of the Golgi system and the T system. Free glycogen particles and glycogenosomes are demonstrated by the PATAg test.Between 7 and 20 hours after ecdysis the T system continues to expand, the fibers subdivide, and the contractile system is degraded. Myofibrils fragment; myofilaments are not enclosed in isolating membranes at the time of their dissolution. The destruction of individual filaments occurs rapidly, with few intermediate stages being seen, while thick filaments tend to disappear earlier than thin filaments and Z-line material. The process is generalized and not confined to specific regions of the fiber. Autophagy destroys cell organelles in apparent synchrony with the first signs of nuclear pycnosis.By 20 to 30 hours after emergence, the fibers are reduced to lamellae of polynucleate sarcoplasm containing no organized contractile material. The sarcoplasm is filled with autophagic vacuoles containing mitochondria, dense lamellar or residual bodies, and ribosome-rich sarcoplasm. The number of mitochondria is drastically reduced at this time.In the final phases of involution (40-49 hours after emergence) shedding of the residual sarcoplasm precedes the expulsion of the pycnotic nuclei into the hemocoele.These results indicate that autophagy is responsible for the selective destruction of mitochondria, glycogen particles, ribosomes, and other organized sarcoplasmic structures. The one exception is the dissolution of the myofilaments, a process which remains undefined but which appears to be independent of lysosomal activity.
    Additional Material: 1 Ill.
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  • 2
    ISSN: 0730-2312
    Keywords: tyrosine phosphorylation ; insulin signaling ; tyrosine kinase ; confocal microscopy ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: The recently identified 53-kDa substrate of the insulin receptor family was further characterized in several retroviral-generated stable cell lines overexpressing the wild type and various mutant forms of the protein. To facilitate the study of its subcellular localization in NIH3T3 cells overexpressing insulin receptor, a myc epitope-tag was added to the carboxy terminus of the 53-kDa protein. Like the endogenous protein in Chinese hamster ovary cells, the expressed myc-tagged 53-kDa protein was found partially in the particulate fraction and was tyrosine phosphorylated in insulin-stimulated cells. Immunofluorescence studies showed for the first time that a fraction of the 53-kDa protein was localized to the plasma membrane. Confocal microscopy of cells double-labeled with antibodies to the insulin receptor and the myc epitope showed the two proteins co-localize at the plasma membrane at the level of light microscopy. Further analyses of the protein sequence of the 53-kDa substrate revealed the presence of a putative SH3 domain and two proline-rich regions, putative binding sites for SH3 and WW domains. Disruption of these three motifs by the introduction of previously characterized point mutations did not affect the membrane localization of the 53-kDa protein, its ability to serve as substrate of the insulin receptor, or its colocalization with the insulin receptor, suggesting these domains are not important in the subcellular targeting of the protein and instead may function in the interaction with subsequent signaling proteins. J. Cell. Biochem. 68:139-150, 1998. © 1998 Wiley-Liss, Inc.
    Additional Material: 7 Ill.
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  • 3
    ISSN: 0003-276X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: The morphology and development of junctional complexes between blastomeres of the preimplantation rabbit embryo were investigated using several approaches. Electron microscopic examination of embryos stained en bloc with uranyl acetate, and the study of junction permeability using horseradish peroxidase and lanthanum nitrate provided information on structure, intermembrane spacing and permeability of the junctional complexes. In addition, the freeze fracture technique was used with day 5 and day 6 blastocysts, since the large size of these embryos facilitated use of this method. These experiments showed that although rudimentary junctions were present between blastomeres of the early cleavage stages, effective tight junctions were not present until the blastocyst stage. Electron microscopic examination of thin sections revealed apical foci of membrane approximation or “fusion” between trophoblast cells by day 4. Freeze fracturing revealed a lattice of interconnecting ridges (on the A face) and grooves (on the B face) in the apical region between trophoblast cells of the day 5 blastocyst. This lattice formed a continuous band along the apical margin of each cell, and therefore constituted a zonula occludens. The zonula occludens of the day 5 blastocyst averaged 2-3 ridges per lattice, while day 6 blastocysts had lattices that averaged 5-6 ridges. Also seen in the freeze fracture replicas from the day 5 and day 6 blastocysts were local accumulations of intramembranous particles on the A face. These particles were often observed in aggregates similar to those of previously described gap junctions. It could not be determined whether these small regions of particles were true gap junctions or a possible primitive form of gap junction because the complementary pitted surfaces (B face pits) were not demonstrated.
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  • 4
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    The @Anatomical Record 189 (1977), S. 499-517 
    ISSN: 0003-276X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: A study of the development of hemopoiesis in the Mongolian gerbil (Meriones unguiculatus) was conducted in order to determine the temporal sequence, the organs involved and the cytology of blood cell formation in this species. Hemopoiesis in the intrauterine life of the gerbil can be divided into four phases based on the site of blood cell formation: (1) the vitelline phase, (2) the hepatic phase, including thymic histogenesis, (3) the splenic phase and (4) the medullary phase, with the development of secondary lymphoid tissues. At the onset of each of these phases a blast-like cell was identifiable in each hemopoietic organ which, because of its morphology and its presumed multipo-tentiality was classified as a “lymphoid cell.” In the yolk sac phase (gestational day 12) two generations of erythrocytes, a primitive and a definitive, are formed. The liver is by day 15 erythropoietic and megakaryopoietic, but later, a few gran-ulocytes are also found in its extravascular compartment. The thymus is exclusively lymphopoietic from the appearance of its earliest cells on day 15. Splenic hemopoiesis is initiated with the presence of lymphoid cells (day 20) followed later by the appearance of morphologically identifiable blood cell lines. Early normoblastic and granulocytic activity begins in the marrow cavities on day 23, though the marrow is not considered to be a source of circulating blood cells during fetal life. Lymph node histogenesis occurs during the last four days of gestation, first in the cervical region and then in other parts of the body. The finding of undifferentiated lymphoid cells in all organs at the initiation of hemopoiesis and in the peripheral blood throughout gestation is discussed in light of the migratory theory of hemopoiesis.
    Additional Material: 2 Tab.
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  • 5
    Electronic Resource
    Electronic Resource
    New York, N.Y. : Wiley-Blackwell
    Journal of Cellular Biochemistry 59 (1995), S. 1-10 
    ISSN: 0730-2312
    Keywords: Breast cancer ; carcinogenesis ; cervical cancer ; chemoprevention ; head and neck cancer ; lung cancer ; skin cancer ; retinoids ; retinoid receptors ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Retinoids, which include natural vitamin A (retinol) and its esters and synthetic analogues, are the best-studied class of agents in chemoprevention. There are more than 4,000 different retinoids which have a wide spectrum of preclinical activities, structures, pharmacological profiles, tissue distributions, receptor specificities, and toxicities. A number of retinoids have significant activity in many in vivo experimental systems including skin, bladder, lung, breast and oral carcinogenesis. In clinical trials, several retinoids have achieved significant activity in the reversal of head and neck, skin, and cervical premalignancy, and in the prevention of second primary tumors associated with head and neck, skin, and non-small cell lung cancer. Since 1984, our group has conducted a series of clinical trials to explore the chemopreventive potential of 13-cis-retinoic acid (13cRA) in the aerodigestive tract. We have conducted two consecutive randomized studies in subjects with premalignant lesions of the oral cavity. These studies showed that high-dose 13cRA alone can achieve significant short-term reversal of oral premalignancy, and that high-dose followed by low-dose 13cRA can maintain suppression of oral carcinogenesis. Three other randomized trials have confirmed significant retinoid activity in this human carcinogenic system. We also developed a randomized, placebo-controlled trial of adjuvant high-dose 13cRA in patients with head and neck cancer. Second primary tumor development was significantly decreased in the 13cRA group, but 13cRA had no impact on primary disease recurrence or survival. This presentation will update the current status of clinical trials and correlative laboratory studies of potential intermediate endpoint biomarkers in retinoid chemoprevention of aerodigestive tract carcinogenesis.
    Additional Material: 2 Ill.
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  • 6
    ISSN: 0002-9106
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: In order to extend the cytological information available on the structure of the interhemal membrane in cricetid rodents, in particular, and myomorph rodents, in general, we have examined the fine structure of the placental labyrinth in five genera of cricetid rodents (Lemmus, Dicrostonyx, Clethrionomys, Microtus and Peromyscus) and one genus of murid rodent (Acomys). Small pieces of labyrinth from near-term placentas were fixed in glutaraldehyde and osmium tetroxide and processed for electron microscopy. The interhemal membranes of all the species examined were hemotrichorial. The outermost layer of trophoblast, bordering the maternal blood spaces, was celular and often contained some patent fenestrae, whereas the middle and inner trophoblastic layers were apparently syncytial. The outermost trophoblastic layer of all the species contained abundant granular endoplasmic reticulum. The intercellular space between the outer and middle layers was variable in width, primarily due to the complex folding of the surface of the middle layer. The surfaces of the middle and inner layers were closely apposed. The middle trophoblastic layer in several species contained filamentous “glomerular bodies” and the innermost layer, particularly of Peromyscus, contained smooth membranous whorls. A basal lamina separated the innermost trophoblastic layer from the fetal capillary endothelium. The endothelial cells in most of the species contained fenestrated regions. These observations are compared to those on other hemochorial placentas, and common features of cricetid (and myomorph) rodent placental fine structure are emphasized.
    Type of Medium: Electronic Resource
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  • 7
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Journal of Cellular Physiology 99 (1979), S. 417-425 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The biosynthesis of NAD has been examined in 3T3 cells. The net synthesis of pyridine nucleotides does not occur when cells are cultured in the absence of performed pyridine ring compounds; however, growth continues normally for up to four cell doublings resulting in cells with a total pyridine nucleotide content that is reduced by as much as 12-fold. The mechanism that adjust the relative amounts of NADP and NAD are also altered such that the amount of NADP relative to NAD increases 5-fold. Both nicotinate and nicotinamide can be used as a precursor for NAD biosynthesis, however nicotinate is utilized less efficiently than nicotinamide. The presence of functional pathways for the biosynthesis of NAD from nicotinate via nicotinate mononucleotide and nicotinate adenine dinucleotide and from nicotinamide via nicotinamide mononucleotide has been demonstrated by identification of biosynthetic intermediates following short term exposure of cells to radiolabelled precursors. When cells are grown in Dulbecco's modified Eagle's medium which contains 33 μM nicotinamide the biosynthesis of NAD proceeds by a single pathway with nicotinamide mononucleotide as the only intermediate. Nicotinamide ribonucleoside which previously has been postulated to be an intermediate in the conversion of nicotinamide to NAD is not an intermediate in NAD biosynthesis.
    Additional Material: 5 Ill.
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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: Minute virus of mice (MVM), a non-defective parvovirus, has been shown to infect cultures of non-pluripotent differentiated teratocarcinoma-derived cells, but pluripotent (and “nullipotent”) embryonal carcinoma cells derived from the same teratocarcinoma resist MVN infection. Somatic cell hybrids between an embryonal carcinoma line and Friend erythroblastic leukemia cells are also resistant to MVM, even though Friend cells are susceptible. Among three blastocyst-derived lines tested, only one, a parietal yolk sac cell line, resists MVM infection. These results suggest that teratocarcinoma cultures may provide useful systems in which to study the cellular factors which mediate susceptibility to this teratogenic and oncolytic virus.
    Additional Material: 1 Ill.
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  • 9
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: To investigate the possible role of mast cells (MC) in regulating leukocyte adhesion to vascular endothelial cells (EC), microvascular and macrovascular EC were exposed to activated MC or MC conditioned medium (MCCM). Expression of intercellular and vascular adhesion molecules (ICAM-1 and VCAM-1) on EC was monitored. Incubation of human dermal microvascular endothelial cells (HDMEC) and human umbilical vein endothelial cells (HUVEC) with activated MC or MCCM markedly increased ICAM-1 and VCAM-1 surface expression, noted as éarly as 4 hr. Maximal levels were observed at 16 hr followed by a general decline over 48 hr. A dose-dependent response was noted using incremental dilutions of MCCM or by varying the number of MC in coculture with EC. At a ratio as low as 1:1,000 of MC:EC, increased ICAM-1 was observed. The ICAM-1 upregulation by MCCM was 〉90% neutralized by antibody to tumor necrosis factor alpha (TNF-α), suggesting that MC release of this cytokine contributes significantly to inducing EC adhesiveness. VCAM-1 expression enhanced by MCCM was partly neutralized (70%) by antibody to TNF-α; thus other substances released by MC may contribute to VCAM-1 expression. Northern blot analysis demonstrated MCCM upregulated ICAM-1 and VCAM-1 mRNA in both HDMEC and HUVEC. To evaluate the function of MCCM-enhanced EC adhesion molecules, T cells isolated from normal human donors were used in a cell adhesion assay. T-cell binding to EC was increased significantly after exposure of EC to MCCM, and inhibited by antibodies to ICAM-1 or VCAM-1. Intradermal injection of allergen in human atopic volunteers known to develop late-phase allergic reactions led to marked expression of both ICAM-1 and VCAM-1 at 6 hr, as demonstrated by immunohistochemistry. These studies indicate that MC play a critical role in regulating the expression of EC adhesion molecules, ICAM-1 and VCAM-1, and thus augment inflammatory responses by upregulating leukocyte binding. © 1995 Wiley-Liss Inc.
    Additional Material: 11 Ill.
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  • 10
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Journal of Cellular Physiology 162 (1995), S. 388-399 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Although basic fibroblast growth factor (bFGF/FGF-2) is found outside cells, it lacks a conventional signal peptide sequence; the mechanism underlying its export from cells is therefore unknown. Using a transient COS-1 cell expression system, we have identified a novel membrane-associated transport pathway that mediates export of FGF-2. This export pathway is specific for the 18-kD isoform of FGF-2, is resistant to the anti-Golgi effects of Brefeldin A, and is energy-dependent. In FGF-2-transfected COS-1 cells, this ER/Golgi-independent pathway appears to be constitutively active and functions to quantitatively export metabolically-labeled 18-kD FGF-2. Co-transfection and co-immunoprecipitation experiments, using a vector encoding the cytoplasmic protein neomycin phosphotransferase, further demonstrated the selectivity of this export pathway for FGF-2. When neomycin phosphotransferase was appended to the COOH-terminus of 18-kD FGF-2, the chimera was exported. However, the transmembrane anchor sequence of the integral membrane glycoprotein (G protein) of vesicular stomatitis virus (VSV) blocked export. The chimeric protein localized to the plasma membrane with its FGF-2 domain extracellular and remained cell-associated following alkaline carbonate extraction. Taken together, the data suggest that FGF-2 is “exported” from cells via a unique cellular pathway, which is clearly distinct from classical signal peptide-mediated secretion. This model system provides a basis for the development and testing of therapeutic agents which may block FGF-2 export. Such an intervention may be of considerable use for the treatment of angiogenesis-dependent diseases involving FGF-2. © 1995 Wiley-Liss, Inc.
    Additional Material: 7 Ill.
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