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  • 1
    ISSN: 0886-1544
    Keywords: myofibrillogenesis ; myosin heavy chain ; myosin light chains ; transfection ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Central to the function of myosin is its ability to assemble into thick filaments which interact precisely and specifically with other myofibrillar proteins. We have established a novel experimental system for studying myofibrillogenesis using transient transfections of COS cells, a monkey kidney cell line. We have expressed both full-length rat α cardiac myosin heavy chain (MHC) and a truncated heavy meromyosin-like α MHC (sHMM) and shown that immunoreactive MHC proteins of the expected sizes were detected in lysates of transfected cells. Surprisingly, the full-length MHC formed large spindle-shaped structures throughout the cytoplasm of transfected cells as determined by immunofluorescence microscopy. The structures were not found in cells expressing the sHMM construct, indicating that their formation required an MHC rod. The spindle-shaped structures ranged in length from approximately 1 μm to over 20 μm in length and were birefringent suggesting that they are ordered arrays of thick filaments. This was confirmed by electron microscopic analysis of the transfected cells which revealed arrays of filamentous structures approximately 12 nm in diameter at their widest point. In addition, the vast majority of transfected MHC did not associate with the endogenous nonmuscle myosin light chains, demonstrating that myosin thick filaments can form in the absence of stoichiometric amounts of myosin light chains. © 1993 Wiley-Liss, Inc.
    Additional Material: 8 Ill.
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    The @Anatomical Record 230 (1991), S. 86-96 
    ISSN: 0003-276X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Although the artery wall consists of three distinct layers, only the structures of the intima and media have been well characterized. The adventitia has generally been overlooked. Our examination focused on the organization of elastin and collagen which are the major components of this tunic. Canine infrarenal aortas were excised, stretched to their in vivo length, then pressure fixed in formalin. Transverse, longitudinal, and frontal sections were prepared with specific elastin and collagen stains. Areas of adventitia in these sections were examined with LM, and interconnections between collagen and elastin were photographed at various magnifications. Subsequently, the slides were fractured for attachment to SEM stubs, and the coverslips were demounted. The identical areas were then examined with SEM using the LM micrographs as a guide to identify elastin and collagen. Whole mount aortic ring preparations were digested in formic acid for 72 and 96 h at 45°C to confirm adventitial elastin architecture. The adventitia was organized in alternating lamellae of collagen and elastin. The elastin lamellae consisted of continuous sheets of elastin with a longitudinal fibrillar substructure. Finer circumferential elastin fibers were also identified. These attached to both longitudinal elastin and adjacent collagen lamellae. Collagen lamellae were arranged in broad corrugated bands of fibrils. The unique architecture of the adventitia may explain some of the visco-elastic properties of the aorta in both normal and pathologic states.
    Additional Material: 30 Ill.
    Type of Medium: Electronic Resource
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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: Lipids tightly bound to cell and tissue proteins (lipoproteins) were investigated with the acetone-sudan procedure in cryostat-cut sections of normal and regenerating limb tissues of the adult newt, Diemictylus viridescens.Nuclear and cytolplasmic membranes of all tissues stained moderately black; the nucleo- and cytoplasm stained less intensely than their membranes. Connective tissue fibers of the dermis, mysial and neural sheaths, and tunics of blood vessels yielded intense responses. In striated muscle of the limb, myofibrils and associated striations responded strongly. Nerve myelin responded weakly.In the preblastemic regenerate, the fibrous adepidermal basement membrane terminated abruptly at the surface of amputation. A distal fibrocellular residue was evident in continuity with the retrograde degeneration of amputation-injured muscle: observations suggest a possible contribution to the fibrocellular reticulum from myofibrils. The regeneration blastema appears isolated from proximal limb tissues by the intervening fibrocellular reticulum. The response of the blastema cell is relatively weaker than that of other cells and tissues. Regenerating muscle is recognized by the appearance of prominently stained myofibrils in myoblastic extensions off the limb stump musculature. The matrix and chondrocytes of regenerating cartilage stain for lipoprotein, with the osteoid of osteogenic centers responding strongly.Polychrome responses were obtained from hyalin-bodies within interphase nuclei, and from chromosomes in mitosis, suggesting that chromosomal lipid has been stained.Supporting the lipidic character of the observed responses is the negative reaction following long term lipid extraction in warm chloroform: methanol.
    Type of Medium: Electronic Resource
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  • 4
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Journal of Cellular Physiology 177 (1998), S. 474-482 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: In this study we examined the regulation of cdk1 expression in normal human epidermal keratinocytes (HEKs) and neoplastic keratinocytes. Keratinocytes were growth-arrested by allowing the cells to grow to confluence or by treating them with interferon-gamma (IFNγ) or 12-O-tetradecanoyl phorbol-13-acetate (TPA). RT-PCR and Western blot analysis demonstrated that cdk1 was profoundly reduced in growth-arrested HEKs when compared with dividing HEKs. In contrast, a squamous carcinoma cell line, SCC25, did not growth-arrest in response to growth inhibitors and did not downregulate cdk1 expression. Transfection of HEKs with a reporter gene driven off a 2.5-kb fragment of the human cdk1 promoter indicated that the downregulation of cdk1 upon growth arrest was transcriptional. Deletion mapping of the cdk1 promoter indicated that a repressor region was located between -949--722 bp. This repressor region was not operative in the SCC25 cells. Examination of DNA:protein binding complexes by gel-shift analysis indicated that nuclear factors from both proliferative and growth-arrested cells bound to the DNA fragment spanning -949--722 bp. Further analysis revealed that this binding could be resolved into a constitutive and growth arrest-specific complex that bound in a similar fashion to regions spanning -892--831 bp and -831--774 bp, respectively. The putative growth arrest-specific complex was not found in contact-inhibited fibroblasts and was found at very low levels in SCC25 cells, indicating that the putative repressor binding was growth arrest-specific and possibly keratinocyte-specific. The binding complexes bound to these two fragments were localized, by competition analysis, to regions -874--853 bp and -830--800 bp. This is the first report of a transcriptional repressor being operative during keratinocyte growth arrest. J. Cell. Physiol. 177:474-482, 1998. © 1998 Wiley-Liss, Inc.
    Additional Material: 8 Ill.
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  • 5
    ISSN: 0148-7280
    Keywords: assisted fertilization ; t-locus mice ; in vitro fertilization ; infertility ; mouse ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Failure of epididymal spermatozoa from T/t mutant mice, but not from t/t individuals, to fertilize oocytes in vitro was partially overcome by opening a small aperture in the zona pellucida with acidified Tyrode's solution to permit direct access of the spermatozoon to the vitellus. This study provides a model system to evaluate requirements for successful zona drilling in the treatment of human infertility and further insights into the effects of the t complex on sperm fertility.
    Additional Material: 1 Ill.
    Type of Medium: Electronic Resource
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  • 6
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    BioEssays 6 (1987), S. 190-190 
    ISSN: 0265-9247
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Type of Medium: Electronic Resource
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  • 7
    ISSN: 0003-276X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Additional Material: 1 Ill.
    Type of Medium: Electronic Resource
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  • 8
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 27 (1994), S. 389-401 
    ISSN: 1059-910X
    Keywords: Biomineralization ; Greigite ; Magnetite ; Pyrite ; Single-magnetic-domain ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Electron microscopic studies on magnetosomes in magnetotactic bacteria have revealed much information on their composition, structure, and even the formation of their mineral phase. The mineral phases of the magnetosomes are of two general types: iron oxides and iron sulfides. Iron oxide-type magnetosomes contain particles of the ferrimagnetic mineral magnetite (Fe3O4) while the iron sulfide-type contain ferrimagnetic greigite (Fe3S4), greigite and non-magnetic pyrite (FeS2), or possibly ferrimagnetic pyrrhotite (Fe7S8). Regardless of their composition, the crystalline particles in magnetosomes have a narrow size range: approximately 35 to 120 nm. Magnetite crystals in this size range are single-magnetic-domains and confer a permanent magnetic dipole moment to the cell. The single-domain size range for greigite is not known but is probably similar to that for magnetite.The morphology of the particles in the bacterial magnetosomes appears to be species-specific. Morphologies of magnetite crystals in different species of magnetotactic bacteria include cubooctahedra, parallelepipedal (truncated hexahedral or octahedral prisms), and tooth- or bullet-shaped (anisotropic). Morphologies of greigite particles include cubo-octahedra and rectangular prismatic. The greigite-pyrite particles are generally pleomorphic with no consistent crystalline morphology. A membrane has been shown to surround the particles in some organisms and may be involved in the formation of the crystalline phase while also providing physical constraints on the size and the shape of the crystal. These results clearly indicate that the biomineralization process involved in the bacterial magnetosome, a good example of a self-assembled structure on a nanometer scale, is highly controlled by the organism. © 1994 Wiley-Liss, Inc.
    Additional Material: 11 Ill.
    Type of Medium: Electronic Resource
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  • 9
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Molecular Reproduction and Development 27 (1990), S. 332-336 
    ISSN: 1040-452X
    Keywords: Caput epididymis ; Micromanipulation ; In vitro fertilization ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Spermatozoa from the caput epididymis are known to be much less capable of fertilization when compared to sperm from more distal segments of the epididymis. The purpose of this study was to determine if two micromanipulative techniques, zona drilling (ZD) and a modification of partial zona dissection (PZD), could be used to enhance fertilization with caput epididymal sperm. A mouse in vitro fertilization model was used. Inseminating oocytes with 500-1,000 sperm/oocyte from the cauda epididymis as a control resulted in fertilization of 98 of 300 (32.6%) oocytes. Of those fertilized, 47 developed to the blastocyst stage (47.9%). Caput sperm fertilized 13 of 116 (11.2%) nonmanipulated oocytes. Only 1 of 13 developed into a blastocyst, while with oocyte ZD, caput sperm fertilized 24 of 144 (16.7%) oocytes, 50% of those fertilized developing to blastocyst (P=0.0129). When modified PZD was performed on oocytes, only one of 23 was fertilized, with no blastocyst development. These results indicate that acid Tyrode ZD enhances both fertilization and early embryonal development when caput epididymal sperm are used for insemination. These mouse studies suggest that ZD or other micromanipulation techniques may prove clinically useful in men with proximal epididymal obstruction where only caput sperm are available.
    Additional Material: 2 Ill.
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  • 10
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Adult newts placed in an atmospheric environment of 85% oxygen, saturated humidity, and at a temperature of 20 ± 1°C survived particularly well a 44-day test period. They did not succumb to “oxygen toxicity” as has been frequently reported for other vertebrate species.Having established the newt's tolerance of high oxygen atmosphere, the effect of oxygen on growth and development in the regenerating newt limb was investigated. Under the atmospheric conditions described above, and under 92% oxygen, the regeneration of adult newt limbs appeared to be retarded during the first 25 days after amputation when compared with regenerating limbs of control animals kept under a normal atmosphere of 21% oxygen (air). Thereafter, little or no difference could be discerned between the regeneration of experimental and control limbs.It is known that molecular oxygen participates directly in the hydroxylation of proline to hydroxyproline in the synthesis of collagen. Sectioned regenerates stained specifically for collagen were examined to determine if collagen synthesis was induced in experimental animals. Two regeneration-inhibited limbs of oxygenated newts showed cicatrical repair of the apical limb stump 25 days after amputation. However, the majority of the experimental animals revealed no obvious increase in collagen fibers.These results contraindicate any marked “oxygen toxicity” affecting the life of the newts, or regeneration of their limbs. It is suggested that a change in collagen fiber type might have been induced by the high-oxygen atmosphere. Investigations to test this hypothesis are currently underway.
    Additional Material: 1 Ill.
    Type of Medium: Electronic Resource
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