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  • 1
    ISSN: 0736-0266
    Keywords: Continuous passive motion ; Nutritional pathways ; Transsynovial transport ; Meniscus ; Life and Medical Sciences
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Thirteen freshly killed immature rabbits were used to study the effect of continuous passive motion (CPM) on regional nutritional pathways of the medial and lateral menisci and the transport of a solute from synovial fluid to the patellar tendon. A bolus of 35SO4 was injected into each knee joint cavity. The right knee underwent CPM for 1 h, whereas the left knee was immobilized (rest extremity). Both knees were then rapidly resected and immediately frozen. The medial and lateral menisci were removed and sectioned into anterior, middle, and posterior thirds for Group 1 animals; in Group 2 animals a portion of patellar tendon was harvested. Radioactivity as counts per minute per milligram of tissue was counted in a scintillation counter. The posterior portion of the lateral meniscus in the rest extremity had significantly higher uptake than the extremity that underwent CPM (p 〈 0.001). In the extremity at rest, the posterior third of the lateral meniscus had a significantly higher uptake than that of the middle third (p = 0.04). In Group 2 rabbits, the patellar tendon of the knee undergoing CPM had significantly higher uptake as compared with the patellar tendon of the knee at rest (p = 0.02). These results indicate that diffusion from synovial fluid to meniscal cells is an important mechanism of transport for low-molecular-weight nutrients such as sulfate; CPM does not facilitate this mechanism. However, even with a decreased amount of 35SO4 left in the CPM-treated joint, the amount taken up by these menisci was still roughly the same as that in the rested knee that had a higher concentration of 35SO4 available in the synovial cavity. Regional differences are small for the 1-h period examined in this study. Transarticular transport of sulfate from the joint space to an extraarticular tissue (patellar tendo) is facilitated by convective flow induced by CPM.
    Additional Material: 2 Ill.
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  • 2
    ISSN: 0749-503X
    Keywords: cell walls ; protease ; β-glucanase ; lysis ; yeast ; antifungal drugs ; glucan ; mannoprotein ; S. cerevisiae ; C. albicans ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The rate of formation of spheroplasts of yeast can be used as an assay to study the structural integrity of cell walls. Lysis can be measured spectrophotometrically in hypotonic solution in the presence of Zymolyase, a mixture of cell wall-digesting enzymes. The optical density of the cell suspension decreases as the cells lyse. We optimized this assay with respect to enzyme concentration, temperature, pH, and growth conditions for several strains of Saccharomyces cerevisiae. The level of variability (standard deviation) was 1-5% between trials where the replications were performed on the same culture using enzyme prepared from the same lot, and 5-15% for different cultures of the same strain. This assay can quantitate differences in cell wall structure (1) between exponentially growing and stationary phase cells, (2) among different S. cerevisiae strains, (3) between S. cerevisiae and Candida albicans, (4) between parental and mutated lines, and (5) between drug- or chemically-treated cells and controls. © 1998 John Wiley & Sons, Ltd.
    Additional Material: 4 Ill.
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  • 3
    ISSN: 0730-2312
    Keywords: monoclonal antibody ; A431 ; EGF receptor ; chromosomal location ; internalization ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: A monoclonal antibody of the IgG class, EGFR1, has been isolated using cells of the epidermoid carcinoma line A431 as immunogen. The A431 antigen recognized by EGFR1 has an apparent molecular weight of approximately 175,000, is a cell-surface molecule which can be specifically cross-linked to EGF, exhibits an EGF-stimulated protein kinase activity, binds to EGFR1 in a number of human cell lines to a degree which parallels EGF binding, and shows EGF-dependent internalization in A431 cells and human fibroblasts. We therefore conclude that EGFR1 is directed against an antigenic site on the human EGF receptor. EGFR1 is not mitogenic for human fibroblasts and does not inhibit EGF binding under a variety of assay conditions. The characterization of EGFR1 has allowed the unambiguous assignment of the structural gene for the human EGF receptor to chromosome 7. Preliminary results suggest that a convenient method for isolating a range of anti-EGF receptor monoclonal antibodies can be developed, based on a hybridoma supernatant screening assay in which positive supernatants bind selectively to a human-mouse cell hybrid containing human chromosome 7.
    Additional Material: 4 Ill.
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  • 4
    ISSN: 0003-276X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: This report describes a relationship between type II pneumonocytes and breaks in continuity in the alveolar septum of the human lung. Breaks in continuity of the septum are defined as gaps in the connective tissue matrix of the alveolar septum, with or without discontinuity of the accompanying alveolar epithelium. Septal connective tissue gaps accompanied by epithelial discontinuity are recognized as interalveolar pores of Kohn. When the discontinuity is confined to the connective tissue matrix, epithelial continuity may be maintained by either a single or a double layer of type I epithelium, by a type II cell, or by both type I and type II epithelial cells.Alveolar septal gaps were studied by electron microscopy on random sections in 26 adult human lung specimens and by serially sectioning and montaging the entire circumference of one alveolus to a depth of 103 μm (approximately one-half a normal alveolus) from one of the specimens. Fixation was by way of the airways in most specimens, but by vascular perfusion in the serially sectioned specimen and in seven others.In lungs studied by random sections, we found that the incidence of septal connective tissue gaps with epithelial continuity per specimen correlated with the incidence of pores (r = .468, P 〈 .016), and also with the incidence of type II cells (r = .422, P 〈 .025) in the specimen. Five precent of all type II cells observed in the random sections in the 26 specimens (103/1,955) occupied septal gaps, and 2.5% (49/1,955) were located at the rim of a pore. In contrast, in the single serially sectioned montaged alveolus, 69% of all type II cells occupied some type of septal gap, with 24% of all type II cells forming part of the rim of a pore. Over half of all pores in this alveolus were associated with a type II cell.We concluded that a relationship between the incidence of type II cells and gaps in the alveolar septum could be demonstrated on random sections in normal human lungs, which was much more obvious in a single serially sectioned hemialveolus. Serial section techniques of whole alveoli may be necessary to establish relationships that may not be apparent on random sections and that require the study of whole cells in continuity with their environment in order to be identified.The findings may be significant in suggesting a possible role of the type II cell in alveolar septal repair.
    Additional Material: 6 Ill.
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  • 5
    ISSN: 0192-253X
    Keywords: Signal transduction ; G-proteins ; adenylyl cyclase ; gene expression ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We have cloned and characterized three genes (CAR1, CAR2, CAR3) encoding potential cell surface, cyclic adenosine 3′:5′ monophosphate (AMP) receptors from Dictyostelium discoideum. The three proteins are predicted to be substantially similar in amino acid sequence throughout most of their transmembrane (TM) and loop domains but are distinctly different in their carboxyl terminal segments. In addition, all three genes possess an intron which interrupts an equivalent codon of TM3.CAR1 is expressed early in development when the cAMP relay system is being established. As development proceeds multiple size forms of CAR1 RNA are detected which apparently result from differences in their 5′-untranslated regions. Late in development levels of CAR1 RNA decrease. In contrast, CAR2 encodes a single sized RNA which is expressed only during postaggregative development. CAR3 expression is ∼10% of CAR1 during early development, is maximal during tight aggregate formation but declines thereafter. Only one size class of CAR3 mRNA is detected throughout development.Because RNA for each of the three genes is present in postaggregative cells, it was of interest to determine the cell type distribution of each RNA. Gene-specific probes were hybridized to RNAs isolated from cells of Percoll gradient-enriched prespore and prestalk fractions and relative levels of hybridization compared. CAR1 and CAR3 show approximately the same pattern of accumulation; a 3-4 fold enrichment in prestalk cells. CAR2, however, is highly enriched in prestalk cells, more than 10 fold relative to prespore cells.
    Additional Material: 5 Ill.
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  • 6
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    BioEssays 18 (1996), S. 955-963 
    ISSN: 0265-9247
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: In mammals, the Y chromosome induces testis formation and thus male sexual development; in the absence of a Y chromosome, gonads differentiate into ovaries and female development ensues. Molecular genetic studies have identified the Y-located testis determining gene SRY as well as autosomal and X-linked genes necessary for gonadal development. The phenotypes resulting from mutation of these genes, together with their patterns of expression, provide the basis for establishing a hierachy of genes and their interactions in the mammalian sex determination pathway.
    Additional Material: 1 Ill.
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  • 7
    ISSN: 0736-0266
    Keywords: Eccentric contractions ; Muscle soreness ; Fluid pressure ; Morphology ; Life and Medical Sciences
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: This investigation illustrates the morphological changes that take place following eccentric exercise and correlates those changes with intramuscular pressure readings. Eight healthy male subjects were asked to exercise their right lower leg anterior compartment eccentrically and their left concentrically. Four hundred submaximal contractions were performed in each exercise regimen over a 20-min period against a load corresponding to 15% of the individual's maximal dorsiflexion torque. Tissue fluid pressures were measured by the slit catheter technique before, during, and after exercise and 48 h later. Needle biopsies of both anterior tibialis muscles were also taken 48 h after completion of the exercise regimens. Overall morphology of the specimens revealed a greater cross-sectional fiber area (both type 1 and type 2) in the eccentrically exercised muscle as compared with the concentrically exercised muscle. Scant evidence of infiammation (only 1 of 8 of the “eccentric” muscle samples) and no fiber necrosis was observed. Fiber type proportions were equal on both sides and type 1 fiber biased (70%). Extremely large type 2 fibers were found in 4 of 8 subjects from the eccentric specimens. This incidence correlated significantly with the length of the time to return to resting pressure after eccentric exercise (r = 0.93, p 〈 0.001). The percentage of water content was significantly higher in the eccentrically exercised muscle. Based on these findings, we conclude that muscle fiber swelling is a predominant feature following eccentric exercise and is directly associated with delayed muscle soreness.
    Additional Material: 4 Ill.
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  • 8
    ISSN: 0192-253X
    Keywords: receptors ; transmembrane signalling ; Dictyostelium ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Using antibodies specific for the 3′, 5′-cyclic AMP (cAMP) cell surface receptor of Dictyostelium discoideum, we have screened γgtll expression libraries and isolated a series of cDNAs derived from cAMP receptor mRNA during early development. The identity of the cDNA clones was verified by multiple criteria: (1) β-galactosidase fusion proteins synthesized by isolated cDNA clones stain intensely with cAMP receptor directed antiserum, (2) these fusion proteins affinity purify antibodies specific for the cAMP receptor, (3) the cDNA probes hybridize to a 2 kb mRNA whose change in relative level of abundance during development parallels that of receptor mRNA as assayed by in vitro translation, (4) the 2 kb mRNA size equals that of receptor mRNA as determined by in vitro translation of size fractionated poly (A)+ RNA, and (5) RNA transcribed in vitro from cDNAs containing the entire protein-coding region produces a polypeptide by in vitro translation with an apparent molecular weight in close agreement with that of nascent cAMP receptor protein produced by in vitro translation of cellular RNA.The DNA sequence predicts an open reading frame of 392 amino acids. The deduced amino acid sequence contains seven domains enriched in hydrophobic residues. A model is proposed in which the cAMP cell-surface receptor traverses the lipid bilayer seven times in a pattern similar to that of other receptors, such as rhodopsin, which interact with G-proteins. The structural similarities suggest a gene family of related surface receptors from such evolutionarily diverse species as Dictyostelium, yeast, and mammals.
    Additional Material: 4 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: Triggering mechanisms for initiating density dependent inhibition of cell division in 3T3 cell monolayers are activated approximately two to three population doublings prior to cessation of cell division at monolayer confluency. This activation occurs at a critical contact cell density of approximately 8 × 103 cells/cm2. During this period there are selective controls on transport and storage of required low molecular weight nutrients. A possible correlation between orthophosphate and rates of cell division has been investigated. We have demonstrated a relationship between cellular concentrations of orthophosphate and initiation of density dependent inhibition of cell division. Prior to critical intercellular contact, the [Pi] in 3T3 cells is 10 mM. During critical contact, this concentration is quickly reduced to approximately 2 mM and remains at this concentration to confluency. Similar alterations do not occur in Py 3T3 cells, which maintain a concentration of approximately 2 mM Pi regardless of cell density. After confluent 3T3 cells are released from inhibition of cell division the [Pi] must increase several-fold before DNA synthesis commences. These are physiological changes in 3T3 cellular [Pi] as a function of cell density, and cannot be attributed to nutrient depletion, altered transport of Pi into the cell, increased [ATP], or increased [PPi] levels. The controlled modulation of [Pi] may regulate glycolysis and coordinate counter-ion changes (Ca++) may regulate mitochondrial activity.
    Additional Material: 2 Ill.
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  • 10
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Journal of Cellular Physiology 113 (1982), S. 8-10 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Trypsinized cells from embryonic chick neural retina redistributed concanavalin A receptors to patches and caps. Between 12 and 16 days of development, the ability to redistribute concanavalin A receptors declined. This restriction in mobility of the receptors was accompanied by changes in susceptibility to the capping-inhibitory drugs colchicine and cytochalasin B.
    Additional Material: 3 Ill.
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