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Biomedical subjects

W T Chen

Publications and source records attributed to W T Chen.

At least 127 records · Page 7Linked to original sources
PubMed1985

Pilomatrixoma of the head and neck in children.

Pilomatrixoma, also known as calcifying epithelioma of Malherbe, is a benign tumor of hair cell origin. It is an unusual, but not particularly rare, tumor. Fifty percent or more of pilomatrixomas occur in the head and neck, and 40% occur in patients under 10 years of age. Pilomatrixomas may be mistaken for cysts, calcified lymph nodes, calcified hematomas or hemangiomas, or parotid gland tumors. We describe our experience with 15 children, 1 1/2-12 years of age, in whom we have excised pilomatrixomas from the head and neck.

Child↗
PubMed1985

Development of cell surface linkage complexes in cultured fibroblasts.

The possible role of a 140K membrane-associated protein complex (140K) in fibronectin-cytoskeleton associations has been examined. The 140K was identified by the monoclonal antibody JG22E. Monoclonal and polyclonal antibodies to the 140K showed identical patterns of binding to the cell membranes of fixed and permeabilized chicken embryonic fibroblasts; localization was diffuse, but with marked concentration in cell-to-extracellular matrix contact sites. Correlative localization with interference reflection microscopy and double-label or triple-label immunofluorescence showed that 140K co-distributed with extracellular fibronectin fibrils and intracellular alpha-actinin in microfilament bundles at extracellular matrix contact sites but tended not to co-localize with tropomyosin present in bundles at sites farther from adhesion sites. In addition, binding of antibodies to 140K, alpha-actinin, and fibronectin was excluded from vinculin-rich focal adhesion sites at the cellular periphery. A progressive development of cell surface alpha-actinin-140K-fibronectin associations was observed in early spreading cells. The anti-140K monoclonal antibody JG22E inhibited the attachment and spreading of both normal and Rous sarcoma virus-transformed chicken embryonic fibroblasts to a fibronectin substratum. However, the anti-140K monoclonal antibody became a positive mediator of cell attachment and spreading if it was adsorbed or cross-linked to the substratum. Our results provide the first description of a membrane-associated protein complex that co-localizes with fibronectin and microfilament bundles, and they suggest that the 140K complex may be part of a cell surface linkage between fibronectin and the cytoskeleton.

Animals↗
PubMed1985

Immunocytochemical localization of 140 kD cell adhesion molecules in cultured chicken fibroblasts, and in chicken smooth muscle and intestinal epithelial tissues.

A monoclonal antibody (JG22 MAb) that was previously raised to a chick embryo myogenic cell preparation had been shown to produce rounding and other morphological changes in myogenic cells in culture, and, in some cases, their detachment from the substratum. In other studies it was shown that the epitope recognized by JG22 was associated with a set of 140 kD cell surface glycoproteins. It is shown that this antigen occurs in a wide variety of cell types; in cultured fibroblasts, it is distributed equally between the dorsal and ventral cell surfaces shortly after plating, but appears to become concentrated on the ventral surface as cell spreading proceeds; by immunoelectron microscopic labeling experiments, it is absent from the focal adhesion contact sites formed by fibroblasts with their substrata and with one another, but is present in clusters at the edge of focal adhesions, and within the close contact sites and extracellular matrix contact sites; in smooth muscle cells, it is absent from the membrane-associated dense plaques, but is located in clusters at adjacent membrane sites; in intestinal epithelium, it is present in clusters at the basolateral membranes, but not at the microvilli or within junctional complexes of the brush border of the cell layers. These and other results are consistent with the suggestion that the antigen recognized by JG22 MAb is important cell adhesion molecules, and performs a characteristic function in a variety of cell-cell contacts and cell adhesions.

Animals↗
PubMed1985

Ultrafiltration hemodynamics in conscious, uremic dogs: effect of a decreasing plasma urea level.

The hemodynamic effects of a decreasing plasma urea level during hemodialysis were studied in acutely uremic, conscious dogs. Each dog was studied twice, during dialytic ultrafiltration against both a urea-free (U-) and a urea-supplemented (U+) dialysate. Plasma osmolality (-13 mosm/kg/H2O +/- 10 SD, p less than 0.01) and urea levels (-43 mg/dl +/- 20, p less than 0.01) decreased when using the (U-) dialysate, but were unchanged with the (U+) dialysate. At the end-point of volume depletion (MAP less than 80 mm Hg) total peripheral resistance index [U (+): 3,376 +/- 743 dyn . s X cm-5 X m2, U(-): 3,091 +/- 537, NS)] and blood volume [U(+): 62.7 +/- 7.4 ml/kg, U(-): 63.7 +/- 7.5, NS)] were comparable whether or not the plasma urea level had been allowed to decrease. The data suggest that an acutely decreasing plasma urea level during dialysis does not impair hemodynamic response to volume depletion in the model described.

Acid-Base Equilibrium↗
PubMed1984

Expression of transformation-associated protease(s) that degrade fibronectin at cell contact sites.

Virus-transformed fibroblasts show an increased production of proteases as well as loss of extracellular adhesive proteins. To determine whether these transformation-associated events are related, we investigated the capacity of Rous sarcoma virus-transformed cells (embryonic chick fibroblasts and mouse BALB/c 3T3) to degrade fibronectin by using a novel cross-linked protein substratum: fluorescence-labeled or radiolabeled fibronectin covalently linked to the surface of a fixed gelatin film. In serum-containing medium, the coupled fibronectin was not released when incubated without cells, and only a small amount was released when incubated with nontransformed cells. However, when transformed cells were seeded on the radiolabeled fibronectin-coupled substratum, there was a threefold increase in the time-dependent release of radioactivity into the medium. The released material was characterized as peptides with molecular sizes of less than 30,000 daltons. Correspondingly, growth of transformed cells on the rhodamine-fibronectin substratum resulted in the appearance of discrete negative fluorescent spots beneath the cells and along their migratory paths, whereas a uniform fluorescent carpet was detected with nontransformed cells. The release of radioactivity was partially inhibited by protease inhibitors, including alpha 2-macroglobulin, leupeptin, and benzamidine, but the negative fluorescent spots appeared unaffected by any of these inhibitors. However, both the release of radiolabeled peptides and the appearance of fluorescence-negative spots were inhibited by 1,10-phenanthroline at concentrations that did not affect cellular attachment and protein synthesis, thus supporting a role for proteases in localized degradation of fibronectin substratum. These fluorescence-negative spots coincided with sites of fibronectin disappearance as judged by indirect labeling with antibodies to cellular fibronectin. In addition, immunofluorescent analyses showed a correlation between vinculin localization and the negative fibronectin spots found under transformed cells, indicating that degradation occurs at cell substratum contact sites. These results can be correlated with other transformation-associated phenotypic changes, and are discussed in terms of the invasion of tumor cells into the extracellular matrix.

Animals↗
PubMed1984

Hemodynamic response to volume depletion in acutely uremic dogs.

Hemodynamic response to volume depletion by isolated ultrafiltration was compared in uremic (U) and nonuremic (N) conscious dogs. Fluid was removed at a constant rate until mean arterial pressure (MAP) decreased to less than 80 mmHg. Initial MAP was higher in the uremic dogs [132 +/- 8.6 (SD) mmHg] than in nonuremic controls (106 +/- 12, P less than 0.001). Initial cardiac index [U 4.97 +/- 0.831 X min-1 X m-2, N 4.44 +/- 0.62] and total peripheral vascular resistance index [(TPRI) U 2,160 +/- 353 dyn X s X cm-5 X m-2, N 1,976 +/- 420] were slightly, but not significantly, higher in uremic animals. Initial central venous pressure, wedge pressure, and plasma norepinephrine level were greater in the uremic dogs. At the end point of volume depletion, both uremic and nonuremic animals had achieved similar levels of TPRI, despite greatly attenuated or absent increases in plasma renin activity in the uremic group. At end point, blood volumes and plasma norepinephrine levels were comparable. The increase in pulse rate was higher in the uremic animals (59 +/- 37 pulses/min) compared with controls (25 +/- 52, P less than 0.05). In an additional group of uremic dogs, cardiovascular responses to hemorrhage and isolated ultrafiltration were compared and found to be similar, after allowance for blood viscosity changes had been made. The data suggest that in acutely uremic conscious dogs, despite reduced renin-angiotensin responses, hemodynamic adaptation to rapid volume depletion is not impaired.

Animals↗
PubMed1982

Acute effects of hemodialysis on echographic-determined cardiac performance: improved contractility resulting from serum increased calcium with reduced potassium despite hypovolemic-reduced cardiac output.

The hemodialysis session leads to reduction in circulating blood volume (TBV) and arterial pressure (BP) plus correction of electrolyte imbalance. The effect of these alterations on cardiac performance was evaluated in 18 patients with end-stage renal disease. Hemodialysis for 5 hours led to significant reduction (p less than 0.001) in weight TBV, and BP. Neither ejection fraction nor percentage fiber shortening was altered, whereas mean velocity of circumferential fiber shortening (mean VCF) and mean systolic ejection rate (MSER) were both significantly increased (1.17 +/- 0.20 to 1.38 +/- 0.28 circ/sec and 2.38 +/- 0.27 to 2.80 +/- 0.40 EDV/sec, respectively; p less than 0.001 for each). Since both venous return and systolic BP were decreased, increase in velocity of ventricular contraction implies enhancement of cardiac performance beyond what would be expected from alterations in ventricular filling and resistance to ejection. This enhancement is possibly related to concomitant reduction in serum potassium (p less than 0.001) and increase in serum calcium (p less than 0.005) achieved by hemodialysis.

Blood Pressure↗
PubMed1982

Immunoelectron microscopic studies of the sites of cell-substratum and cell-cell contacts in cultured fibroblasts.

Our object was to obtain information about the molecular structures present at cell-substratum and cell-cell contact sites formed by cultured fibroblasts. We have carried out double immunoelectron-microscopic labeling experiments on ultrathin frozen sections cut through such contact sites to determine the absolute and relative dispositions of the three proteins fibronectin, vinculin, and alpha-actinin with respect to these sites. (a) Three types of cell-substratum and cell-cell contact sites familiar from plastic sections could also be discriminated in the frozen sections by morphological criteria alone, i.e., the gap distances between the two surfaces, and the presence of submembranous densities. These types were: (i) focal adhesions (FA); (ii) close contacts (CC); and (iii) extracellular matrix contacts (ECM). This morphological typing of the contact sites allowed us to recognize and assign distinctive immunolabeling patterns for the three proteins to each type of site on the frozen sections. (b) FA sites were immunolabeled intracellularly for vinculin and alpha-actinin, with vinculin labeling situated closer to the membrane than alpha-actinin. Fibronectin was not labeled in the narrow gap between the cell surface and the substratum, or between two cells, at FA sites. Control experiments showed that this could not be ascribed to inaccessibility of the FA narrow gap to the immunolabeling reagents but indicated an absence or severe depletion of fibronectin from these sites. (c) CC sites were labeled intracellularly for alpha-actinin but not vinculin and were labeled extracellularly for fibronectin. (d) ECM sites were characterized by large separations (often greater than 100 nm) between the cell and substratum or between two cells, which were connected by long cables of extracellular matrix components, including fibronectin. In late (24-36 h) cultures, ECM contacts predominated over the other types. ECM sites appeared to be of two kinds, one labeled intracellularly for both alpha-actinin and vinculin, the other for alpha-actinin alone. (e) From these and other results, a coherent but tentative scheme is proposed for the molecular ultrastructure of these contacts sites, and specific functional roles are suggested for fibronectin, vinculin, and alpha-actinin in cell adhesion and in the linkage of intracellular microfilaments to membranes at the different types of contact sites.

Actinin↗
PubMed1982

Rapid cellular translocation is related to close contacts formed between various cultured cells and their substrata.

Interference-reflection microscopy combined with time-lapse cinemicrography was used to examine the relationship between cell-to-substratum contact patterns and the speeds of translocation for a variety of cell types. Rapid translocation of amphibian leukocytes (average speed = 9.0 micron/min), amphibian epidermal cells (7 micron/min) and teleost epidermal cells (7 micron/min) was found to correlate with patterns of broad grey close contacts. Similar contact patterns were found under freshly seeded (2 h) chick heart fibroblasts (moving 1-3 micron/min), the rapidly advancing (1-5 micron/min) margin of spreading human WI-38 fibroblasts, and isolated MDCK canine epithelial cells (0.5-1.0 micron/min). Conversely, numerous dark streaks of focal contact were found associated with the slow rate of translocation displayed by older cultures (72 h) of chick fibroblasts (less than 0.1 micron/min), well-spread WI-38 cells (less than or equal to 0.3 micron/min) and confluent MDCK cells (less than 0.01 micron/min). It is concluded that close contacts, but not focal contacts, are associated with rapid cellular translocation, and that the build-up of focal contacts is associated with reduced cellular translocation and maintenance of the spread cell shape.

Animals↗
PubMed1982

Masking of extracellular collagen and the co-distribution of collagen and fibronectin during matrix formation by cultured embryonic fibroblasts.

Embryonic chicken heart fibroblasts (CHF) were examined by double immunofluorescence in order to study the interactions of collagen and fibronectin. Double-labelling studies of permeabilized whole mounts of CHF, cultured for 15 min to 2 days after plating, detected extracellular collagen at very early times (15 min to 1 h) but not at times later than 3 h. Abundant fibronectin was seen at all times. These observations suggested that the collagen/procollagen antigen was being "masked' by matrix components at the later times. This possibility was tested in two ways: (1) immunolabelling of frozen sections of multilayered 4-day CHF cultures, which showed extensive extracellular collagen superimposable with fibronectin. (2) Limited trypsinization of 4-day CHF cultures, which allowed collagen to be immunolabelled concomitantly with a partial decrease in the degree of fibronectin labelling. The results are discussed in terms of the secretion and assembly of collagen and fibronectin in cultures of embryonic fibroblasts.

Animals↗
PubMed1981

Mechanism of retraction of the trailing edge during fibroblast movement.

Retraction of the taut, trailing portion of a moving chick heart fibroblast in vitro is an abrupt dynamic process. Upon retraction, the fibroblast tail always ruptures, leaving a small amount of itself attached to the substratum by focal contacts. Time-lapse cinemicrography shows that retraction produces a sudden, massive movement of both surface and cytoplasmic material toward a cluster of focal contacts near the main body of the cell. The appearance of folds on the upper cell surface at this time and the absence of endocytotic vesicles are consistent with this forward movement. Retraction of the trailing edge, either occurring naturally or produced artificially with a microneedle, consists of an initial fast component followed and overlapped by a slow component. Upon artificial detachment in the presence of iodoacetate, dinitrophenol, and sodium fluoride, and at 4 degrees C, the slow component is strongly inhibited and the fast one only slightly inhibited. Moreover of the bundles of microfilaments oriented parallel to the long axis of the tail seen in TEM. Most of the birefringence is lost during the fast phase and the rest during the slow phase of retraction. Concurrently, the bundles of microfilaments disappear during the fast phase of retraction and are replaced by a microfilament meshwork. All of these results are consistent with the hypothesis that the initial fast component of retraction is a passive elastic recoil, associated with the oriented bundles of microfilaments, and that the slow component of retraction is an active contraction, associated with a meshwork of microfilaments.

Adenosine Triphosphate↗
PubMed1981

Surface changes during retraction-induced spreading of fibroblasts.

Retraction of the trailing edge of an embryonic chick heart fibroblast results in an abrupt increase in protrusive activity at the leading edge of the cell. This increase was studied with time-lapse cinemicrography and scanning electron microscopy. Increased spreading following retraction results primarily from an increase in the duration of the extension phase of lamellipodial spreading. Much ruffling accompanies this increased spreading, particularly during its earliest phase. Upon retraction of the trailing edge, folds appear on the surface of the retracted tail and adjacent cell body and, soon after, microvilli-like structures appear as well. Once the moving cell has fully respread, however, the upper surface is once again smooth and free of folds and microvilli. Artificial detachment of a spreading lamella with a microneedle, and its consequent retraction, also causes increased protrusive activity of the remaining lamellae of the cell. These results are consistent with the hypothesis that retraction of one part of the cell makes surface membrane and cytoplasm available for forming protrusions elsewhere.

Animals↗