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L Terracio

Publications and source records attributed to L Terracio.

At least 73 records · Page 4Linked to original sources
PubMed1988

Expression of platelet-derived growth factor receptors is induced on connective tissue cells during chronic synovial inflammation.

The tissue distribution of the receptor for platelet-derived growth factor (PDGF) was investigated by immunohistochemistry on frozen sections from normal and inflamed synovial tissue using monoclonal antibodies to the receptor. Non-inflamed synovial tissue showed no staining, indicating that PDGF receptor expression is low or absent in normal tissue. In contrast, tissue from synovitis with prominent neovascularization showed a strong staining in the tunica media of the proliferating blood vessels as well as on connective tissue cells in the stroma. Tissue from synovitis with prominent proliferation of synovial lining showed intense staining for PDGF receptors in fibroblast-like cells of the lining and a less intense staining on vascular and connective tissue cells deeper in the stroma. Staining for PDGF receptors was also intense in the pannus tissue close to infiltrated bone and cartilage. In all these forms of synovitis, PDGF receptor staining was associated with increased HLA-DR staining and infiltration of macrophages and T lymphocytes. The finding that PDGF receptor expression is induced in conjunction with the chronic synovial inflammation associated with rheumatoid arthritis and some other forms of arthritides suggests that PDGF may play a role in the stimulation of mesenchymal cell proliferation that often accompanies chronic inflammatory disease.

Chronic Disease↗
PubMed1987

Transformation of rabbit arterial smooth muscle cells with simian virus 40.

In vitro studies were carried out to induce viral transformation of vascular smooth muscle cells. Cultured rabbit arterial smooth muscle cells were infected with simian virus 40 (SV 40), and transformed cultures were produced that exhibit altered morphology, increased growth rate and plating efficiency, growth on semi-solid substrate, and chromosomal abnormalities. Nuclear SV 40 T-antigen was detected in all cells of these cultures. Muscle-specific actin was identified by a specific monoclonal antibody suggesting retention of smooth muscle cell characteristics by the transformed cells. Significant cytoplasmic lipid accumulation occurred in transformed cells incubated with beta-very low density lipoprotein, as revealed both by chemical analyses and Nile Red lipid staining of the culture. The transformed smooth muscle cells grow permanently in cell culture. Our investigations show that arterial smooth muscle cells transformed with SV 40 virus exhibit altered phenotypic properties distinct from that of normal arterial smooth muscle cells.

Animals↗
PubMed1986

Direct stimulatory effects of insulin-like growth factor-I on Leydig cell steroidogenesis in primary culture.

Insulin-like growth factor-I (IGF-I) in concentration as low as 10 ng/ml significantly increased basal testosterone formation and 100 ng/ml of IGF-I increased testosterone production more than two fold in primary cultures of purified mature Leydig cells. IGF-I also markedly potentiated hCG-induced testosterone formation in a dose-dependent manner. Furthermore, IGF-I enhanced 8-bromo cyclic AMP induced steroidogenesis and hCG-stimulated cyclic AMP formation. The binding of 125I-IGF-I to purified Leydig cells was linear with a binding affinity of 0.56 +/- 0.07 X 10(9) M-1 and a capacity of 167 +/- 10.2 fmol/mg protein. Insulin and multiplication-stimulating activity were less potent than IGF-I in competing the binding of 125I-IGF-I to purified Leydig cells. This suggests that Leydig cells contain specific type I IGF receptor and IGF-I could modulate Leydig cell steroidogenesis.

Animals↗
PubMed1986

Identification of fibroblasts as a major site of albumin catabolism in peripheral tissues.

Rat serum albumin has been labeled with dilactitol-125I-tyramine, (125I-DLT) a radioactive tracer which remains entrapped within lysosomes following cellular uptake and degradation of the carrier protein. Similar kinetics of clearance from the rat circulation were observed for albumin labeled conventionally with 125I or 125I-DLT-albumin, both proteins having circulating half-lives of approximately 2.2 days. In contrast, the recovery of whole body radioactivity had half-lives of approximately 2.2 and 5.1 days, respectively, for the two protein preparations, indicating substantial retention of degradation products derived from catabolism of 125I-DLT-albumin. Measurement of total and acid-soluble radioactivity in tissues 2 or 4 days after injection of 125I-DLT-albumin revealed that skin and muscle accounted for the largest fraction (50-60%) of degradation products in the body. Fibroblasts were identified by autoradiography as the major cell type containing radioactive degradation products in skin and muscle. Fibroblasts were isolated from skin by collagenase digestion, followed by density gradient centrifugation. The amount of acid-soluble radioactivity recovered in these cells was in excellent agreement with that predicted based on acid precipitation of solubilized whole skin preparations. These studies demonstrate for the first time that fibroblasts are a major cell type involved in the degradation of albumin in vivo.

Animals↗
PubMed1986

Transformation of prostatic epithelial cells and fibroblasts with cadmium chloride in vitro.

Primary cultures of fibroblasts and epithelial cells were established from rat ventral prostate (RVP), canine (CP), baboon (BP), and human (HP) prostates, and were used in an assay system to evaluate cadmium chloride (CdCl2) cytotoxicity in vitro. Fibroblasts were always more susceptible to CdCl2 cytotoxicity than the epithelial cells of the same species. There was a distinct species variability to CdCl2 cytotoxicity, with RVP cells being greater than 200 times more susceptible than HP. Primary cultures treated with CdCl2 were subcultivated to establish cell lines. Only RVP fibroblast and epithelial cells resulted in permanent cell lines. Two fibroblast and two epithelial cell lines were derived from CdCl2-treated RVP cell cultures. The epithelial cell lines possessed tonofilaments, desmosomes and keratin. All four cell lines were resistant to CdCl2, had different karyotypes and an excess of chromosome 13. These results demonstrate the transforming potential of cadmium on prostate cells. The role of metallothionein and the significance of extra chromosomes 13 are discussed as possible factors of cadmium resistance.

Animals↗
PubMed1986

Characterization of insulin and insulin-like growth factor I receptors of purified Leydig cells and their role in steroidogenesis in primary culture: a comparative study.

Characterization of insulin and type I insulin-like growth factor (IGF-I) receptors and the effects of insulin and IGF-I on steroidogenesis were evaluated by using purified adult Leydig cells from Sprague-Dawley rats. Purified Leydig cells were found to contain both high and low affinity binding sites for insulin, with Ka values of 1.08 X 10(9) and 1.1 X 10(7) M-1, respectively. Using affinity cross-linking of [125I]iodoinsulin to plasma membrane insulin receptor, several bands were identified by autoradiography under nonreduced conditions with mol wt of 230,000, 280,000, and 300,000. After reduction with 50 mM dithiothreitol, only one band was identified with a mol wt of 130,000, consistent with the alpha-subunit of insulin receptor. Purified Leydig cells also contain specific type I IGF receptors with estimated binding affinity of 0.6 X 10(9) M-1. Multiple high mol wt bands (greater than 250,000) were identified under nonreduced conditions by affinity cross-linking. Under reduced conditions, one band with an approximate mol wt of 135,000 was identified. Purified Leydig cells (10(5) cells/ml) were cultured in Dulbecco's Modified Eagle's Medium-Ham's F-12 Nutrient Mixture (1:1) containing 0.1% fetal calf serum at 37 C in a humidified atmosphere of 5% CO2-95% air. Insulin and IGF-I stimulated testosterone formation as early as 3 h after administration, and their effects were completely blocked by the addition of a protein synthesis inhibitor, cycloheximide (1 microgram/ml). Insulin and IGF-I also significantly potentiated hCG-and 8-bromo-cAMP-induced testosterone formation. Furthermore, insulin and IGF-I potentiated hCG-stimulated cAMP formation. This suggests that insulin and IGF-I have effects at both the LH receptor sites and the steps beyond adenylate cyclase. The ED50 values of insulin and IGF-I-stimulated testosterone formation were comparable (25 ng/ml). In conclusion, we found that Leydig cells contain specific insulin and type I IGF receptors, and both insulin and IGF-I are capable of modulating Leydig cell steroidogenesis.

Animals↗
PubMed1985

Immunofluorescent localization of an androgen-dependent isoenzyme of prostatic acid phosphatase in rat ventral prostate.

Isoenzymes of rat ventral prostate (RVP) acid phosphatase were isolated and partially purified by ultracentrifugation, Sephadex G-100 column chromatography, and isoelectric focusing. Antisera were raised to the isoenzymes of prostatic acid phosphatase by immunization of New Zealand white rabbits. Rabbit antisera reacting specifically to homologous but not heterologous isoenzymes of acid phosphatase were then reacted with a variety of tissues using indirect immunofluorescence. The tissues included prostate, spleen, bone marrow, liver, kidney, salivary gland complex, small intestine, and adrenal glands. An antiserum against a RVP acid phosphatase isoenzyme with a pI of 4.5 (A-PAP) localized acid phosphatase only in the supranuclear region of rat ventral prostate epithelial cells, and did not react with acid phosphatase in any of the other organs tested. A-PAP did not localize acid phosphatase in the ventral prostate from rats 14 days after castration. A-PAP did localize acid phosphatase in the ventral prostate from castrated animals that were treated with testosterone. These results indicate the A-PAP localized an androgen-dependent isoenzyme of acid phosphatase in RVP epithelial cells that may be secretory in nature. This antiserum should prove to be an ideal marker for studies involving hormonal regulation of prostatic epithelial function in vivo and in vitro.

Acid Phosphatase↗
PubMed1985

Adhesion of cardiac myocytes to extracellular matrix components.

The interaction of ECM components with adult cardiac myocytes is not well understood, but is of physiological importance. Most physiological studies are conducted on myocytes in suspension yet in vivo the cells are attached to each other and to the ECM. In this paper, we further define the interaction of isolated adult myocytes with the ECM substrates. Of interest is not only the short-term attachment of cells to ECM substrates but also the ability of ECM components to support the long-term maintenance of cardiac myocytes in cultures.

Animals↗
PubMed1985

Extracellular matrix components influence the survival of adult cardiac myocytes in vitro.

Calcium-tolerant myocytes were isolated from adult rat hearts by collagenase perfusion and plated on various substrates in serum-free medium and their adhesion to various extracellular matrix (ECM) components was determined. The myocytes attached readily to dishes coated with collagen type IV (C-IV), laminin (LN), and to fetal bovine serum (FBS) in a manner dependent on the concentration of the components. Substantially fewer myocytes adhered to dishes coated with fibronectin (FN) or to uncoated plastic dishes. Cells adhered equally well to dishes coated with C-IV, LN and FBS within 1-4 h. However, when examined after 2 weeks in culture it was found that only C-IV and LN could support survival of the attached myocytes, and when cultured on C-IV or LN the myocytes were spread and had formed a dense monolayer. The actin filaments had at this time reorganized linearly along the long axis of the cell and the myocytes contracted spontaneously. Rabbit antibodies were raised against myocyte membranes and their ability to inhibit attachment to ECM components was studied. Purified IgG inhibited attachment to C-IV, while having only a minor effect on attachment to LN. These data are compatible with the presence of a specific cell surface component(s) that interacts with ECM substrates and influences cell shape and possibly thereby influences cellular functions.

Animals↗
PubMed1984

Pulmonary type II cell lamellar body ultrastructure preserved by rapid freezing and freeze drying.

Lamellar body ultrastructure was examined in cultured type II alveolar epithelial cells processed by a method of rapid freezing and freeze drying in the absence of both chemical fixation and solvent dehydration. This method of specimen preparation was chosen to optimize the retention of soluble substances within the type II cell. The use of cultured cell aggregates in which type II cells line the free surface facilitated the effectiveness of rapid freezing for the preservation of lamellar body fine structure. Lamellar bodies of frozen/frozen dried type II cells showed none of the often profound lipid extraction artifact produced by conventional processing. Instead they exhibited a substructure with noteworthy characteristics in common with lamellar bodies processed by resin dehydration lipid retention methods (Stratton, 1976). Importantly, the lamellae of frozen/frozen dried lamellar bodies were contiguous, with no interlamellar space, as is commonly observed in solvent-processed (extracted) specimens. The dimensions of lamellar components in frozen/frozen dried lamellar bodies were, however, different from published values for resin-dehydrated lipid-retained specimens. Lamellar width and the widths of component phospholipid head and fatty acid tail regions in frozen/frozen dried lamellar bodies were approximately 35% smaller than values reported for resin-dehydrated lamellar bodies. This difference was attributed to shrinkage of lamellar components as water was removed from the unfixed tissue during the freeze-drying process. Lamellar bodies preserved by rapid freezing/freeze drying to optimize the in situ retention of intracellular components possess closely adherent concentric membranous lamellae. This supports the contention that the widely appreciated lamellar pattern of the pulmonary lamellar body represents the in vivo molecular organization of intracellular surfactant phospholipids.

Animals↗
PubMed1984

Cell surface modifications in the epithelium of rat ventral prostate during adaptation to in vitro conditions: an ultrastructural study.

Sequential changes in epithelial cells of collagenase-dissociated rat ventral prostate were studied by thin-section and freeze-fracture electron microscopy. Epithelial cells did not attach to the substrate for 48 h. Pelleted cells obtained 1, 24, and 48 h after dissociation were assigned to three categories depending on morphology and cellular associations. (a) Solitary epithelial cells degenerated as determined by extensive vacuolization in the cytoplasm and aggregation of intramembranous particles (IMP). (b) Epithelial clusters consisted of a homogeneous population of well-maintained, closely packed cells. Aggregation of IMP was minimal. Tight junctions that formed between cells at the periphery of the clusters appeared normal and provided an effective permeability barrier demonstrated by the exclusion of ruthenium red tracer. (c) Tissue fragments were comprised of varying combinations of epithelial, endothelial, and smooth muscle cells as well as fibroblasts and erythrocytes. Maintenance of tissue fragments was variable. Plasma membranes often displayed aggregated IMP and proliferated tight junctional strands. An effective permeability barrier was absent. After the 48 h "latent period," epithelial cells in the clusters lost interdependence, disassociated from one another, and attached to the substrate. These isolated cells, which did not display aggregated IMP, retained the ability to form an effective permeability barrier upon reaching confluency. During the first 48 h, epithelial cells did not tolerate solitary existence, yet as participants in clusters they were well maintained. After this interval, they no longer required interactions with neighbors in order to survive. These results indicate that under our experimental conditions, an adaptation period is required by prostatic epithelial cells. The enhanced quality of maintenance associated with epithelial clusters suggests that control over the internal microenvironment, provided by a tight junctional barrier, may be important during the initial period of adaptation in vitro.

Adaptation, Physiological↗
PubMed1982

Primary epithelial cell cultures derived from canine prostate: isolation, culture, and characterization.

Epithelial-cell-enriched primary cultures were established from canine prostate. Minced tissue was dissociated with 750 units/ml of collagenase in F12K tissue culture medium containing 1% fetal bovine serum. This treatment resulted in the gradual removal of stromal elements from the base of the epithelial cells. After 30 minutes of digestion, aggregates of epithelial cells free of stroma were dislodged from the minced pieces of prostate. These aggregates were washed and plated at high density in F12K plus 10% fetal bovine serum. After 12-16 hours in vitro the unattached cellular aggregates were removed from the culture dishes, washed, and reinoculated into new culture vessels containing fresh medium. After 48 hours in vitro, the aggregates had attached to the culture vessels and spread out to yield discrete patches of epithelial cells. By 120 hours in vitro the patches of cells had grown and coalesced to form a confluent monolayer of epithelial cells. Ultrastructural examination of these cultures indicated that adjacent cells were joined by desmosomes and tight junctions and had tonofilaments and microvilli, giving the cells an epithelial appearance. The cells contained rough endoplasmic reticulum, Golgi apparatus, and secretory granules similar to those of the epithelial cells in the intact organ. In addition, intracellular "blebs" containing acid phosphatase were observed in the monolayers and were found to increase in size and number with time in vitro. Differentiated function of the cultures was demonstrated by the presence of ornithine decarboxylase and acid phosphatase and the ability of the cultures to metabolize testosterone to primarily 5 alpha-reduced metabolites.

Acid Phosphatase↗
PubMed1982

Densitometric and morphometric evaluation of growth in primary cultures of rat ventral prostate epithelial cells.

This study describes a method for establishing primary cultures in 24-well culture vessels and evaluating the effects of serum, hormones, and other factors on cell growth using densitometry. Primary cultures of rat ventral prostate epithelial cells were grown in 24-well culture vessels containing F12K culture medium supplemented with various concentrations of the following substances: fetal bovine serum (FBS), horse serum (HS), testosterone (T), dihydrotestosterone (DHT), hydrocortisone (HC), zinc (Zn), transferrin (TR), cysteine (CYS), glutamine (GLT), selenium (SEL), and ascorbic acid (AC). The effect of each supplement on cell growth was evaluated on fixed and stained cultures using a photovoltaic cell densitometer designed to read the total culture surface of a 16-mm well. Increasing concentrations of both HS and FBS resulted in an increase in cell growth. T, Zn, HC, TR, and AC each had a stimulatory effect on cell growth. CYS, GLT, and DHT had little effect on cell growth, while SEL was inhibitory to cell growth. This data compares favorably with that obtained by other methods, such as morphometric analysis and ornithine decarboxylase production. These results indicate that densitometry is a useful method for determining the effect of supplements on cell growth in primary culture.

Animals↗
PubMed1980

Isolation, culture and characterization of epithelial cells derived from rat ventral prostate.

Epithelial-cell enriched primary cultures have been established from rat ventral prostate (RVP). Minced ventral prostates were dissociated with 0.5% collagenase in F12K tissue culture medium containing 1% fetal bovine serum. This treatment resulted in the gradual removal of stromal elements from the base of the epithelial cells. After 60 minutes of digestion the aggregates of epithelial cells were washed and plated at high density in F12K plus 10% horse serum. After 48 hours in vitro the unattached cells were removed from the culture dishes, washed, and reinoculated into new culture vessels containing fresh medium. After 96 hours in vitro, the aggregates had attached to the culture vessels and spread out to yield discrete patches of epithelial cells. By 144 hours in vitro the patches of cells had grown and coalesced to form a semi-confluent monolayer of epithelial cells. Ultrastructrual examination of these cultures indicated that adjacent cells were joined by desmosomes and tight junctions and had formed "lumen-like structures" into which projected microvilli. In addition, the cells contained secretory granules and tonofilaments, giving them a morphological appearance similar to prostate epithelial cells in the intact organ. The primary cultures also retained histochemical activities for acid phosphatase, beta-glucuronidase, and succinic dehydrogenase that were similar to the intact organ.

Animals↗