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Query: UNIPROT:P06889 (Mol)
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Cryosections of normal colon (NC), tubular and villous adenomas (TA, VA), and variably differentiated colon adenocarcinomas (CA) were immunostained with monoclonal antibodies to alpha 1-6 and alpha v, and beta 1-4 integrin subunits; select samples were stained for cytokeratin (Ck) 20 and villin. In NC, alpha 2 staining was strongest in crypt cells; alpha 1,3 and alpha v, and beta 1,3 and beta 4, and Ck 20 and villin predominated in superficial enterocytes. In TA and VA, monolayered glands showed integrin, Ck 20 and villin patterns that differed slightly from both crypt and superficial enterocytes. Complex glands in VA showed decreased integrin staining and basal polarization; Ck 20 and villin were strong only in luminal cells. CA showed overall weaker integrin staining than adenomas. Regardless of invasion depth, well formed malignant glands mimicked TA; pleomorphic glands mimicked VA with focal basal integrin polarization and solid clusters displayed scanty integrins, uneven Ck 20, and villin in occasional cells. Diverse integrins in crypt compared with superficial enterocytes reflect changing adhesive requirements as cells migrate and terminally differentiate. Decreasing expression and altered distribution of integrins, Ck 20 and villin noted in TA, VA, and in CA of increasing grade indicate that certain adhesive and cytoskeletal features more closely relate to glandular architecture than to depth of invasion.
Virchows Arch B Cell Pathol Incl Mol Pathol 1993
PMID:Immunolocalization of integrins in the normal and neoplastic colonic epithelium. 768

The expression of cytokeratin (CK) 17 was studied in 28 primary transitional cell carcinomas (TCCs) of the human urinary tract using CK 17-specific monoclonal antibody E3. While CK 17 was not detectable at all or only present in some areas of basal cells in normal--appearing urothelium, a certain subpopulation of cells of all G1 and G1/G2 TCCs examined (9 cases) stained positive for CK 17. These latter cells were either restricted to the basal compartment or located also in suprabasal layers exhibiting a decreasing intensity of immunoreactivity. CK 17 was seen in practically all cells in G2 and G2/G3 tumors (7 cases). In contrast, G3 TCCs and anaplastic carcinomas showed a highly variable CK 17 staining pattern ranging from completely negative to completely positive with several intermediate phenotypes. Our results indicate that CK 17 could be a useful marker for the progression of urinary tumors.
Virchows Arch B Cell Pathol Incl Mol Pathol 1993
PMID:Immunohistochemical localization of cytokeratin 17 in transitional cell carcinomas of the human urinary tract. 769 61

To study the molecular structure of Mallory body (MB) proteins we applied infrared spectroscopy of the isolated MBs from livers obtained from autopsied patients with alcoholic cirrhosis and griseofulvin-fed (GF-fed) mice. Liver frozen sections were extracted with detergent and digested with deoxyribo- and ribonuclease and collagenase. MB-enriched fractions were then separated out using the aqueous two-phase polymer system. Immunohistochemical and electron microscopic examination showed that the MB composition was virtually identical in human and mouse livers. Infrared spectra of both MB samples showed that the MBs had more numerous and stronger intermolecular hydrogen bonding than did the background control fractions as well as the cytoskeletal fraction from control and GF-fed mice. This may explain why the proteins in MBs are aggregated. The relative amount of beta-sheets was increased compared to the alpha-helices in the MBs, indicating that conformational changes in the cytokeratin peptides of the MBs had occurred. This may explain why the antigenic sites observed in MB proteins show changes in affinity for antibodies to cytokeratins as observed by immunohistochemical staining of MBs.
Exp Mol Pathol 1993 Dec
PMID:Molecular structural changes in Mallory body proteins in human and mouse livers: an infrared spectroscopy study. 813 2

Alveolar type II cells proliferate to restore the alveolar epithelium after lung injury and differentiate into type I epithelial cells. A variety of factors promote rat type II cell DNA synthesis in vitro; however, only low levels of proliferation occur when type II cells are cultured at high density. We plated type II cells at low density to determine if those growth factors that stimulate thymidine incorporation also stimulate low density proliferation. Type II cells were plated at 1 x 10(3) cells/cm2 in Dulbecco's modified Eagle's medium containing 2% fetal bovine serum, cholera toxin, insulin, epidermal growth factor, acidic fibroblast growth factor (aFGF), and concentrated bronchoalveolar lavage fluid from normal rats. By 7 days, numerous colonies had grown out that exhibited an epithelial morphology and stained positively for cytokeratin. The cell number at day 7 in the presence of the combined factors was 5.9 x 10(3) (+/- 0.6 x 10(3)) cells/cm2 (n = 4). There was no colony formation in the absence of fetal bovine serum. The addition of linoleic acid to serum-free medium containing all the growth supplements was found to partially restore colony formation. When aFGF or lavage fluid was omitted from the culture medium, colony formation was dramatically reduced. The colonies lacked characteristics of differentiated type II cells, which was anticipated since these cells were cultured on tissue culture plastic. To see if these cells could express differentiated functions, we maintained the colonies under growth conditions, removed them from the plastic substratum, and then replated them on EHS matrix.(ABSTRACT TRUNCATED AT 250 WORDS)
Am J Respir Cell Mol Biol 1993 Jul
PMID:Proliferation of rat alveolar epithelial cells in low density primary culture. 833 78

We report here the establishment and characterization of a corpus cavernosum cell line (DS-1) from human penile tissue. This is the first cell line of its type derived from cavernosum tissue. DS-1 cells have become immortalized in culture, and show growth in monolayers. These cells have a doubling time of about 45 h in in vitro culture. Cytogenetic analysis by G-banding demonstrated a diploid karyotype with a model chromosome number of 46. The chromosome constitution of DS-1 cells was found to be male (XY), in 28/30 cells scored. Two of the 30 cells showed an extra structurally rearranged "marker" chromosome, that appeared to be a derivative of chromosome 18 with excessive chromosome on the short arm. Ploidy analysis revealed that the majority of DS-1 cells had a DNA index of one. About 35% cells were found to be in G-1 phase and 52% cells in S phase. Light and electron microscopy of DS-1 cells and original penile tissue showed typical characteristics of this tissue. Immunocytochemistry studies using antibodies to smooth muscle actin, desmin, vimentin and cytokeratin (LP34, CAM5.2) showed that the DS-1 cell line had predominantly smooth muscle cells, as these cells were positive for smooth muscle actin, desmin and vimentin.
Biochem Mol Biol Int 1993 Jul
PMID:Phenotypic and cytogenetic characterization of a human corpus cavernosum cell line (DS-1). 840 13

Previous attempts to culture mouse alveolar type II (ATII) cells have been hampered by limited purity and cell recovery. We have now obtained culturable ATII cells from female C57BL/6 mice at a purity of 92% +/- 3 (mean +/- SD; n = 20), with viabilities of 96% +/- 2 and total yields of 5.1 +/- 0.7 X 10(6) cells per mouse. Crude lung cell suspensions were prepared by intratracheal instillation of Dispase and agarose followed by mechanical disaggregation of the lungs. Crude cell suspensions were purified by negative selection using a biotinylated-antibody, streptavidin-coated biomagnetic particle system. Cell purities were determined by Pap staining and confirmed ultrastructurally. Purified ATII cells were cultured on fibronectin-coated chamber slides and maintained for up to 5 days in DMEM with 10% fetal bovine serum. Cultures exhibited minimal contamination by Clara cells, mesenchymal cells, or endothelial cells, and the epithelial nature of the cultures was confirmed by positive cytokeratin staining in at least 97% of the cells through day 5. Day 3 cultures demonstrated osmium tetroxide/tannic acid-stained granules consistent with lamellar bodies in 76% +/- 3.6 of the cells. The cultures displayed features distinct from those previously described for adult rat ATII cells, including irregularly-shaped cells and the formation of numerous cytoplasmic projections in direct contact with other cells. These studies indicate that excellent yields of highly purified, culturable ATII cells can be obtained from genetically defined mice. These techniques may provide powerful new models for the study of parenchymal lung disease in vitro.
Am J Respir Cell Mol Biol 1996 Apr
PMID:Isolation and primary culture of murine alveolar type II cells. 860 Sep 33

The ability to induce proliferation by temporary duct ligation suggested an hypothesis that retrovirus-mediated gene transfer into cells of the biliary tract could be accomplished. The time course of histologic changes, incorporation of 3H-thymidine and immunofluorescent staining with a monoclonal antibody to cytokeratin-19 (a marker for differentiated bile ducts) was studied in male Fischer F344 rats. A recombinant Gibbon ape leukemia virus (GALV), containing a gene encoding Escherichia coli beta-galactosidase was next introduced into 24 hr obstructed bile ducts. Gene transfer was maximal when virus was exposed to the obstructed duct for 12 hr (approximately 0.1%). The majority of X-gal positive cells were in cytokeratin-19 negative peribiliary tissues, which had the appearance of newly forming bile ducts. The data suggest that cells targeted by retroviral infection of the obstructed rat bile duct may be a precursor of mature, fully differentiated biliary epithelium.
Somat Cell Mol Genet 1996 Jan
PMID:Targeted retroviral gene transfer into the rat biliary tract. 864 91

The supply of fresh bronchial tissue from human donors for in vitro culture is limited. Routine fiberoptic bronchoscopy offers a safe and easy procedure for obtaining minor biopsies and we wanted to see if the material provided could be used for organ culture by using a simple liquid overlay technique. Bronchial biopsies were cut into fragments 400-500 microns and kept immersed in a standard serum-supplemented medium for 40 days. An agar base prevented adhesion of the tissue. By light and electron microscopy it was shown that the tissue fragments had a differentiated epithelium at their surface throughout the culture period. An outgrowth of epithelial cells on the scaffold of the exposed stroma, covering the surface of the whole fragment, occurred within the first 5 days of culture. This epithelium was partly ciliated, 2-4 cell layers thick with squamous and cuboidal cells and expressed epithelial markers (cytokeratin and Ber-Ep4). The amount of cilia increased during the first 15 days of culture. The epithelium rested on a neosynthesized basement membrane as visualized by electron microscopy and immunohistochemistry with antibodies directed against collagen IV, laminin, and fibronectin. The central stroma consisted of loose connective tissue with fibroblasts. This simple tissue culture model combines maintenance and neoformation of bronchial epithelium on top of a living natural substrate, thus enabling direct biological studies on clinical biopsy material under perfectly viable conditions.
Am J Respir Cell Mol Biol 1996 Aug
PMID:Nonadhesive stationary organ culture of human bronchial mucosa. 870 75

Infection with the wild type SV40 virus was used to transform primary cultures of human tracheal gland serous (HTGS) cells. Over 80 different cell lines were obtained, but the majority had lost some of their epithelial and secretory features. However, one of these cell lines, MM-39, was shown to have conserved the physiologic characteristics of the genuine HTGS cells-i.e., the presence of cytokeratin, expression of cystic fibrosis transmembrane conductance regulator mRNA, a level of secretory leukocyte proteinase inhibitor secretion comparable to that of the native cells (25 +/- 3 ng/10(6) cells/h), and the responsiveness to pharmacological agonists: carbachol (+260 +/- 40%), isoproterenol (+260 +/- 40%), and adenosine 5'-triphosphate (+280 +/- 30%). These characteristics describe a transformed cell line of human tracheal gland cells which has retained the features of the native serous cells. As a result, this cell line appears to be a useful tool for large-scale physiologic and pharmacologic studies of bronchial secretion at the cellular level.
Am J Respir Cell Mol Biol 1996 Oct
PMID:A transformed human tracheal gland cell line, MM-39, that retains serous secretory functions. 887 86

A spontaneously established porcine granulosa cell line (PGC-2) was cloned through the continuous culturing of primary granulosa cells collected from equine chorionic gonadotropin (eCG)-treated prepubertal gilts. This established cell line has undergone approximately 100 passages and shows contact-inhibition of growth. PGC-2 stained with a monoclonal antibody (mAb) directed against cytokeratin, indicating its epithelial nature, but not with a mAb directed against vimentin, suggesting that it is not fibroblast-derived. Immunoblotting revealed that PGC-2 expresses cadherin, an epithelial Ca+2-dependent cell adhesion molecule. The cells were dependent on serum for growth and had a doubling time of approximately 20 hr when cultured with 10% fetal bovine serum. The cell line was examined for the presence of FSH receptors, cAMP responses, and steroidogenic capabilities. The cell line lacks FSH receptors as assessed by radiolabelled-ligand binding, and no transcripts for FSH receptor were detected by Northern blotting of total cellular RNA. Neither FSH nor cholera toxin (0.5 ng/mL) stimulated increases in cAMP levels in these cells, whereas forskolin (10 microM) induced a fivefold increase in cAMP production. When a higher concentration of cholera toxin (300 ng/mL) was used, however, cAMP levels doubled by 2 hr. Despite a lack of responsiveness to purified of SH or oLH, the cells were capable of progesterone and estradiol production when provided with the appropriate substrates. We conclude that PGC-2 display properties that are similar to immature granulosa cells and may provide a suitable in vitro model for the study of granulosa cell function.
Mol Reprod Dev 1996 Nov
PMID:Steroidogenic properties of a spontaneously established porcine granulosa cell line (PGC-2). 891 40


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