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At least 1,171 records · Page 65Linked to original sources
PubMed1992

Differential colchicine effects on the transport of membrane and secretory proteins in rat hepatocytes in vivo: bipolar secretion of albumin.

We carried out a comparative investigation on the effects of colchicine (25 mumoles/100 gm body wt) on the intracellular transport, processing and discharge by secretion or proteolytic processing of a membrane protein (i.e., the polymeric IgA receptor) and a secretory protein (i.e., albumin) in rat hepatocytes. The results obtained indicated the following: (a) the transport and processing of polymeric IgA receptor is strongly inhibited and delayed, but the appearance of secretory component in the bile is not arrested; (b) polymeric IgA receptor reaches the sinusoidal plasmalemma in colchicine-treated specimens, as it does in controls; (c) albumin discharge into the plasma is strongly inhibited and markedly delayed in colchicine-treated as compared with control animals; (d) the reverse applies for albumin secretion in the bile, which is increased by a large factor; (e) newly synthesized albumin secreted directly from hepatocytes in control and in colchicine-treated animals is the major source of bile albumin; and (f) colchicine affects in different ways the polymeric IgA receptor and albumin arrival at the sinusoidal front and especially at the biliary front of the hepatocyte.

Animals↗
PubMed1993

Endocrine cells in colorectal adenocarcinomas: incidence, hormone profile and prognostic relevance.

The occurrence of endocrine cells in 350 cases of colorectal adenocarcinoma was studied by immunohistochemistry for chromogranin A (CGA). The hormone profile of endocrine tumor cells, the correlation between endocrine differentiation and presence of other colorectal epithelial-cell lineages and the prognostic relevance of endocrine differentiation in colorectal cancer were investigated. CGA-positive tumor cells were found in 30% of cases, 21% showing moderate positivity and 9.0% extensive positivity. Of CGA-positive tumors, 70% additionally produced neurohormones, mainly indigenous to normal colorectal epithelium: 55% showed immunoreactivity for glucagon-like substances, 20% for serotonin and 10% for somatostatin, PYY and HCG. No immunoreactivity was found for various neurohormones not normally produced by colorectal endocrine cells. CGA-positive tumors tended to be more aggressive than CGA-negative tumors. Especially, tumors with extensive CGA positivity showed shorter survival, which was most apparent within Dukes' stage C. In multivariate analysis, extensive CGA positivity was an independent indicator of poor prognosis. CGA immunoreactivity significantly correlated with mucin production, but not with expression of secretory component (SC), a columnar-cell marker. Mucin production significantly correlated with SC expression. Tumors positive for CGA but not for mucin and/or SC showed the worst prognosis. SC expression was a relatively favorable feature, and mucin-producing tumors showed intermediate behavior.

Adenocarcinoma↗
PubMed1994

Expression of APO-1 (CD95), a member of the NGF/TNF receptor superfamily, in normal and neoplastic colon epithelium.

APO-1 is a 48-kDa cell-membrane protein identical to the Fas antigen now designated CD95. It is a member of the NGF/TNF receptor superfamily. Anti-APO-1 monoclonal antibody induces apoptosis in a variety of cell types expressing this antigen. We immunohistochemically investigated APO-1 expression in normal colon mucosa, 20 adenomas, 258 colon carcinomas and 10 liver metastases and carried out in vitro studies using a panel of colon-carcinoma cell lines. Immunohistochemically, APO-1 was regularly expressed at the basolateral membrane of normal colon epithelia. In a minor fraction of colon adenomas and in 39.1% of colon carcinomas APO-1 expression was diminished and in 48.1% of carcinomas, predominantly of the non-mucinous type, APO-1 expression was completely abrogated. The normal level of APO-1 in carcinomas was correlated with the mucinous type. Reduced/lost APO-1 expression was more frequent in rectal carcinomas. Complete loss of APO-1 was more frequent in tumors that had already metastasized. APO-1 expression in liver metastases essentially corresponded to that of the primary tumors. Comparative analysis with data from previous studies revealed that the mode of APO-1 expression is correlated with that of HLA-A,B,C./beta 2m, HLA-DR, HLA-D-associated invariant chain and of the secretory component. Surface expression of APO-1 was heterogeneous in colon-carcinoma cell lines; SW480 expressed considerable amounts of APO-1 on all cells, while HT-29 constitutively did less so and only in a minority of cells. Surface density of APO-1 and the fraction of positive cells in HT-29 was enhanced by interferon-gamma (IFN-gamma) and, additively, by tumor necrosis factor-alpha (TNF-alpha), whereas in SW480 APO-1 expression was not modulated by these cytokines. We conclude that neoplastic transformation of colon epithelium often leads to a loss of the physiologic, high level of surface APO-1 by giving rise either to a stable lack of APO-1 or to an IFN-gamma/TNF-alpha-sensitive phenotype of inducible APO-1 expression.

Adenoma↗
PubMed2006

Polyethersulfone dead-end tube as a scaffold for artificial lacrimal glands in vitro.

Polyethersulfone (PES) dead-end tubes were fabricated by means of a phase inversion technique, and then were used as scaffolds for artificial lacrimal glands. The wall of the dead-end tubes could allow nutrients such as ascorbic acid, L-tryptophan, and glucose to pass through, but prevents rat IgG from passing through. Lacrimal acinar epithelial cells of Sprague-Dawley rats were cultured in vitro, and cell-associated secretory component was detected with an immunofluorescence technique to identify the acinar cells. The second passage of the cells showed high degree of cellular differentiation, and was used to seed on the PES tubes. The results showed that the PES tube could support the attachment, the growth, and the proliferation of the rat lacrimal acinar cells. Thus, PES is a substrate for the growth of lacrimal acinar cells and may be a useful scaffolding biomaterial for tissue engineering, such as a scaffold for artificial lacrimal glands.

Animals↗
PubMed1989

Sodium arsenite induced alterations in bilirubin excretion and heme metabolism.

The acute administration of sodium arsenite (AsIII) to rats resulted in a biphasic alteration of the hepatic cytosolic "free" heme pool. The first stage was an increase in the cytosolic "free" heme without significant effects on the content of cytochrome P-450 or on bilirubin excretion. The second stage consisted of a continuous fall of the cytosolic "free" heme and of the content of cytochrome P-450. These changes were concurrent with an eight-fold increase in heme oxygenase activity and associated with marked elevations in the biliary excretion of bilirubin. The bile was collected from chronically cannulated rats to avoid artifacts related to anesthesia or post anesthetic effects. The rapid increase in biliary excretion of labeled heme degradation products indicated an increased breakdown of newly synthesized heme. Immunoelectrophoresis of bile proteins showed an altered pattern of bile protein excretion. The increased biliary haptoglobin suggested some hemolysis, while the reduction in the free immunoglobulin A (IgA) secretory component showed an AsIII-related decreased protein transport across hepatocytes to bile. Further research is required to assess the direct role of an increased heme degradation in the genesis of the hepatotoxic effects of AsIII.

Animals↗
PubMed1991

Secretion of mutant leucine-specific binding proteins with internal deletions in Escherichia coli.

The leucine-specific binding protein, encoded by the livK gene, is located in the periplasm of E. coli. The present study is an attempt to identify intragenic regions that determine the efficiency of its secretion into the periplasm. C-terminal deletions or fusions of the livK gene to trpA (encoding the alpha subunit of tryptophan synthetase) were secreted with little loss of efficiency [1]. A series of deletions was constructed at the unique Sphl site within livK, near the 5' end of the region coding for the mature protein. Between 16 and 113 amino acids were deleted in the amino-terminal one-third of the protein. A few of these deletions were located within a few amino acids of the signal sequence processing site. Deletions extending within thirteen residues of the processing site were processed and secreted more slowly than normal. Secondary structure predictions suggested that the alpha-helical core region of the signal sequence extends into the mature protein in the case of the slow processing mutants, perhaps interfering with the recognition site for leader peptidase or other secretory components. These results suggest that the conformation around the signal processing site may be a critical factor in determining the efficiency of secretion. During the course of this study, it was found that the difference in molecular weight between precursor and mature forms of some binding protein mutants, as judged by SDS-PAGE, was much greater than could be accounted for by processing of the signal sequence. This anomalous mobility on gels, however, could be eliminated by performing SDS-PAGE in the presence of 6 M urea.

Amino Acid Sequence↗
PubMed1992

Biochemical and morphological differentiation of the human colonic epithelial cell line SW620 in the presence of dimethylsulfoxide.

In vitro models of intestinal cell differentiation provide an important adjunct for studying normal and abnormal intestinal epithelial cell differentiation. The studies reported herein describe morphologic and biochemical changes in the colonic epithelial cell line SW620 following dimethylsulfoxide (DMSO) incubation. Cells cultured in the presence of DMSO showed striking changes in morphology characterized by enlargement, elongation, and formation of process-like structures by light microscopy and a propensity to form microvillus-like structures by electron microscopy. These changes were accompanied by significant differences in the expression of the cell surface markers CD4 (HIV gp120 receptor), CD44 (hyaluronate receptor), and KS1 (adenocarcinoma/epithelial specific antigen). There was a marked decrease in CD4 expression (38% to 2%), an increase in CD44 expression (4% to 50%) and a decrease in KS1 expression (98% to 66%) as detected by flow cytometry following incubation of SW620 cells in DMSO. Parallel changes in the expression of these markers were seen by metabolic and surface labeling studies. Although SW620 cells were infected by HIV-1, DMSO-treated SW620 cells could not be infected. DMSO-induced changes in surface expression of CD4, CD44, and KS-1 were reversible over time upon removal of DMSO from the culture medium. Secretory component, sucrase, neuron-specific enolase, chromogranin-A, and mucin were not detectable in SW620 cells with or without DMSO treatment. SW620 cells provide a useful model for studying specific biochemical and molecular events involved in intestinal epithelial cell differentiation and function.

Antigens, Neoplasm↗
PubMed1992

Cytoskeleton and other differentiation markers in the colon.

Differentiation of intestinal epithelial cells involves a complex process of establishment of cell polarity, commitment to cell lineage, and inhibition of cell division. Polarized epithelial cells are characterized by specific junctional complexes and cytoskeletal proteins which produce specific membrane domains. Intestinal cytoskeletal proteins are often preserved in neoplastic colonic tissues, and can be used to identify the cell of origin of poorly differentiated cancers. In this context, these proteins are markers of organ-specific differentiation. In addition, since loss of cytoskeletal polarity commonly occurs in transformed cells, aberrant expression of these proteins may be used as a marker of neoplasia in the colon. Normal polarization of basolateral proteins (secretory component) and apical proteins such as brush border hydrolases, cytoskeletal proteins (villin, fodrin), and carcinoembryonic antigen can become disrupted in adenomas and cancers. Cytoskeletal intermediate filaments (cytokeratins) demonstrate increased immunoreactivity in villous adenomas and cancers compared with normal colonic crypts. Altered actin bundles are found in preneoplastic mucosa such as colon from patients with familial polyposis coli. Molecular mechanisms responsible for altered cytoskeletal structures remain unclear; however, altered protein phosphorylation most likely plays a role. For example, the phosphorylation status of cytoskeletal and junctional complex proteins appears to influence their solubility and interactive properties, which may result in altered cell polarity. Markers of altered cytoskeletal structure and polarity can identify neoplastic colonocytes; however, the extent to which they can be used as intermediate markers of colonic neoplasia remains to be determined.

Actins↗
PubMed1987

Post-testicular change in the reptile sperm surface with particular reference to the snake, Natrix fasciata.

Sperm surface changes occurring in the reptile Wolffian duct have been explored with particular references to the snake, Natrix fasciata. In the snake Wolffian duct there are several proteins not present in serum, the pattern of which changes in concert with the seasonal testicular cycle. Whereas testicular spermatozoa did not bind antibody to duct secretions, all Wolffian duct spermatozoa did so over both head and tail, according to immunofluorescence patterns. Thus, on entering the Wolffian duct, the entire surface of N. fasciata spermatozoa acquires one of more of the duct's secretory components. As indicated by immunofluorescence, immunoelectrophoresis, and immunodiffusion, epitopes on at least some molecules that bind to spermatozoa or that remain free in the duct fluid are shared with those in other Natrix species, but not in more distant reptiles (turtle, anole lizard), nor chicken, rat, or rabbit. In regard to glycoproteins, one prominent con A-reactive band was present in polyacrylamide gel electrophoresis (PAGE) of snake fluid and more were evident in fluid collected from the turtle duct. However, such lectin-reactive elements did not bind to spermatozoa as judged by an absence of any change in snake, turtle and lizard sperm lectin-binding patterns in passing from the testis into and through the Wolffian duct. In all, evidence from these and other species studied begins to suggest that the nature of the post-testicular sperm surface modification displayed in most vertebrates that fertilize internally may differ in sub-therian and therian groups, respectively. There appears to be a relative emphasis on glycosyl-rich surface elements in the latter. The possible significance of these changes for sperm function in the different groups is discussed briefly in terms of sperm survival/storage, as well as capacitation and sperm binding to the zona.

Animals↗
PubMed1991

Zinc-protein from rat prostate fluid binds epididymal spermatozoa.

The detection and the isolation of a zinc-protein from the secretion of the rat dorsolateral prostate is described. The purification procedure, based on gel filtration and cationic exchange chromatography, allowed to separate a minor protein (Mr approximately 66,000) from free zinc ions and other secretory components. Two zinc ions were estimated to be associated with one molecule of isolated protein. The zinc-protein was labelled with 125I and then incubated at 37 degrees C with spermatozoa from rat epididymal cauda. Time-dependent in vitro binding of the radioactive protein to sperm cells was demonstrated. This binding was not affected by the presence of proteins from the seminal vesicle during the incubation, while it was blocked in the presence of an excess of unlabelled zinc-protein. After binding, the labelled spermatozoa were treated with a buffer containing 0.5% sodium deoxycholate and 40 mM EDTA; only very small amounts of label were removed from the cells, thus suggesting that the zinc-proteins were kept on the plasma membrane by interactions which do not involve merely hydrophobic bonds.

Animals↗
PubMed1989

Adenoid cystic carcinoma of the tracheobronchial tree: clinicopathology and immunohistochemistry.

Adenoid cystic carcinoma of the tracheobronchial tree in five patients was treated surgically and the clinicopathologic manifestations and histogenesis were examined in detail. Symptoms such as cough, dyspnea, hemoptysis, and atelectasis on chest X-ray were present in four patients, and the other patient was asymptomatic. Histologically, growth patterns were classified as tubular, cribriform, and solid. The solid pattern was the most aggressive with extensive perineural invasion. Immunohistochemically, secretory component, lactoferrin, and epithelial membrane antigen were present in the cells lining the gland-like lumen of tissues with the tubular and cribriform patterns, but was rare in those with a solid pattern. Desmin and S-100 protein were detected in the nonlining cells of tissues with all three patterns. These findings suggest that this tumor originates from the myoepithelial cells of the bronchial gland and that the solid pattern was the most poorly differentiated form.

Adult↗
PubMed1986

The occurrence and clinicopathological significance of serotonin immunoreactive cells in large bowel carcinoma.

We studied the incidence, clinicopathological relevance and prognostic significance of serotonin immunoreactive cells in a clinically well-documented series of 300 large bowel cancer patients. Serotonin immunoreactive cells were detected in 8 per cent of the carcinomas, occurring either as focal clusters (4.7 per cent) or as occasional single cells (3.3 per cent). Both types frequently displayed presence of mucin and/or immunoreactivity for secretory component as well. In respect of localization, stage or tumour extension and histological grade tumours with serotonin immunoreactivity showed no significant differences in comparison with carcinomas lacking this feature. In terms of survival, however, the tumours with serotonin immunoreactivity demonstrated a more aggressive clinical course in comparison with tumours without these cells. This phenomenon reached borderline statistical significance. It is therefore concluded that the study of serotonin immunoreactivity in large bowel carcinomas enables the identification of a subpopulation of colorectal carcinomas with a relatively poor prognosis.

Colonic Neoplasms↗
PubMed1990

Detection and enumeration of colonic mucosal cells in amniotic fluid using a colon epithelial-specific monoclonal antibody.

Since its introduction, prenatal diagnosis of chromosomal and metabolic disorder by midtrimester amniocentesis has relied upon the use of a mixture of fetal cells obtained from amniotic fluid. Little knowledge has been gained in the sorting of these cells for diagnosis of tissue-specific disorders. In an attempt to determine the contribution of fetal colonic mucosal cells to the overall amniocyte population, we used the colonic epithelial-specific monoclonal antibody (MC-Ab) 7E12H12, IgM isotype. Specimens of the small intestine, colon, buccal mucosa, kidney, urinary bladder, and umbilical cord were obtained from electively aborted normal fetuses of 12-28 weeks' gestation. All of these specimens were examined with 7E12H12 by the immunoperoxidase technique. The MC-Ab reacted with the colonic epithelial cells but not with any of the other tissues. In addition, 40 amniotic fluid samples obtained from women between 16 and 18 weeks of gestation, who underwent amniocentesis because of advanced maternal age, were tested using a fluorescent activated cell sorter. Among the amniotic fluid specimens examined, 18.4 +/- 10.3 per cent cells reacted with 7E12H12. Double immunofluorescence studies revealed that all Mc-Ab-stained cells contained secretory component, confirming that they were epithelial in origin. All fetuses whose amniotic fluid was analysed had normal karyotypes and amniotic fluid alpha-fetoprotein levels that were also normal. This study demonstrates that cell-specific Mc-Ab can be used to detect colon cells in the amniotic fluid and that colon cells contribute significant numbers in the mixture of amniotic fluid cells.(ABSTRACT TRUNCATED AT 250 WORDS)

Amniotic Fluid↗
PubMed1982

Age-dependent changes in the carbohydrate pattern of human prostatic epithelium as determined by peroxidase-labeled lectins.

Peroxidase-conjugated lectins (peanut agglutinin: PNA; Ricinus communis agglutinin: RCA) were used for histochemical demonstration of the respective carbohydrate binders (PNA: beta-D-galactosyl-(1-3)-N-acetyl-galactosamine; RCA: D-galactose). The study of prostatic specimens from four different age groups showed marked differences in cytoplasmic PNA binding of prostatic epithelium. The basal cells of the glandular epithelium displayed cytoplasmic PNA binding only in the infantile stage and during early puberty. With the onset of puberty a number increasing with age of secretory cells developed cytoplasmic binding of PNA, mostly appearing as apically located vacuoles. Intraluminal secretion, however, was never stained. During senile involution the prostatic epithelium lost rapidly its PNA binding capacity. Since no RCA binding of the epithelium was observed, the carbohydrate moiety responsible for PNA binding presumably represents a galactosyl-N-actyl-galactosamine containing molecule, most likely a glycoprotein, which is present in actively functioning prostatic epithelial cells. It is uncertain whether or not it represents a structural or a secretory component of the prostatic cell.

Adolescent↗
PubMed1983

Effects of androgens on the ultrastructure and subcellular zinc distribution in the prostatic epithelium of castrated rats.

Sprague Dawley rats were maintained on testosterone propionate or 5 alpha-dihydrotestosterone for 3 days following bilateral orchidectomy for a 7-day period. Ultrastructural examination showed only partial recovery of the prostatic epithelium with testosterone propionate while 5 alpha-dihydrotestosterone caused the lateral and ventral lobes to revert to the appearance of control tissues. The latter metabolite induced greater stimulation of the prostate evidenced by increased mitotic division of the epithelial cells and an increased number of basal cells exhibiting ciliary formation was observed. Zinc concentrations in subcellular regions of both lateral and ventral prostate lobes were affected by the two androgens. Testosterone propionate was most effective in elevating zinc in the lateral lobe, particularly within the secretory components. In the ventral lobe both androgens caused an increase in subcellular zinc concentrations above control levels. The increase of nuclear and nucleolar zinc was related to the increase in nuclear activity and cellular response to the androgen administration.

Androgens↗
PubMed1998

A monoclonal antibody that recognizes a common carbohydrate epitope shared by various glycoproteins in human secretions.

A monoclonal antibody (mAb; a mouse IgM referred to as 1CF11) recognizing various human glycoproteins was obtained. While the immunoreaction of glycoproteins from human secretions including milk, saliva, and bronchus was demonstrated as a typical dose-responded S-shaped reaction curve on ELISA, no reaction was detected with milks and sera of animal origin as well as human serum. In the constituting polypeptides of the human milk secretory IgA molecule, only the secretory component was recognized by this mAb. Among various chemical treatments of the purified human milk lactoferrin (Lf), only either periodate or mild alkaline treatment abolished the immunoreactivity of the glycoprotein. A recombinant human Lf was not immunoreactive. Finally, the immunoreactive fragments were isolated from human milk Lf, which remained reactive with PAS reagent while lacking the previously reported N-glycans. These results strongly suggest that the mAb 1CF11 recognizes a new glycan O-glycosidically linked to glycoproteins in human secretions.

Animals↗
PubMed2001

Selective secretion of protein kinase C isozymes by thrombin-stimulated human platelets.

The protein kinase C (PKC) was secreted from thrombin-stimulated human platelets in a time- and dose-dependent manner. The PKC specific inhibitors Ro31-8220 (0.05 microM) and GF 109203X (0.5 microM) totally inhibited the secreted kinase activity. Western blot analysis of the secretory components showed reactivity to PKCalpha, PKCbetaII, and PKCdelta antibodies, but not to PKCbetaI, and p42/44 MAPK, although they were present in lysed platelets. The fractionation of platelets secreted components showed that PKC activity increased in both soluble and microparticle fractions after thrombin treatments. This is the first report demonstrating that activated human platelets selectively secrete protein kinase C isozymes. Protein kinase C secreted by platelets in this unique manner may have an extracellular role in the plasma, and may regulate cellular functions, including remodeling of vascular endothelial cells.

Blood Platelets↗
PubMed2001

Hybrid IgA2/IgG1 antibodies with tailor-made effector functions.

Immunoglobulin (Ig) A and IgG are the principal immune effector molecules at mucosal surfaces and in blood, respectively. Mucosal IgA is polymeric and bound to secretory component, whereas serum IgG is monomeric. We have now produced IgA2/IgG1 hybrid antibodies that combine the properties of IgA and IgG. Antibodies with Calpha3 at the end of the IgG H chain resemble IgA and form polymers with J chain that bind the polymeric Ig receptor. Like IgG, the hybrid proteins activated complement and bound FcgammaRI and protein A. Though the hybrid proteins contained both Cgamma2 and Cgamma3, they have a short in vivo half-life. Surprisingly, this decreased half-life correlated with a higher avidity than that of IgG for murine FcRn. Interestingly, antibodies with Calpha1 replacing Cgamma1 were resistant to extremes of pH, suggesting that Calpha1 increases antibody stability. These results provide insights into engineering antibodies with novel combinations of effector functions.

Animals↗