concanavalin-a and Pancreatic-Neoplasms

The need for analytical devices for detecting cancer at early stages has motivated research into nanomaterials where synergy is sought to achieve high sensitivity and selectivity in low-cost biosensors. In this study, we developed a film architecture combining self-assembled monolayer (SAM) and layer-by-layer (LbL) films of polysaccharide chitosan and the protein concanavalin A, on which a layer of anti-CA19-9 antibody was adsorbed. Using impedance spectroscopy with this biosensor, we were capable of detecting low concentrations of the antigen CA19-9, an important biomarker for pancreatic cancer. The limit of detection of 0.69U/mL reached is sufficient for detecting pancreatic cancer at very early stages. The selectivity of the biosensor was inferred from a series of control experiments with samples of cell lines that were tested positive (HT29) and negative (SW620) for the biomarker CA19-9, in addition to the lack of changes in the capacitance value for other analytes and antigen that are not related to this type of cancer. The high sensitivity and selectivity are ascribed to the very specific antigen-antibody interaction, which was confirmed with PM-IRRAS and atomic force microscopy. Also significant is that used information visualization methods to show that different cell lines and commercial samples containing distinct concentrations of CA19-9 and other analytes can be easily distinguished from each other. These computational methods are generic and may be used in optimization procedures to tailor biosensors for specific purposes, as we demonstrated here by comparing the performance of two film architectures in which the concentration of chitosan was varied.

Topics: Antigens, Tumor-Associated, Carbohydrate; Biomarkers, Tumor; Cell Line, Tumor; Chitosan; Concanavalin A; Dielectric Spectroscopy; Electric Capacitance; Fatty Acids; Gold; Humans; Pancreatic Neoplasms; Spectrophotometry, Infrared; Sulfhydryl Compounds

The mechanisms of immunonutrition on reducing infectious complications are still poorly understood. This prospective randomized study was designed to determine whether immunonutrition influences the following factors: cell-mediated immunity, differentiation of T helper type 1 (Th1) and Th2 cells, interleukin (IL)-17-producing CD4(+) helper T (Th17) cell response, and infectious complication rate after pancreaticoduodenectomy.. Thirty patients who underwent pancreaticoduodenectomy were divided into 3 groups. Ten patients in the perioperative group received immune-enhancing diets enriched with arginine, omega-3 fatty acids, and RNA for 5 days before operative resection, which was prolonged after operative resection by enteral infusion. Ten patients in the postoperative group received early postoperative enteral infusion of the same enriched formula with no artificial nutrition before operative resection. Ten patients in the control group received total parenteral nutrition postoperatively. The primary endpoint was immune responses; the secondary endpoint was the rate of infectious complications.. Concanavalin A (Con A)- or phytohemagglutinin (PHA)-stimulated lymphocyte proliferation and natural killer cell activity were significantly higher in the perioperative group than in the other groups. Messenger RNA (mRNA) expression levels of T-bet, interferon-gamma (IFN-gamma), related orphan receptor gammat (RORgammat), and interleukin-17F (IL-17F) were significantly higher in the perioperative group than in the other groups. In the perioperative group, the rate of infectious complications was significantly reduced compared with that in the other groups.. Perioperative immunonutrition reduced stress-induced immunosuppression after a major stressful operative resection. The modulation of Th1/Th2 differentiation and Th17 response may play important roles in this immunologic effect.

Topics: Aged; Aged, 80 and over; Arginine; Bile Duct Neoplasms; Cell Differentiation; Concanavalin A; Diet; Enteral Nutrition; Fatty Acids, Omega-3; Female; Humans; Immunity, Cellular; Immunosuppression Therapy; Interferon-gamma; Intraoperative Care; Lymphocyte Activation; Male; Neoplasm Staging; Pancreatic Neoplasms; Pancreaticoduodenectomy; Phytohemagglutinins; Preoperative Care; RNA; RNA, Messenger; Th1 Cells; Th2 Cells

We report a rare case of pyloric gland-type tubular adenoma of the main pancreatic duct. It was a grossly visible polypoid nodule and was composed of closely packed pyloric-type glands. This adenoma was present within an intraductal papillary mucinous adenoma (IPMA). In this IPMA lesion, aggregations of pyloric-type glands were occasionally observed, and most of the cells including ductal lining cells expressed pyloric gland-type mucin. The IPMA of the present case showed more extensive pyloric gland metaplasia or differentiation than commonly noted in IPMAs. We consider this pyloric gland-type tubular adenoma to be derived from a selective growth of IPMA cells showing pyloric gland metaplasia.

Topics: Adenoma; Aged; Antibodies, Monoclonal; Concanavalin A; DNA, Neoplasm; Exocrine Glands; Gallstones; Gastric Mucosa; Genes, ras; Humans; Immunohistochemistry; Male; Metaplasia; Mucins; Mutation; Neoplasms, Second Primary; Pancreatic Ducts; Pancreatic Neoplasms; Polymerase Chain Reaction

Previous investigations have suggested that expression of matrix metalloproteinases (MMPs) may be related to increased invasiveness of various tumors. This study evaluated a possible relation between pancreatic tumor cell invasiveness and MMPs.. A Matrigel invasion assay was performed with pancreatic tumor cell line SUIT-2 and its sublines S2-007, S2-013, S2-020, and S2-028. The degree of invasiveness of stimulated and unstimulated cell lines was correlated with MMP gene expression measured by RT-PCR and MMP protein product measured by gelatin zymography. Cell lines were stimulated by 12-O-tetradecanoylphorbol-13-acetate (TPA), concanavalin (Con-A), and polymerized collagen type I gel (Vitrogen).. For SUIT-2, S2-007, S2-013, S2-020, and S2-028, 3.2, 1.0, 4.1, 6.4, and 0.4%, respectively, of the cells invaded the Matrigel membrane. TPA, Con-A, and Vitrogen resulted in the up-regulation of MMP-2 in S2-020. TPA and Vitrogen resulted in up-regulation of MMP-9 in each of the cell lines, while Con-A could up-regulate MMP-9 expression only in SUIT-2. There was no constitutive expression of either MMP-2 or MMP-9 in SUIT-2 or its sublines. There was a positive relationship between Matrigel invasiveness and up-regulation of MMP-2 and MMP-9 expression.. These data suggest that, while MMP-2 and MMP-9 are not constitutively expressed in pancreatic carcinoma cell lines, they may be up-regulated by TPA, Con-A, and Vitrogen. Since MMP-2 and MMP-9 expression correlated with degree of tumor cell invasiveness, the ability to up-regulate MMP-2 and MMP-9 expression may play a role in facilitating pancreatic tumor cell invasion.

Topics: Biocompatible Materials; Carcinoma; Collagen; Concanavalin A; Drug Combinations; Extracellular Matrix; Gels; Gene Expression Regulation, Neoplastic; Humans; Laminin; Matrix Metalloproteinase 2; Matrix Metalloproteinase 9; Matrix Metalloproteinases, Membrane-Associated; Metalloendopeptidases; Neoplasm Invasiveness; Pancreatic Neoplasms; Proteoglycans; Reverse Transcriptase Polymerase Chain Reaction; Tetradecanoylphorbol Acetate; Tissue Inhibitor of Metalloproteinases; Tumor Cells, Cultured

Alterations of the N-linked carbohydrate core structure of cell surface glycoproteins (beta 1-6 branching) can be detected by phytohemagglutinin (PHA-L) lectin binding and has been linked to tumor progression and K-ras activation in colon cancer. The purpose of this study was to determine the prevalence of this carbohydrate alteration and its relationship to K-ras activation in pancreatic cancer. Nine human pancreatic cancer cell lines and 4 colon lines as controls were grown under standard tissue culture conditions. K-ras genome analysis was performed by polymerase chain reaction amplification and sequencing. The proportion of cellular p21-ras bound to GTP (ras-GTP level) was determined using immunoprecipitation of 32P-labeled cell lysates followed by thin layer chromatography and phosphoimaging analysis. Lectin blot analysis was performed on crude membrane preparations. Sensitivity to lectins was assessed with cell culture thymidine incorporation. Of 9 pancreatic cancer lines tested, 3 had wild type K-ras, 2 had heterozygous and 4 had homozygous mutations in codon 12 of K-ras. These genotypes correlated strongly with the level of ras-GTP measured. K-ras mutants had increased levels of ras-GTP compared to wild-type cell lines. PHA-L binding to cell membranes correlated positively with ras-GTP levels in 7 out of 9 cell lines. PHA-L toxicity was greatest in cells with positive PHA-L reactivity on Western blotting. A positive correlation between the presence of K-ras mutation, increased ras-GTP level, and increased cell surface beta 1-6 N-linked carbohydrate exists in pancreatic cancer cell lines.

Topics: Concanavalin A; Genes, ras; Guanosine Triphosphate; Humans; Membrane Glycoproteins; Mutation; Oncogene Protein p21(ras); Pancreatic Neoplasms; Phytohemagglutinins; Tumor Cells, Cultured

Topics: Adult; Aged; Antigens, Tumor-Associated, Carbohydrate; Biomarkers, Tumor; Concanavalin A; Female; Humans; Male; Middle Aged; Pancreatic Neoplasms

5'-Nucleotidase of a human pancreatic tumor cell line (PaTu II) has been purified to homogeneity after extraction with detergent followed by two affinity chromatographic steps. Sodium dodecyl sulphate polyacrylamide gel electrophoresis of purified 5'-nucleotidase revealed a single polypeptide band of 67 kDa. The Western blotted enzyme can be overlaid with concanavalin A proving its glycoprotein nature. After treatment with endoglycosidase F the deglycosylated 5'-nucleotidase exhibits an apparent molecular mass of 58 kDa. The kinetic properties of the solubilized enzyme have been determined (Km (AMP) of 4.0 microM; Vmax (AMP) = 8.6 muMOL/min.mg). Adenosine 5'-[alpha,beta-methylene]diphosphate is a competitive inhibitor of 5'-nucleotidase, whereas concanavalin A inhibits the enzymatic activity in a non-competitive manner. Polyclonal antibodies against purified 5'-nucleotidase of PaTu II have been produced which inhibit its enzymatic activity. Polyclonal antibodies raised against the enzyme purified from rat liver or bull seminal plasma also recognize 5'-nucleotidase of PaTu II cells, whereas polyclonal and monoclonal antibodies against the enzyme derived from chicken gizzard show no cross-reactivity. 5'-Nucleotidase appears to be concentrated in the plasma membrane of PaTu II cells as judged by cell fractionation and indirect immunofluorescence studies.

Topics: 5'-Nucleotidase; Adenosine Diphosphate; Antigen-Antibody Complex; Cell Line; Chromatography, Affinity; Concanavalin A; Electrophoresis, Polyacrylamide Gel; Fluorescent Antibody Technique; Humans; Kinetics; Molecular Weight; Pancreatic Neoplasms

Profiles of concanavalin A (Con A) and lentil agglutinin (LCH) affinity immunoelectrophoresis were compared for serum alpha-fetoprotein (AFP) from patients with yolk sac tumors and carcinomas of the gastrointestinal tract, in order to find some correlations between peaks of AFP subfractions detectable by two different lectins, and to investigate whether or not it is possible to prove that the binding of AFP to LCH is weakened to some extent if a fucosylated sugar chain has, in addition, a bisect N-acetylglucosamine (GlcNAc) attached to the beta-linked mannose. The results obtained with our improved techniques tend to indicate that a Con A-reactive AFP subfraction (peak a) corresponds to an LCH strongly reactive AFP (peak A), while a Con A-nonreactive AFP (peak b) corresponds to an LCH weakly reactive AFP (peak B). the authors consider the present data sufficient to support the above explanation.

Topics: alpha-Fetoproteins; Concanavalin A; Dysgerminoma; Female; Humans; Immunoelectrophoresis; Lectins; Mesonephroma; Ovarian Neoplasms; Pancreatic Neoplasms; Plant Lectins; Stomach Neoplasms

Techniques have been studied which distinguish two variants of human alpha-fetoprotein (AFP) on the basis of characteristics of the carbohydrate moiety of this glycoprotein. AFP in serum samples from six children with tumors of yolk sac origin showed little concanavalin-A (Con A) binding. In contrast, Con A binding of AFP was almost complete in serum samples from 14 other subjects with elevated AFP, including two with liver-cell tumors, eight with neonatal cholestasis, and four normal newborn infants. Differences were confirmed by immunoelectrophoretic studies. Thus, AFP from cells of yolk sac origin can be distinguished from AFP from liver cells or from tumors of hepatic cell origin.

Topics: alpha-Fetoproteins; Bile Ducts; Carcinoma, Hepatocellular; Child; Child, Preschool; Cholestasis; Chromatography, Affinity; Concanavalin A; Female; Hepatitis; Humans; Immunoelectrophoresis, Two-Dimensional; Infant; Infant, Newborn; Jaundice, Neonatal; Liver Neoplasms; Male; Mesonephroma; Pancreatic Neoplasms; Teratoma

The binding of concanavalin A to the plasmalemma of acinar carcinoma cells was characterized by electron microscopy utilizing horseradish peroxidase. Heavy labeling due to specific concanavalin A binding was detected on the plasmalemma of undifferentiated carcinoma cells lacking zymogen maturation, neoplastic cells of intermediate differentiation with only occasional zymogen granules, and highly differentiated acinar carcinoma cells containing numerous cytoplasmic zymogen granules. The plasmalemma of acinar carcinoma cells was also compared to the normal pancreatic acinar cell plasmalemma by measurement of specific 125I-labeled concanavalin A binding. Although only about one-third of pancreatic acinar carcinoma cells demonstrate mature zymogen differentiation, the acinar carcinoma had a full complement of normal plasmalemma receptors for 125I-labeled concanavalin A. It is concluded that, unlike normal pancreas, the presence of concanavalin A receptors on the plasmalemma of acinar carcinoma cells is not a specific membrane marker for differentiated cells containing zymogen granules.

Topics: Animals; Cell Differentiation; Concanavalin A; Kinetics; Microscopy, Electron; Neoplasms, Experimental; Pancreas; Pancreatic Neoplasms; Rats; Receptors, Concanavalin A

A method is described for using a fluorescein isothiocyanate concanavalin A conjugate to stain human cell membranes in formalin fixed paraffin embedded tissue. 57 neoplastic and normal tissue sites were examined. In 54 malignant tumours, bright green fluorescence was confined to the cell membranes while in 23 benign tumours and normal tissue sites, the membranes were unstained or showed a diminished level of fluorescence. The distinction between malignant and hyperplastic or normal cells was clear cut and definite.

Topics: Breast Neoplasms; Concanavalin A; Female; Fluorescein-5-isothiocyanate; Fluoresceins; Fluorescent Antibody Technique; Humans; Liver Neoplasms; Lung Neoplasms; Neoplasms; Pancreatic Neoplasms; Skin; Skin Neoplasms; Thiocyanates

The hypertrophic duct epithelium of the pancreas, including the pyloric gland metaplasia, mucous cell hypertrophy and ductal papillary hyperplasia were studied clinico-pathologically and histochemically to examine their precancerous character. A total of 180 surgical and autopsy specimens (90 pancreata with cancer and 90 pancreata without cancer) were analysed. The overall incidence of these three types of hypertrophic epithelium in the pancreas cancer was much higher than that in the pancreas without cancer. These hypertrophic lesions appeared most frequently in the interlobular duct. The histochemical study revealed the presence of a new type of glycoprotein in these hypertrophic duct epithelia, however, this substance was not detected in the cancer cells nor in the normal epithelium. This suggests that these hypertrophic lesions may not be the precursors of cancer but rather the coexistent lesions of pancreas cancer.

Topics: Adult; Aged; Concanavalin A; Cytoplasm; Epithelium; Glycoproteins; Glycosaminoglycans; Histocytochemistry; Horseradish Peroxidase; Humans; Hypertrophy; Middle Aged; Mucoproteins; Pancreatic Diseases; Pancreatic Ducts; Pancreatic Neoplasms; Precancerous Conditions; Staining and Labeling

Topics: Agglutination; Animals; Cell Membrane; Concanavalin A; Microvilli; Neoplasm Transplantation; Pancreas; Pancreatic Neoplasms; Rats; Receptors, Concanavalin A; Secretin

Topics: Antigens, Neoplasm; Concanavalin A; Electrophoresis, Polyacrylamide Gel; Glycoproteins; Humans; Immunodiffusion; Immunoelectrophoresis; Molecular Weight; Neuraminidase; Pancreas; Pancreatic Neoplasms; Papain; Pepsin A; Perchlorates; Trypsin

The use of Con A-Sepharose affinity chromatography for preparation of CEA from two metastatic liver tumors resulted in a separation of two species of CEA. One is concanavalin A-reactive CEA (CEA-M): the other is Con A-nonreactive CEA (CEA-P). Both CEA-M and CEA-P were glycoproteins and have identical antigenicity. However, 4 samples of CEA-M and CEA-P subfractions differed in their protein:carbohydrate ratios. The yield of CEA-M was greater than that of CEA-P. In electrophoresis, both CEA-M and CEA-P migrated at the region of beta-globulin of human blood serum. The isoelectric points of 4 samples of CEA-M and CEA-P subfractions from two different tumor sources differed from each other. In these preparations CEA-M usually showed a larger value of isoelectric point than CEA-P. Ultracentrifugal analysis of these four preparations revealed only a single peak, except CEA-P in case 1. Antigenic activity of CEA-M was almost completely destroyed by beta-N-acetylhexosaminidase treatment but only partly digestion with pronase. A possibility was suggested that N-acetylglucosamine at nonreducing terminal(s) is essential for the antigenic determinant groups of CEA molecule.

Topics: Binding Sites; Carbohydrates; Carcinoembryonic Antigen; Chromatography, Affinity; Concanavalin A; Hexosamines; Humans; Immunodiffusion; Immunoelectrophoresis; Liver Neoplasms; Molecular Weight; Neoplasm Metastasis; Pancreatic Neoplasms; Protein Binding; Rectal Neoplasms

Monosaccharide compositions of Con A-reactive CEA (CEA-M) and Con A-nonreactive CEA (CEA-P) separated from two different samples of CEA were analysed by gas liquid chromatography. It was revealed that all CEA subfractions possessed N-acetylglucosamine, fucose, and galactose residues. One out of 4 subfractions did not contain sialic acid and another one lacked glucose in its carbohydrate moiety. N-acetylgalactosamine was not detected in measurable amount in any of the 4 subfractions. A large amount of mannose was found in CEA-M, but only a small amount in CEA-P.

Topics: Acetylgalactosamine; Acetylglucosamine; Binding Sites; Carcinoembryonic Antigen; Chromatography, Affinity; Concanavalin A; Fucose; Galactose; Glucosamine; Humans; Liver Neoplasms; Mannose; Neoplasm Metastasis; Pancreatic Neoplasms; Protein Binding; Rectal Neoplasms; Sialic Acids