muramidase and Colonic-Neoplasms

Enzymological alterations in functional disturbances and in diseases of the intestine are reviewed. Examples are given for diagnostic significance (e.g. in Hirschsprung's and Crohn's diseases), for pathogenetic considerations (e.g. in hypolactasia and in celiac disease), and for secondary involvement of the liver (e.g. in intestinal tumors and after bypass surgery) and are discussed in more detail.

Topics: Acetylcholinesterase; Adult; Alcoholism; Animals; Antineoplastic Agents; Black People; Celiac Disease; Child; Child, Preschool; Colonic Neoplasms; Diarrhea; Enteritis; Enteropeptidase; Humans; Infant; Intestinal Diseases; Isoenzymes; Jejunum; L-Lactate Dehydrogenase; Lactose; Malabsorption Syndromes; Megacolon; Microbial Collagenase; Microvilli; Muramidase; Rats; Rectum; White People

Topics: Animals; Autoimmune Diseases; Biopsy; Colitis, Ulcerative; Colonic Diseases; Colonic Neoplasms; Diagnosis, Differential; Eye Manifestations; Humans; Lactose Intolerance; Liver Diseases; Milk; Muramidase; Prognosis; Psychophysiologic Disorders; Radiography; Sigmoidoscopy

The goal of this study was to determine the unique characteristics of Enterococcus faecium MC-5, a probiotic bacteria isolated from the intestine of a fish, Cyprinus carpio specularis, collected from Dal Lake in Srinagar, Kashmir, India. For this, the important valuable probiotic attributes, some functional properties, and safety assessments were analyzed in-vitro for the strain MC-5. The strain E. faecium MC-5 exhibited high resistance to low pH, high bile salt, lysozyme, and phenol. The strain MC-5 showed excellent auto- and co-aggregation properties and displayed remarkable hydrophobicity towards various tested hydrocarbons which suggested that the strain possesses venerable adhesion properties. Apart from these, the cell-free supernatant (CFS) of strain MC-5 exhibited phenomenal antimicrobial activity against the tested pathogens. A scanning electron microscope (SEM) image revealed strain MC-5 finely adhered to human colon adenocarcinoma cells (HCT-15 cells). The strain MC-5 showed high bile salt hydrolase activity and excellent cholesterol removal ability of 70.27%. The intact cells of strain MC-5 also showed strong DPPH scavenging activity. The EPS produced by E. faecium MC-5 inhibited the adhesion of Listeria monocytogenes, Staphylococcus aureus, and Salmonella enterica on HCT-15 cells with maximum inhibition rates of 41.82, 40.34, and 55.51%, respectively for displacement assay, which was higher as compared to exclusion (26.06, 26.11, and 39.23%) and competition assays (30.06, 26.7, and 41.20%). Strain MC-5 did not exhibit hemolysis and was also found susceptible to vancomycin and other clinically important antibiotics. When evaluating all the results from the present study, it is propounded that strain MC-5 has enviable probiotic characteristics and thus can be used as bio-protective cultures and/or bio-shield in food and pharmaceutical industries.

Topics: Adenocarcinoma; Animals; Anti-Bacterial Agents; Bile Acids and Salts; Carps; Cell Line, Tumor; Cholesterol; Colonic Neoplasms; Enterococcus faecium; Humans; Muramidase; Phenols; Probiotics; Vancomycin

Chitosan derivatives such as N,N,N-trimethylated chitosan (TMC) are currently being investigated for the delivery of drugs, vaccines and genes. However, the influence of the extent of N-acetylation of these polymers on their enzymatic degradability and biological properties is unknown. In this study, TMCs with a degree of acetylation (DA) ranging from 11 to 55% were synthesized by using a three-step method. First, chitosan was partially re-acetylated using acetic anhydride followed by quantitative dimethylation using formaldehyde and sodium borohydrate. Then, in presence of an excess amount of iodomethane, TMC was synthesized. The TMCs obtained by this method showed neither detectable O-methylation nor loss in acetyl groups ((1)H NMR) and a slight increase in molecular weight (GPC) with increasing degree of substitution, implying that no chain scission occurred during synthesis. The extent of lysozyme-catalyzed degradation of TMC, and that of its precursors chitosan and dimethyl chitosan, was highly dependent on the DA and polymers with the highest DA showed the largest decrease in molecular weight. On Caco-2 cells, TMCs with a high DA ( approximately 50%), a DQ of around 44% and with or without O-methylated groups, were not able to open tight junctions in the trans-epithelial electrical resistance (TEER) assay, in contrast with TMCs (both O-methylated and O-methyl free; concentration 2.5mg/ml) with a similar DQ but a lower DA which were able to reduce the TEER with 30 and 70%, respectively. Additionally, TMCs with a high DA ( approximately 50%) demonstrated no cell toxicity (MTT, LDH release) up to a concentration of 10mg/ml.

Topics: Acetylation; Adenocarcinoma; Animals; Biopolymers; Cell Line, Tumor; Cell Survival; Chickens; Chitosan; Colonic Neoplasms; Electric Impedance; Epithelial Cells; Humans; Hydrolysis; L-Lactate Dehydrogenase; Methylation; Molecular Weight; Muramidase; Nuclear Magnetic Resonance, Biomolecular; Tight Junctions

Colon cancers develop after accumulation of multiple genetic and epigenetic alterations in colon epithelial cells. To shed light on global changes in gene expression of colon cancers and to gain further insight into the molecular mechanisms underlying colon carcinogenesis, we have conducted a comprehensive microarray analysis of mRNA using a rat colon cancer model with the food-borne carcinogen, 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP). Of 8749 genes or ESTs on a high density oligonucleotide microarray, 27 and 46 were over- and underexpressed, respectively, by > or =3-fold in colon cancers in common in two rat strains with distinct susceptibility to PhIP carcinogenesis. For example, genes involved in inflammation and matrix proteases and a cell cycle regulator gene, cyclin D2, were highly expressed in colon cancers. In contrast, genes encoding structural proteins, muscle-related proteins, matrix-composing and mucin-like proteins were underexpressed. Interestingly, a subset of genes whose expression is characteristic of Paneth cells, i.e. the defensins and matrilysin, were highly overexpressed in colon cancers. The presence of defensin 3 and defensin 5 transcripts in cancer cells could also be confirmed by in situ mRNA hybridization. Furthermore, Alcian blue/periodic acid Schiff base (AB-PAS) staining and immunohistochemical analysis with an anti-lysozyme antibody demonstrated Paneth cells in the cancer tissues. AB-PAS-positive cells were also observed in high grade dysplastic aberrant crypt foci, which are considered to be preneoplastic lesions of the colon. Our results suggest that Paneth cell differentiation in colon epithelial cells could be an early morphological change in cryptic cells during colon carcinogenesis.

Topics: Alcian Blue; Animals; Carcinogens; Cell Differentiation; Cell Line, Tumor; Colonic Neoplasms; Cyclin D2; Cyclins; Defensins; Expressed Sequence Tags; Gene Expression Regulation, Neoplastic; Humans; Imidazoles; Immunohistochemistry; Muramidase; Nucleic Acid Hybridization; Oligonucleotide Array Sequence Analysis; Oligonucleotides; Paneth Cells; Rats; Rats, Inbred F344; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger

The presence of lysozyme protein in some gastric adenomas and adenocarcinomas has been well documented. There have been relatively few studies investigating the presence of lysozyme in tumours of the large intestine and they show contrasting results. We aim to investigate the cellular source and expression of lysozyme in colonic adenomas and adenocarcinomas.. We randomly selected 29 and 27 colonic adenomas and adenocarcinomas, respectively. Using in-situ hybridization (ISH) and immunohistochemistry (IHC), we found an up-regulation of lysozyme in the dysplastic epithelium of all the adenomas studied, with more than 80% of cases expressing moderate to strong signals. Although the up-regulation of lysozyme was also observed in adenocarcinomas, only 30% of the cases showed moderate to strong signals, mostly with an uneven distribution. Down-regulation of lysozyme in the severely dysplastic and invasive foci were noted in some cases of adenoma with malignant transformation. Normal colonic glands were consistently negative for lysozyme at both the mRNA and the protein level, but inflamed and immature regenerative colonic epithelium at the crypt base showed positive signals in a similar pattern to those observed in the dysplastic epithelium of the adenomas.. Our results confirm that colonic epithelium can produce lysozyme and its expression is up-regulated in the dysplastic epithelium in adenomas and in invasive cancer cells. It is interesting that regenerative colonic epithelium showed a similar pattern of lysozyme expression as in adenomas. The loss of lysozyme secreting phenotype in most of the invasive tumours suggests that lysozyme may not confer an advantage to tumour progression.

Topics: Adenocarcinoma; Adenoma; Colonic Neoplasms; Humans; Immunohistochemistry; In Situ Hybridization; Intestinal Mucosa; Muramidase; Random Allocation; RNA, Messenger; Up-Regulation

alpha 1-Antitrypsin (alpha 1-AT) is an acute phase plasma protein predominantly derived from the liver which inhibits neutrophil elastase. Previous studies have suggested that alpha 1-AT is also expressed in human enterocytes because alpha 1-AT mRNA could be detected in human jejunum by RNA blot analysis, and alpha 1-AT synthesis could be detected in a human intestinal adenocarcinoma cell line Caco2, which spontaneously differentiates into villous-like enterocytes in tissue culture. To definitively determine that the alpha 1-AT gene is expressed in human enterocytes in vivo, we examined tissue slices of human jejunum and ileum by in situ hybridization. The results demonstrate specific hybridization to enterocytes from the bases to the tips of the villi. Although there was no hybridization to enterocytes in most of the crypt epithelium, there was intense specific hybridization in one region of the crypt. Double-label immunohistochemical studies showed that alpha 1-AT and lysozyme co-localized to this region, indicating that it represented Paneth cells. Finally, there was a marked increase in hybridization to alpha 1-AT mRNA in villous enterocytes and Paneth cells in Crohn's disease. The results of this study provide definitive evidence that alpha 1-AT is expressed in human jejunal and ileal enterocytes in vivo, and show that alpha 1-AT is also a product of Paneth cells. Together with the results of other studies, these data raise the possibility that alpha 1-AT detected in fecal alpha 1-AT clearance assays for diagnosing protein-losing enteropathies is predominantly derived from sloughed enterocytes.

Topics: Adenocarcinoma; alpha 1-Antitrypsin; alpha 1-Antitrypsin Deficiency; Antisense Elements (Genetics); Colonic Neoplasms; Epithelial Cells; Epithelium; Fluorescent Antibody Technique; Gene Expression; Humans; Ileum; Immunohistochemistry; In Situ Hybridization; Jejunum; Liver; Muramidase; RNA, Messenger; Tumor Cells, Cultured

An immuno-histological study of metastatic adenocarcinoma has revealed the following results. Metastatic adenocarcinomas of the lymph-node of pulmonary and colonic origin were positive for CEA and negative for lysozymes, and those from gastric, pancreatic, and gallbladder tumors were positive CEA and lysozymes, and those from gastric and pancreatic tumors were positive for the secretory component. The prostate specific antigen was exclusively positive for metastatic prostatic adenocarcinoma with a low frequency and prostate acid phosphatase had many false positive results. Thyroglobulin was found to be positive only to colloid. Lactalbumin showed no specificity to metastatic breast adenocarcinoma. For achieving the final diagnosis of a primary tumor, its location in lymph nodes, the clinical history and the results of other examinations must also be taken into consideration.

Topics: Adenocarcinoma; Biomarkers, Tumor; Carcinoembryonic Antigen; Colonic Neoplasms; Humans; Keratins; Lactalbumin; Lung Neoplasms; Lymphatic Metastasis; Muramidase; Neoplasms, Unknown Primary; Secretory Component

Topics: alpha 1-Antitrypsin; Animals; Cat Diseases; Cats; Colonic Neoplasms; Female; Immunoenzyme Techniques; Muramidase; Sarcoma

After diluting faecal samples with a solution of Brij and saline and subsequently ultrafiltrating the faecal mixtures, lysozyme concentration can be reproducibly measured in the obtained faecal fluids, using a turbidimetric method. Measuring faecal lysozyme concentration enables discrimination normal individuals and patients with irritable bowel syndrome between patients with inflammatory bowel disease and colonic cancer. Lysozyme distribution in stools appears to be homogeneous. Faecal lysozyme concentration is stable when samples are stored during at least 1 wk at 6 degrees C. It appears that the lysozyme activity is directly correlated with the clinical status and severity of the disease. Faecal lysozyme may thus serve as an important tool both in diagnosis and in follow-up in the out-patients clinic for gastroenterology.

Topics: Adenocarcinoma; Adult; Aged; Aged, 80 and over; Colonic Diseases, Functional; Colonic Neoplasms; Feces; Gastrointestinal Diseases; Humans; Immunodiffusion; Middle Aged; Muramidase; Nephelometry and Turbidimetry; Reference Standards

Six of a total of 14 human colon carcinoma cell lines produce and secrete lysozyme in vitro. Three also produce the enzyme when propagated in vivo in athymic mice. None of the lysozyme positive cells stained in a manner typical of Paneth cells. Additionally, lysozymes from all six colon lines possess identical molecular weights (approximately 14,000 daltons).

Topics: Animals; Cell Line; Colonic Neoplasms; Histocytochemistry; Humans; Immunoenzyme Techniques; Mice; Mice, Inbred BALB C; Muramidase; Neoplasm Transplantation

Peripheral monocyte migration, phagocytosis and lysozyme production has been investigated in 46 patients with colorectal malignancies. The diagnosis of colorectal carcinoma was based on clinical, radiological, and endoscopic examinations and confirmed histopathologically after removal of the tumour. The results were compared with a group of 36 normal healthy controls. Monocytes from patients with colorectal malignancies displayed reduced migration towards the chemotactic agent (P less than 0.001), random migration (P less than 0.001), and phagocytic activity (P less than 0.01) compared to normal controls. Lysozyme production by monocytes from patients with colorectal malignancies was significantly increased (P less than 0.01) compared to control subjects. The impairment in monocyte function in patients with colorectal carcinoma was not due to direct serum factors or to abnormal blood biochemistry, suggesting that the defect is intrinsic to the monocytes. These results suggest that an impairment in monocyte function may play an important role in the control of malignancies by cellular immune processes.

Topics: Adult; Aged; Blood Specimen Collection; Cell Movement; Cell Separation; Colonic Neoplasms; Female; Humans; Immunity, Cellular; Male; Middle Aged; Monocytes; Muramidase; Phagocytosis; Rectal Neoplasms

A protein-free synthetic cell-growth medium has been defined that permits long-term survival (greater than 120 days) of an established human colon tumor cell line, HT-29. Viability is dependent upon both the concentration of L-glutamine in the medium and the cell density at the time of initial transfer into it. Cell proliferation is minimal, thus obviating the necessity for subculturing. HT-29 adenocarcinoma cells maintained in large-scale culture with this medium continue to secrete the established colon tumor marker carcinoembryonic antigen as well as growth factors and lysozyme. These and, potentially, other important tumor-derived products can therefore be generated continuously in such cultures so that they can be isolated from a conditioned medium free of contaminating serum and protein supplements.

Topics: Carcinoembryonic Antigen; Carcinoma; Cell Line; Colonic Neoplasms; Culture Media; Fibrosarcoma; Glutamine; Growth Substances; Humans; Lung Neoplasms; Muramidase; Proteins

One of the major proteins secreted by an established human colon adenocarcinoma cell line has been isolated in 25% yield from the serum-free medium in which the cells were grown and identified as lysozyme. Its purification was achieved by sequential steps of acidification, cation-exchange chromatography, and reversed-phase high-performance liquid chromatography. It was recognized to be a human lysozyme on the basis of its molecular weight (14 000), isoelectric point (10.5), amino acid composition, and enzymatic activity. Its identity with previously characterized human lysozymes was established by amino-terminal sequence, peptide composition, immunological properties, NMR, and crystallography. A 4-day, 7-L collection of conditioned medium contained 20.3 mg of secreted protein of which 4.9 mg or approximately 24% of the total was tumor-derived lysozyme. The intracellular level of lysozyme was approximately 18 ng per 10(6) carcinoma cells. The possible significance of these findings in regard to the malignant process and tumor maintenance is discussed.

Topics: Adenocarcinoma; Amino Acid Sequence; Cell Line; Colonic Neoplasms; Crystallization; Culture Media; Humans; Immunodiffusion; Molecular Weight; Muramidase; Peptide Fragments

Five thousand seven hundred seventy-eight adenomas or adenomas containing carcinoma from 3215 patients were examined by routine histologic methods for the presence of epithelial metaplasias. Three forms of epithelial metaplasia were encountered: squamous cell metaplasia (0.44%), Paneth cell metaplasia (0.20%), and melanocytic metaplasia (0.017%). In several instances multiple forms of metaplasia were encountered in the same polyp. In those cases in which the paraffin blocks were available, a Grimelius stain was performed. Grimelius-positive cells were present in 63% of the adenomas containing a metaplastic cell type. All cases with Paneth cell differentiation were immunoreactive for lysozyme; all lesions containing areas of squamous differentiation were immunoreactive for keratin except 2. The histopathologic features of these cases are discussed, and it is concluded that rather than representing a true metaplastic process, Paneth cell, squamous cell, and melanocyte differentiation represent the full range of cellular differentiation that endodermally derived tissues can exhibit, particularly when they undergo neoplastic alterations.

Topics: Adenoma; Adult; Age Factors; Aged; Cell Differentiation; Colonic Neoplasms; Female; Histocytochemistry; Humans; Intestinal Polyps; Intestine, Large; Keratins; Male; Melanocytes; Metaplasia; Middle Aged; Muramidase; Rectal Neoplasms; Retrospective Studies; Sex Factors

The light microscopic, electron microscopic and histochemical features of a highly malignant colonic tumor resected from a 39 year old man are presented. The tumor was composed predominantly of undifferentiated cells with focally admixed neuroendocrine, exocrine and squamous cells, occasionally arranged in an organoid manner. Histochemically the tumor contained argyrophilic cells as well as cells that reacted positively with the antibodies to alpha-1-antitrypsin, alpha-1-antichymotrypsin, carcinoembryonic antigen and lysozyme. The term "stem cell carcinoma of the intestine" is proposed for this highly malignant tumor composed of undifferentiated cells exhibiting only focally their multidirectional developmental capacity.

Topics: Adult; alpha 1-Antichymotrypsin; alpha 1-Antitrypsin; Carcinoembryonic Antigen; Carcinoma; Chymotrypsin; Colonic Neoplasms; Histocytochemistry; Humans; Immunologic Techniques; Keratins; Male; Microscopy, Electron; Muramidase; Staining and Labeling

Topics: Adenocarcinoma, Mucinous; alpha 1-Antitrypsin; Colonic Neoplasms; Humans; Muramidase; Rectal Neoplasms

Topics: Aged; Colonic Neoplasms; Feces; Humans; Middle Aged; Muramidase; Rectal Neoplasms

Lysozyme, alpha 1-Antichymotrypsin and alpha 1-Antitrypsin were demonstrated by an immunoperoxidase technique (PAP) in malignant cells of adenocarcinomas of the stomach but not of the large intestine. Lymph-node metastases showed identical immunoreactivity to that of the primary tumour. Neoplasms arising from the cardia, the body and the pyloric antrum of the stomach showed different immunostaining reactions. It seems that these differences partly reflect the distribution of lysozyme, alpha 1-Antichymotrypsin and alpha 1-Antitrypsin in the normal gastric mucosa. The usefulness of our findings in the identification of the primary tumour in cases of lymph node metastases of unknown origin, is also discussed.

Topics: Adenocarcinoma; alpha 1-Antichymotrypsin; alpha 1-Antitrypsin; Cardia; Chymotrypsin; Colonic Neoplasms; Gastric Mucosa; Histocytochemistry; Humans; Immunoenzyme Techniques; Lymphatic Metastasis; Muramidase; Pyloric Antrum; Stomach Neoplasms

The determination of lysozyme has been shown to be more relevant than assumed until now. It can be used as a marker in the therapy of acute and chronic urinary tract infections. The determination of lysozyme in cerebro spinal fluid and blood serum are helpful to differentiate between bacterial and aseptic meningitides or infections. Elevated fecal lysozyme excretion in adolescents are an indicator for a chronic inflammatory bowel disease. Control of fecal lysozyme excretion can be used as a marker for a relapse and to monitor therapeutic efficiency in patients with inflammatory bowel disease. A consistent high level of fecal lysozyme excretion in adults over the age of 40 is an indicator for possible colorectal tumors and warrants further thorough investigation.

Topics: Adolescent; Adult; Bacterial Infections; BCG Vaccine; Colonic Neoplasms; Crohn Disease; Diagnosis, Differential; Feces; Humans; Macrophages; Meningitis; Monocytes; Muramidase

Topics: Adolescent; Colonic Neoplasms; Crohn Disease; Humans; Middle Aged; Muramidase; Rectal Neoplasms

Topics: Colonic Neoplasms; Humans; Muramidase; Rectal Neoplasms

The serum muramidase levels were measured in 128 patients with primary or metastatic colorectal cancer, 166 tumour-free patients after resection of a colorectal cancer, and 172 controls. Muramidase levels over 10 mug/ml were detected in 30%-39% of the tumour-bearing patients, in 8.2% of the tumour free, and in only 1.7% of the controls (normal level 6.68 +/- 1.42 mug/ml). Long-term follow up indicated that raised levels may occur as a transient phenomenon in recurrent or metastatic disease. The likely relation of abnormal serum muramidase activity and stimulation of the reticuloendothelial system is discussed.

Topics: Adenocarcinoma; Carcinoembryonic Antigen; Colonic Neoplasms; Creatinine; gamma-Glutamyltransferase; Humans; Kidney Failure, Chronic; Leucyl Aminopeptidase; Liver Neoplasms; Muramidase; Neoplasm Metastasis; Rectal Neoplasms; Recurrence

Topics: Animals; Appendicitis; Biological Evolution; Cell Division; Colitis, Ulcerative; Colonic Neoplasms; Crohn Disease; Ethionine; Gastritis; Germ-Free Life; Histocytochemistry; Humans; Intestinal Neoplasms; Intestines; Metaplasia; Microscopy, Electron; Muramidase; Peutz-Jeghers Syndrome; Puromycin; Rats; Stomach Neoplasms; Stomach Ulcer; Triparanol; Whipple Disease; Zinc