Chitin, a polymer of N-acetyl-D-glucosamine (GlcNAc), is a major structural component in chitin-containing organism including crustaceans, insects and fungi. Mammals express two chitinases, chitotriosidase (Chit1) and acidic mammalian chitinase (AMCase). Here, we report that pig AMCase is stable in the presence of other digestive proteases and functions as chitinolytic enzyme under the gastrointestinal conditions. Quantification of chitinases expression in pig tissues using quantitative real-time PCR showed that Chit1 mRNA was highly expressed in eyes, whereas the AMCase mRNA was predominantly expressed in stomach at even higher levels than the housekeeping genes. AMCase purified from pig stomach has highest activity at pH of around 2-4 and remains active at up to pH 7.0. It was resistant to robust proteolytic activities of pepsin at pH 2.0 and trypsin and chymotrypsin at pH 7.6. AMCase degraded polymeric chitin substrates including mealworm shells to GlcNAc dimers. Furthermore, we visualized chitin digestion of fly wings by endogenous AMCase and pepsin in stomach extract. Thus, pig AMCase can function as a protease resistant chitin digestive enzyme at broad pH range present in stomach as well as in the intestine. These results indicate that chitin-containing organisms may be a sustainable feed ingredient in pig diet.
- MeSH
- chitin metabolismus MeSH
- chitinasy genetika izolace a purifikace metabolismus MeSH
- chymotrypsin metabolismus MeSH
- dieta * MeSH
- Drosophila chemie MeSH
- endopeptidasy metabolismus MeSH
- gastrointestinální trakt metabolismus MeSH
- křídla zvířecí chemie MeSH
- messenger RNA genetika metabolismus MeSH
- orgánová specificita MeSH
- pepsinogen A metabolismus MeSH
- prasata genetika MeSH
- rozpustnost MeSH
- substrátová specifita MeSH
- Tenebrio MeSH
- tkáňové extrakty MeSH
- trypsin metabolismus MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
AIM: The first aim of this study was to search for new biomarkers to be used in gastric cancer diagnostics. The second aim was to verify the findings presented in literature on a sample of the local population and investigate the risk of gastric cancer in that population using a multivariant statistical analysis. PATIENTS AND METHODS: We assessed a group of 36 patients with gastric cancer and 69 healthy individuals. We determined carcinoembryonic antigen, cancer antigen 19-9, cancer antigen 72-4, matrix metalloproteinases (-1, -2, -7, -8 and -9), osteoprotegerin, osteopontin, prothrombin induced by vitamin K absence-II, pepsinogen I, pepsinogen II, gastrin and Helicobacter pylori for each sample. RESULTS: The multivariate stepwise logistic regression identified the following biomarkers as the best gastric cancer predictors: CEA, CA72-4, pepsinogen I, Helicobacter pylori presence and MMP7. CONCLUSION: CEA and CA72-4 remain the best markers for gastric cancer diagnostics. We suggest a mathematical model for the assessment of risk of gastric cancer.
- MeSH
- antigeny sacharidové asociované s nádorem krev MeSH
- dospělí MeSH
- Helicobacter pylori imunologie MeSH
- imunoglobulin G krev MeSH
- infekce vyvolané Helicobacter pylori imunologie MeSH
- karcinoembryonální antigen krev MeSH
- lidé středního věku MeSH
- lidé MeSH
- matrixová metaloproteinasa 7 krev MeSH
- multivariační analýza MeSH
- nádorové biomarkery krev MeSH
- nádory žaludku krev diagnóza MeSH
- pepsinogen A krev MeSH
- protilátky bakteriální krev MeSH
- rizikové faktory MeSH
- senioři nad 80 let MeSH
- senioři MeSH
- teoretické modely * MeSH
- Check Tag
- dospělí MeSH
- lidé středního věku MeSH
- lidé MeSH
- senioři nad 80 let MeSH
- senioři MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Isolated porcine pepsinogen A was used for the preparation of polyclonal rabbit and polyclonal chicken anti-pepsinogen A antibodies. Immunochemical properties of both immunoglobulin fractions were compared. The rabbit anti-serum was further purified using immobilized porcine pepsinogen A on magnetic cellulose beads and the resulting anti-pepsinogen A fraction proved to be applicable for the separation and the determination of porcine pepsinogen A. In contrary, antibodies prepared from chicken eggs by the same way have been found not suitable for the evaluation of the pepsinogen A level. Unexpectedly, the pre-immune fraction of chicken antibodies showed reactivity against porcine pepsinogen A and the affinity separation of specific polyclonal chicken anti-pepsinogen A antibodies on immobilized porcine pepsinogen A did not result in an enrichment of anti-pepsinogen A antibodies.
- MeSH
- ELISA MeSH
- imunohistochemie MeSH
- králíci MeSH
- kur domácí MeSH
- nádory žaludku imunologie metabolismus MeSH
- pepsinogen A imunologie metabolismus MeSH
- prasata MeSH
- protilátky imunologie MeSH
- skot MeSH
- žaludeční sliznice imunologie metabolismus patologie MeSH
- zvířata MeSH
- Check Tag
- králíci MeSH
- skot MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- MeSH
- dospělí MeSH
- ELISA MeSH
- endoskopie MeSH
- gastritida diagnóza etiologie patologie MeSH
- Helicobacter pylori patogenita MeSH
- lidé středního věku MeSH
- lidé MeSH
- pepsinogen A krev MeSH
- senioři MeSH
- žaludeční sliznice patologie MeSH
- Check Tag
- dospělí MeSH
- lidé středního věku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- senioři MeSH
- ženské pohlaví MeSH
- Publikační typ
- přehledy MeSH
- srovnávací studie MeSH
- MeSH
- finanční podpora výzkumu jako téma MeSH
- imunoblotting metody MeSH
- imunochemie metody MeSH
- lidé MeSH
- pepsin A krev MeSH
- pepsinogen A krev MeSH
- protilátky enzymologie MeSH
- žaludeční sliznice MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- přehledy MeSH
- MeSH
- dospělí MeSH
- dvanáctníkové vředy farmakoterapie krev mikrobiologie MeSH
- Helicobacter pylori izolace a purifikace MeSH
- kombinovaná farmakoterapie MeSH
- lidé středního věku MeSH
- lidé MeSH
- pepsinogen A krev MeSH
- senioři MeSH
- výsledek terapie MeSH
- Check Tag
- dospělí MeSH
- lidé středního věku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- senioři MeSH
- ženské pohlaví MeSH
