A tight link exists between dietary factors and irritable bowel syndrome (IBS), one of the most common functional syndromes, characterized by abdominal pain/discomfort, bloating and alternating bowel habits. Amongst the variety of foods potentially evoking "food sensitivity", gluten and other wheat proteins including amylase trypsin inhibitors represent the culprits that recently have drawn the attention of the scientific community. Therefore, a newly emerging condition termed non-celiac gluten sensitivity (NCGS) or non-celiac wheat sensitivity (NCWS) is now well established in the clinical practice. Notably, patients with NCGS/NCWS have symptoms that mimic those present in IBS. The mechanisms by which gluten or other wheat proteins trigger symptoms are poorly understood and the lack of specific biomarkers hampers diagnosis of this condition. The present review aimed at providing an update to physicians and scientists regarding the following main topics: the experimental and clinical evidence on the role of gluten/wheat in IBS; how to diagnose patients with functional symptoms attributable to gluten/wheat sensitivity; the importance of double-blind placebo controlled cross-over trials as confirmatory assays of gluten/wheat sensitivity; and finally, dietary measures for gluten/wheat sensitive patients. The analysis of current evidence proposes that gluten/wheat sensitivity can indeed represent a subset of the broad spectrum of patients with a clinical presentation of IBS.
Amylases ; Biomarkers ; Cross-Over Studies ; Diagnosis ; Glutens* ; Humans ; Irritable Bowel Syndrome* ; Triticum ; Trypsin Inhibitors

In canine acute pancreatitis (AP), inappropriate release and activation of zymogen proteases within the pancreas results in the consumption of serum antiproteases. The aim of this study was to examine whether the serum concentrations of α₂-macroglobulin (A2MG), α₁-antitrypsin (A1AT), and C-reactive protein (CRP) differ between dogs with AP and healthy dogs. Twenty healthy dogs and 20 dogs with AP were included in this study. Concentrations of A2MG, A1AT, and CRP were measured in the sera of healthy dogs and dogs diagnosed with AP. Serum A2MG and A1AT concentrations were significantly lower in dogs with AP than in healthy dogs, whereas the serum CRP concentration was significantly higher. In addition, the concentrations of A2MG and A1AT were significantly higher in AP survivors than in AP non-survivors, while the CRP concentration was significantly lower. However, in both AP survivors and non-survivors, the CRP concentrations showed a negative correlation with A2MG concentrations but not with A1AT. These findings indicate that serum antiproteases and CRP concentrations might be associated with the mortality rate of AP in dogs.
Animals ; C-Reactive Protein ; Dogs ; Humans ; Mortality ; Pancreas ; Pancreatitis ; Peptide Hydrolases ; Protease Inhibitors ; Survivors ; Trypsin

BACKGROUND: Urinary trypsin inhibitors (UTI) have been widely used for the treatment of diseases including disseminated intravascular coagulation, shock, and pancreatitis. Since UTI synthesis is likely to be reduced in patients who have undergone liver resection, the incidence of inflammatory reactions may be increasing accordingly. For such patients, the liver enzyme increases after the operation can reflect liver damage. The purpose of this study was to examine if ulinastatin can inhibit liver enzyme increases after liver resection. METHODS: After receiving Institutional Review Board approval, a retrospective chart review was performed on 201 patients who underwent hepatic resection from 2006 to 2010. We divided the records into the control (n = 69) and ulinastatin (n = 132) groups according to the use of intraoperative ulinastatin and compared the preoperative and postoperative laboratory test results. The number of patients who had > 400 U/L elevation of aspartate transaminase (AST) level after surgery was compared between the 2 groups. RESULTS: The mean AST, alanine transaminase (ALT), and total bilirubin levels after liver resection were significantly lower in the ulinastatin group than in the control group. The number of patients who showed an AST > 400 U/L after liver resection was significantly higher in the control group (odds ratio = 3.02). CONCLUSIONS: Ulinastatin attenuates the elevation of hepatic enzymes and bilirubin after liver resection.
Alanine Transaminase ; Aspartate Aminotransferases ; Bilirubin ; Disseminated Intravascular Coagulation ; Ethics Committees, Research ; Glycoproteins ; Hepatectomy ; Humans ; Incidence ; Liver ; Liver Function Tests ; Pancreatitis ; Retrospective Studies ; Shock ; Trypsin ; Trypsin Inhibitors

BACKGROUND: Recently, much attention has been focused on the role of protease inhibitors such as acid stable trypsin inhibitor (ASTI) in the invasive growth of malignant tumors. However, there are no report on expression of ASTI from premalignant and/or malignant skin tumors. OBJECTIVES: In the present study the expression of ASTI was investigated in the different type of premalignant and/or, malignant skin tumors in attempt to clarify the relation between the expression of the ASTI and malignancy. METHODS: For the detection of ASTI in the tumor tissue, the immunoperoxidase techniques that used mouse antibody raised against highly purified ASTI. The degree of ASTI immunoreactivity was semiquantitatively assessed for staining intensity as the percentage of ASTI-positive cells. RESULTS: ASTI immunoreactivity was detected in most of the premalignant and malignant skin tissues. Especially, ASTI expression was present widespread in squamous cell carcinoma(SCC) with strong cytoplasmic membrane, where as in normal epidermis they were primarily present in the horny layer. The strong staining was the SCC, keratoacanthoma, Bowen's disease, actinic keratosis, basal cell epithelioma, Paget's disease in decreasing order. The significant difference in the staining intensity was observed between SCC and other groups. CONCLUSION: Results of immunohistochemical studies suggest that the tumor cells themselves could produce ASTI. Considering the suggestion that ASTI is a self protector, inhibitor to proteolytic protease as well as growth-stimulating factor. The present findings may indicate that ASTI expressed in malignant cells may play a role possibly closely associated with tumor development.
Animals ; Bowen's Disease ; Carcinoma, Basal Cell ; Cell Membrane ; Epidermis ; Immunoenzyme Techniques ; Keratoacanthoma ; Keratosis, Actinic ; Mice ; Protease Inhibitors ; Skin* ; Trypsin*

BACKGROUND: Eosinophilic leukocytes are prominent cellular participants in the pathogenesis of allergic disease and asthma. Chemotaxis is still a very useful method in evaluating the response of human eosinophil to novel modulators. Degranulated mast cells and activated T lymphocytes are responsible for the pathophysiology of asthma and tryptase is one of most important proteases released after activation of mast cells. The purpose of this study was to investigate the actions of trypsin and chymotrypsin on eosinophils in terms of chemotaxis and activation. METHOD: Eosinophils were isolated by negative immunoselection from the peripheral blood of atopic donors. Chemotaxis was studied by using micro-Boyden chambers and ECP release was assayed by fluoroimmunoassay. RESULTS: Eosinophil showed a chemotactic response to trypsin. Maximal chemotactic response was with 1000microg/ml trypsin (56.52 +/- 14.50/HPF) which was comparable to PAF. But chymotrypsin showed no significant chemotactic response to eosinophils. Trypsin at the concentration of 10, 100,1000microg/ml induced secretion of ECP, which at the concentration of 10microg/ml represented about 2.7 times of the spontaneous rate of release. Soybean protease inhibitor reduced trypsin induced ECP release. CONCLUSION: Trypsin can induce chemotactic response to eosinophils and activation of eosinophils that can induce secretion of ECP. On the contrary, chymotrypsin showed no direct effect on eosinophils. We propose a role of trypsin on the chemotaxis and activation of eosinophils.
Asthma ; Chemotaxis* ; Chymotrypsin ; Eosinophils* ; Fluoroimmunoassay ; Humans ; Leukocytes ; Mast Cells ; Peptide Hydrolases ; Protease Inhibitors ; Soybeans ; T-Lymphocytes ; Tissue Donors ; Trypsin* ; Tryptases

SKPI (shrimp Kunitz-type protease inhibitor) from Marsupenaeus japonicus is a member of serine protease inhibitors which play an important role in the arthropod immunity. To fully understand its function in the innate immunity of shrimp, the skpi gene was cloned into a modified pPIC9K vector with a 6-His tag and expressed by Pichia pastoris GS115. The secretory SKPI was purified from the medium with high purity by using Ni Sepharose High Performance. This results also indicated that the purified SKPI could inhibit the activity of trypsin specifically.
Animals ; Aprotinin ; biosynthesis ; genetics ; isolation & purification ; Pandalidae ; chemistry ; Pichia ; genetics ; metabolism ; Recombinant Proteins ; biosynthesis ; genetics ; isolation & purification ; Serine Proteinase Inhibitors ; biosynthesis ; genetics ; Trypsin Inhibitors

CpTI (Cowpea Trypsin Inhibitor) is a widely used insect resistance gene in the plant genetic engineering for its high insecticidal activity and the minimal ability of the insects to evolve resistance to it. To facilitate the safety assessment of genetically modified foods (GMFs) with CpTI protein, we need to produce gram quantities of this protein in microbes. With the pGEX fusion expression system, we expressed the GST-CpTI protein in E. coli BL21, which accounted for approximately 40% of germ proteins. By Glutathione Sephrose 4B affinity chromatography, GST-CpTI was obtained with the purity up to 90%. Overnight incubate the fusion proteins with Thrombin protease, we got the CpTI proteins cleavage of GST tag. Both of the GST-CpTI and CpTI proteins showed notable trypsin inhibitor activity. Immunization of rabbits with purified fusion protein generated high titer antibodies (> 20000), measuring by ELISA. Western Blotting also showed specific Ag-Ab binding band between the antiserum and the CpTI proteins no matter in the whole supersonic germ proteins or purified from the column. All these made a good ground for the further safety assessment of CpTI protein.
Animals ; Blotting, Western ; Chromatography, Affinity ; Enzyme-Linked Immunosorbent Assay ; Escherichia coli ; genetics ; metabolism ; Rabbits ; Recombinant Proteins ; genetics ; metabolism ; Trypsin Inhibitors ; genetics ; metabolism

OBJECTIVETo study the effects of ulinastatin on coagulation in children who underwent open-heart surgery with cardiopulmonary bypass (CPB).

METHODSFifty children who underwent open-heart surgery for ventricular septal defect were randomly divided into two groups: ulinastatin treatment and control. Before CPB, ulinastatin (1.0×10(4) U/kg) was added to CPB priming fluid only in the ulinastatin treatment group. Activated partial thromboplasin time (APTT), prothrombin time (PT), thrombin time (TT), fibrinogen and international normalized ratio (INR) were measured both before and at 1 hr, 6 hrs and 24 hrs after CPB.

RESULTSThe PT in the ulinastatin group was more prolonged than in the control group at 1 hr after CPB (18.7 ± 0.7 s vs 15.5 ± 0.5 s) and 6 hrs after CPB (17.5 ± 0.6 s vs 15.0 ± 0.6 s). The APTT in the ulinatatin group was also significantly more prolonged than in the control group at 6 hrs after CPB (38.7 ± 3.1 s vs 35.3 ± 3.1 s) and 24 hrs after CPB (34.2 ± 3.0 s vs 31.1 ± 2.6 s).

CONCLUSIONSUlinastatin may prolong PT and APTT after CPB, and thus affects coagulation in children.

Blood Coagulation ; drug effects ; Cardiac Surgical Procedures ; Cardiopulmonary Bypass ; Female ; Glycoproteins ; pharmacology ; Humans ; Infant ; Male ; Partial Thromboplastin Time ; Prothrombin Time ; Trypsin Inhibitors ; pharmacology

OBJECTIVETo compare the ileal digestibility of protein and amino acids in parental rice and rice genetically modified with sck gene.

METHODSSix experimental swines were surgically fixed with a simple T-cannula at the terminal ileum and fed with parental rice and rice genetically modified with sck gene alternately. The ileum digesta were collected and analyzed for determination of apparent and true digestibility of protein and amino acids.

RESULTSThe apparent and true digestibility of protein was similar in these two types of rice. Except for the apparent digestibility of lysine, there was no difference in the apparent and true digestibility of the other 17 amino acids.

CONCLUSIONThe digestibility of protein and amino acids is not changed by the insertion of foreign gene, so it can meet the request of "substantial equivalence" in digestibility of protein and amino acids.

Amino Acids ; metabolism ; Animals ; Digestion ; Fabaceae ; Ileum ; metabolism ; Male ; Oryza ; genetics ; Phytic Acid ; metabolism ; Plants, Genetically Modified ; Proteins ; metabolism ; Swine ; metabolism ; Trypsin Inhibitors ; genetics

The study was purposed to investigate the apoptosis of HL-60 cells induced by recombinant common buckwheat trypsin inhibitor (rBTI) and its mechanism. The inhibition rate of rBTI on HL-60 cells was detected by MTT (3-[4, 5-dimethylthiazol-2-yl]-2, 5-diphenyltetrazolium bromide); the morphology of HL-60 nuclei was observed by fluorescence microscopy; the apoptosis cells of HL-60 detected by agarose gel electrophoresis and the changes of apoptosis rate was assayed by flow cytometry (FCM), when the HL-60 cells were treated with different concentration of rBTI for 24 hours. The results showed that the growth of HL-60 cells was inhibited evidently after treatment with rBTI in a dose-dependent manner, but there were minimal effects on normal human peripheral blood mononuclear cells (PBMNCs). The nuclei of HL-60 cells showed the characteristics of apoptosis, the analysis by flow cytometry indicated that the apoptosis rate of HL-60 cells was 52% after treatment with rBTI (100 microg/ml), DNA analyzed by agarose gel electrophoresis showed "ladder" pattern. It is concluded that rBTI obviously inhibits growth of HL-60 and induces its apoptosis which provides a foundation for use of recombinant common buckwheat trypsin inhibitor to cure the acute myeloid leukemia.
Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; Cell Proliferation ; drug effects ; Fagopyrum ; chemistry ; HL-60 Cells ; Humans ; Recombinant Proteins ; biosynthesis ; genetics ; pharmacology ; Trypsin Inhibitors ; biosynthesis ; genetics ; pharmacology