alpha-chymotrypsin and Inflammation

Tissue damage of all types, such as surgical or accidental injuries, fractures, and burns, stimulates a well-orchestrated, physiological process of healing, which ultimately leads to structural and functional restoration of the damaged tissues. The tissue repair process can be broadly divided into four continuous and overlapping phases-hemostasis and coagulation, inflammation, proliferation, and remodeling. If the process is interrupted or halted during any stage, it leads to impaired healing and formation of a chronic wound. Chronic wounds are associated with significant morbidity, mortality, and poor quality of life. Therefore, prompt and effective management of acute tissue injury is necessary to prevent it from progressing to a chronic wound. Proteolytic enzymes have been used to facilitate tissue repair since ancient times. Trypsin:chymotrypsin is an oral proteolytic enzyme preparation which has been in clinical use since the 1960s. It provides better resolution of inflammatory symptoms and promotes speedier recovery of acute tissue injury than several of the other existing enzyme preparations. This review article revisits the role and clinical utility of trypsin:chymotrypsin combination in tissue repair.. Torrent Pharmaceuticals Limited.

Topics: Burns; Chymotrypsin; Drug Combinations; Humans; Inflammation; Peptide Hydrolases; Quality of Life; Trypsin; Wound Healing; Wounds and Injuries

All acute and/or chronic pathological processes resulting in tissue damage and destruction lead to an inflammatory response. The purpose of this response is homeostatic and is made up of local and systemic events, the signs of which, taken as a whole, constitute the acute phase syndrome. Among the metabolic changes occurring in this syndrome, the rise in the plasma concentration of a group of heterogenous proteins known as the "Acute Phase Reactant Proteins" ( APRP ) is a very reliable and sensitive indicator of the presence of pathology. Most of these proteins are involved in the mechanisms initiating and controlling the inflammatory response. Some of them seem to establish a link between the body's specific and non-specific defence mechanisms. APRP are indicators of the inflammatory response without being specific to a particular etiology. The degree of variation in APRP levels is not generally speaking a good indicator of the severity or spread of the tissue lesions. On the other hand, they are very useful for detecting and monitoring infectious and/or inflammatory states, whether these are being treated or not. Thus during anti-infectious or anti-inflammatory treatment the return of APRP levels to their physiological baseline is a very good argument in favour of the efficacy of such treatment.

Topics: Acute-Phase Proteins; alpha 1-Antichymotrypsin; alpha 1-Antitrypsin; Animals; Blood Proteins; C-Reactive Protein; Ceruloplasmin; Chymotrypsin; Complement C3; Haptoglobins; Humans; Immune Tolerance; Infections; Inflammation; Orosomucoid; Serum Amyloid A Protein

Topics: alpha 1-Antichymotrypsin; alpha 1-Antitrypsin; alpha-Macroglobulins; Animals; Chymotrypsin; Endopeptidases; Humans; In Vitro Techniques; Inflammation; Macrophages; Mucus; Neutrophils; Oxygen; Phagocytes; Protease Inhibitors

Topics: Adrenal Cortex Hormones; alpha 1-Antitrypsin; alpha-Macroglobulins; Animals; Anti-Inflammatory Agents; Aprotinin; Arthritis, Experimental; Arthritis, Rheumatoid; Beta-Globulins; Carrageenan; Chymotrypsin; Dimethylnitrosamine; Humans; Inflammation; Kallikreins; Leukocytes; Liver; Lysosomes; Macrophages; Protease Inhibitors; Protein Binding; Proteins; Synovial Fluid

Topics: Cataract Extraction; Chymotrypsin; Deoxyribonucleases; Fibrinolysin; Humans; Inflammation; Peptide Hydrolases; Streptodornase and Streptokinase; Trypsin; Wounds and Injuries

Topics: Animals; Chymotrypsin; Clinical Trials as Topic; Dimethyl Sulfoxide; Drug Evaluation; Drug Evaluation, Preclinical; Enzymes; Heparin; Humans; Hyaluronoglucosaminidase; Inflammation; Iontophoresis; Trypsin

Topics: Chymotrypsin; Clinical Trials as Topic; Edema; Episiotomy; Female; Hematoma; Humans; Inflammation; Pain; Placebos; Postoperative Complications; Pregnancy; Tablets, Enteric-Coated; Wound Healing

Faecal biomarkers have emerged as important tools in managing of inflammatory bowel disease [IBD], which includes Crohn's disease [CD] and ulcerative colitis [UC].. To identify new biomarkers of gut inflammation in the stools of IBD patients using a proteomic approach.. Proteomic analysis of stools was performed in patients with both active CD and CD in remission and in controls by 2-DIGE and MALDI-TOF/TOF MS. An ELISA was used to confirm results in a second cohort of IBD patients and controls.. 2-DIGE analysis detected 70 spots in the stools of patients with active CD or patients in remission CD and in controls. MALDI-TOF/TOF MS analysis identified 21 proteins with Chymotrypsin C, Gelsolin and Rho GDP-dissociation inhibitor 2 [RhoGDI2] best correlating with the levels of intestinal inflammation. Results were confirmed in a second cohort of IBD patients and controls [57 CD, 60 UC, 31 controls]. The identified faecal markers significantly correlated with the severity of intestinal inflammation in IBD patients [SES-CD in CD, Mayo endoscopic subscore in UC] [CD; Chymotrypsin-C: r = 0.64, p < 0.001; Gelsolin: r = 0.82, p < 0.001; RhoGDI2: r = 0.64, p < 0.001; UC; Chymotrypsin-C: r = 0.76, p < 0.001; Gelsolin: r = 0.75, p < 0.001; RhoGDI2: r = 0.63, p < 0.001]. Moreover, ROC analysis showed that Gelsolin [p < 0.0002] and RhoGDI2 [p < 0.0001] in CD, and RhoGDI2 [p = 0.0004] in UC, have higher sensitivity and specificity than faecal calprotectin in discriminating between patients and controls.. We show for the first time that 2-DIGE is a reliable method to detect proteins in human stools. Three novel faecal biomarkers of gut inflammation have been identified that display good specificity and sensitivity for identifying IBD and significantly correlate with IBD severity.

Topics: Biomarkers; Chymotrypsin; Colitis, Ulcerative; Crohn Disease; Feces; Gelsolin; Humans; Inflammation; Inflammatory Bowel Diseases; Intestinal Mucosa; Leukocyte L1 Antigen Complex; Proteomics; rho Guanine Nucleotide Dissociation Inhibitor beta; Severity of Illness Index

A study was carried out to appraisal the function of methionine on intestinal digestion and the health of grass carp (Ctenopharyngodon idella) fry (initial weight 0.36 ± 0.01 g). The fry were fed graded dietary methionine levels (0.33%-1.20% dry matter) in 18 recirculatory tanks (180 L). After an 8-week breeding experiment, the results revealed that 0.71%-1.20% dietary methionine levels markedly upregulated the mRNA levels of intestinal digestion including trypsin, amylase, chymotrypsin and AKP, and 0.71%-0.87% dietary methionine level significantly increased intestinal trypsin activities compared with the 0.33% dietary methionine level. For inflammation, 0.71%-1.20% dietary methionine levels downregulated the mRNA levels of NF-κBp65, IL-1β, IL-6, IL-8, IL-15 and IL-17D, whereas upregulated the mRNA levels of anti-inflammatory cytokines, including IL-4/13B, IL-10 and IL-11. In terms of antioxidants, although dietary methionine levels had no significant effect on the expression of most core genes of the Nrf2/ARE signaling pathway, such as Nrf2, Keap 1, GPx4, CAT, Cu/Zn-SOD. Furthermore, dietary methionine levels had no significant effect on the expression of p38MAPK, IL-12p35, TGF-β2 and IL-4/13A. 0.71%-1.20% dietary methionine levels still increased the mRNA levels of GPx1α, GSTR and GSTP1. Furthermore, higher intestinal catalase activity and glutathione contents were also observed in fry fed 0.71%-1.20% diets. In summary, 0.71%-1.20% dietary methionine levels played a positive role in improving the intestinal digestion capacity of digestion, anti-inflammatory reaction and oxidation resistance of grass carp fry. This study provided a theoretical basis for improving the survival rate and growth of grass carp fry.

Topics: Aeromonas hydrophila; Amylases; Animal Feed; Animals; Carps; Catalase; Chymotrypsin; Dietary Supplements; Digestion; Fish Diseases; Fish Proteins; Glutathione; Inflammation; Interleukin-10; Interleukin-11; Interleukin-12 Subunit p35; Interleukin-15; Interleukin-27; Interleukin-4; Interleukin-6; Interleukin-8; Methionine; NF-E2-Related Factor 2; RNA, Messenger; Superoxide Dismutase; Transforming Growth Factor beta2; Trypsin

Inflammation and oxidative stress play a major role in the development of sepsis and its associated complications, leading to multiple organ failure and death. The lungs, liver, and kidneys are among the early affected organs correlated with mortality in sepsis. Alpha-chymotrypsin (α-ch) is a serine protease that exerts anti-inflammatory, anti-edematous, and anti-oxidant properties.. This study was undertaken to elucidate if the anti-inflammatory and anti-oxidant effects of α-ch observed in previous studies can alleviate lung, liver, and kidney injuries in a cecal ligation and puncture (CLP)-induced sepsis model, and thus decrease mortality.. Septic animals were given α-ch 2 h post CLP procedure. Sepsis outcomes were assessed in the lungs, liver, and kidneys. Separate animal groups were investigated for a survival study.. CLP resulted in 0% survival, while α-chymotrypsin post-treatment led to 50% survival at the end of the study. Administration of α-chymotrypsin resulted in a significant attenuation of sepsis-induced elevated malonaldehyde (MDA) and total nitrite/nitrate (NOx) levels. In addition, there was a significant increase in reduced glutathione (GSH) content and superoxide dismutase (SOD) activity in the lungs, liver, and kidneys. Administration of α-ch reduced elevated tissue expression of toll-like receptor-4 (TLR4), nuclear factor kappa-B (NF-κB), myeloperoxidase (MPO), and inducible nitric oxide synthase (iNOS). Alpha-chymotrypsin resulted in a significant reduction in serum levels of tumor necrosis factor-alpha (TNF-α), interleukin-1 beta (IL-1β), and interleukin-6 (IL-6). Alpha-chymotrypsin attenuated the rise in serum creatinine, cystatin C, blood urea nitrogen (BUN), alanine aminotransferase (ALT), and aspartate aminotransferase (AST) levels that was observed in the septic group. In addition, α-ch significantly reduced the lung wet/dry weight ratio, total protein content, and leukocytic counts in bronchoalveolar lavage fluid (BALF). Histopathological examination of the lungs, liver, and kidneys confirmed the protective effects of α-ch on those organs.. α-ch has protective potential against sepsis through lowering tissue expression of TLR4, NF-κB, MPO, and iNOS leading to decreased oxidative stress and inflammatory signals induced by sepsis. This effect appeared to alleviate the damage to the lungs, liver, and kidneys and increase survival in rats subjected to sepsis.

Topics: Animals; Anti-Inflammatory Agents; Antioxidants; Chymotrypsin; Inflammation; Kidney; Liver; Lung; NF-kappa B; Pneumonia; Punctures; Rats; Sepsis; Toll-Like Receptor 4

Mesotrypsin is a low-abundance human trypsin isoform with a unique evolutionary mutation that conferred resistance to trypsin inhibitors and restricted substrate specificity. Mesotrypsin degrades the serine protease inhibitor Kazal type 1 (SPINK1) and thereby might increase risk for pancreatitis. Here, we report a mouse model designed to test the role of mesotrypsin in pancreatitis. We introduced the human mesotrypsin evolutionary signature mutation into mouse cationic trypsinogen (isoform T7), resulting in a Gly to Arg change at the corresponding position 199. In biochemical experiments using purified proteins, the p.G199R T7 mutant recapitulated all salient features of human mesotrypsin. T7G199R mice developed normally with no spontaneous pancreatitis or other obvious phenotypic changes. Cerulein-induced acute pancreatitis in C57BL/6N and T7G199R mice showed similar severity with respect to inflammatory parameters and acinar cell necrosis while plasma amylase activity was higher in T7G199R mice. Neither SPINK1 degradation nor elevated intrapancreatic trypsin activation was apparent in T7G199R mice. The results indicate that in T7G199R mice the newly created mesotrypsin-like activity has no significant impact on cerulein-induced pancreatitis. The observations suggest that human mesotrypsin is unimportant for pancreatitis; a notion that is consistent with published human genetic studies.

Topics: Animals; Ceruletide; Chymotrypsin; Disease Models, Animal; Gene Expression Regulation; Glycoproteins; Humans; Inflammation; Mice; Mice, Inbred C57BL; Mutation; Pancreatitis; Prostatic Secretory Proteins; Trypsin; Trypsin Inhibitor, Kazal Pancreatic; Trypsinogen

Hypertensive cardiac remodeling is a major cause of heart failure. The immunoproteasome is an inducible form of the proteasome and its catalytic subunit β5i (also named LMP7) is involved in angiotensin II-induced atrial fibrillation; however, its role in deoxycorticosterone-acetate (DOCA)-salt-induced cardiac remodeling remains unclear. C57BL/6 J wild-type (WT) and β5i knockout (β5i KO) mice were subjected to uninephrectomy (sham) and DOCA-salt treatment for three weeks. Cardiac function, fibrosis, and inflammation were evaluated by echocardiography and histological analysis. Protein and gene expression levels were analyzed by quantitative real-time PCR and immunoblotting. Our results showed that after 21 days of DOCA-salt treatment, β5i expression and chymotrypsin-like activity were the most significantly increased factors in the heart compared with the sham control. Moreover, DOCA-salt-induced elevation of blood pressure, adverse cardiac function, chamber and myocyte hypertrophy, interstitial fibrosis, oxidative stress, and inflammation were markedly attenuated in β5i KO mice. These findings were verified in β5i inhibitor PR-957-treated mice. Moreover, blocking of PTEN (the gene of phosphate and tensin homolog deleted on chromosome ten) markedly attenuated the inhibitory effect of β5i knockout on DOCA-salt-induced cardiac remodeling. Mechanistically, DOCA-salt stress upregulated the expression of β5i, which promoted the degradation of PTEN and the activation of downstream signals (AKT/mTOR, TGF-β1/Smad2/3, NOX, and NF-κB), which ultimately led to cardiac hypertrophic remodeling. This study provides new evidence of the critical role of β5i in DOCA-salt-induced cardiac remodeling through the regulation of PTEN stability, and indicates that the inhibition of β5i may be a promising therapeutic target for the treatment of hypertensive heart diseases.

Topics: Animals; Cardiomegaly; Chymotrypsin; Desoxycorticosterone Acetate; Fibrosis; Hypertension; Inflammation; Male; Mice, Inbred C57BL; Mice, Knockout; Oxidative Stress; Proteasome Endopeptidase Complex; Protein Subunits; PTEN Phosphohydrolase; Signal Transduction; Up-Regulation; Ventricular Remodeling

The authors previously demonstrated that unconjugated bilirubin (UCB) may inhibit the activities of various digestive proteases, including trypsin and chymotrypsin. The digestive proteases in the lower gut are important in the pathogenesis of inflammatory bowel diseases. The effects of UCB on the inflammation and levels of digestive proteases in feces of rats with colitis have not yet been revealed. The present study investigated the effect of UCB on the inflammatory status and levels of trypsin and chymotrypsin in the feces of rats with trinitrobenzenesulfonic acid (TNBS)‑induced colitis. The data indicated that treatment with TNBS resulted in a marked reduction in weight gain, which was significantly alleviated in UCB‑treated rats. Furthermore, UCB treatment alleviated the inflammation induced by TNBS, detected via macroscopic damage and microscopic inflammation scores, and pro‑inflammatory markers including myeloperoxidase (MPO), tumor necrosis factor (TNF)‑α and interleukin (IL)‑1β. Furthermore, rats with colitis demonstrated significant increases in fecal trypsin and chymotrypsin levels, whereas UCB treatment significantly alleviated these increases. A significant positive correlation was additionally revealed among the pro‑inflammatory markers (MPO, TNF‑α and IL‑1β) and fecal digestive proteases (trypsin and chymotrypsin) in colitis. The results of the present study demonstrated that UCB ameliorated the inflammation and digestive protease increase in TNBS-induced colitis.

Topics: Animals; Bilirubin; Biomarkers; Chymotrypsin; Colitis; Colon; Cytokines; Digestion; Endopeptidases; Feces; Inflammation; Inflammation Mediators; Male; Rats, Sprague-Dawley; Trinitrobenzenesulfonic Acid; Trypsin; Weight Loss

To compare liver proteolysis and proteasome activation in steatotic liver grafts conserved in University of Wisconsin (UW) and Institut Georges Lopez-1 (IGL-1) solutions.. Fatty liver grafts from male obese Zücker rats were conserved in UW and IGL-1 solutions for 24 h at 4 °Cand subjected to ". Our comparison of these two preservation solutions suggests that IGL-1 helps to prevent ATP breakdown more effectively than UW and subsequently achieves a higher UPS inhibition and reduced liver proteolysis.

Topics: Adenosine; Allopurinol; Animals; Apoptosis; Autophagy; Chromatography, High Pressure Liquid; Chymotrypsin; Fatty Liver; Glutathione; Graft Survival; Homozygote; Inflammation; Insulin; Liver; Liver Transplantation; Male; Mitochondria; Organ Preservation; Organ Preservation Solutions; Perfusion; Proteasome Endopeptidase Complex; Proteolysis; Raffinose; Rats; Rats, Zucker

Postoperative pancreatic fistula (POPF) is the most serious and challenging complication following gastroenterological surgery. Activated pancreatic juice leaking from the organ remnant contains proteases that attack the surrounding tissue, potentially leading to severe inflammation, tissue necrosis, and fistula formation. However, it is difficult to observe pancreatic leakage during surgery and to evaluate the protease activity of leaked fluid at the patient's bedside. This report describes a protein nanocage-based protease ratiometric sensor comprising a pancreatic protease-sensitive small heat-shock protein (HSP) 16.5, which is a naturally occurring protein in Methanococcus jannaschii that forms a spherical structure by self-assembly of 24 subunits, and a chemically conjugated donor-acceptor dye pair for Förster resonance energy transfer (FRET). The HSP-FRET probe was constructed by subunit exchange of each dye-labeled engineered HSP, resulting in a spherical nanocage of approximately 10 nm in diameter, which exhibited very high stability against degradation in blood plasma and no remarkable toxicity in mice. The efficiency of FRET was found to depend on both the dye orientation and the acceptor/donor ratio. Pancreatic proteases, including trypsin, α-chymotrypsin, and elastase, were quantitatively analyzed by fluorescence recovery with high specificity using the HSP-FRET nanoprobe. Furthermore, the HSP-FRET nanoprobe was sufficiently sensitive to detect POPF in the pancreatic juice of patients using only the naked eye within 10 min. Thus, this novel nanoprobe is proposed as an effective and convenient tool for the detection of POPF and the visualization of activated pancreatic juice during gastroenterological surgery.

Topics: Animals; Chymotrypsin; Fluorescence Resonance Energy Transfer; Gastrointestinal Tract; General Surgery; Heat-Shock Proteins, Small; Humans; Inflammation; Methanocaldococcus; Mice; Nanostructures; Pancreatic Fistula; Postoperative Complications; Quantum Dots

Elastase-like enzymes are involved in important diseases such as acute pancreatitis, chronic inflammatory lung diseases, and cancer. Structural insights into their interaction with specific inhibitors will contribute to the development of novel anti-elastase compounds that resist rapid oxidation and proteolysis. Proteinaceous Kunitz-type inhibitors homologous to the bovine pancreatic trypsin inhibitor (BPTI) provide a suitable scaffold, but the structural aspects of their interaction with elastase-like enzymes have not been elucidated. Here, we increased the selectivity of ShPI-1, a versatile serine protease inhibitor from the sea anemone Stichodactyla helianthus with high biomedical and biotechnological potential, toward elastase-like enzymes by substitution of the P1 residue (Lys(13)) with leucine. The variant (rShPI-1/K13L) exhibits a novel anti-porcine pancreatic elastase (PPE) activity together with a significantly improved inhibition of human neuthrophil elastase and chymotrypsin. The crystal structure of the PPE·rShPI-1/K13L complex determined at 2.0 Å resolution provided the first details of the canonical interaction between a BPTI-Kunitz-type domain and elastase-like enzymes. In addition to the essential impact of the variant P1 residue for complex stability, the interface is improved by increased contributions of the primary and secondary binding loop as compared with similar trypsin and chymotrypsin complexes. A comparison of the interaction network with elastase complexes of canonical inhibitors from the chelonian in family supports a key role of the P3 site in ShPI-1 in directing its selectivity against pancreatic and neutrophil elastases. Our results provide the structural basis for site-specific mutagenesis to further improve the binding affinity and/or direct the selectivity of BPTI-Kunitz-type inhibitors toward elastase-like enzymes.

Topics: Animals; Aprotinin; Cattle; Chymotrypsin; Cloning, Molecular; Crystallography, X-Ray; Humans; Hydrogen Bonding; Inflammation; Models, Molecular; Mutagenesis, Site-Directed; Pancreatic Elastase; Protein Binding; Protein Conformation; Serine Endopeptidases; Serine Proteases; Serine Proteinase Inhibitors; Swine; Trypsin

The purpose of this study was to investigate the mechanism(s) of interleukin (IL)-8 suppression by Treponema denticola, one of the major periodontal pathogens, in gingival epithelial cells. Immortalized human gingival epithelial HOK-16B cells were infected with wild-type (WT), dentilisin-deficient (K1) or flagellin-deficient (flgE) T. denticola in the presence or absence of 2% human serum for 24 h. The levels of IL-8 expression were measured with real-time reverse transcription PCR and ELISA. In the absence of human serum, the WT and flgE, but not K1, substantially reduced not only the levels of IL-8 protein but also of IL-8 mRNA. Such downregulation of IL-8 mRNA was independent of bacterial invasion. Degradation of cytokine mixture by the WT, K1 and flgE revealed dentilisin-dependent preferential degradation of tumor necrosis factor (TNF)-α, an IL-8-inducing cytokine. WT and flgE significantly decreased the levels of TNFα secreted by HOK-16B cells, suggesting modulation of IL-8 through dentilisin-mediated degradation of TNFα. The addition of human serum to the culture potentiated the suppressive effect of T. denticola, resulting in substantial reductions of IL-8 and TNFα levels, even by K1. The serum-dependent effects of T. denticola were attributed to its ability to suppress the accumulation of intracellular reactive-oxygen species (ROS), a group of ubiquitous signaling molecules. Pretreatment with an antioxidant suppressed TNFα-induced IL-8 expression, confirming the role of ROS in TNFα signaling. Collectively, T. denticola targeted a key inflammatory cytokine and its signaling molecule to modulate the host innate immune response, which provides a new insight into modulation of host immunity by a periodontal pathogen.

Topics: Bacterial Proteins; Cell Line; Chymotrypsin; Gene Expression Regulation; Gingiva; Humans; Inflammation; Interleukin-8; Keratinocytes; Peptide Hydrolases; Proteolysis; Reactive Oxygen Species; Treponema denticola; Treponemal Infections; Tumor Necrosis Factor-alpha

Golden retriever muscular dystrophy (GRMD) is a genetic myopathy corresponding to Duchenne muscular dystrophy (DMD) in humans. Muscle atrophy is known to be associated with degradation of the dystrophin-glycoprotein complex (DGC) via the ubiquitin-proteasome pathway. In the present study, we investigated the effect of bortezomib treatment on the muscle fibers of GRMD dogs. Five GRMD dogs were examined; two were treated (TD- Treated dogs) with the proteasome inhibitor bortezomib, and three were control dogs (CD). Dogs were treated with bortezomib using the same treatment regimen used for multiple myeloma. Pharmacodynamics were evaluated by measuring the inhibition of 20S proteasome activity in whole blood after treatment and comparing it to that in CD. We performed immunohistochemical studies on muscle biopsy specimens to evaluate the rescue of dystrophin and dystrophin-associated proteins in the muscles of GRMD dogs treated with bortezomib. Skeletal tissue from TD had lower levels of connective tissue deposition and inflammatory cell infiltration than CD as determined by histology, collagen morphometry and ultrastructural analysis. The CD showed higher expression of phospho-NFκB and TGF-β1, suggesting a more pronounced activation of anti-apoptotic factors and inflammatory molecules and greater connective tissue deposition, respectively. Immunohistochemical analysis demonstrated that dystrophin was not present in the sarcoplasmic membrane of either group. However, bortezomib-TD showed higher expression of α- and β-dystroglycan, indicating an improved disease histopathology phenotype. Significant inhibition of 20S proteasome activity was observed 1 hour after bortezomib administration in the last cycle when the dose was higher. Proteasome inhibitors may thus improve the appearance of GRMD muscle fibers, lessen connective tissue deposition and reduce the infiltration of inflammatory cells. In addition, proteasome inhibitors may rescue some dystrophin-associated proteins in the muscle fiber membrane.

Topics: Animals; Boronic Acids; Bortezomib; Chymotrypsin; Collagen; Dogs; Dystroglycans; Dystrophin-Associated Protein Complex; Gene Expression; Immunohistochemistry; Inflammation; Muscle, Skeletal; Muscular Dystrophy, Animal; Muscular Dystrophy, Duchenne; Proteasome Endopeptidase Complex; Pyrazines

Protease-activated receptor 2 (PAR2) is implicated in the pathogenesis of chronic inflammatory diseases, including periodontitis; it can be activated by gingipain and produced by Porphyromonas gingivalis and by neutrophil protease 3 (P3). PAR2 activation plays a relevant role in inflammatory processes by inducing the release of important inflammatory mediators associated with periodontal breakdown. The effects of periodontal treatment on PAR2 expression and its association with levels of proinflammatory mediators and activating proteases were investigated in chronic periodontitis patients. Positive staining for PAR2 was observed in gingival crevicular fluid cells and was reflective of tissue destruction. Overexpression of PAR2 was positively associated with inflammatory clinical parameters and with the levels of interleukin-6 (IL-6), IL-8, tumor necrosis factor alpha, matrix metalloprotease 2 (MMP-2), MMP-8, hepatocyte growth factor, and vascular endothelial growth factor. Elevated levels of gingipain and P3 and decreased levels of dentilisin and the protease inhibitors secretory leukocyte protease inhibitor and elafin were also associated with PAR2 overexpression. Healthy periodontal sites from individuals with chronic periodontitis showed diminished expression of PAR2 mRNA and the PAR2 protein (P < 0.05). Furthermore, periodontal treatment resulted in decreased PAR2 expression and correlated with decreased expression of inflammatory mediators and activating proteases. We concluded that periodontal treatment resulted in decreased levels of proteases and that proinflammatory mediators are associated with decreased PAR2 expression, suggesting that PAR2 expression is influenced by the presence of periodontal infection and is not a constitutive characteristic favoring periodontal inflammation.

Topics: Adhesins, Bacterial; Adult; Bacterial Proteins; Chymotrypsin; Cysteine Endopeptidases; Elafin; Female; Gingipain Cysteine Endopeptidases; Gingival Crevicular Fluid; Hepatocyte Growth Factor; Humans; Inflammation; Inflammation Mediators; Interleukin-6; Interleukin-8; Male; Matrix Metalloproteinase 2; Matrix Metalloproteinase 8; Middle Aged; Myeloblastin; Peptide Hydrolases; Periodontal Pocket; Periodontitis; Porphyromonas gingivalis; Receptor, PAR-2; RNA, Messenger; Secretory Leukocyte Peptidase Inhibitor; Tumor Necrosis Factor-alpha; Vascular Endothelial Growth Factor A; Young Adult

In this study, 22 new betulinic acid (BA) derivatives were synthesized and tested for their inhibition of the chymotrypsin-like activity of 20S proteasome. From the SAR study, we concluded that the C-3 and C-30 positions are the pharmacophores for increasing the proteasome inhibition effects, and larger lipophilic or aromatic side chains are favored at these positions. Among the BA derivatives tested, compounds 13, 20, and 21 showed the best proteasome inhibition activity with IC(50) values of 1.42, 1.56, and 1.80 μM, respectively, which are three to fourfold more potent than the proteasome inhibition controls LLM-F and lactacystin.

Topics: Antineoplastic Agents; Betulinic Acid; Chymotrypsin; Cysteine Proteinase Inhibitors; Drug Design; Inflammation; Molecular Structure; Molecular Targeted Therapy; Neoplasms; Pentacyclic Triterpenes; Proteasome Inhibitors; Signal Transduction; Structure-Activity Relationship; Triterpenes

Recent evidence indicates that shock is accompanied by a failure of the mucosal barrier in the intestine and entry of pancreatic digestive enzymes into the wall of the intestine. To investigate the formation of cytotoxic mediators produced by enzymatic digestion of the intestine, we applied homogenates of rat small intestinal wall to human neutrophils and used flow cytometry measurements of propidium iodide uptake to determine cytotoxicity. We show that homogenates of the small intestine after ischemia by occlusion of the superior mesenteric and celiac arteries for 3 h, but not without ischemia, are cytotoxic. Digestion of homogenates of nonischemic intestinal wall with purified trypsin, chymotrypsin, or elastase, proteases normally present in the intestinal lumen, yielded cytotoxic mediators. Before cell death, we saw cell damage in the form of bleb formation and flow cytometry measurements of cell size changes due to blebbing. Cytotoxicity was prevented by serine protease inhibition with phenylmethylsulfonyl fluoride (PMSF) before, but not after proteolytic digestion of the wall homogenates, indicating that enzymatic action of proteases on the homogenate is necessary for cytotoxicity. Cytotoxicity of wall homogenates digested by enzymes in the fluid collected from the lumen of the intestine was greater than digests by the individual purified proteases. Cytotoxicity is undetectable if digestive enzymes in the luminal fluid are inhibited with a combination of enzyme inhibitors PMSF and 6-amidino-2-naphthyl p-guanidinobenzoate dimethanesulfonate before addition of wall homogenates. Passage of digested intestinal wall homogenates across a hydrophobic glass-fiber filter reduced cytotoxicity. Furthermore, we found that luminal fluid itself may be cytotoxic, possibly because of digestion of ingested food. To test whether digested food can be cytotoxic, we homogenized rat food and digested it in vitro with chymotrypsin or endogenous enzymes in luminal fluid. Cytotoxicity was significantly increased after digestion of food by luminal fluid compared with luminal fluid or undigested food. These results indicate the presence of a previously unknown mechanism for hemorrhagic necrosis in shock.

Topics: Animals; Apoptosis; Chymotrypsin; Cytotoxins; Flow Cytometry; Humans; Inflammation; Intestines; Ischemia; Male; Neutrophils; Pancreas; Rats; Rats, Wistar; Shock

Lipopolysaccharide (LPS) is a major structural component of all Gram-negative organisms and has been implicated in Gram-negative sepsis and septic shock. In the present study, Affymetrix microarray analysis of RNA derived from murine macrophages treated with LPS in the absence or presence of the proteasome inhibitor lactacystin revealed that the vast majority of genes regulated by LPS is under control of the proteasome. Analysis of the data has revealed that the products of these genes participate in 14 distinct signaling pathways. This represents a novel approach to the identification of signaling pathways that are both toll-like receptor 4- and proteasome-dependent and may lead to the development of new drug targets in Gram-negative sepsis and septic shock.

Topics: Acetylcysteine; Animals; Cell Survival; Chymotrypsin; Gene Expression Regulation, Enzymologic; Inflammation; Lipopolysaccharides; Macrophages; Mice; Oligonucleotide Array Sequence Analysis; Proteasome Endopeptidase Complex; Shock, Septic; Signal Transduction; Toll-Like Receptor 4

Alpha-1-antitrypsin (alpha1-AT) is a member of the serine protease inhibitor family regulating numerous proteolytic processes. The genetic disorder, alpha1-AT deficiency, is well known as a cause of hereditary pulmonary emphysema and liver cirrhosis. To create an animal model of human alpha1-AT deficiency, we disrupted the major murine isoform PI2, which is similar to human alpha1-AT and is one of 7 alpha1-AT isoforms found in the mouse. The ability of the serum to inhibit the activities of human leukocyte elastase (HLE) and human chymotrypsin (CYT) was significantly lower in heterozygous mice (alpha1-AT/PI2 -/+) than wild-type (alpha1-AT/PI2 +/+) mice (73.2% vs. 100% for HLE and 67.8% vs.100% for CYT, respectively; P<0.05). The distribution of genotypes among F(2) progeny was not in accordance with Mendelian distribution (P<0.01), as the percentages of wild-type, heterozygotes and homozygotes were 47.8%, 37.3% and 14.9%, respectively. Thus, it is likely that impairment of the protease inhibitor had a critical effect on fetus development. The alpha1-AT/PI2 deficient mouse will be a useful animal model for elucidating the function of alpha1-AT in fetal development, studying the mechanisms of chronic inflammatory disease and evaluating therapeutic candidates for the treatment of inflammatory disease.

Topics: alpha 1-Antitrypsin; alpha 1-Antitrypsin Deficiency; Animals; Chronic Disease; Chymotrypsin; Disease Models, Animal; Female; Fetal Development; Inflammation; Leukocyte Elastase; Male; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Proteins; Serpins

Shock is associated with a dramatic rise in the level of inflammatory mediators found in plasma. The exact source of these mediators has remained uncertain. We recently examined a previously undescribed mechanism for production of inflammatory mediators in shock involving pancreatic digestive enzymes. The current in vitro study was designed to identify particular pancreatic enzymes and organs that may potentially produce inflammatory mediators. A selection of different organs from the rat (heart, liver, brain, spleen, pancreas, intestine, diaphragm, kidney, and lung) were homogenized and incubated with purified trypsin, chymotrypsin, elastase, lipase, nuclease, or amylase and the supernatant was incubated with fresh naïve leukocytes for 15 min. The level of leukocyte activation in the form of pseudopod formation and the fraction of cell death were measured. Without the addition of purified enzymes, only the homogenate of the pancreas yielded enhanced cell activation. Organs incubated with physiological concentrations of trypsin also stimulated significantly higher levels of pseudopod formation as compared with the undigested organs or enzymatic controls. Lipase and chymotrypsin were able to elicit cellular activation from selected organs such as the heart, intestine, liver and diaphragm. Undigested pancreatic homogenates were capable of producing substantial cell death, as compared with all other undigested organs. Intestinal digests with elastase, lipase, trypsin and chymotrypsin also stimulated significant cell mortality. Lipase-treated heart, liver, intestine, diaphragm, kidney, and lung stimulated cell death as well. We conclude that the intestine, as well as several other organs, may serve as a major source of inflammatory mediators during shock if exposed to digestive enzymes.

Topics: Amylases; Animals; Cell Death; Chymotrypsin; Flow Cytometry; Humans; Inflammation; Leukocytes; Male; Neutrophils; Pancreas; Pancreatitis; Rats; Rats, Wistar; Shock; Time Factors; Tissue Distribution

On the basis of their amino acid sequences, tryptases are just another group of serine proteinases related to trypsin that happen to be expressed and stored in mast cells rather than the pancreas. On the basis of their biochemical and biological features, however, tryptases show little family likeness to trypsin and most other trypsin-like proteases. The intriguing discrepancies have been explained by the crystal structure of the tryptase tetramer. It is now clear how tryptases, by forming tetramers, have gained the ability to prevail enzymatically active in tissues, but, at the cost of an unusual narrow substrate specificity. The tryptase tetramer thus became both a (neuro)peptidase and a long-lasting initiator that orchestrates responses by the cleavage of a few key proteins, the activation of other proteases with broader specificity, and the stimulation of cellular responses. With the support of these performers, tryptase drives a variety of processes contributing to chronic inflammation and tissue remodeling, the diversity of which is still emerging.

Topics: Animals; Aprotinin; Asthma; Bronchi; Bronchoconstriction; Cell Degranulation; Chronic Disease; Chymotrypsin; Dogs; Humans; Inflammation; Inflammation Mediators; Mast Cells; Mice; Muscle, Smooth; Pancreatic Elastase; Peptide Hydrolases; Protease Inhibitors; Proteinase Inhibitory Proteins, Secretory; Proteins; Serine Endopeptidases; Serine Proteinase Inhibitors; Sheep; Species Specificity; Substrate Specificity; Tryptases

There is increasing evidence that the ligation of adhesion molecules such as L-selectin can activate phagocytes to their full inflammatory potential. Sulfatide has been established as ligand for L-selectin and shown to trigger intracellular signals in human neutrophils. However, it remains unclear whether the ligation of L-selectin with sulfatide affects neutrophil phagocytosis. We studied the effects of sulfatide upon Fc gamma R- and CR3-mediated human neutrophil phagocytosis. Adhesion of the cells to a sulfatide-coated surface resulted in a dose-dependent enhancement of phagocytosis mediated via Fc gamma R or CR3, or both receptors. Galactocerebroside, but not glucocerebroside, also enhanced phagocytosis by neutrophils; therefore, galactose residue is thought to be required on ceramide molecules for the activation. Chymotrypsin-treated neutrophils, from which most L-selectin had been removed, reacted with sulfatide and galactocerebroside to enhance phagocytosis. These results suggest that an unidentified receptor for these cerebrosides exists on neutrophils and participates in the enhancement of phagocytosis.

Topics: Cell Adhesion; Cell Membrane; Cell Separation; Cells, Cultured; Ceramides; Cerebrosides; Chymotrypsin; Dose-Response Relationship, Drug; Flow Cytometry; Galactosylceramides; Glucosylceramides; Humans; Inflammation; L-Selectin; Macrophage-1 Antigen; Neutrophils; Phagocytes; Phagocytosis; Receptors, IgG; Sulfoglycosphingolipids

We purified a new trypsin-chymotrypsin inhibitor, designated tessulin, from the rhynchobdellid leech Theromyzon tessulatum. This 9-kDa peptide was purified to apparent homogeneity by gel-permeation and anion-exchange chromatographies followed by reverse-phase HPLC. The structure of tessulin was determined by reduction, S-beta-pyridylethylation, trypsin digestion, automated Edman degradation and matrix-assisted laser desorption mass spectrometry (m/z 8985 Da). The 81-amino-acid peptide possesses 16 cysteines and exhibits a 16% sequence similarity with antistasin-type inhibitors. Tessulin inhibits trypsin (Ki 1 pM) and chymotrypsin (Ki 150 pM) and exhibits no activity with thrombin, factor Xa, cathepsin G and elastase. This is the first trypsin-chymotrypsin inhibitor isolated from leeches that does not inhibit elastase or cathepsin G, except for cytin and therin. Furthermore, tessulin, in conjunction with other serine-protease inhibitors isolated from Theromyzon (therin, theromin), significantly diminishes the level of human granulocyte and monocyte activation induced by lipopolysaccharides (10 microg). The combined level of inhibition is higher than that of aprotinin, another serine-protease inhibitor used biomedically. Thus, tessulin may be clinically significant in reducing inflammatory events.

Topics: Adjuvants, Immunologic; Amino Acid Sequence; Animals; Binding Sites; Chymotrypsin; Electrophoresis, Capillary; Inflammation; Leeches; Leukocytes; Lipopolysaccharides; Molecular Sequence Data; Peptides; Proteins; Sequence Analysis; Serine Proteinase Inhibitors; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Trypsin Inhibitors

In this study, we show that three proteolytic enzymes of different specificity-pronase, chymotrypsin, and trypsin-induced a dramatic stimulation of neutrophil apoptosis as shown by morphologic characteristics, analysis of cell DNA content, and presence of a characteristic "ladder" pattern of DNA fragmentation. The action of either chymotrypsin or trypsin was completely prevented by the serine protease inhibitor aprotinin, indicating that the proteolytic activity of the enzymes accounts for apoptosis induction. Stimulation of neutrophil apoptosis by proteases was observed in culture medium supplemented with either inactivated fetal calf serum (0.1-50%), autologous serum (0.1-50%), bovine serum albumin (0.1%), or in protein-free medium. Other cell types such as human peripheral blood monocytes and lymphocytes, human leukemic cells from THP-1, HL-60 and K562 lines, murine L929 fibroblasts, and unstimulated murine macrophages harvested from the peritoneal cavity were not induced to undergo apoptosis after the treatment with proteases. In an attempt to determine whether neutrophil serine proteases could induce apoptosis as chymotrypsin and trypsin do, the effect of elastase was assessed. A significant increase in the percentage of apoptotic cells was observed in elastase-treated neutrophils. We propose that the selective stimulation of neutrophil apoptosis by proteolytic enzymes may play an important role in the normal resolution of inflammation by limiting the autotoxic potential of the neutrophil.

Topics: Animals; Apoptosis; Aprotinin; Cattle; Chymotrypsin; Culture Media; DNA; Humans; In Vitro Techniques; Inflammation; Leukocyte Elastase; Neutrophils; Pancreatic Elastase; Peptide Hydrolases; Pronase; Serine Proteinase Inhibitors; Serum Albumin, Bovine; Trypsin

The glycoprotein (GP) L-selectin initiates adhesive interactions between leukocytes and endothelial cells (EC). It functions as a lymphocyte-lectin homing receptor recognizing carbohydrate determinants of the peripheral lymph node addressing on high endothelial venules. It also mediates neutrophil rolling, the earliest interaction of neutrophils with acutely inflamed venules. Neutrophil L-selectin presents sialyl-LewisX (sLe(X)) as a ligand to P- and E-selectin in vitro, and we have proposed that this is a major mechanism of L-selectin-mediated rolling in vivo. In contrast, the contribution of neutrophil L-selectin as a receptor protein recognizing one (or more) ligand(s) on inflamed EC is unclear. To address this question, an sLe(X)-negative murine pre-B cell line, L1-2, that can neither bind vascular selectins nor roll in inflamed rabbit venules, was transfected with human L-selectin cDNA. L-selectin expression in stable transfectants was sufficient to confer significant rolling in vivo. Rolling was unaffected by neuraminidase treatment but completely blocked by anti-L-selectin monoclonal antibody (MoAb) DREG-56. Thus, L-selectin can initiate leukocyte interactions with EC determinants potentially through recognition of endothelial carbohydrates. In contrast, when human neutrophils were tested, rolling was reduced, but not abolished, by MoAb DREG-56. Likewise, treatment with neuraminidase or anti-sLe(X) MoAbs decreased, but did not abrogate, neutrophil rolling, consistent with residual EC recognition via L-selectin. Combination of MoAb DREG-56 and neuraminidase resulted in almost complete loss of rolling, as did removal of glycosylated L-selectin by chymotrypsin. Together with the demonstrable rolling of L-selectin transfectants, our results support the concept of a bidirectional interaction between L-selectin bearing sLe(X) on neutrophils and activated EC in vivo. These findings also suggest that L-selectin may mediate rolling of lymphocytes that lack carbohydrate ligands for E- or P-selectin, although probably less efficiently than through bidirectional recognition.

Topics: Animals; Cell Adhesion; Cell Adhesion Molecules; Chymotrypsin; Endothelium, Vascular; Humans; In Vitro Techniques; Inflammation; L-Selectin; Lewis X Antigen; Mice; Microscopy, Electron; Neuraminidase; Neutrophils; Transfection; Venules

Antibody blocking studies in the mouse suggest that the MEL-14 antigen is involved in neutrophil-endothelial cell interactions and may be important in neutrophil extravasation to sites of inflammation in vivo. We recently showed that chemotactic factor activation causes a rapid (within minutes) shedding of a large fragment of the MEL-14 antigen from the surface of neutrophils. We report here that chymotrypsin, at low doses (0.1 units/1 x 10(6) cells), but not trypsin, elastase, or collagenase, causes an activation-independent rapid loss (greater than 90%) of the MEL-14 antigen from the surface of murine neutrophils. Under the same treatment conditions chymotrypsin has no effect on the expression of four other neutrophil surface antigens, including the Mac-1 adhesion protein. Chymotrypsin treatment has no effect on neutrophil adhesion to plastic, migration to C5a, regulation of the Mac-1 antigen, but causes a greater than 95% reduction in neutrophil binding to high endothelial venules (HEV) in peripheral lymph nodes measured in the ex vivo frozen section HEV binding assay. The level of inhibition of neutrophil adhesion to HEV was comparable to that seen with the MEL-14 antibody. This experimental system allows us for the first time to specifically examine the consequences of removing the MEL-14 antigen from the surface of neutrophils on function in vivo. We show that treatment with chymotrypsin blocks greater than 85% of the ability of neutrophils injected back into the animal to home to the inflamed peritoneum. In similar in vivo experiments the MEL-14 antibody blocks neutrophil homing by 60-70%. These results further support the importance of the MEL-14 antigen in neutrophil extravasation in vivo and indicate that chymotrypsin could be useful in examining the molecular mechanisms involved in extravasation of leukocytes into a variety of diverse tissue sites of inflammation.

Topics: Animals; Bone Marrow Cells; Cell Adhesion; Cell Adhesion Molecules; Chemotaxis, Leukocyte; Chymotrypsin; Dose-Response Relationship, Drug; Flow Cytometry; Inflammation; Macrophage-1 Antigen; Mice; Mice, Inbred BALB C; Neutrophils; Plastics; Receptors, Lymphocyte Homing

Myofibrillar protease activity and activity of inhibitors toward trypsin, chymotrypsin, elastase, and the myofibrillar protease were determined in human skeletal muscle. The protease activity was found to increase in patients with acute and chronic inflammation as well as in patients with metastatic and nonmetastatic tumors. The inhibitory activity directed against trypsin and chymotrypsin was not affected by acute inflammation, while the inhibition of elastase and the myofibrillar protease was increased. Chronic inflammation did not affect the ability of the muscle cytosol to inhibit trypsin and elastase, but increased the inhibition of chymotrypsin and the myofibrillar protease. Nonmetastatic tumors produced an increase in the activity of inhibitors toward trypsin, chymotrypsin, and elastase, while patients bearing metastatic tumors had a high level of cytosolic inhibitors of all the tested proteolytic activities. These results indicate that the myofibrillar protease and the cytosolic protease inhibitors in human skeletal muscle are differentially affected by catabolic conditions.

Topics: Abdominal Muscles; Chymotrypsin; Digestive System Diseases; Digestive System Neoplasms; Humans; Inflammation; Muscles; Pancreatic Elastase; Peptide Hydrolases; Protease Inhibitors; Trypsin

The anti-inflammatory activity of a proteinase inhibitor on carrageenin-induced inflammation was studied by using N-(2,4-dinitrophenyl)-benzisothiazolinone-1,1-dioxide, a selective inhibitor of elastase, cathepsin G and chymotrypsin. The selective inhibitor suppressed leukocyte chemotaxis in vivo and in vitro, vascular permeability and development of granulation tissue. These results suggest that a selective inhibitor of elastase, cathepsin G and chymotrypsin is an effective agent for suppression of induction and development of carrageenin-induced inflammation in rats.

Topics: Animals; Anti-Inflammatory Agents; Carrageenan; Cathepsin G; Cathepsins; Cell Membrane Permeability; Chemotaxis, Leukocyte; Chymotrypsin; Inflammation; Male; Neutrophils; Pancreatic Elastase; Rats; Rats, Inbred Strains; Serine Endopeptidases

Topics: Acute Disease; alpha 1-Antichymotrypsin; alpha 1-Antitrypsin; C-Reactive Protein; Chymotrypsin; Female; Fibrinogen; Haptoglobins; Humans; Inflammation; Orosomucoid; Pregnancy; Proteins; Trypsin Inhibitors

The composition of serum seromucoid, the protein fraction of serum not precipitated by 0.6 M perchloric acid, has been shown to vary with the technique of preparation. Immunochemical examination revealed that 91.5% of the protein present in the seromucoid fraction of serum was alpha 1 acid glycoprotein, the remainder consisting of alpha 1 antitrypsin, alpha 1 antichymotrypsin, beta 2 glycoprotein, haemopexin, albumin, and pre-albumin. Serum alpha 1 acid glycoprotein concentration determined by radial immunodiffusion correlated well with serum seromucoid concentration although the former was usually 0.4 g/l lower. The determination of serum alpha 1 acid glycoprotein by an immunological method is more precise than the seromucoid method and is not subject to interference from other proteins.

Topics: Adolescent; Adult; Albumins; alpha 1-Antichymotrypsin; alpha 1-Antitrypsin; Chymotrypsin; Female; Hemopexin; Humans; Immunodiffusion; Immunoelectrophoresis; Inflammation; Intestinal Diseases; Male; Middle Aged; Orosomucoid; Prealbumin; Trypsin Inhibitors

The effect of trypsin or chemotrypsin on the levels of benzylpenicillin, ampicillin, oxacillin, kanamycin or tetracycline in the peritoneal inflammation fluid, as well as the effect of trypsin on penetration of these antibiotics into the granulemas of 2 types was studied. It was found that intramuscular administration of the enzymes to animals in a dose of 10 mg/kg resulted in increased levels of penicillins and kanamycin in the inflammation peritoneal fluid. An analogous effect of trypsin was observed with respect to experimental granulemas. The levels of tetracycline in restricted inflammatory foci increased only after intraperitoneal administration of trypsin to rats. Increased penetration of benzylpenicillin into the peritoneal fluid and connective tissue granulema under the effect of the enzymes was observed. The effect of trypsin on penetration of benzylpenicillin into granulema according to Selie did not differ from the effect of the enzyme on penetration of semisynthetic penicillins and kanamycin into it.

Topics: Animals; Anti-Bacterial Agents; Ascitic Fluid; Biopharmaceutics; Blood; Chymotrypsin; Drug Interactions; Granuloma; Inflammation; Injections, Intramuscular; Peptide Hydrolases; Rats; Trypsin

Topics: Animals; Chymotrypsin; Humans; Inflammation; Rabbits; Staphylococcal Infections

Topics: Adolescent; Adult; Chymotrypsin; Episiotomy; Female; Genital Diseases, Female; Humans; Hysterectomy; Inflammation; Middle Aged; Parametritis; Pelvic Inflammatory Disease; Postoperative Care; Trypsin

Topics: Aspirin; Chymotrypsin; Cresols; Evaluation Studies as Topic; Female; Genital Diseases, Female; Humans; Inflammation; Trypsin

Topics: Animals; Chymotrypsin; Cortisone; Drug Synergism; Granuloma; Hyaluronoglucosaminidase; Inflammation; Penicillin G; Penicillin G Procaine; Rats

Topics: Animals; Chymotrypsin; Immunization, Passive; Immunochemistry; Immunodiffusion; Inflammation; Mice; Papain; Rabbits; Trypsin

Topics: Animals; Chymotrypsin; Drug Synergism; Exudates and Transudates; Granuloma; Inflammation; Penicillin G; Rats; Staphylococcus; Stimulation, Chemical; Turpentine

Topics: Acute Disease; Adult; Aged; Bronchiectasis; Chymotrypsin; Female; Humans; Inflammation; Lung Abscess; Lung Diseases; Male; Middle Aged; Pneumonia; Trypsin

Topics: Bone Diseases; Chymotrypsin; Drug Synergism; Humans; Inflammation; Ointments; Phenylbutazone; Postoperative Complications

Topics: Animals; Chlorides; Chymotrypsin; Drug Synergism; Dry Ice; Guinea Pigs; Histamine; Inflammation; Injections, Intramuscular; Injections, Intraperitoneal; Lithium; Muramidase; Prednisolone; Rats; Salicylates

Topics: Arthus Reaction; Chromatography; Chymotrypsin; Cysteine; Electrophoresis; Humans; Hydrogen-Ion Concentration; Inflammation; Injections, Intradermal; Leukocytes; Peptide Hydrolases; Spectrum Analysis; Trypsin

Topics: Aged; Anti-Inflammatory Agents; Bromelains; Chymotrypsin; Deoxyribonucleases; Female; Fibrinolysin; Genital Diseases, Female; Humans; Inflammation; Mastitis; Middle Aged; Oxyphenbutazone; Pregnancy; Uterine Cervical Neoplasms

Topics: Acid Phosphatase; Aminocaproates; Animals; Antifibrinolytic Agents; Aprotinin; Carbon Tetrachloride Poisoning; Chymotrypsin; Cyclohexanecarboxylic Acids; Edema; Female; Fibrinolysin; Guinea Pigs; Inflammation; Kallikreins; Microbial Collagenase; Pancreatic Elastase; Peptides; Protease Inhibitors; Rats; Ribonucleases; Trypsin

Topics: Aminopeptidases; Animals; Chymotrypsin; Connective Tissue; Esterases; Inflammation; Muscles; Ovalbumin; Phosphoric Monoester Hydrolases; Rats

Topics: Anti-Infective Agents; Anti-Inflammatory Agents; Chymotrypsin; Drug Synergism; Humans; Inflammation; Mouth Diseases; Tetracycline; Trypsin

Topics: Anti-Infective Agents; Anti-Inflammatory Agents; Chymotrypsin; Drug Synergism; Humans; Inflammation; Mouth Diseases; Tetracycline; Trypsin

Topics: Anti-Inflammatory Agents; Bromelains; Chymotrypsin; Humans; Indomethacin; Inflammation; Mouth Diseases; Oxyphenbutazone; Streptokinase

Topics: Animals; Carotid Arteries; Cats; Chymotrypsin; Collagen Diseases; Dura Mater; Endopeptidases; Inflammation; Injections, Spinal; Muscles; Necrosis; Nerve Degeneration; Subarachnoid Space

Topics: Adrenalectomy; Animals; Bradykinin; Chymotrypsin; Deoxyribonucleases; Edema; Fibrinolysin; Granuloma; Inflammation; Male; Muramidase; Pancreatin; Peptide Hydrolases; Rats; Serratia; Trypsin

Topics: Adult; Chymotrypsin; Drug Hypersensitivity; Humans; Inflammation; Injections, Intramuscular; Male

Topics: Adult; Chymotrypsin; Edema; Hematoma; Humans; Inflammation; Ligaments; Male; Mandibular Fractures; Trypsin

Topics: Anaphylaxis; Animals; Arthus Reaction; Chemical Precipitation; Chemistry Techniques, Analytical; Chromatography; Chymotrypsin; Electrophoresis; Endopeptidases; Enzyme Inhibitors; Inflammation; Metabolism; Papain; Protease Inhibitors; Rabbits; Research; Skin; Trypsin; Ultracentrifugation

Topics: Chymotrypsin; Edema; Glucuronidase; Glycoside Hydrolases; Inflammation; Lyases; Pharmacology; Rats; Research

Topics: Anti-Inflammatory Agents; Antitoxins; Chymotrypsin; Heparin; Inflammation; Pharmacology; Phenylbutazone; Rats; Research; Staphylococcus; Toxicology; Toxins, Biological

Topics: Anti-Bacterial Agents; Antibiotics, Antitubercular; Chymotrypsin; Drug Therapy; Humans; Hydrolases; Infections; Inflammation; Pharmacology; Protein Synthesis Inhibitors; Tetracycline; Trypsin

Topics: Blood Protein Electrophoresis; Chymotrypsin; Inflammation; Irritants; Pharmacology; Pleurisy; Rats; Research; Toxicology

Topics: Animals; Birds; Chymotrypsin; Hematologic Tests; Heparin; Humans; Inflammation; Pharyngitis; Stomatitis

Topics: Balsams; Chymotrypsin; Female; Guaiacol; Gynecology; Humans; Inflammation; Pancreatic Extracts; Pelvic Inflammatory Disease; Pelvis; Peritonitis; Trypsin

Topics: Accidents; Accidents, Occupational; Chymotrypsin; Ecchymosis; Edema; Humans; Inflammation; Occupational Medicine; Wounds and Injuries

Topics: Chymotrypsin; Edema; Humans; Inflammation; Postoperative Complications; Rifamycins; Suppuration; Surgical Wound Infection; Thrombophlebitis; Trypsin

Topics: Animals; Chymotrypsin; Exercise; Granuloma; Hematologic Tests; Inflammation

Topics: Chymotrypsin; Hematologic Tests; Inflammation

Topics: Chymotrypsin; Hematologic Tests; Inflammation

Topics: Chymotrypsin; Female; Gynecology; Humans; Inflammation; Parametritis; Pelvic Inflammatory Disease; Quinine

Topics: Abscess; Chymotrypsin; Hematologic Tests; Inflammation; Medicine; Peptide Hydrolases; Psychotherapy; Trypsin; Wounds and Injuries

Topics: Administration, Oral; Chymotrypsin; Episiotomy; Female; Gynecology; Humans; Inflammation; Obstetrics; Pregnancy; Trypsin

Topics: Chymotrypsin; Female; Gynecology; Humans; Inflammation; Ribonucleases; Trypsin

Topics: Aprotinin; Chymotrypsin; Humans; Inflammation; Peptides

Topics: Chymotrypsin; Hematologic Tests; Inflammation; Surgical Procedures, Operative

Topics: Anti-Inflammatory Agents; Chymotrypsin; Hematologic Tests; Inflammation; Trypsin

Topics: Chymotrypsin; Hematologic Tests; Inflammation; Ribonuclease, Pancreatic; Ribonucleases; Trypsin

Topics: Chymotrypsin; Gynecology; Hematologic Tests; Inflammation; Pelvis; Trypsin; Urinary Tract

Topics: Acridines; Chymotrypsin; Edema; Hematologic Tests; Inflammation; Ointments; Trypsin

Topics: Anti-Inflammatory Agents; Chymotrypsin; Inflammation

Topics: Chymotrypsin; Humans; Inflammation

Topics: Chymotrypsin; Endopeptidases; Granuloma; Hydrolases; Inflammation; Peptide Hydrolases; Trypsin