Titers of antibodies against antigen O of E. coli O14 in patients with inflammatory diseases of the large intestine

The content of serum antibodies to E. coli O14 O-antigen in patients with inflammatory diseases of the large intestine was studied in the passive hemagglutination test. These antibodies were detected in 77% of patients with nonspecific ulcerous colitis and in all patients with Crohn's disease; of these, 93.5% had antibody titers of 1:32 and greater, while none of the patients with ulcerous colitis showed such titers. This study has led to the conclusion that the detection of these antibodies in high titers can be one of the criteria for the differential diagnosis of nonspecific ulcerous colitis and Crohn's disease.     

Characteristics of lymphoid subpopulations isolated from healthy subjects and those suffering from nonspecific ulcerative colitis

Results of the studies of functional activity of lymphocyte subpopulations T+G, T-G, "O" subpopulations isolated peripheral blood of healthy persons and those with nonspecific ulcerative colitis (NUC) have been presented in this paper. The functional activity of the cells was assessed by means of local xenogeneic graft-vs-host reaction performed on CBA mice and by determination of nonspecific esterase enzyme. It has been established that analogous lymphocyte subpopulations influence the graft-vs-host reaction development in different ways depending on the condition of cell donor state: health/pathology. "O" subpopulations isolated from healthy persons hampered the graft-vs-host reaction development, and on the contrary stimulated it in patients with NUC. The assay of nonspecific esterase in lymphocytes showed that the least activity was noted in T+G and "O" cells in nonspecific ulcerative colitis.     

X-ray colonometry in the diagnosis of colonic dyskinesias and inflammatory diseases

Clinical and X-ray studies were made in 316 patients, which revealed intestinal dyskinesia, chronic colitis, and nonspecific ulcerative colitis in 105, 133, and 78 patients, respectively. Irrigoscopy (administration of a contrast enema, examination of the mucosal contour, and double contrasting) was performed in all the patients, by analyzing X-ray planimetric indices. X-ray colonoplanimetry makes it possible to objectify the interpretation of the X-ray pattern in chronic inflammatory diseases of the large bowel and colonic dyskinesia.     

The characteristics of the phenotype and lymphocyte function in patients with a mucosal lesion of the large intestine

The results of studies of functional activity of lymphocyte subpopulations: theophylline-sensitive (thphs-1) and theophylline-resistant (thphr-1), separated from peripheral blood of healthy people and patients with nonspecific ulcerative colitis (NUC) are represented in this paper. The evaluation of the separated subpopulations was performed by local xenogeneic graft-VS-host reaction, study of distribution of nonspecific esterase in cells and determination of relative amount of Fc gamma-R positive lymphocytes among thphs-1 and thphr-1. It was established that lymphocytes of patients with NUC had an alternative form of action on development of xenogeneic graft-VS-host reaction in contrast with cells of donors, that is inhibited the reaction. Among thphr-1 patients with NUC the amount of EC gamma-R cells and cells with diffuse distribution of enzyme with small degree activity was positively increased.     

The protective effect of nigeglanine on dextran sulfate sodium-induced experimental colitis in mice and Caco-2 cells

Ulcerative colitis (UC) was a nonspecific inflammatory disease. The treatment of UC is imperative. The present study aimed to investigate the effect of nigeglanine on dextran sulfate sodium-induced UC in experimental mice and Caco-2 cells and define the underlying mechanism. The nigeglanine was shown a significant protective effect on the colon, significantly reduced the weight and colon length loss and inhibited intestinal epithelial cell damage. Nigeglanine also reduced proinflammatory factors and increased anti-inflammatory factor production. The results indicate that nigeglanine suppresses the nuclear factor kappa B and mitogen-activated protein kinases pathways in addition to NLRP3 inflammasome action, inhibiting colon epithelial cell pyroptosis. Surprisingly, ZO-1 and occludin protein levels increased with nigeglanine treatment, suggesting that nigeglanine plays a protective role in barrier integrity, reducing colitis progression. The present study suggests that dietary therapy with nigeglanine may be a useful treatment for prophylaxis and palliative UC.     

Antimicrobial Agent-Associated Colitis and Diarrhea

Although antimicrobial agent-associated colitis has been recognized as a clinicopathologic entity for years, the cause of this disease has been determined only recently. Virtually all cases of pseudomembranous colitis and some cases of antimicrobial agent-associated nonspecific colitis or diarrhea have been shown to be caused by a toxin of Clostridium difficile. Methods for cultivating C difficile from feces and for detecting the toxin have been developed. Oral administration of vancomycin has proved to be effective for the treatment of C difficile-induced colitis, although isolated instances of relapse after treatment have been documented.The discovery of C difficile as a human intestinal pathogen has provided an explanation for some, but not all cases of antimicrobial agent-associated diarrhea. The epidemiology, pathogenesis and means of prevention of C difficile toxin-induced diarrhea remain to be determined.     

Roxadustat protect mice from DSS-induced colitis in vivo by up-regulation of TLR4

BackgroundThe incidence of inflammatory bowel disease (IBD) is growing in the population. At present, the etiology of inflammatory bowel disease remains unclear, and there is no effective and low-toxic therapeutic drug. The role of the PHD-HIF pathway in relieving DSS-induced colitis is gradually being explored.MethodsWild-type C57BL/6 mice were used as a model of DSS-induced colitis to explore the important role of Roxadustat in alleviating DSS-induced colitis. High-throughput RNA-Seq and qRT-PCR methods were used to screen and verify the key differential genes in the colon of mice between normal saline (NS) and Roxadustat groups.ResultsRoxadustat could alleviate DSS-induced colitis. Compared with the mice in the NS group, TLR4 were significantly up-regulated in the Roxadustat group. TLR4 KO mice were used to verify the role of TLR4 in the alleviation of DSS-induced colitis by Roxadustat.ConclusionRoxadustat has a repairing effect on DSS-induced colitis, and may alleviate DSS-induced colitis by targeting the TLR4 pathway and promote intestinal stem cell proliferation.     

ICMSF methods studies. XII. Comparative study for the enumeration of Clostridium perfringens in feces.

As the second phase of an international comparative study for the enumeration of Clostridium perfringens, four methods were compared for "total" and spore counts of C. perfringens in fecal specimens: the SFP (Shahidi-Ferguson perfringens) agar (A), TSC (tryptose-sulfite-cycloserine) agar (B), SC (sulfite-cycloserine) agar (C), and neomycin blood agar (D) methods. In both the total and spore count procedures, the confirmed C. perfringens counts in method D were lower than in methods A, B, and C. Little differences among methods were found in the percentages of presumptive colonies confirmed as C. perfringens. The nonspecific counts in methods A and D were generally greater than in B and C, but nonspecific microorganisms did not interfere in the enumeration of C. perfringens spores by any of the four methods. In overall performance, methods B and C were superior to A and D. The mean C. perfringens spore count was only 0.17 log lower than the mean total count. Spore counts alone are, therefore, adequate in investigations of C. perfringens outbreaks.     

Hyperbaric oxygenation and drug therapy in treatment of nonspecific ulcerative colitis and Crohn's disease]

Patients with nonspecific ulcerative colitis and Crohn's disease were treated with drug therapy (prednisolone, sulphasalazine, metronidazole per os and hydrocortisone per rectum) and subjected to 12 sessions of HBO. Every 10-14 months the HBO course was repeated. After 7 years of such a treatment a gradual partial recovery of large intestine mucosa and in some cases even its absolute recovery were observed. It was most difficult to get good results when treating lesion of distal colon segments. HBO produced a good effect when the disease was diagnosed at the early stage, when the intestine injury was accompanied by hepatobiliary system disease and in teenagers.     

A computerized protein–protein interaction modeling study of ampicillin antibody specificity in relation to biosensor development

Nonspecific interactions between immobilized biomolecules and interfering proteins significantly impede biosensor development and commercialization. Advances in bioinformatics and computer technology have facilitated a greater understanding of biological interactions. We employed two different protein–protein docking programs to simulate the nonspecific interaction between ampicillin antibody and potential interfering proteins (human serum albumin and ovalbumin). To evaluate the contact and probability of association with the active site of the antibody, different amino acid chains from human serum albumin (HSA) and ovalbumin (OVA) were modeled in the simulation. In addition, a well-known specific immune complex, lysozyme and lysozyme antibody, was simulated for comparison. The results demonstrated that the cluster density of nonspecific interactions was smaller than the specific interaction between lysozyme and antibody, and that the dock scores were scattered. However, the active site of ampicillin antibody was prone to nonspecific protein interactions. The strength of interaction was different for specific binding and nonspecific binding. These results provide a platform for detecting the probability of nonspecific interactions and for improving methods of biosensor detection construction with reduced nonspecific adsorption.     

Sulfasalazine-induced colitis complicating idiopathic ulcerative colitis.

A diagnosis of idiopathic ulcerative colitis was made in a previously healthy 9-year-old boy. Symptoms persisted despite therapy with sulfasalazine, 50 mg/kg daily, but they eventually responded to treatment with parenteral nutrition and prednisone, 40 mg daily. Metronidazole was also given to eradicate persistent Dientamoeba fragilis from the stools. The symptoms resolved over 3 weeks, and the daily dose of prednisone was tapered. On two subsequent occasions a challenge with sulfasalazine caused an immediate recurrence of loose, blood-streaked stools and of nonspecific histologic features of ulcerative colitis, which resolved when the sulfasalazine was discontinued.     

Heme Oxygenase-1 Ameliorates Dextran Sulfate Sodium-induced Acute Murine Colitis by Regulating Th17/Treg Cell Balance

Inflammatory bowel disease (IBD), including ulcerative colitis and Crohn''s disease, is a group of autoimmune diseases characterized by nonspecific inflammation in the gastrointestinal tract. Recent investigations suggest that activation of Th17 cells and/or deficiency of regulatory T cells (Treg) is involved in the pathogenesis of IBD. Heme oxygenase (HO)-1 is a protein with a wide range of anti-inflammatory and immune regulatory function, which exerts significantly protective roles in various T cell-mediated diseases. In this study, we aim to explore the immunological regulation of HO-1 in the dextran sulfate sodium-induced model of experimental murine colitis. BALB/c mice were administered 4% dextran sulfate sodium orally; some mice were intraperitoneally pretreated with HO-1 inducer hemin or HO-1 inhibitor stannum protoporphyrin IX. The results show that hemin enhances the colonic expression of HO-1 and significantly ameliorates the symptoms of colitis with improved histological changes, accompanied by a decreased proportion of Th17 cells and increased number of Tregs in mesenteric lymph node and spleen. Moreover, induction of HO-1 down-regulates retinoic acid-related orphan receptor γt expression and IL-17A levels, while promoting Treg-related forkhead box p3 (Foxp3) expression and IL-10 levels in colon. Further study in vitro revealed that up-regulated HO-1 switched the naive T cells to Tregs when cultured under a Th17-inducing environment, which involved in IL-6R blockade. Therefore, HO-1 may exhibit anti-inflammatory activity in the murine model of acute experimental colitis via regulating the balance between Th17 and Treg cells, thus providing a possible novel therapeutic target in IBD.     

Characteristics of lymphoid subpopulations of peripheral blood of patients with shigellosis]

This work presents some investigation results of lymphoid subpopulation functional activity: Tg+1, Tg-1, Theophylline-resistant and Theophylline-sensitive lymphocytes and O-1 isolated from the peripheral blood of patients with bacterial dysentery (BD) in local xenogenic GVHD. It has been established that Tg+1 and Tg-1 have stimulating effect on local xenogenic GVHD, but O-1 are inert though the quantity of Fcg+--receptor carrying lymphocytes among O-1 in BD and nonspecific ulcerative colitis is increased compared with donors. Theophylline-resistant and Theophylline-sensitive lymphocytes demonstrated inhibiting effect on GVHD formation which was evident in the latter case.     

Demonstration of myoglobin and CK-M in myocardium. Comparison of five fixation methods and three immunohistochemical techniques

The results of immunohistochemical staining vary depending on the tissue, fixative, antigen-antibody system, and immunohistochemical staining methods used. The purpose of this study was to evaluate the effect of different methods of fixation, different antigen-antibody systems, and different immunohistochemical methods on immunohistochemical staining of myocardium. Samples of normal fresh canine myocardium from six dogs were fresh frozen and fixed in 10% neutral buffered formalin, Bouin's, Bayley's and Carnoy's fixatives. Immunohistochemical staining for myoglobin and creatine kinase M was performed using the ABC (avidin-biotin complex) and indirect peroxidase-antiperoxidase (PAP) techniques. Tissues fixed in formalin showed the most intense specific staining for both antigens with the least background and nonspecific staining. All other fixation methods and frozen section techniques gave a more variable degree of specific positive staining and substantial background staining and/or nonspecific staining. ABC and PAP techniques gave similar results with both antigen-antibody systems and with each fixation method. Thus, no differences in specificity or sensitivity were observed between ABC and PAP techniques. Differences in staining intensity and pattern were related primarily to differences in fixation methods.     

Method for Extracting Viral Hemagglutination-Inhibiting Antibodies from the Nonspecific Inhibitors of Serum

Various methods are used to remove nonspecific inhibitors from sera before titering viral hemagglutination-inhibiting antibodies. These methods have several undesirable features; some are tedious and time-consuming, some remove antibody along with nonspecific inhibitors, and different techniques are usually required to remove the nonspecific inhibitors for different viruses. This communication describes a single method that uses diethylaminoethyl-Sephadex to extract the immunoglobulin G antibodies for several viruses from nonspecific inhibitors. The procedure is fast, simple to perform, and removed the nonspecific inhibitors for influenza, Western equine encephalitis, dengue-2, and rubella viruses.     

ICMSF methods studies. VIII. Comparative study for the enumeration of Clostridium perfringens in foods.

Four methods were compared in an international comparative study for the enumeration of Clostridium perfringens: the SFP (Shahidi-Ferguson perfringens) agar (A), TSC (tryptose-sulfite-cycloserin) agar (B), SC (sulfite-cycloserine) agar (C), and neomycin blood agar (D) methods. The confirmed C. perfringens counts were slightly lower for D than for A-C. The percentages of presumptive colonies confirmed as C. perfringens were essentially the same in each method. The relative numbers of nonspecific colonies were the lowest in C, followed by B, D, and A. The methods were also compared for simplicity and for aspects associated with the recognition and selection of presumptive colonies.     

Demonstration of Cholinesterase in Teeth

Human teeth have been studied by treatment with copper thio-choline, the method developed by Koelle for demonstrating activity of both specific and nonspecific cholinesterases. Freshly extracted teeth were collected and immediately sectioned on a cutting machine designed for calcified tissues. One series of teeth was sectioned sufficiently thin for microscopic study. Another series of teeth was bisected to expose the pulp chambers to the reagents. These teeth were divided into 5 experimental groups. The first group was treated with 10^-6M di-isopropylfluorophosphate (DFP) for 30 min at 37°C and then incubated with acetylthiocholine (AThCh) for 16 to 20 hr at 37°C in order to reveal the sites of activity of the specific enzyme, AChEase. The second group was incubated in a substrate of butyrylthiocholine (BuThCh) for 12 to 16 hr at 37°C to indicate the sites of the nonspecific ChEase. The third group was incubated in AThCh for 16 to 20 hr at 37°C without previous treatment by an inhibitor in order to reveal the presence and location of both the specific and nonspecific ChEase. The fourth and fifth groups were utilized as controls. Group 4 tissues were incubated without a substrate while those of group 5 were treated with DFP and then incubated with BuThCh. The specimens then were treated with ammonium sulfide to outline the sites of ChEase activity. The thin sections were mounted directly but the series of halved teeth were decalcified, embedded in paraffin, sectioned and then mounted. By these methods the presence and location of both specific and nonspecific ChEase activity were observed in human teeth. Concentration of specific ChEase was observed along the coronal aspect of the pulp chamber and along the course of the pulpal nerves. The nonspecific ChEase was observed throughout the pulpal tissue and appeared to be concentrated along the nerves and blood vessels. Neither series of control tissues exhibited any staining in the pulp tissue.     

Comparative proteomic studies on the pathogenesis of human ulcerative colitis

Ulcerative colitis (UC) is a chronic inflammatory disorder primarily affecting the colon mucosa. Its etiology and pathogenesis remain unclear. We used 2-DE and MS to identify differentially expressed proteins among the UC active, UC inactive, nonspecific colitis, and normal colon mucosa. Thirteen down-regulated and six up-regulated proteins were identified. Of the down-expressed proteins, eight (heat-shock protein 90 (HSPA9B), heat-shock protein 60 (HSPD1), H+-transporting two-sector ATPase (ATP5B), prohibitin (PHB), mitochondrial malate dehydrogenase (MDH2), voltage-dependent anion-selective channel protein 1 (VDAC1), thioredoxin peroxidase (PRDX1), and thiol-specific antioxidant (PRDX2)) were mitochondrial proteins, three (ATP5B, MDH2, triosephosphate isomerase) were involved in energy generation, three (PRDX1, PRDX2, SELENBP1) were cellular antioxidants, and six (HSPD1, HSPA9B, PRDX1, PRDX2, PHB, VDAC1) were stress-response proteins. Transmission electron microscopy revealed pathological alterations of mitochondrial ultrastructures even before the global colonocyte changes in the UC colon mucosa. PHB, an essential mitochondrial component protein, was down-expressed in the disease active as well as inactive colon mucosa from the patients of UC, indicative of an early event of mitochondrial changes during UC development. In contrast, aberrant activation of NFAT and ectopic expression of potential immunogenic proteins (tumor rejection antigen 1 and poliovirus receptor related protein 1) were found in the UC-diseased colon mucosa. Our findings suggest the implications of colonocyte mitochondrial dysfunction and perturbed mucosa immune regulation in the pathogenesis of UC and provide potential targets for the development of a new therapy.     

Mortality of captive tortoises due to viviparous nematodes of the genus Proatractis (Family Atractidae)

Between September 1982 and January 1984, verminous colitis was diagnosed post mortem in eight red-footed tortoises (Geochelone carbonaria) and three leopard tortoises (Geochelone pardalis) from the reptile collection of the National Zoological Park. This represented 69% of 16 tortoise necropsy accessions for that period. Etiology was determined to be a viviparous pinworm-like nematode of the genus Proatractis (Family Atractidae). Clinical signs were either nonspecific, consisting of anorexia, lethargy, and depression, or were absent. Limited trials with piperazine citrate and fenbendazole appeared to be ineffectual against the parasite and supportive therapy was unsuccessful. Post mortem examination revealed roughening and thickening of the mucosa of the cecum and colon, and in severe cases myriads of tiny (0.5-1.0 cm) nematodes were evident on the mucosal surface. In six tortoises, worms were found also in the small intestine. Histopathologic features in severe cases included mucosal necrosis with parasites and mixed inflammatory cells extending into the tunica muscularis. Focal to diffuse lymphoplasmacytic infiltrates were present consistently in the submucosa of the cecum and colon, and similar but milder lesions occasionally occurred in the small intestine.     

Alpha 1-antitrypsin as an endogenous marker of protein-losing enteropathies]

A novelty of the present studies is the use of alpha 1-antitrypsin (A-1--AT) as an endogenous marker of enteric protein loss. Enteric clearance of alpha 1-antitrypsin was determined in 10 patients with the symptoms of PLE, and in 6 healthy individuals. Alpha 1-Antitrypsin concentration has been assayed in single, random samples of feces collected from 42 patients and 12 healthy individuals (normal values: 1.31 +/- 0.72 mg/g of feces). Markedly increased enteric clearance and A-1-AT concentrations in single, random samples of feces have been found in patients with enteric lymphangiectasis, Crohn's disease, ulcerative colitis, and constrictive pericarditis, slightly lower in coeliac, chronic diarrhoea, nonspecific hemorrhagic colitis, esophagitis, lambliasis, hypogammaglobulinemia, Wiskott-Aldrich syndrome, Rendu-Osler-Weber syndrome, hepatitis in newborn, and Gilbert's disease. Statistically significant positive clearance has been noted (r = 0.997; p less than .001). A single assay of A-1-AT in feces is simple, repeatable, and sensitive technique in the diagnosis and evaluation of these diseases in which the symptoms of enteric protein loss are seen.