Anti-TNF-alpha antibody (infliximab) therapy supports the recovery of eNOS and VEGFR2 protein expression in endothelial cells.

The aim of this study is to investigate the effect of sera obtained from patients of Crohn's disease treated by anti-TNF-alpha antibody (Infliximab) on the expression of endothelial nitric oxide synthase (eNOS) and vascular endothelial growth factor receptor-2 (VEGFR2) protein in human umbilical vein endothelial cells (HUVEC) cultured in vitro. HUVEC was cultured in the presence of sera derived from patients before and after treatment, or from healthy individuals. Effects of sera on the expression of eNOS and VEGFR2 were monitored by determination of mRNA and protein levels using real time quantitative PCR and Western blot analysis, respectively. The serum of Crohn's patients contained elevated levels of TNF-alpha (34±1.80 pg/mL), which resulted in a decrease in the protein level of eNOS in HUVEC with a simultaneous induction of VEGFR2. Infliximab treatment normalized the expression level of these proteins by decreasing TNF-alpha level, particularly in those cases when clinical healing was also recorded, and it also conferred restitution of the level of angiogenic cytokines. Results suggest that altered angiogenesis possibly contributes to the initiation and perpetuation of inflammatory processes in inflammatory bowel disease (IBD). Endothelial dysfunction, a selective feature of Crohn's disease is beneficially affected by intravascular TNF-alpha neutralization.

ATG16L1 and IL23 receptor (IL23R) genes are associated with disease susceptibility in Hungarian CD patients.

BACKGROUND: North American and European genome-wide association scans have identified ATG16L1 and IL23R as novel inflammatory bowel disease (IBD) susceptibility genes and subsequent reports confirmed these findings in large independent populations. The aims of this study were to investigate the association and examine genotype-phenotype relationships in a Hungarian IBD cohort. METHODS: 415 unrelated IBD patients (CD: 266, age: 35.2+/-12.1 years, duration: 8.7+/-7.5 years and UC: 149, age: 44.4+/-15.4 years, duration: 10.7+/-8.9 years) and 149 healthy subjects were investigated. IL23R Arg381Gln (R381Q, rs11209026) and ATG16L1 Thr300Ala (T300A, rs2241880) polymorphisms were tested using LightCycler allele discrimination method. Detailed clinical phenotypes were determined by reviewing the medical charts. RESULTS: The association between IL23R rs11209026, ATG16L1 rs2241880 and CD was confirmed (OR(IL23R381Q): 0.38, 95% CI: 0.16-0.87; OR(ATG16L1300AA): 1.86, 95% CI: 1.04-3.40). No difference was found between patients with UC and either controls or CD. In CD, IL23R 381Gln heterozygosity was associated with inflammatory disease (70% vs. 34%, p=0.037), while disease restricted to the colon was more prevalent in patients with the ATG16L1 300Ala/Ala homozygosity (33.3% vs. 21.1%, p=0.036). In addition, carriage of the variant alleles did not predict response to steroids, infliximab or need for surgery. CONCLUSIONS: We confirmed that ATG16L1 and IL23R are susceptibility loci for CD in Hungarian CD patients. Further studies are needed to confirm the reported phenotype-genotype associations found in this study.

Interaction between seroreactivity to microbial antigens and genetics in Crohn's disease: is there a role for defensins?

Antibodies against different microbial epitopes are associated with disease phenotype, may be of diagnostic importance and may reflect a loss of tolerance in Crohn's disease (CD). Recently, an association was reported between the presence of these antibodies and mutations in pattern receptor genes. Our aim was to investigate whether mutations in various genes other than NOD2/CARD15 or TLR4 associated with CD (NOD1/CARD4, DLG5 and DEFB1) may influence the presence of antibodies against bacterial proteins and carbohydrates in a Hungarian cohort of CD patients. Three hundred and seventy-six well-characterized, unrelated, consecutive CD patients (male/female: 191/185, age at onset: 29.1 +/- 12.9 years, duration: 7.9 +/- 11.7 years) were investigated. Sera were assayed for anti-Omp, anti-Saccharomyces cerevisiae antibodies (ASCAs) immunoglobulin (Ig) A and IgG, and antibodies against a mannan epitope of S. cerevisiae (gASCA), laminaribioside (ALCA), chitobioside (ACCA), and mannobioside (AMCA). NOD1/CARD4, DLG5 and DEFB1 variants were tested by polymerase chain reaction-restriction fragment length polymorphism, and DEFB1 was genotyped in a subgroup of 160 patients. Detailed clinical phenotypes were determined by reviewing the patients' medical charts. The carriage of DEFB1 20A variant alleles less frequently led to antiglycan positivity compared with patients without (29.6% vs 46.2%, OR: 0.49, 95% CI: 0.25-0.97), regardless of disease location or behavior. Similar tendency was observed for DEFB1 44G (present: 21.6% vs absent: 10.2%, P = 0.06) and ALCA. A gene or serology dosage effect was not observed. However, no association was found between the DEFB1 G52A, DLG5 R30Q, and NOD1/CARD4 E266K variants and any of the serology markers. We found that variants in human beta-defensin 1 gene are inversely associated with antiglycan antibodies, further confirming an important role for innate immunity in the pathogenesis of CD.

Esophageal replacement using cryopreserved tracheal graft.

Esophageal replacement using artificial material is not a new concept. Prior experiences with prostheses, allografts and composite grafts have not proved particularly successful. The aim of this study is to investigate whether cryopreserved animal trachea is suitable for the replacement of the esophagus. In 12 beagle dogs a 6-cm-long segment of the cervical esophagus was removed, and was replaced with cryopreserved trachea, which had been stored for 21 days on -86 degrees C. The proximal and distal ends of the esophagus were joined together with the graft by simple continuous suture (Biosyn 3/0) and covered with the sternohyoid flap. Postoperatively 16 hematological parameters were measured. The dogs were planned to be euthanized at random on days 28, 42 and 56 after the operation. Tests for air leak were performed and the inner diameter of the graft was measured to detect shrinkage. The microscopic structure of the graft was analyzed using haematoxylin and eosin staining. There was no indication of insufficiency. Based on the air leak test the sutures withheld properly. The inner diameter of the graft narrowed from an average 19 mm (+/- 1 mm) to 15.8 mm (+/- 0.6 mm). In length, the graft shortened from an average 60 mm to 47 mm (+/- 3 mm). No feeding difficulty was observed. In two cases wound suppuration was found involving only the cutaneous and subcutaneous layers. Concerning the laboratory parameters, only the fibrinogen level and white blood cell count showed temporary although significant changes. Histology findings on the 56th day showed absolute integration of the trachea with the esophagus, with disintegration of the tracheal cartilages. Cryopreserved trachea seems to be suitable for the replacement of a 5-6-cm-long esophageal segment.

NOD1 gene E266K polymorphism is associated with disease susceptibility but not with disease phenotype or NOD2/CARD15 in Hungarian patients with Crohn's disease.

BACKGROUND: NOD1/CARD4, a member of the pattern-recognition receptor family, is a perfect candidate as a susceptibility gene for Crohn's disease. Since only limited and conflicting data are available on G796A polymorphisms in inflammatory bowel disease patients, we set out to study the effect of this polymorphism on the susceptibility and course of Crohn's disease in the Hungarian population. METHODS: Four hundred thirty-four unrelated Crohn's disease patients (age at presentation: 28.6+/-9.6 years, female/male: 210/224, duration of Crohn's disease: 8.2+/-6.9 years) and 200 healthy subjects (blood donors) and 136 non-inflammatory bowel disease gastrointestinal controls with chronic gastritis were investigated. NOD1 G796A was detected by using polymerase chain reaction/restriction fragment length polymorphism. Detailed clinical phenotypes were determined by reviewing the medical charts. RESULTS: The frequencies of the variant alleles of NOD1 G796A differed significantly between the Crohn's disease patients and both healthy (GG 49.5% vs. 67%; AG 41.5% vs. 28%; and AA 9.0% vs. 5.2%; p<0.0001) and non-inflammatory bowel disease controls with chronic gastritis. Carriage of the single nucleotide polymorphism of NOD1 G796A proved to be a highly significant risk factor for Crohn's disease compared to both healthy (p<0.0001, OR: 2.1, 95% CI: 1.5-2.9) and non-inflammatory bowel disease controls with chronic gastritis (p=0.008). Significant associations were not found between the different genotypes and the demographic data on the patients or the clinical characteristics of Crohn's disease. The different polymorphisms of pattern-recognition receptors (e.g. NOD2/CARD15 SNP8, SNP12 and SNP13 mutations, the TLR4 D299G polymorphism and NOD1 G796A) did not reveal a mutual basis. CONCLUSIONS: Our results suggest that carriage of the NOD1 G796A mutation increases susceptibility for Crohn's disease in the Hungarian population.

[Hematologic aspects of inflammatory bowel diseases]. / A gyulladásos bélbetegségek hematológiai vonatkozásai.

Anaemia, thrombocytosis are common secondary changes in inflammatory bowel disease (IBD), reflecting the clinical severity of the IBD cases, too. On the other hand, increased platelet function, fibrinolytic abnormalities, hypercoagulation of IBD patients predispose to thromboembolic events, and they may as well contribute to the local microcirculatory alterations leading to IBD itself. Reduced FXIII levels have been observed in IBD, which seems to be correlated with mucosal repair and might have therapeutic importance, too. Genetic thrombophilia received much attention recently, however, much less is known how frequent they are in IBD, what their clinical significance is, do they modify the clinical course itself. A short, concise review about links between haematology and IBD is given.

Evaluation of hemostatic changes and blood antioxidant capacity in acute and chronic pancreatitis.

BACKGROUND/AIMS: We examined changes in hemostasis, in levels of total antioxidant capacity, and pancreatic enzymes (amylase, lipase) in patients with pancreatitis 1, 3 and 7 days after admission to the clinic, in order to evaluate the inflammatory processes in acute and chronic pancreatitis and to identify new prognostic markers. METHODOLOGY: The rate of CD62 expression--a marker of platelet hyperactivity--and the rate of platelet-leukocyte aggregates were measured by flow cytometry. The connection between the parameters measured and the severity of pancreatitis and also the differences of the parameters in acute and chronic pancreatitis were investigated. RESULTS: On the basis of previous studies it was assumed, that there is a connection between the level of parameters measured and the inflammatory process in the pancreas, and also between the defending processes of the body against free radicals. CONCLUSIONS: Based on our results, we suggest to extend the laboratory measurements to the investigation of hemostatic parameters. The measurement of plasma level of fibrinogen, von Willebrand factor and the rate of platelet activation is especially important.

[Diagnostic possibilities in gastric leiomyoma in relation to two of our cases]. / A gyomor leiomyomájának diagnosztikai lehetöségeiröl--két esetünk kapcsán.

Two cases of rare, benignant gastric tumors are reported. The suggest that while in the diagnosis of tumors with a mucous membrane involvement endoscopy has doubtless a leading role, tumors not infiltrating the mucous membrane are usually better recognizable by radiological (ultrasonography, computer tomography and double contrast x-ray) methods. An appropriate diagnosis followed by surgical removal of the tumor might result in a complete healing of the patient.

[Detection of Helicobacter pylori in tissue samples of stomach cancer]. / A Helicobacter pylori kimutatása gyomorcarcinomás szöveti mintákban.

The role of Helicobacter pylori in the carcinogenesis of the stomach has been recognised both in intestinal and diffuse forms. The occurrence of the bacterium was studied in this report, with various methods in biopsy samples from the cancerous stomach, as well as the presence of associated gastritis and metaplasia related to the histological type. Retrospective histological examination were performed on endoscopic biopsy samples from 124 patients with distal stomach cancer using haematoxillin-eosin and Giemsa staining and immunohistochemical tests. Out of the 124 samples 69 (55.64%) was positive: 48 with Giemsa staining and further 21 samples showed immunohistochemical positivity on atrophic gastritis samples despite negative Giemsa staining. In view of the presence of gastritis and metaplasia significant difference (p < 0.001) was found between the positive and negative cases. The ratio of the Helicobacter pylori positive samples was high both for intestinal and diffuse type carcinomas. Our results suggest that the presence of Helicobacter pylori infection is important in the development of both types of carcinoma, nevertheless, the hystological type of the tumor is also decisively influenced by the onset of action of other more direct local eliciting factors.

[Current status of the management of upper gastrointestinal hemorrhage at a specialized hospital unit]. / Felsö tápcsatornai vérzések korszerü ellátása erre specializált gastrointestinalis részlegen.

The prevalence of bleeding of the gastrointestinal tract is around 100/100,000 adults/year. These events need special diagnostic and therapeutic approach, which previously was located mostly to surgical departments. At the beginning of 1996 a specialized ward ("gastrointestinal bleeding unit, GBU") was created at the 2nd Dept. Medicine of the University Medical School of Debrecen. The authors give an account about their experiences with the first 250 cases, try to establish the optimal and necessary conditions and analyse the consequences of their newly developed "risk-score" system. The overall mortality was 9% during this period and surgical intervention proved to be necessary in only 10 cases. On the basis of the collected experiences, they are convinced that the described and elaborated model can well be used for the proper, up-to-date management of gasrointestinal bleeding disorders. As next they suggest an overall, regional organisation of such units, together with the correct determination and provision of the financial background.

[The role and significance of the most important known cytokines in inflammatory bowel diseases]. / A legfontosabb, ismert citokinek szerepe és jelentösége gyulladásos bélbetegségekben.

Several theories have already been postulated in connection with the pathogenesis of inflammatory bowel diseases, yet none of them has been approved. Recently increasing attention has been payed to different cytokines, playing central role in the development of inflammatory processes. In the intestinal mucosa of patients suffering from inflammatory bowel diseases increasing amounts of interleukin-1 (IL-1), tumor necrosis factor (TNF) and platelet activating factor (PAF) could be measured. On the other hand, antiinflammatory cytokines seem to be ineffective, or being present in insufficient amount (IL-4 and IL-10 respectively). It is therefore probable, that altered ratios of cytokines, or pathologic regulation of their production lead to progression of inflammation in IBD. Influence of cytokine production may open new therapeutic approach, e.g. IL-10 enema proved to be effective in the treatment of some cases of steroid-resistant ulcerative colitis, while intravenous administration was useful in Crohn's disease. A brief, comprehensive review of our present knowledge about cytokines in IBD is given.

Effect of L-arginine on in vitro plasmin-generation and fibrinogenolysis.

L-arginine ahs received much attention in numerous aspects of the regulation of vascular tone and haemostasis. L-arginine seems to be capable to bind to plasminogen, too. The aim of the present paper is to investigate the action of L-arginine on in vitro plasmin generation and fibrino(geno)lysis by chromogenic, kinetic plasmin generation assay and electrophoretic analysis. The acceleration of tPA-induced plasmin generation in the presence of low concentration of L-arginine, along with augmentation of in vitro fibrinogenolysis have been documented. L-arginine may have a role in the modification of fibrinogenolysis, and this role should be considered if arginine is used as an element of some novel antithrombotic agents.

[Nitrogen monoxide (NO) in the function of the gastrointestinal tract and the liver]. / Nitrogén-monoxid (NO) a gyomor-bél csatorna és a máj müködésében.

This review is concerning with the endothel derived relaxing factor (EDRF) discovered in 1980 and identified as NO in 1987 in the function and activities of the gastrointestinal tract and the liver. It is of importance, that NO seems to basic mediator of the so called nonadrenerg-noncholinerg inhibitory innervation of the bowel, and also takes part in the regulation of mucosal blood supply and secretory function. In portal hypertension elevated cGMP is found in the urine, as evidence of increased nitrogen oxide synthase (NOS) activity. NO may act both as hepatoprotective and cytotoxic factor in the free radical reactions of several liver disease.

[Effect of paracetamol on the acid hemolysis of erythrocytes, with special reference to the intracellular glutathione level]. / Paracetamol hatása a vörösvérsejtek savi haemolysisére, különös tekintettel az intracellularis glutathion szintre.

The authors give account of their study concerning the effect of paracetamol on the acid haemolysis, and glutathione content of human erythrocytes. In vitro, paracetamol decreased intracellular glutathione content in a dose dependent manner. This may lead to an increased sensitivity of erythrocytes to the haemolytic effect of hydrochloric acid, what may explain the dose dependent decrease of time of haemolysis, presented in form of dynamic haemolysis curve. In vivo four hours following the oral intake of 500 mg paracetamol, although the intracellular glutathione content did not decrease significantly, shortening of the time of acid haemolysis could be demonstrated in more than half of the persons studied. No haemolysis was caused by the given dose of the drug.

[Newest method for the detection of chronic intestinal ischemia: color Doppler sonography]. / Krónikus intestinalis ischaemia igazolásának újabb lehetösége: color-Doppler vizsgálat.

Authors describe the case of a patient with chronic abdominal pain due to the occlusion of the superior mesenteric and the significant stenosis of the inferior mesenteric artery, discovered by color-Doppler ultrasonography. The role of this new non-invasive method in the diagnose of chronic intestinal ischaemia is emphasized. It seems to be suitable for the replacement of angiography also in the postoperative period.

The effect of silibinin (Legalon) on the the free radical scavenger mechanisms of human erythrocytes in vitro.

The effect of Legalon was investigated parallel with that of Adriblastina (doxorubicin) and paracetamol on some parameters characterizing the free radical scavenger mechanisms of human erythrocytes in vitro and on the time of acid hemolysis performed in aggregometer. Observations suggest that Adriblastina enhances the lipid peroxidation of the membrane of red blood cells, while paracetamol causes significant depletion of intracellular glutathione level, thus decreasing the free radical eliminating capacity of the glutathione peroxidase system. Legalon on the other hand, is able to increase the activity of both superoxide dismutase and glutathione peroxidase, which may explain the protective effect of the drug against free radicals and also the stabilizing effect on the red blood cell membrane, shown by the increase of the time of full haemolysis.

[Effect of SIN 1 on the interactions of endothelial cells and thrombocytes]. / Einfluss von SIN 1 auf Wechselwirkungen von Endothelzellen und Thrombozyten.

A new method has been developed to investigate the direct effect of endothelial cells on platelet aggregation in the presence of cultured confluent human EC monolayers. The inside of the disk shaped cuvettes are covered by human umbilical vein ECs. The cuvettes rotate in the light beam of a photometer. In these cuvettes the effect of different aggregation inducers was inhibited in a concentration-dependent manner, e. g. for collagen at 0.5 microgram/ml, ADP at 5 x 10(-7) M, epinephrine at 5 x 10(-7) M and thrombin at 0.05 U/ml final concentration. (Platelet count 5 x 10(5)/microliters). Higher concentrations of the inducers led to irreversible platelet aggregation and were used for testing of antiplatelet drugs. In this system we detected a synergism between the antiaggregatory effect of the EC monolayer and that of SIN 1 (Cassella, Frankfurt/Main) the main metabolite of Molsidomine. 10 micrograms/ml PRP of SIN 1 alone partially inhibited platelet aggregation induced by 1 microgram/ml collagen and 10(-6) M ADP respectively in uncovered aggregation cuvettes. In the presence of an EC monolayer complete inhibition of platelet aggregation was obtained at a 5-fold final concentration of both inducers. After pretreatment of ECs with 1 mmol ASA over 30 min. the antiaggregatory effect of SIN 1 decreased, but was more pronounced (50%) than in uncovered cuvettes (25%).

Changes of spontaneous and induced platelet aggregation in the presence of a human endothelial cell monolayer.

A new, simple method has been developed for the investigation of platelet aggregation in the presence of a cultured confluent human endothelial cell monolayer, in disk shaped rotating cuvettes, without magnetic stirring. The endothelial cells inhibited the aggregating effect of several inducers in a concentration dependent manner. At a platelet count of 5 x 10(5)/microliter in PRP e.g. the aggregating effect of 1 microgram/microliter thrombin was completely abolished. Spontaneous aggregation was also prevented by the EC-monolayer. A correlation could be established between the inhibitory effect of ECs and the number of platelets. In PRP with a platelet count of 7 x 10(-5)/microliters the inhibitory effect of the endothelial cell monolayer significantly decreased.

[Protein C and apoprotein levels in ischemic heart disease diagnosed by coronarography]. / Protein-C és apolipoprotein-szintek vizsgálata coronarographiával igazolt ischaemiás szívbetegségben.

The levels of protein C and different lipid parameters were measured in 22 patients, younger than 50 years, suffering from coronary heart disease, proved previously by angiography. The severity of coronariasclerosis showed certain correlation with the concentration of apolipoprotein B and triglycerids, while low levels of protein C were also mainly observed in those with severe coronary heart disease.