Increased gastric and intestinal permeability in patients with Crohn's disease

OBJECTIVES: Patients with Crohn's disease exhibit marked changes in intestinal permeability that can be assessed by lactulose and mannitol. Sucrose is a novel marker for gastric permeability. We combined these three sugars to investigate whether patients with Crohn's disease demonstrate changes in gastric permeability and if so, whether these changes are matched with altered intestinal permeability. METHODS: Fifty patients with Crohn's disease and 30 healthy subjects each drank a solution containing 20 g of sucrose, 10 g of lactulose, and 5 g of mannitol. Patients' and subjects' 5-h sugar urinary excretion levels were determined by high performance liquid chromatography and an enzymatic method (sucrose). Furthermore, patients with Crohn's disease underwent endoscopy of the upper GI tract and were grouped according to endoscopic and histological findings. RESULTS: Patients with Crohn's disease showed higher gastric and intestinal permeability compared with healthy control subjects. Gastric permeability was correlated with intestinal permeability. Patients with granuloma had more pronounced changes in both gastric and intestinal permeability than patients with various endoscopic and histological lesions. Patients with normal mucosa had normal permeability. CONCLUSIONS: Alterations in gastric mucosa caused by Crohn's disease are reflected by changes in gastric permeability and can be used to noninvasively screen for Crohn's disease involvement of the upper GI tract.     

In situ interleukin 5 gene expression in pediatric Crohn's disease

BACKGROUND: Eosinophils contribute to the intestinal inflammatory infiltrate in Crohn's disease (CD). Eosinophilic infiltration occurs early in Crohn's recurrences, and a release of eosinophil cationic proteins has been observed in active CD. The proliferation, differentiation, and activation of eosinophils are highly dependent on the cytokine interleukin 5 (IL5). In the present study, we used in situ hybridization (ISH) to investigate the expression of the IL5 gene in intestinal specimens from patients with CD. METHODS: We studied 14 intestinal samples from eight children who had undergone ileocolectomy for advanced CD. The samples were examined for the intensity of the inflammatory infiltrate. Normal pediatric intestine specimens served as controls. In situ hybridization was performed on frozen tissue using radiolabeled IL5 mRNA probes. RESULTS: Positive signal with the IL5 antisense probe was observed within numerous cells infiltrating the specimens involved with CD. The number of IL5-expressing cells correlated with the histological grade of inflammation. Most of the labeled cells were eosinophils, characterized by their bilobed nuclei. Rare IL5-positive cells were detected in the control tissues. No positive signal was obtained with the IL5 sense probe. CONCLUSION: These results suggest that IL5 can be produced by eosinophils at the sites of inflammation in active CD and could be involved in the immune response by activating eosinophils, at least in part through an autocrine pathway, and perhaps by interacting with B and T cells.     

Increased expression of costimulatory molecules on peripheral blood monocytes in patients with Crohn's disease

BACKGROUND: Activation of T lymphocytes and monocytes/macrophages has been implicated in the pathogenesis of Crohn's disease (CD). Costimulatory molecules play important roles in optimal T-cell activation. METHODS: With flow cytometric analysis we have investigated the expression of the costimulatory molecules B7-1 (CD80), B7-2 (CD86), and CD18 and the intercellular adhesion molecule-1 (ICAM-1) on peripheral blood monocytes and the expression of the activation markers HLA-DR and IL-2R (CD25) on peripheral blood T lymphocytes from 31 CD patients, 17 ulcerative colitis (UC) patients, and 10 healthy controls. RESULTS: In CD patients the percentage of activated T cells (CD3+ HLA-DR+ and CD3+ IL-2R+) was significantly increased compared with those of controls and UC patients (P < 0.05). Most monocytes from all three groups expressed B7-2, CD18, and ICAM-1 molecules (all > 79%), but only a few positive cells expressed B7-1 molecules (< 5%). No significant differences were detected in the percentage positivity of all costimulatory molecules tested among CD, UC, and controls. The mean fluorescence intensity (MFI) of B7-1 in all three groups was very weak and not significantly different. However, in CD patients there was a significantly increased MFI of B7-2, CD18, and ICAM-1 molecules compared with UC and controls (P < 0.05). On the other hand, both the percentage positivity and MFI of HLA-DR molecules on monocytes of UC patients were significantly lower than those of CD patients and controls (P < 0.05). CONCLUSIONS: Expression of the costimulatory molecules B7-2, CD18, and ICAM-1 on peripheral blood monocytes of CD patients is increased. In CD patients activation of peripheral T lymphocytes may correlate with increased expression of these costimulatory molecules on peripheral blood monocytes.     

Interleukin-12 and perforin mRNA expression is augmented in blood mononuclear cells in multiple sclerosis

Cytokines are suggested to orchestrate an abnormal immune response in multiple sclerosis (MS). The regulatory cytokine interleukin (IL)-12 induces T-helper (Th) cell switch to the Th1 type and the production by cytotoxic T cells of perforin, a cell lysis-inducing factor. It has been suggested that Th1-like cytokines may promote the development of MS, and the production of perforin to induce oligodendrocyte damage. In-situ hybridization with radiolabelled synthetic oligonucleotide probes was used to detect and enumerate mononuclear cells (MNC) expressing IL-12 and perforin mRNA in blood and cerebrospinal fluid (CSF) from patients with MS and controls. Plasma and CSF levels of IL-12 (p70) were evaluated by ELISA. Higher numbers of IL-12 and perforin mRNA-expressing MNC were registered in blood in MS and also in controls with aseptic meningoencephalitis (AM) compared to healthy subjects. There were a few patients with other non-inflammatory neurological diseases who also had high levels of IL-12 or perforin mRNA expressing blood MNC. A parallel elevation was observed for IL-12 (p70) concentrations in plasma. In the MS patients' CSF, there was a further augmentation of IL-12 mRNA expressing MNC. To evaluate autoantigen-induced IL-12 and perforin mRNA expression, blood MNC were cultivated +/- myelin basic protein (MBP), a candidate autoantigen in MS. Higher numbers of MBP-reactive IL-12 and perforin mRNA expressing blood MNC were detected in MS than controls. The augmentation of both IL-12 and perforin in MS might reflect ongoing inflammatory processes in MS and could represent targets for future treatments.     

Intestinal expression of human heat shock protein 90 in patients with Crohn's disease and ulcerative colitis

Heat shock proteins are induced by several stress factors and are potential antigens in autoimmune disorders. Expression of heat shock protein 90 (HSP 90) was investigated in patients with inflammatory bowel disease and normal controls. We combined western blot analysis with laser densitometry for quantitation. Localization of HSP 90 was investigated by immunohistochemistry. Western blots showed a significant mucosal expression of HSP 90, which was comparable in patients and controls. There was also no difference between normal and inflamed mucosa in inflammatory bowel disease. In immunohistochemical staining studies, HSP 90 was detected in epithelial cells, mononuclear cells, giant cells, nerve cells, and endothelial cells of small vessels. There was no difference in the intensity of staining or localization in patients with inflammatory bowel disease compared to controls. These findings render a potential protective or immunogenic function of HSP 90 in inflammatory bowel disease unlikely.     

Sialosyl Tn antigen expression is associated with the prognosis of patients with advanced gastric cancer

BACKGROUND: Several studies have revealed a correlation between sialosyl Tn antigen (STN) and certain clinicopathologic features of various cancers, and that STN is an independent prognostic factor. However, the clinical significance of the expression of STN in gastric cancer has not been reported. Thus, the purpose of this study was to evaluate immunohistochemically the clinical significance of expression of STN in gastric cancer. METHODS: The expression of STN in surgically resected specimens of human gastric cancer was evaluated immunohistochemically using a monoclonal antibody (TKH-2), in 60 patients whose serum STN levels were measured and in 54 patients with advanced cancer who had been followed for more than 5 years after gastrectomy. The correlations between the level of STN expression and clinicopathologic factors were analyzed. The staining intensity was graded as follows: (-), less than 5% of the cancer cells expressed STN; (+), 5-50%; (++), more than 50%. RESULTS: Sialosyl TN antigen staining was detected mainly on the cell membrane, in the cytoplasm, and in the luminal contents, and 57.2% of the 60 specimens expressed STN, whereas the corresponding value for positive serum levels was 15%. A higher percentage of advanced tumors expressed STN than did the early cases, but the difference was not statistically significant. All cases with strong staining, the (++) cases, were advanced cases either with lymph node metastases or with cancer invading in or beyond the muscle layer proper. The expression of STN appeared to be related to the clinical stage, the extent of cancer invasion, and the presence of lymph node metastases. Sialosyl TN antigen was detected in the serum in less than 6% of the patients whose tumors were (-) or (+) for STN expression, and in 86.7% of the patients whose tumors expressed high levels of STN (++). The estimated 5-year survival in advanced cases (Stage III) was significantly better in those with negative STN expression than in those with positive STN expression (P < 0.01). CONCLUSIONS: These results suggest that STN may be a useful marker associated with the prognosis of patients with advanced gastric cancer.     

Free protein S deficiency in patients with Crohn's disease

Multifocal intestinal infarctions, due to thrombosis in small vessels, might be a pathogenetic mechanism for Crohn's disease (CD). Deficiency of free protein S may contribute to the development of such thrombotic occlusions. In the present study free protein S was measured in 54 patients with CD. In 31 patients (57.4%) the plasma concentrations of free protein S were below the lower normal range. The mean value of free protein S in CD patients was 72.2%, as compared with 97.5% in healthy subjects (p < 0.01). The concentrations of C4b-binding protein and protein C were similar in the two groups. Free protein S levels were not correlated to disease activity, previous surgery or complications, extraintestinal manifestations, or current medical therapy. The impairment of the protein S/protein C/thrombomodulin system found in patients with CD favours coagulation and might be of importance for both the development of CD and its thromboembolic complications.     

25-hydroxycholecalciferol serum levels in patients with Crohn's disease

In 37 patients with Crohn's disease the 25-hydroxycholecalciferol (25-HCC) serum level, serum concentration of calcium and inorganic phosphate, and the enzyme activity of alkaline phosphatase were measured. Furthermore the activity index of Crohn's disease was determined in every patient. There was no statistically significant difference of 25-HCC serum levels in these patients compared to a healthy control group. Correspondingly most patients showed normal alkaline phosphatase enzyme activity and normal serum concentration of calcium and inorganic phosphate. No correlation between 25-HCC concentration and site of inflammation or activity index was found.     

Adenylate cyclase of gastric mucosa in patients with chronic renal failure

It has been shown previously that antisecretory response of famotidine is altered in patients with renal failure. To evaluate the underlying mechanism(s) of this clinical observation we obtained biopsy specimens of fundic mucosa from 3 groups of patients with variable renal function (group 1 normal renal function (n = 16); group 2 chronic renal failure (n = 16), CLCR > or = 5 < 90 ml/min; group 3 hemodialysis therapy (n = 16)) (matched for age, sex, and Helicobacter pylori (Hp) status. In the homogenized samples adenylate cyclase (AC) activity was assessed and the influence of uremia on this second messenger system involved in gastric acid secretion was tested. AC activity was measured as the formation of cAMP, which was determined by RIA. The mean basal AC activity was 150 in group 1, 190 in group 2, and 120 pmol cAMP/mg protein/20 min in group 3. There was a dose-dependent stimulation by histamine (1 microM-1 mM). Emax of cAMP formation ranged between 230 and 403 pmol cAMP/mg protein/20 min and EC50 between 5.9 and 20.1 microM histamine, dependent on Hp status. Histamine-stimulated AC activation was reduced to about 50% by 0.1 mM famotidine. The sensitivity of AC to histamine seems to decrease in patients undergoing hemodialysis. Similarly, the colonization with Hp may result in decreased maximal response of the AC system towards histamine.     

Pathology of olfactory mucosa in patients with Alzheimer''s disease

Radiographs of hand and hip joints of 15 hemodialysis patients taken annually over at least 15 years were evaluated in this retrospective study. We looked for signs of A beta 2M-amyloidosis such as lucencies, erosions, and soft tissue swelling. The femoral head-soft tissue distance (FHSTD) was used to estimate the soft tissue swelling of the hips. The number and size of lucencies, the number of erosions, and the FHSTD increased during dialysis. Both hyperparathyroidism and failure of renal allograft transplantation influenced the development of lucencies and erosions after 15 years. The FHSTD at the start of the dialysis appeared to be an independent prognostic factor for the lucencies of the hips and hands. We conclude that the radiological signs of A beta 2M-amyloidosis are influenced not only by the duration of dialysis, but also by age, failure of renal transplantation, hyperparathyroidism and the FHSTD at the start of dialysis.     

Insulin resistance in cancer patients is associated with enhanced tumor necrosis factor-alpha expression in skeletal muscle

The roles of tumor necrosis factor (TNF)-alpha and the facilitative glucose transporter (GLUT) 4 on the induction of insulin resistance in peripheral tissues of cancer patients was examined by quantitative competitive PCR on biopsies of abdominal rectal muscle from patients with gastrointestinal cancer. The degree of insulin resistance in these patients was measured by the euglycemic hyperinsulinemic glucose clamp using a high physiologic insulin concentration (100 microU/ml). Quantitative competitive PCR was carried out using DNA competitors constructed by deleting 20-30 bp between the two primer annealing sites. Decreased glucose uptake (M value) in peripheral tissues was accompanied by a significantly increased TNF-alpha mRNA in skeletal muscle (r=0.867, p=0.0025). GLUT4 mRNA, however, was positively correlated with M values (r=0.739, p=0.015). The amounts of mRNAs for TNF-alpha and GLUT4 in skeletal muscle were not correlated. Serum TNF-alpha concentrations remained below the limit of detection. These findings suggest that the insulin resistance in peripheral tissues of cancer patients is in part due to the induction TNF-alpha mRNA and the down regulation of GLUT4 mRNA in peripheral tissues.     

Interleukin-6 expression in cord blood of patients with clinical chorioamnionitis

BACKGROUND: The occurrence of hepatocellular neoplasms is frequently reported in young females. The role of oral contraceptives in the development of these tumors is considered. METHODS: A 14-cm tumor was diagnosed in a 24-year-old female who had been taking the contraceptive pill for the last 3 years. Numerous blocks of this lesion were histologically and immunohistochemically analyzed. Some fragments of the lesion were snap-frozen in order to search for sex hormonal receptors and hepatitis B and C virus. RESULTS: On histological examination, the tumor was found to be a hepatocellular, multilobular adenoma with small foci of hepatocellular carcinoma. Neither hepatitis B virus nor hepatitis C virus was found in serum or in the tumor. Nuclear estrogen receptors and progesterone receptors were detected by immunohistochemical analysis in both the adenoma and the carcinoma, but only progesterone receptors were detected by a radio-ligand binding assay in the tumor. CONCLUSION: This finding suggests that the contraceptive pill may stimulate the growth of hepatocellular, multilobular adenomas through the binding of hormonal compounds to their specific receptors within tumoral cells.     

Usefulness of Doppler ultrasound in the evaluation of patients with active Crohn's disease

OBJECTIVES: To assess the usefulness of Doppler ultrasound in the evaluation of the vascular changes in the splanchnic circulation and bowel wall described in patients with active Crohn's disease (ACD). DESIGN: We analyzed prospectively with Doppler ultrasound the mean velocity of portal flow, the resistive index (RI) of the superior mesentery artery (SMA) and we looked for vessels within the bowel wall. PATIENTS: 50 patients with ACD and 30 normal individuals. RESULTS: In comparison with normal individuals, patients with ACD showed a statistically significant difference (p < 0.001) in the mean velocity of the portal flow and in the RI of the SMA. In all patients with ACD, vessels could be seen within the bowel wall using the color Doppler ultrasound. CONCLUSION: Doppler ultrasound can be used as a non-invasive method to evaluate the vascular changes which develop in the splanchnic circulation and bowel wall of patients with ACD.     

Severe expression of corpus gastritis is characteristic in gastric cancer patients infected with Helicobacter pylori

BACKGROUND: Cancer of the skin may be difficult to diagnose clinically if there are minimal skin findings on examination. OBJECTIVE: To remind physicians that the symptoms of squamous cell carcinoma (SCC) may precede physical signs of the tumor. METHODS: Perineural invasion of SCC was confirmed by histologic examination of surgical specimens. RESULTS: We present two patients whose initial presentation of SCC was facial pain due to extensive perineural invasion. Their diagnoses were delayed until skin lesions eventually became evident months later. CONCLUSION: Early detection can be lifesaving, thus emphasizing the importance of including skin cancer in the differential for facial pain or paresthesia.     

Interleukin-12 gene-expression of macrophages is regulated by nitric oxide

PURPOSE: To determine the origin of subclavian vein catheter and lead dysfunction. MATERIALS AND METHODS: Cineradiography was performed on 10 patients with subclavian venous catheter dysfunction and three patients with pacemaker or defibrillator lead dysfunction. The leads and catheters were removed and replaced with use of a fluoroscopically guided technique; the needle entered the vein lateral to the first rib. Repeat cine examinations were performed following placement of new catheters. RESULTS: The cause of the dysfunction of all 10 catheters was shown to be pinch by the subclavicular musculotendinous tissues as the catheter passed below the clavicle toward its entry into the vein. All three leads were entrapped in the subclavicular tissues and stretched during abduction. The abnormal motion and clinical problems were eliminated after replacement. CONCLUSION: Subclavian catheter and lead malfunction is not due to compression between the first rib and the clavicle. It is due to entrapment in the subclavius muscle-costoclavicular ligament complex, which binds or compresses the device during movements. These problems can be avoided by employing fluoroscopically guided puncture techniques that enter the vein lateral to the first rib.     

Analysis of cell damage and proliferation in Helicobacter pylori-infected human gastric mucosa from patients with gastric adenocarcinoma

Helicobacter pylori (HP) infection, a cause of multifocal atrophic gastritis, is considered an important factor related to the evolution of the human gastric mucosa from normal to intestinal-type adenocarcinoma. We examined cell proliferation and both double and single strand DNA damage in situ in 35 patients undergoing gastrectomy for adenocarcinoma with HP-infected gastric mucosa by immunolocalization of Ki-67, terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling, and in situ nick translation. We also studied the distribution of intraepithelial neutrophils by elastase immunolocalization. HP infection was confirmed in all cases by serum anti-HP antibodies, ureas testing, and histopathological examination. HP-infected gastric mucosa was classified according to the degree of inflammation and intestinal metaplasia. Ki-67, terminal deoxynucleotidyl transferase-mediated labeling, in situ nick translation, and intraepithelial neutrophil indices all increased with the progression of gastritis and were highest in glands with incomplete intestinal metaplasia. All indices were lowest in gastric glands with complete intestinal metaplasia. Significant positive correlations were observed among these markers. Increased proliferative activity in HP-associated chronic gastritis in response to cell damage or injury was clearly demonstrated, suggesting that both HP-associated toxins and intraepithelial neutrophils are important in HP-related gastric epithelial injury. Increased cell turnover associated with incomplete intestinal metaplasia may result in DNA instability and subsequent development of intestinal-type gastric adenocarcinoma in HP-infected mucosa.     

Heterotopic gastric mucosa in the ileum with perforated ulcer

Heterotopic gastric mucosa (HGM) in the small bowel, other than in the Meckel's diverticulum or other congenitally anomalous bowel, is exceedingly rare. A patient with HGM of the small intestine with perforation of the ileum due to ulceration of the adjacent mucosa is presented. The authors remark the importance of the histological criteria to differentiate between true HGM from metaplastic changes, two entities with different behavior and prognostic implications.