Residues of the minimal sequence of G0S2 collectively contribute to ATGL inhibition while C-and N-terminal extensions promote binding to ATGL.

The protein encoded by the G0/G1 switch gene 2 (G0S2) is a potent inhibitor of adipose triglyceride lipase (ATGL) and thus an important regulator of intracellular lipolysis. Since dysfunction of lipolysis is associated with metabolic diseases including diabetes and obesity, inhibition of ATGL is considered a therapeutic strategy. G0S2 interacts with ATGL's patatin-domain to mediate non-competitive inhibition, however atomic details of the inhibition mechanism are incompletely understood. Sequences of G0S2 from higher organisms show a highly conserved N-terminal part, including a hydrophobic region covering amino acids 27 to 42. We show that predicted G0S2 orthologs from platypus, chicken and Japanese rice-fish are able to inhibit human and mouse ATGL, emphasizing the contribution of conserved amino acid to ATGL inhibition. Our site directed mutagenesis and truncation studies give insights in the protein-protein interaction on a per-residue level. We determine that the minimal sequence required for ATGL inhibition ranges from amino acids 20 to 44. Residues Y27, V28, G30, A34 G37, V39 or L42 within this sequence play a substantial role in ATGL inhibition. Furthermore, we show that unspecific interactions of the N-terminal part (amino acids 20-27) of the minimal sequence facilitate the interaction to ATGL. Our studies also demonstrate that full-length G0S2 shows higher tolerance to specific single amino acid exchanges in the hydrophobic region due to the stronger contributions of unspecific interactions. However, exchanges of more than one amino-acid in the hydrophobic region also result in the loss of function as ATGL inhibitor even in the full-length protein.

Computed tomography virtual bronchoscopy: normal variants, pitfalls, and spectrum of common and rare pathology.

A broad spectrum of pathologies that involve the laryngotracheobronchial airway and imaging plays a crucial role in evaluating these abnormalities. Computed tomography with virtual bronchoscopy has been found to be very helpful in defining the location, extent, and nature of these lesions, and is increasingly being used even in patients with contraindications for fiberoptic bronchoscopy and laryngoscopy. Ionizing radiation, associated with virtual bronchoscopy, can be minimized by using low-dose multidetector computed tomography and hybrid iterative reconstruction techniques. Furthermore, retrospectively generated virtual bronchoscopy from a routinely acquired computed tomography data set eliminates additional cost and radiation. In the future, virtual bronchoscopy assisted with advanced navigational techniques will broaden the diagnostic and therapeutic landscape. This article presents the characteristic features of common and rare laryngotracheobronchial pathologies seen with virtual bronchoscopy.

Anatomical variation of pulmonary venous ostium and its relationship with atrial arrhythmia in the Saudi population.

PURPOSE: To assess the anatomical variation of the ostial pattern of pulmonary veins observed on coronary computed tomography (CT) angiogram and to estimate the relationship of atrial arrhythmia with similar ostial variants in the Saudi population. MATERIALS AND METHODS: Thin-section (0.625 mm) contrast enhanced CT scans of 151 consecutive patients obtained over a span of a two-year period (January 2009-January 2011) for the diagnosis of ischemic heart disease, aortic regurgitation, infective endocarditis, aortic aneurysm, fibrosing mediastinitis, and pericardial defect. The retrospective study was focused on the identification of the number of venous ostia on either side of the left atrium and the drainage patterns of pulmonary veins. The frequency of each pattern was tabulated, and significance of their relationship with atrial arrhythmias was assessed with the X(2) and Fisher's exact tests. RESULTS: Out of 151 patients analyzed, 26 patients (17.2%) had anatomical variation of the pulmonary venous ostia. Atrial arrhythmia was recorded in 16 of 26(61.5%) patients (p = 0.000). The association of higher anatomical variation of pulmonary venous ostia was recorded as higher (p = 0.034) in the female group (n = 15, 57.7%). CONCLUSION: The association between anatomically varied venous ostia and atrial arrhythmia was significant (p = 0.000) with a significantly higher anatomical variation of pulmonary venous ostia in the female Saudi population (p = 0.034).

Genetic enhancement of host plant-resistance of the Lalat cultivar of rice against bacterial blight employing marker-assisted selection.

To incorporate durable resistance against bacterial blight, a major disease rice, three resistance genes, xa 5, xa13 and Xa21, from IRBB 60 were transferred through marker-assisted backcrossing using RG 556, RG 136 and pTA248 markers linked to the three genes to supplement the Xa4 gene present in Lalat, a popular rice cultivar. Effective selection enabled the transfer in three back-crosses and a generation of selfing and background selection employing morphological and grain quality traits and molecular markers, led to >90 % recovery of the recurrent parental genome. The gene pyramids exhibited high levels of resistance against the pathogen in multi-location evaluation trials conducted over several locations of bacterial blight in India. IL-2 (CRMAS2621-7-1), a gene pyramid, was identified as being promising for several endemic regions of bacterial blight and was released as Improved Lalat in one of the identified regions. The success of the study demonstrates the vast potential of marker-assisted selection for gene stacking and recovery of the parental genome with high precision.

Patients with inflammatory bowel disease may have a transforming growth factor-beta-, interleukin (IL)-2- or IL-10-deficient state induced by intrinsic neutralizing antibodies.

Ulcerative colitis (UC) and Crohn's disease (CD) are considered to be immunologically mediated disorders that share certain features with murine models of colitis. Whether any of these models are physiologically relevant to the human condition remains controversial. The hypothesis is that increased amounts of antibodies neutralizing transforming growth factor (TGF)-beta, interleukin (IL)-2 or IL-10 create a relative immunodeficient state in inflammatory bowel disease (IBD) that predisposes to disease. To evaluate this, serum samples from patients with UC or CD and from normal healthy individuals were studied by enzyme-linked immunosorbent assays. Antibodies recognizing TGF-beta were most prevalent in UC (P<0.01); anti-IL-10 antibodies were elevated in CD (P<0.05), while anti-IL-2 antibodies were the same for all three groups. Importantly, the percentage of IBD patients with at least one of the antibody levels greater than any control value was 30% for UC and 33% for CD. To verify the presence of these antibodies, immobilized TGF-beta was exposed to UC sera and the attached proteins identified by Western blot assay. The proteins proved to be exclusively immunoglobulin (Ig) G. To evaluate the neutralizing activity of these antibodies, cytokine-specific IgG from subjects in each group of patients was incubated with TGF-beta, IL-2 or IL-10 before addition to a bioassay with changes in viability determined by a colorimetric analysis. Antibodies from most individuals in all three groups neutralized the action of each cytokine. This study shows that about one-third of IBD patients may have a relative deficiency of TGF-beta, IL-2 or IL-10 due to an increase in neutralizing antibodies in their sera.

Non-response to infliximab may be due to innate neutralizing anti-tumour necrosis factor-alpha antibodies.

Infliximab is a chimeric anti-tumour necrosis factor (TNF)-alpha antibody that is therapeutic in many patients with inflammatory bowel disease. What causes certain patients not to respond is unknown. The question posed is whether innate anti-TNF-alpha antibodies play any role in the response to infliximab. Blood was drawn prior to the initial dose of infliximab. Serum anti-TNF-alpha antibodies were quantitated by enzyme-linked immunosorbent assay (ELISA). Affinity-purified anti-TNF-alpha antibodies were isolated from serum immunoglobulin G using TNF-alpha-coated beads. The ability of these antibodies to induce apoptosis of macrophages was measured by annexin and propidium iodide staining. Changes in TNF receptor type 2 (TNFR2) expression and release were determined by immunofluorescence and ELISA respectively. TNF-alpha-neutralization was assessed by the reversal of the lytic actions of TNF-alpha on WEHI cells. The amounts of innate anti-TNF-alpha antibodies in the serum from infliximab responders versus non-responders were the same. Apoptosis of monocytes increased with infliximab and by several of the purified anti-TNF-alpha antibodies, but these findings did not vary with the patients' responses to infliximab. Effects of the anti-TNF-alpha antibodies on the expression of TNFR2 on monocytes and their release of soluble TNFR2 did not vary with the patients' responses to infliximab. However, the neutralizing capacity of these antibodies differed, with responders having antibodies that reduced only 47 +/- 4% of the TNF-alpha activity while those from non-responders reduced 70 +/- 5% of the TNF-alpha activity (P < 0.01). Non-responders have innate anti-TNF-alpha antibodies with greater neutralizing activity than antibodies from responders. Any TNF-alpha-mediated disease process would be neutralized by intrinsic antibodies, so that the disease is likely to be driven by non-TNF-alpha-mediated events.

Coronary artery dissection with rupture of aortic valve commissure following type A aortic dissection: the role of 64-slice MDCT.

A rare case of bilateral coronary artery dissection with rupture of aortic valve commissure following type A aortic dissection is described. 64-slice multidetector computed tomography (MDCT) was able to demonstrate both this findings along with involvement of other neck vessels. TEE demonstrated the severity and mechanisms of aortic valve damage and assisted the surgeon in valve repair. MDCT has played an invaluable role in the diagnosis of the abnormal details of such life-threatening vascular complications.

Contrast-enhanced 64-section coronary multidetector CT angiography versus conventional coronary angiography for stent assessment.

PURPOSE: To prospectively evaluate the accuracy of 64-section computed tomography (CT) for diagnosis of stent restenosis, by using conventional coronary angiography as the reference standard. MATERIALS AND METHODS: The ethics committee granted permission for the study; patients gave written consent. Contrast material-enhanced coronary CT angiography was performed in 53 patients (45 men, eight women; mean age, 54 years +/- 9 [standard deviation]) suspected of having stent restenosis. Coronary CT angiographic findings were compared with conventional coronary angiographic findings. Two physicians analyzed coronary CT angiographic data sets with multiplanar reformatted images and three-dimensional reformations by using a volume-rendering technique and looked for stent detectability, low-attenuation in-stent filling defects, and grades of restenosis. Conventional coronary angiographic results were interpreted by one of several observers in consensus for stent restenosis; they were blinded to coronary CT angiographic data. Statistical software and general estimating equations were used for data analysis. RESULTS: One hundred ten stents were identified in 53 patients. Sensitivity, specificity, positive predictive value, negative predictive value, and accuracy of coronary CT angiography in detection of in-stent restenosis were 96.9%, 88.0%, 77.5%, 98.5%, and 91%, respectively. Coronary CT angiography depicted in-stent low-attenuation filling defects with an accuracy of 91% and negative predictive value of 98.5% (95% confidence interval: 90.9, 99.9). Coronary CT angiography depicted the status of 97 of 107 stents. There was no significant difference between in-stent lumen visibility and stent diameter (P = .104). Coronary CT angiography helped diagnose 15 of 18 stent restenoses with less than 50% narrowing, five of five stent restenoses with 50%-74% narrowing, and nine of nine (100%) stent restenoses with 75% or greater narrowing or total occlusion of the stent lumen. CONCLUSION: Coronary CT angiography can depict in-stent low-attenuation filling defects, which appear to be a reliable sign of stent restenosis, and 64-section CT depicts such defects with a high degree of accuracy.

K-ras mutations and cell kinetics in Helicobacter pylori associated gastric intestinal metaplasia: a comparison before and after eradication in patients with chronic gastritis and gastric cancer.

BACKGROUND: Helicobacter pylori related gastric intestinal metaplasia (IM) is considered to be a precancerous lesion. AIMS: To identify the effects of H pylori eradication on K-ras mutations, cell kinetics in IM and histological changes in patients with and without gastric cancers in a one-year prospective study. METHODS: Patients included group A (n = 39), chronic gastritis, and group B (n = 53), intestinal-type early gastric cancer patients who had all undergone endoscopic mucosal resection (n = 25) or surgical resection (n = 28). K-ras codon 12 mutations in IM were examined, followed by DNA sequencing analysis. Proliferating and apoptotic cells were detected with anti-Ki-67 antibody and using the TUNEL method, respectively. RESULTS: The incidence of K-ras mutations in the cancer was only 3.8%. The mutant K-ras in IM was observed more frequently in group A (46.2%) than in group B patients (1.9%) (p<0.005). After eradication, the K-ras mutations significantly declined to 12.8% in group A (p<0.005). The mutation pattern of K-ras codon 12 before eradication was that GGT was mainly changed to AGT (50%) in group A. AGT transformation was not affected by treatment. Apoptosis in IM showed an increase after H pylori eradication in both groups (p<0.05 in group A) although no histological improvement in IM was observed. The monocyte score was significantly higher in group A than in group B (p<0.05); the score improved significantly after eradication. CONCLUSIONS: K-ras mutations in IM do not always play a role in gastric carcinogenesis but cell kinetics, especially apoptosis, in IM may contribute to it. There are early events in K-ras mutations which are influenced by H pylori infection; some mutations may also be selected by eradication. These unstable K-ras mutations in IM may be related to lymphocyte infiltration caused by H pylori infection.

Intracardiac shunting of ruptured Sinus of Valsalva aneurysm in a patient presented with acute myocardial infarction: role of 64-slice MDCT.

Rupture of Sinus of Valsalva aneurysm (SVA) may be either congenital or acquired. This report describes a case of intracardiac shunting of a ruptured SVA of atherosclerotic origin to the right atrium, presenting with acute myocardial infarction. The sinus of Valsalva aneurysm and the intracardiac shunt track into the right atrium was not defined by the two-dimensional echocardiography could be seen by 64-slice multi detector computed tomography (MDCT).

Anomalous origin of the three coronary arteries from the right aortic sinus Valsalva: role of MDCT coronary angiography.

BACKGROUND: Monocuspal origin of all three coronary arteries through separate ostia from the right aortic sinus (RCS) is a rare occurrence. To date, the use of multidetector computed tomography (MDCT) for imaging of congenitally abnormal coronary arteries has been discussed only in few individual case reports. OBJECTIVE: To describe the role of MDCT coronary angiography in the evaluation of two rare cases of monocuspal origin of all three coronary from RCS. PATIENTS AND METHODS: We had a retrospective review of clinical information and imaging studies for two patients presented with chest pain. Both patients underwent conventional coronary angiography followed by noninvasive imaging with MDCT. RESULTS: Both patients had anomalous origin of the all three coronary arteries from the RCS. In one case the LAD took an intramural course in between the aorta and the right ventricular outflow tract (RVOT) while it passed anterior to the RVOT in the other patient. In the first patient, there was also associated coronary fistula to the right ventricle along with right coronary artery (RCA) and left anterior descending coronary artery (LAD) narrowing. Both the stenosed segments were successfully stented and were demonstrated to be patent in the subsequent MDCT. CONCLUSION: Monocuspal origin of all three coronary artery from the RCS is a rare anomaly, can be reliably diagnosed by MDCT. CT angiogram is a convenient complementary tool for the coronary arteriography.

Activation antigens on colonic T cells in inflammatory bowel disease: effects of IL-10.

Activated T cells that express activation antigens are termed nonprofessional antigen-presenting cells (T-APCs). This study evaluates the ability of lamina propria lymphocytes (LPLs) in inflammatory bowel disease (IBD) to become T-APCs. LPLs were stained by two-colour immunofluorescence to determine the expression of activation antigens on T cells. Those from actively inflamed IBD mucosa expressed greater amounts of MHC class II (DR) and CD86 than did LPL T cells from disease controls or normal individuals. After culture in IL-2 with or without IL-10, the ability of the T-APCs from IBD colon to stimulate allogeneic peripheral blood T cell proliferation was measured. The T-APCs from IBD stimulated an allogeneic mixed lymphocyte reaction, particularly through their expression of DR and CD86, as demonstrated by antibody blocking. Normal LPLs acquired these properties only if repeatedly stimulated with allogeneic peripheral blood lymphocytes (PBLs) used as cell lines in the presence of IL-2. Addition of IL-10 reduced expression of activation antigens and the stimulatory ability of LPLs from either IBD patients or from these cell lines. In summary, LPLs from active IBD, but not from disease controls, express activation antigens that stimulate naïve T cells, a process that is reduced by IL-10. This may contribute to perpetuation of the inflammation.

Gastric intestinal metaplasia as detected by a monoclonal antibody is highly associated with gastric adenocarcinoma.

BACKGROUND: Some forms of gastric intestinal metaplasia (GIM) may be precancerous but the cellular phenotype that predisposes to gastric carcinogenesis is not well characterised. Mucin staining, as a means of differentiating GIM, is difficult. A monoclonal antibody, mAb Das-1 (initially called 7E(12)H(12)), whose staining is phenotypically specific to colon epithelium, was used to investigate this issue. METHODS: Using mAb Das-1, by a sensitive immunoperoxidase assay, we examined histologically confirmed GIM specimens from two countries, the USA and Japan. A total of 150 patients comprised three groups: group A, GIM (fields away from the cancer area) from patients with gastric carcinoma (n=60); group B, GIM with chronic gastritis (without gastric carcinoma) (n=72); and group C, chronic gastritis without GIM (n=18). RESULTS: Fifty six of 60 (93%) patients with GIM (both goblet and non-goblet metaplastic cells) from group A reacted intensely with mAb Das-1. Cancer areas from the same 56 patients also reacted. In contrast, 25/72 (35%) samples of GIM from patients in group B reacted with mAb Das-1 (group A v B, p<0.0001). None of the samples from group C reacted with the mAb. CONCLUSIONS: Reactivity of mAb Das-1 is clinically useful to simplify and differentiate the phenotypes of GIM. The colonic phenotype of GIM, as identified by mAb Das-1, is strongly associated with gastric carcinoma.

Cellular immune response against tropomyosin isoform 5 in ulcerative colitis.

We have reported an autoantibody response in ulcerative colitis (UC) against human tropomyosin isoform 5 (hTM5), the predominant colonic epithelial cell hTM isoform. In this report, we determined the number of IFN-gamma-secreting cells (spot-forming cells, SFC) against hTM5 by an enzyme-linked immunospot (ELISPOT) assay. Another cytoskeletal protein, caldesmon, CaD40, was used as a control antigen. Peripheral blood mononuclear cells were separated by a Ficoll density gradient from 28 patients with UC, 13 patients with Crohn's disease (CD), and 9 healthy subjects (HS). The mean (+/-SEM) SFC values against hTM5 in UC, CD, and HS were 48.8 +/- 8.1, 18.6 +/- 4.6, and 20.8 +/- 8.6, respectively. The value in UC was significantly higher than those in CD (P < 0.005) and HS (P < 0.025). SFC values in CD did not differ from those in HS. None of the 50 samples (except 1 UC) reacted to the CaD40 antigen. This study demonstrates, for the first time, a defined colon epithelial cell antigen, hTM5, that is capable of inducing a significant T cell response in UC but not in CD.

Colon epithelial cellular protein induces oral tolerance in the experimental model of colitis by trinitrobenzene sulfonic acid.

Rectal administration of trinitrobenzene sulfonic acid (TNBS) produces chronic colitis in experimental animals. However, the role of epithelial cellular protein(s) in this model is unknown. We examined whether oral tolerance can be induced in this model with colon epithelial cell proteins and whether it is organ specific. Rats were fed five times with extracts of LS-180 human colon cancer cells or HT 1080 human fibroblast cells. Syngeneic normal rat colon or small intestinal extracts were fed to separate groups of rats. After oral feedings, each rat received TNBS by enema. Rats were killed 15 days later, and the following were measured: gross and histologic disease score, weight, thickness, and myeloperoxidase values of colon and serum interferon-gamma (IFN-gamma) and transforming growth factor-beta (TGF-beta) levels. Rectal TNBS alone produced severe colitis with a 26% mortality rate. Rats fed LS-180 or rat colon extract before TNBS enema were protected, as evidenced by reductions in mortality rate, disease scores, and myeloperoxidase values. However, rats fed HT 1080 or small intestine extract lacked such protection. To examine the possible mechanism of the oral tolerance, T lymphocytes from mesenteric lymph nodes and spleen of LS-180 extract-fed rats were passively transferred to naive rats, and this was followed by TNBS enema. These rats showed clear protection. Protected animals had low IFN-gamma and high TGF-beta levels. This study demonstrates that cellular protein(s) from human colon epithelial cells, but not from human fibroblasts, can induce oral tolerance in experimental colitis. This oral tolerance is mediated by primed mesenteric and splenic T lymphocytes.

A phenotypic change of small intestinal epithelium to colonocytes in small intestinal adenomas and adenocarcinomas.

OBJECTIVE: Using a novel monoclonal antibody (mAb Das-1) that specifically reacts with colon epithelium, we examined if there is a phenotypic change of small intestinal enterocytes toward colonocytes in small intestinal neoplastic tissue. METHODS: Tissue sections of the small intestine consisting of adenomas (n = 20, five with histories of familial polyposis), adenocarcinomas (eight primary and one metastatic from colon). carcinoids (n = 2), and hyperplastic polyps (n = 3) were examined by a sensitive immunoperoxidase assay using mAb Das-1 (IgM isotype). Normal jejunal (n = 10) and colonic (n = 10) biopsy specimens were also included as additional controls. RESULTS: mAb Das-1 reacted with normal colonic epithelium but not with jejunal mucosa. However, mAb Das-1 reacted strongly with each of the five adenomas (100%) from patients with histories of familial polyposis, but only five of 15 (33%) of the adenomas from nonfamilial polyposis patients, and each of the eight (100%) adenocarcinomas of the small intestine (p < 0.001). The reactivity with the adenomas from nonfamilial polyposis patients was very focal, whereas in the adenomas with familial polyposis the reactivity was more extensive. Each of the eight carcinomas reacted strongly with mAb Das-1. Adjacent normal small intestinal mucosa did not react. Hyperplastic polyps and the carcinoids did not react with mAb Das-1. CONCLUSION: These data demonstrate a phenotypic change in small intestinal epithelium toward the colonic phenotype, particularly in familial polyposis and in adenocarcinomas. mAb Das-1 may be clinically useful in identifying small intestinal adenomas with "high risk" for malignancy, such as in familial polyposis.

Oral administration of unmodified colonic but not small intestinal antigens protects rats from hapten-induced colitis.

Colonic administration of a hapten, 2,4,6-trinitrobenzene sulphonic acid (TNBS) has been shown to induce colitis in rats. We are using this model to investigate the role of colonic antigens in the immunopathology. In this study, we show that colitis can be suppressed by oral administration of haptenized colonic antigens prior to the TNBS enema. Moreover, our data suggest that haptenization of the colonic antigens is not essential because oral feeding of non haptenized colonic antigens too protects rats from TNBS-induced colitis. Thus, unmodified colonic antigens may be involved in the induction of oral tolerance, and possibly in the pathogenesis in this model of colitis. Further, we show that the protective immunity or oral tolerance induced by non haptenized colonic antigens can be passively transferred to naïve rats by mesenteric T lymphocytes. Interestingly, oral feeding of small intestinal antigens, haptenized and non haptenized, does not protect rats from colitis, suggesting a specific role for colonic antigens. These data underscore the usefulness of this rat model in the identification of pathogenic antigens in colitis and in the development of therapeutic strategies based on oral tolerance.

Phenotype of Barrett's esophagus and intestinal metaplasia of the distal esophagus and gastroesophageal junction: an immunohistochemical study of cytokeratins 7 and 20, Das-1 and 45 MI.

The pathogenesis of short segment Barrett's esophagus (SSBE) and intestinal metaplasia (IM) of the gastroesophageal junction (IMGEJ) are poorly understood. Also, these conditions are difficult to distinguish from one another based solely on endoscopic and pathologic criteria. Therefore, the aim of this study was to evaluate the immunophenotypic features of SSBE and IMGEJ and to compare the results with lesions of known etiologies: long segment BE (LSBE) caused by reflux disease and Helicobacter pylori-induced IM of the gastric antrum (IMGA). Routinely processed mucosal biopsy specimens from 11 patients with LSBE, 17 with SSBE, 10 with IMGEJ, 16 with IMGA, 17 with a normal nonmetaplastic GEJ, and 7 patients with a normal gastric antrum were immunohistochemically stained with monoclonal antibodies to: Das1, an antibody shown to react specifically with colonic goblet cells; 45M1, an antibody that recognizes the M1 gastric mucin antigen; and cytokeratin (CK) 7 and 20, antibodies that have previously been reported to show specific staining patterns in BE versus IMGA. Also evaluated was nonintestinalized mucinous epithelium from LSBE, SSBE, and also the normal GEJ and gastric antrum. LSBE, SSBE, and IMGEJ showed similar prevalences of Das1 (91% versus 88% versus 100%) and 45M1 reactivity (100% versus 100% versus 100%), and a similar pattern of CK7/20 reactivity (diffuse strong CK7 staining of the surface and crypt epithelium, and strong surface and superficial crypt CK20 staining) (91% versus 94% versus 90%). In contrast, although 45M1 reactivity in IMGA (93%) was similar to that of the other three groups, IMGA showed a significantly lower prevalence of Das positivity (13%, p < 0.001), and only a 14% prevalence of the CK7/20 staining pattern that was predominant in the other three groups (p < 0.001). Das1, 45M1, and CK7/20 staining were similar in nonintestinalized "cardia-type" mucinous epithelium from LSBE, SSBE, and the GEJ, but all were distinct from the normal gastric antrum. In summary, the immunophenotypic features of SSBE and IMGEJ are similar and closely resemble those seen in classic LSBE, but are distinct from IMGA. This may indicate that IM in LSBE, SSBE and at the GEJ have similar biologic properties. Based on our data, SSBE and IMGEJ cannot be distinguished on the basis of their immunophenotype.

Autoimmunity in ulcerative colitis (UC): a predominant colonic mucosal B cell response against human tropomyosin isoform 5.

We set out to examine if the IgG-producing cells in the colonic mucosa in UC are committed to tropomyosin isoform 5 (hTM5), a putative autoantigen in UC. Lamina propria mononuclear cells (LPMC) were isolated from colonoscopic biopsy specimens from recto-sigmoid and proximal colon. Twenty-three patients with UC, eight with Crohn's colitis (CC), and 10 non-inflammatory bowel disease (non-IBD) controls were included. The ELISPOT assays were used to quantify lamina propria B cells producing total immunoglobulin (IgA, IgG, IgM), IgG, IgA, as well as IgG against hTM5 isoform. The median value of percentage of total IgG-producing lymphocytes was similar in UC (12%) and CC (11%), but was significantly (P < 0.0002) higher than non-IBD controls (6%). However, in UC, but not in CC and non-IBD, a large number of lamina propria B cells produced IgG against hTM5 (median values: UC 42%, CC 2.5%, non-IBD 0%). This difference in UC when compared with CC and non-IBD was highly significant (P < 0.00001). Twenty-one of 23 (91%) patients with UC had percentage of anti-hTM5 IgG-producing immunocytes more than 2 s. d. above the mean for non-UC patients. In UC but not in CC and non-IBD controls, the increased number of IgG-producing cells are largely committed to produce IgG against hTM5-related epitope(s).