Marker For Inflammatory Bowel Disease Research Articles

Is faecal calprotectin useful in the screening of inflammatory bowel disease in children? - a three year review of practice

Aims Calprotectin is a neutrophil intracellular calcium and zinc binding protein that has been shown to be associated with intraluminal intestinal inflammation when detected in stool. Its use as a non-invasive marker of inflammatory bowel disease (IBD) is well validated in children and adults. Levels of faecal calprotectin (FC) >60 μg/g is considered abnormal, however a FC >100 μg/g has a sensitivity of 98% and specificity of 91% in detecting children with IBD. The authors carried out a study to evaluate the current use of FC as screening tool for IBD, in a tertiary paediatric gastroenterology unit. Methods All FC requests sent over a 3 year period between January 2008 and February 2011 in children aged 1 to 16 years of age (n=68) were reviewed. Additionally clinical notes were analysed for indication for testing, clinical impression on first consultation and subsequent change in management following FC result. Results Of the 68 FC requests, 47 were in children suspected to have inflammatory bowel disease, 7 were used to monitor known inflammatory bowel disease and 14 were for other indications. FC was >100 μg/g in 23 of 68 children (30%). Of the 47 children with suspected IBD at referral, 35 (74%) had normal FC and did not subsequently undergo an endoscopy. 4 children with FC 5 children with suspected IBD with a FC>100 μg/g had abnormal endoscopy. 1 child with a FC>100 μg/g, had no endoscopy as campylobacter was isolated through stool culture, and responded well to treatment. There were however 2 children with elevated FC, but with normal histology following endoscopy. Conclusion Review of our practice over the last 3 years has demonstrated that when FC was measured ie when IBD was possible but children seemed clinically well, IBD was only diagnosed in those with a raised FC. During the study period no child with symptoms consistent with IBD, with a normal FC was subsequently diagnosed with IBD. In well children a normal FC could reduce the need for diagnostic endoscopy. FC is a useful additional screening tool.

Deficiency of N-acetylgalactosamine in O-linked oligosaccharides of IgA is a novel biologic marker for Crohnʼs disease

Ideal biomarkers are required to be developed for the diagnosis and prediction of the treatment of inflammatory bowel disease (IBD). We have reported that alteration of N-linked oligosaccharides of immunoglobulin (Ig) G is a novel diagnostic marker of IBD. Oligosaccharide alterations of IgA, however, have not been investigated in IBD patients. N- and O-linked oligosaccharides of serum IgA purified from 32 patients with Crohn's disease (CD), 30 patients with ulcerative colitis (UC), and 30 healthy volunteers (HV) were analyzed with high-performance liquid chromatography and mass spectrometry. Enzymes related to oligosaccharide attachment were investigated. N-linked oligosaccharides of IgA were not different between IBD and HV. In contrast, the number of N-acetylgalactosamines per hinge glycopeptide (GalNAc/HP) in the O-linked oligosaccharides of IgA was significantly decreased in patients with CD compared with UC and HV. GalNAc/HP had high sensitivity and specificity for discriminating between CD and HV based on receiver operating characteristic analysis. Lower GalNAc/HP was associated with more severe disease activity of CD. Changes in GalNAc/HP levels in 6 weeks after treatment with infliximab were associated with the clinical activity of CD at 30 weeks. GalNAc transferase expression of naïve B cells and extent of GalNAc attachment in IgA were significantly decreased by interleukin-21 in vitro. The number of GalNAc attached in the IgA O-linked glycans of CD patients was significantly decreased, and strongly correlated with the clinical activity. Alterations of GalNAc attachment in IgA could be useful as a novel diagnostic and prognostic marker of CD.

Development and analytic validation of an immunoassay for the quantification of canine S100A12 in serum and fecal samples and its biological variability in serum from healthy dogs

S100A12 (calgranulin C) is a Ca 2+-binding protein that has been proposed to play a central role in both innate and acquired immune responses. In humans, S100A12 has been reported to be increased in serum and/or plasma in patients with various inflammatory disorders, and this protein has been suggested to be a sensitive and specific marker for inflammatory bowel disease (IBD). An immunoassay for S100A12 is currently available for use in humans, but antibodies against the human protein do not cross-react with canine S100A12 (cS100A12). Both sensitive and specific markers for canine patients with systemic or localized inflammatory diseases are currently lacking, thus the aim of this study was to develop and analytically validate a radioimmunoassay (RIA) for the quantification of cS100A12 in serum and fecal specimens and to determine the biological variation of cS100A12 in serum from healthy dogs. A competitive liquid-phase RIA was developed and analytically validated by determining assay working range, dilutional parallelism, spiking recovery, and intra- and inter-assay variability. Reference intervals for serum and fecal concentrations of cS100A12 were established from 124 and 65 healthy dogs, respectively, and components of variation for serum cS100A12 were determined from 11 dogs over 2.6 months. The working range of the assay was 0.6–432.7 μg/L. No cross-reactivity was observed with the cS100A8/A9 protein complex, the closest structural analogues available. Observed-to-expected ratios (O/E) for the serial dilution of serum and fecal extracts ranged from 97.2 to 146.8% and from 75.3 to 129.8%, respectively. O/E for spiking recovery for serum and fecal extracts ranged from 87.8 to 130.4% and from 84.8 to 143.8%, respectively. Coefficients of variation (CV) for intra- and inter-assay variability for sera were ≤8.1% and ≤7.8%, respectively, and were ≤7.8% and ≤8.7%, respectively, for fecal extracts. Reference intervals for serum and fecal cS100A12 were 33.2–225.1 μg/L and <24−745 ng/g, respectively. For biological variability testing, analytical, intra-individual, inter-individual, and total CV were 5.7, 29.2, 31.2, and 66.0%, respectively, yielding an index of individuality of 0.95 and a minimum critical difference ( p < 0.05) for sequential values of 84.9%. The RIA for cS100A12 measurement described here is analytically sensitive and specific, linear, accurate, precise, and reproducible, and will facilitate further research into the clinical utility of quantifying serum and fecal cS100A12 in canine patients with inflammatory diseases. Moderate changes in serum cS100A12 concentrations may be clinically relevant; however, the use of a population-based reference interval may require caution.

Update of fecal markers of inflammation in inflammatory bowel disease

The diagnosis, prognosis, and assessment of disease activity of inflammatory bowel disease (IBD) require investigating clinical, radiological, and histological criteria, as well as serum inflammatory markers. However, a range of fecal inflammatory markers now appears to have the potential to greatly assist in these processes. Calprotectin, a prominent neutrophil protein, was identified two decades ago as a potentially revolutionary marker for IBD. Following this discovery, numerous additional markers, including S100A12, lactoferrin, and M2-pyruvate kinase, have also been suggested as novel markers of IBD. In the present study, we provide an up-to-date review of fecal markers of IBD, and further, provide a novel analysis of each of these fecal markers in severe ulcerative colitis and compare their expression pattern in contrast to calprotectin.

Activated thrombin activatable fibrinolysis inhibitor (TAFIa) is associated with inflammatory markers in inflammatory bowel diseases: TAFIa level in patients with IBD

Background Thrombin activatable fibrinolysis inhibitor (TAFI) has been reported to be involved in the pathogenesis and progression of inflammatory bowel disease (IBD). Activated TAFI (TAFIa) attenuates fibrinolysis by cleaving C-terminal lysine residues thus down-regulating plasminogen activation. To date, no reports on TAFIa in IBD have been published. Methods Plasma levels of TAFIa were measured using a functional assay in 55 consecutive patients with ulcerative colitis (UC) and 50 with Crohn's disease (CD). Associations of TAFIa with disease activity, hemostatic variables and inflammatory markers were assessed. Results Plasma TAFIa was higher in CD patients than in those with UC. The disease activity correlated positively with TAFIa levels in the UC group, but not in the CD group. In UC patients, there were positive correlations of TAFIa with white blood cells, C-reactive protein and fibrinogen and an inverse correlation with albumin. In the CD group, a positive correlation was shown for C-reactive protein, fibrinogen and platelet count, while a negative correlation was noted for albumin. Conclusions This study is the first to show that TAFIa is increased in CD patients compared with UC and its levels are associated with inflammatory markers in both forms of IBD. These findings fit in the hypothesis that TAFIa may be a marker of active IBD, and in particular of active UC.

Current and future role of biomarkers in Crohn's disease risk assessment and treatment

Background:Crohn’s disease (CD), a chronic inflammatory bowel disease (IBD), occurs in genetically susceptible individuals who develop aberrant immune responses to endoluminal bacteria. Recurrent inflammation increases the risk of several complications. Despite use of a traditional “step-up” therapy with corticosteroids and immunomodulators, most CD patients eventually require surgery at some time in their disease course. Newer biologic agents have been remarkably effective in controlling severe disease. Thus, “top-down,” early aggressive therapy has been proposed to yield better outcomes, especially in complicated disease. However, safety and cost issues mandate the need for careful patient selection. Identification of high-risk candidates who may benefit from aggressive therapy is becoming increasingly relevant. Serologic and genetic markers of CD have great potential in this regard. The aim of this review is to highlight the clinical relevance of these markers for diagnostics and prognostication.Methods:A current PubMed literature search identified articles regarding the role of biomarkers in IBD diagnosis, severity prediction, and stratification. Studies were also reviewed on the presence of IBD markers in non-IBD diseases.Results:Several IBD seromarkers and genetic markers appear to be associated with complex CD phenotypes. Qualitative and quantitative serum immune reactivity to microbial antigens may be predictive of disease progression and complications.Conclusion:The cumulative evidence provided by serologic and genetic testing has the potential to enhance clinical decision-making when formulating individualized IBD therapeutic plans.

The role of red cell distribution width as a marker in inflammatory bowel disease

To determine whether red cell distribution width could be used for the assessment of disease activity in patients with inflammatory bowel disease. Materials and methods: A total of 165 patients with inflammatory bowel disease (105 ulcerative colitis; 60 Crohn's disease) and 43 healthy blood donors were included in this retrospective study in a tertiary care setting. The medical records of the patients were reviewed to note clinical activity indices, red cell distribution width, serum C reactive protein, erythrocyte sedimentation rates, leukocyte, and platelet counts. Results: Red cell distribution width, serum C reactive protein, erythrocyte sedimentation rate, platelet count, and the proportion of individuals with increased red cell distribution width were all significantly elevated in the patients compared to the controls (P < 0.05). Red cell distribution width and the proportion of patients having elevated red cell distribution width were higher in patients with Crohn's disease compared with ulcerative colitis (P < 0.05). Red cell distribution width was significantly correlated with erythrocyte sedimentation rate, endoscopic activity index (in ulcerative colitis), and platelet count (P < 0.05). For a red cell distribution width cut-off of 14.45 sensitivity for differentiation of Crohn's disease from ulcerative colitis was 70% and specificity was 56%. Conclusion: Red cell distribution width, as a cost-effective tool, could be an additional parameter to assess disease activity in inflammatory bowel disease and an adjunctive marker in the differentiation between ulcerative colitis and Crohn's disease.

Effect of an Extract Based on the Medicinal Mushroom Agaricus blazei Murill on Expression of Cytokines and Calprotectin in Patients with Ulcerative Colitis and Crohn’s disease

An immunomodulatory extract (AndoSan™) based on the medicinal mushroom Agaricus blazei Murill (AbM) has shown to reduce blood cytokine levels in healthy volunteers after 12 days' ingestion, pointing to an anti-inflammatory effect. The aim was to study whether AndoSan™ had similar effects on cytokines in patients with ulcerative colitis (UC) and Crohn's disease (CD). Calprotectin, a marker for inflammatory bowel disease (IBD), was also measured. Patients with CD (n = 11) and with UC (n = 10) consumed 60 ml/day of AndoSan™. Patient blood plasma was harvested before and after 6 h LPS (1 ng/ml) stimulation ex vivo. Plasma and faecal calprotectin levels were analysed using ELISA and 17 cytokines [IL-2, IFN-γ, IL-12 (Th1), IL-4, IL-5, IL-13 (Th2), IL-7, IL-17, IL-1β, IL-6, TNF-α, IL-8, MIP-1β, MCP-1, G-CSF, GM-CSF and IL-10] by multiplex assay. After 12 days' ingestion of AndoSan™, baseline plasma cytokine levels in UC was reduced for MCP-1 (40%) and in LPS-stimulated blood for MIP-1β (78%), IL-6 (44%), IL-1β (41%), IL-8 (30%), G-CSF (29%), MCP-1 (18%) and GM-CSF (17%). There were corresponding reductions in CD: IL-2 (100%), IL-17 (55%) and IL-8 (29%) and for IL-1β (35%), MIP-1β (30%), MCP-1 (22%), IL-8 (18%), IL-17 (17%) and G-CSF (14%), respectively. Baseline concentrations for the 17 cytokines in the UC and CD patient groups were largely similar. Faecal calprotectin was reduced in the UC group. Ingestion of an AbM-based medicinal mushroom by patients with IBD resulted in interesting anti-inflammatory effects as demonstrated by declined levels of pathogenic cytokines in blood and calprotectin in faeces.

Serological markers for inflammatory bowel disease in AIDS patients with evidence of microbial translocation.

BackgroundBreakdown of the gut mucosal barrier during chronic HIV infection allows translocation of bacterial products such as lipopolysaccharides (LPS) from the gut into the circulation. Microbial translocation also occurs in inflammatory bowel disease (IBD). IBD serological markers are useful in the diagnosis of IBD and to differentiate between Crohn's disease (CD) and ulcerative colitis (UC). Here, we evaluate detection of IBD serological markers in HIV-infected patients with advanced disease and their relationship to HIV disease markers.MethodsIBD serological markers (ASCA, pANCA, anti-OmpC, and anti-CBir1) were measured by ELISA in plasma from AIDS patients (n = 26) with low CD4 counts (<300 cells/µl) and high plasma LPS levels, and results correlated with clinical data. For meta-analysis, relevant data were abstracted from 20 articles.ResultsIBD serological markers were detected in approximately 65% of AIDS patients with evidence of microbial translocation. An antibody pattern consistent with IBD was detected in 46%; of these, 75% had a CD-like pattern. Meta-analysis of data from 20 published studies on IBD serological markers in CD, UC, and non-IBD control subjects indicated that IBD serological markers are detected more frequently in AIDS patients than in non-IBD disease controls and healthy controls, but less frequently than in CD patients. There was no association between IBD serological markers and HIV disease markers (plasma viral load and CD4 counts) in the study cohort.ConclusionsIBD serological markers may provide a non-invasive approach to monitor HIV-related inflammatory gut disease. Further studies to investigate their clinical significance in HIV-infected individuals are warranted.

Functional analysis of agalactosyl IgG in inflammatory bowel disease patients

Agalactosyl immunoglobulin (Ig) G is increased in inflammatory bowel disease (IBD) similarly to rheumatoid arthritis (RA). The lectin complement pathway is shown to be activated through association of agalactosyl IgG with mannan-binding lectin (MBL) in RA. Functional changes of IgG agalactosylation in IBD, however, have not yet been clarified. The ratio of the agalactosyl/non-agalactosyl fraction in fucosylated IgG oligosaccharides (G0F/G2F) and serum MBL levels were analyzed in 59 patients with Crohn's disease (CD), 64 ulcerative colitis (UC), and 39 healthy volunteers (HV). The MBL levels associated with serum IgG were analyzed by enzyme-linked immunosorbent assay. MBL expression in the intestinal mucosa was analyzed by immunohistochemistry. Phagocytosis of sheep red blood cells (SRBC) reacted with either an agalactosyl or non-agalactosyl SRBC-specific IgG antibody was determined by flow cytometry. The serum MBL levels were not significantly different among CD, UC, or HV. In patients with CD, the serum MBL levels were negatively correlated with the Crohn's Disease Activity Index (CDAI). The levels of MBL associated with agalactosyl IgG were not different from those associated with non-agalactosyl IgG. Immunoreactivity to MBL was less in the inflamed mucosa compared with the noninflamed mucosa. Phagocytic activity of SRBC was significantly higher in the presence of agalactosyl IgG compared to non-agalactosyl IgG. Agalactosyl IgG oligosaccharides enhanced antibody-dependent phagocytosis in vitro but did not activate the lectin complement pathway. Oligosaccharide alterations of IgG are not only a marker of IBD but also functionally modulate the immune function of IBD.

Faecal calprotectin—a useful screening test for detecting gastrointestinal inflammation in children

<h3>Aims</h3> Faecal calprotectin is a neutrophil cytosolic protein which is validated as a reliable non-invasive marker of gastrointestinal inflammation in adults as well as children. The authors carried out an audit to assess the utility of faecal calprotectin in evaluation of children with chronic gastrointestinal symptoms. <h3>Methods</h3> Stool samples from 104 children aged 1–16 years who presented to a medium sized district general hospital with gastrointestinal symptoms were analysed using a commercially available kit. The normal range of faecal calprotectin is &lt; 50 mcg/g. <h3>Results</h3> Faecal calprotectin was outside the normal range in 37 out of 104 patients (36%). There were a total of 20 children with inflammatory bowel disease (14—newly diagnosed, 6—relapse) and all of them except one had elevated faecal calprotectin of greater than 200.The likelihood of organic bowel pathology was high when faecal calprotectin was greater than 200. Faecal calprotectin as a stool marker for inflammatory bowel disease (IBD) had an overall sensitivity of 95% and specificity of 79%. The specificity of the test in predicting IBD increased to 98.8% when calprotectin measured greater than 200. The positive predictive value was 95% and negative predictive value was 98% when faecal calprotectin value was &gt; 200. High faecal calprotectin did not co-relate well with other inflammatory markers in children diagnosed with inflammatory bowel disease. <h3>Conclusion</h3> Raised faecal calprotectin of greater than 200 should prompt urgent further assessment in children as IBD can be a likely diagnosis however these levels can be found in non-IBD organic GI disease. Faecal calprotectin can be used as a screening test reliably in evaluation of a child with chronic gastrointestinal symptoms and faecal calprotectin value outside the normal range of 50 mcg/g should involve prompt review to decide whether further investigations are needed.

Anticarbohydrate antibodies as markers of inflammatory bowel disease in a Central European cohort

The study discusses the role of antichitobioside carbohydrate antibody (ACCA), antilaminaribioside carbohydrate antibodies (ALCA), and antimannobioside carbohydrate antibodies (AMCA) in Central European patients with inflammatory bowel disease (IBD). Twohundred and seventy-two serum samples were used - 116 Crohn's disease (CD), 84 ulcerative colitis, and 72 healthy control samples. All samples were evaluated using enzyme-linked immunosorbent assay for the following four anticarbohydrate assays: ACCA, ALCA, AMCA, and anti-Saccharomyces cerevisiae antibodies (gASCA). gASCA antibodies showed the highest sensitivity (67%) for a CD diagnosis, followed by AMCA (31%), ACCA (27%), and ALCA (25%). Positivity of at least one of the four assays increased the overall sensitivity of antibody testing in CD up to 85.5%. Mean serum gASCA levels were significantly higher in CD patients who were younger at diagnosis and had a longer disease duration before blood sampling (P<0.001). In nonstricturing, nonpenetrating CD, serum gASCA levels were lower than in patients with stricturing and/or penetrating behavior (P<0.05). The strongest association of gASCA was found with ileocolonic CD and with upper gastrointestinal disease (P<0.001). No association between anticarbohydrate (AMCA, ACCA, and ALCA) antibodies and CD location, behavior, age at onset, and disease duration was found; however, that sample size of some of our subgroups was probably too small to make firm conclusions on associations with all CD phenotypes. None of the assessed anticarbohydrate assays was predictive of colonic CD in patients in whom the distinction between CD and ulcerative colitis is not obvious using routine diagnostic methods. There was no relationship between the presence or concentration of anticarbohydrate antibodies and the inflammation measured by C-reactive protein levels. The use of a panel of anticarbohydrate antibodies may provide additional help in distinguishing IBD from non-IBD disease patterns. The addition of AMCA, ALCA, and ACCA assays as IBD serology markers improves the overall sensitivity of immunological examinations in IBD; however, anticarbohydrate assays are not helpful for predicting CD behavior.

Oral manifestations of inflammatory bowel disease: a review based on the observation of six cases

Inflammatory bowel disease (IBD) comprises two chronic, tissue-destructive, clinical entities: Crohn's disease (CD) and ulcerative colitis (UC), both immunologically based. Bowel symptoms are predominant, but extra-intestinal complications may occur, including involvement of the oral cavity. Oral involvement during IBD includes several types of lesions: the most common are aphthae; uncommon lesions include, among others, pyostomatitis vegetans and granulomatous lesions of CD. Starting with a presentation of six patients with oral manifestations, which were crucial for the final diagnosis of IBD, a review on the subject is presented. Oral involvement in IBD may be previous or simultaneous to the gastrointestinal symptoms. However, in the majority of cases, bowel disease precedes the onset of oral lesions by months or years. In many patients, the intestinal symptoms may be minimal and can go undetected; thus, most authors believe that the bowel must be thoroughly examined in all patients with suspected IBD even in the absence of specific symptoms. Usually, the clinical course of oral lesions is parallel to the activity of IBD; therefore, oral manifestations are a good cutaneous marker of IBD.

Diagnostic Role and Clinical Association of ASCA and ANCA in Brazilian Patients with Inflammatory Bowel Disease

Anti-Saccharomyces cerevisae antibody (ASCA) and perinuclear anti-neutrophil cytoplasmatic antibody (pANCA) remain the most well-established markers in inflammatory bowel disease (IBD), and both may be associated with disease phenotype. To determine the utility of ASCA and pANCA as markers in a Brazilian cohort of IBD patients. A total of 90 patients with ulcerative colitis (UC), 77 patients with Crohn's disease (CD), and 57 healthy individuals were included in the study. ASCA was determined by enzyme-linked immunosorbent assay (ELISA) and pANCA by immunofluorescence assay. In support of diagnosis of UC, the sensitivity and specificity of pANCA were 51% and 100%, respectively. ASCA (IgA or IgG isotypes) presented sensitivity of 62% and specificity of 93% for CD. The combination of ASCA negativity and pANCA positivity (ASCA-/pANCA+) displayed sensitivity of 43% and specificity of 100% for diagnosis to UC. In CD patients, ASCA+/pANCA- presented sensitivity and specificity of 57% and 93%, respectively. Additionally, ASCA positivity correlated with early age at disease onset and ileal location in CD patients. In UC patients, pANCA positivity was correlated with pancolitis or left colitis. The results evidenced that low sensitivity of ASCA and pANCA markers limits their use in IBD screening in the general population; however, their specificity may contribute to differentiation between CD and UC in IBD patients. Our study lends further support to the suggestion that serologic assessment identifies different subtypes of IBD.

Characterization of single-nucleotide polymorphisms relevant to inflammatory bowel disease in commonly used gastrointestinal cell lines

The era of genome-wide association studies (GWAS) has led to the identification of many inflammatory bowel disease (IBD)-associated single-nucleotide polymorphisms (SNPs) with unknown function. The next step would be to identify the functional consequences of these polymorphisms in order to target them efficiently for therapeutic purposes. One way to study this type of genetic variation is the use of cell line models. However, to characterize the functional effect of a SNP, it is important to know if the selected cell line model itself carries the studied genetic variation. Here, we genotyped 50 IBD markers across 32 susceptibility genes in 9 commonly used gastrointestinal cell lines. We used Sequenom, TaqMan, and DNA sequencing for the genotyping. To determine the expression profile of the selected genes, we conducted real-time RT-PCR. We found variant SNPs in all analyzed cell lines. Almost every minor allele was carried by at least one of the tested cell lines. We analyzed the effect of 4 SNPs in more detail using quantitative real-time RT-PCR (qRT-PCR) comprising genes ATG16L1, CD14, MDR1, and OCTN2. According to our data, only 2 of the commonly studied SNPs in MDR1 and CD14 have an impact on gene expression. We have identified genotype variants in all analyzed cell lines. Some of them are functional and alter the response to drugs (MDR1) or affect bacterial recognition (TLR4, NOD2). Our results highlight that the genotype should not be neglected in experimental design when using model cell lines.

Rhodanese, but not cystathionine-γ-lyase, is associated with dextran sulfate sodium-evoked colitis in mice: A sign of impaired colonic sulfide detoxification?

Clinical studies suggest that colonic luminal hydrogen sulfide (H 2S), produced by sulfate-reducing bacteria or through other pathways, might be involved in the pathogenesis of inflammatory bowel disease (IBD). Nonetheless, this hypothesis has been poorly investigated by basic studies using laboratory animals. We thus focused on two enzymes, cystathionine-γ-lyase (CSE) that generates H 2S from l-cysteine, and rhodanese that directly or indirectly detoxifies H 2S, particularly in relation to the colitis induced by dextran sulfate sodium (DSS) in mice. CSE was a major H 2S-forming enzyme in colonic and renal homogenates from mice and rats, and the rhodanese activity was also detectable in both tissues. Colitis-related symptoms including decreased body weight gain, diarrhea, hematochezia and shortening of colon length were observed in the mice drinking DSS. Those symptoms were not or only slightly attenuated by repeated administration of a CSE inhibitor. CSE activity and protein levels in the colonic tissue did not notably change in the mice with colitis. In contrast, the activity and protein/mRNA levels of rhodanese in the colon, but not kidney, significantly decreased nearly in parallel with the development of colitis, followed by elevation of rhodanese activity in red blood cells (RBCs). These data show that rhodanese, but not CSE, is associated with DSS-induced colitis in mice, leading to a hypothesis that impaired detoxification of H 2S due to down-regulation or suppression of colonic rhodanese is involved in IBD. The delayed enhancement of rhodanese activity in RBCs, a possible compensative event, might be available as a disease marker for IBD.

Biological markers in inflammatory bowel disease: Practical consideration for clinicians

The biomarkers are important in the Inflammatory Bowel Disease (IBD) to gain an objective measurement of disease activity and severity, as well as prognostic indicator and outcome of therapy. And they can be helpful to avoid invasive procedures. The ideal biomarker does not exist for IBD and it is likely that more than one biomarker will be needed. Biological markers potentially useful in IBD include acute-phase proteins, fecal markers, several antibodies and novel genetic determinants. The C-reactive protein (CRP) is the most studied and has been shown to be an objective marker of inflammation. CRP is a good marker of measuring disease activity in Crohn's disease (CD) and its levels can be used to guide therapy. The fecal markers (calprotectin and lactoferrin) may be helpful in differentiating patients with IBD from those with functional disorders and to predict clinical relapse. The panel of serologic markers (anti-Saccharomyces cerevisiae antibody, perinuclear anti-neutrophil cytoplasmic antibody, anti-OmpC and anti-I2 and antiglycan antibodies) for IBD can be used to stratify IBD patients into more homogeneous subgroups with respect to disease progression. Correlating serologic markers with genotypes and clinical phenotypes should enhance our understanding of the pathophysiology of IBD. The development of biomarkers in IBD will be very important in the future with the increasing utilization of novel methodological approaches like genomics and proteomics.

S1660 Deficiency of Galactosyltransferase, Indispensable Enzyme for IgG Galactosylation, Ameliorates Murine Dextran Sodium Sulfate (DSS)-Induced Colitis

Background and aims: Immunoglobulin (Ig) G carries N-linked oligosaccharides at the Cgamma2 domain.We have reported that agalactosyl fraction in the fucosylated IgG oligosaccharides was increased in inflammatory bowel disease (IBD) and that agalactosyl IgG is an effective serological marker in IBD, especially in Crohns disease (CD) (Am J Gastroenterol. 2008). The mRNA level and activity of beta-1,4-galactosyltransferase I (Beta4GalT I), which is indispensable for attaching galactose to IgG oligosaccharides, were significantly lower in B cells and plasma cells of patients with CD than in those of ulcerative colitis patients. To clarify the pathophysiology of agalactosyl IgG elevation in IBD, we investigated the role of oligosaccharide alterations in colitis by using Beta4GalT I deficient mice. Methods: Beta4GalT I hetero-type mice (HT) and littermate wild-type mice (WT) were used because Beta4GalT I null mice were semi-lethal and Beta4GalT I activity in HT was reported to be half in WT. Both HT and WT were administered with 3% dextran sodium sulfate (DSS; MW 3500050000) ad libitum for 7 days. Solution was replaced to normal water for 3 days and mice were sacrificed at day 10. The severity of colitis was evaluated by the colon length, the body weight loss and the histological examination. CD4+, CD11b+ and CD19+ cells were separated from mononuclear cells from the spleen, mesenteric lymph nodes and lamina propria by MACS system. Cytokine productions in the culture supernatants of these cells were measured by ELISA. Results: Agalactosyl oligosaccharide structures of serum IgG were increased in Beta4GalT I HT compared to WT littermates and the IgG oligosaccharides in HT showed similar pattern as CD patients in terms of agalactosylation. When DSS colitis was induced in HT and WT, HT showed significantly less shortening in the colon length and less body weight loss than WT. Furthermore, histological score was significantly lower in HT than in WT. Thus, DSS-induced colitis was significantly ameliorated in HT compared to WT. Production of IL-10 in splenocytes of HT, especially in CD11b+ macrophages, was increased compared to that in WT. IgG oligosaccharide structures were not altered by DSS administration. Conclusions: Decrease in Beta4GalT I activity ameliorated murine DSS colitis. Agalactosyl IgG might not be the cause of colitis and is suggested to play a protective role for the colonic inflammation.

Two cases of pyodermatitis‐pyostomatitis vegetans

Pyodermatitis-pyostomatitis vegetans (PPV) is a rare chronic pustular and vegetating mucocutaneous dermatosis. The oral lesions present as multiple, friable and yellowish pustules, which form "snail tracts" and rupture easily. The cutaneous lesions begin as crusted erythematous papulopustules that coalesce to form large vegetating plaques, usually in the axillae, genital area and scalp. Cutaneous lesions usually develop at the same time as the oral lesions or thereafter. PPV is usually considered as a specific marker for inflammatory bowel disease since the concurrence of PPV and inflammatory bowel disease has been reported in approximately 70% of cases. We report two patients who showed typical clinical and histopathological features of PPV, but were not accompanied by inflammatory bowel disease.

Questions and answers on the role of fecal lactoferrin as a biological marker in inflammatory bowel disease

Among the available fecal biomarkers for the diagnosis and monitoring of inflammatory bowel disease (IBD), only calprotectin and lactoferrin have translated into useful clinical tools. Lactoferrin can be detected using simple and cheap techniques and it has excellent stability in feces over a long period of time. Fecal lactoferrin has a good diagnostic precision for separating organic and functional intestinal disease. However, a negative fecal lactoferrin test should be interpreted merely as the absence of significant neutrophilic intestinal inflammation. The mean sensitivity and specificity of the fecal lactoferrin determination for the diagnosis of IBD is 80% and 82%, respectively. Some studies have suggested a lower accuracy of lactoferrin when compared with calprotectin for the diagnosis of IBD, indicating that more studies on this topic are necessary. A parallel between fecal lactoferrin levels and IBD activity estimated with clinical, endoscopic, and histological parameters has been confirmed. However, this correlation seems to be lower in Crohn's disease than in ulcerative colitis, mainly when Crohn's disease patients with purely ileal disease are considered. Fecal lactoferrin determination may be useful in predicting impending clinical relapse in IBD patients. Fecal lactoferrin may be a helpful noninvasive diagnostic tool for monitoring therapeutic efficacy, mainly on mucosal healing, as a decreasing concentration of lactoferrin can be interpreted as a marker of therapeutic response. Finally, in patients with Crohn's disease who have undergone ileocolonic resection, those with higher lactoferrin fecal levels might be more prone to postsurgical recurrence.