Levels In Crohn's Disease Patients Research Articles

Gut Microbiome-Generated Phenylacetylglutamine from Dietary Protein is Associated with Crohn's Disease and Exacerbates Colitis in Mouse Model Possibly via Platelet Activation.

Our aims were to better understand the interplay of diet and gut microbiota in Crohn's disease [CD], taking advantage of a new-onset treatment-naïve CD cohort. We focus on phenylacetylglutamine [PAGln], a diet-derived meta-organismal prothrombotic metabolite. We collected faecal and serum samples from a CD cohort [n = 136] and healthy controls [n = 126] prior to treatment, and quantified serum PAGln using LC-MS/MS. Diet was assessed using food-frequency questionnaires. Mice [C57BL/6] were fed high/low-protein diets and administered dextran sodium sulphate [DSS] to examine plasma PAGly, thrombosis potential, and colitis severity. PAGly or saline was administered to DSS-induced colitis mice, and colitis severity and colonic tissue gene expression were examined. P-selectin and CD40L expression were determined in human platelet-rich plasma [n = 5-6] after exposure to platelet agonists following PAGln priming. Bioinformatic analysis and bacterial culturing identified the main contributor of PAGln in CD. PAGln, a meta-organismal prothrombotic metabolite, is associated with CD. Administration of PAGly exacerbated colitis in a mouse model and upregulated coagulation-related biological processes. Antiplatelet medicine, dipyridamole, attenuated PAGly-enhanced colitis susceptibility. PAGln enhanced platelet activation and CD40L expression in platelet-rich plasma ex vivo. Further study revealed that high dietary protein intake and increased abundance of phenylacetic acid [PAA]-producing Proteobacteria mediated by phenylpyruvate decarboxylase act in concert to cause the elevated PAGln levels in CD patients. Taken together, ppdc-carrying Proteobacteria-generated PAGln from dietary protein is associated with CD and exacerbates colitis possibly via platelet-induced coagulation and inflammation These results suggest that PAGln is a potential early diagnostic marker and therapeutic target of CD.

Correlation of serum lipid profile and disease activity in patients with inflammatory bowel disease

To analyze the correlation between lipid profile and disease activity in patients with inflammatory bowel disease (IBD).A total of 307 Crohn's disease (CD) patients, 232 ulcerative colitis (UC) patients and 165 healthy subjects from the same geographic region were included. Total cholesterol (TC), triglyceride (TG), high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C) and lipoprotein(a)[Lp(a)] were retrieved from their medical records. Crohn disease activity index (CDAI) and Mayo scores were calculated as measurement of disease severity for CD and UC separately. Patients with CD and UC had lower TC, TG, HDL-C and LDL-C levels than those in control group (P<0.05). Additionally, CDAI was negatively associated with TC, HDL-C and LDL-C levels (r=-0.218, -0.210, -0.176, P<0.05), while TG level was not associated with CDAI. Mayo scores was not significantly associated with TC, HDL-C, LDL-C and TG. Patients with CD had higher Lp(a) levels than those in UC and control group (P<0.05). Furthermore, patients with active CD had higher Lp (a) levels than those with inactive disease (P<0.05).The Lp(a) levels in CD patients were positively associated with CDAI (r=0.151, P<0.05), while Lp(a) level in UC group was nor assocriated with Mayo score. Patients with IBD have dyslipidemia and lipid profile is associated with disease activity in CD patients.

Association of serum interleukin-6 and soluble interleukin-2-receptor levels with disease activity status in patients with inflammatory bowel disease: A prospective observational study.

Crohn’s disease (CD) and ulcerative colitis (UC) are characterized by overexpression of proinflammatory cytokines. We determined the association of serum levels of interleukin (IL)-6, soluble-IL-2-receptor (sIL-2R) and CRP as well as of faecal calprotectin (FC) values with disease activity in CD and UC patients. This prospective study included 145 CD and 84 UC patients. Serum proinflammatory biomarkers and FC levels were measured and demographic, clinical and endoscopic characteristics were collected. Uni- and multivariate statistical analyses were performed. Serum IL-6 and CRP levels as well as FC values of CD patients were associated with clinical and endoscopic remission. In multivariate analysis serum IL-6 levels remained significantly associated with clinical and endoscopic remission. FC levels were also associated with endoscopic remission in CD patients. CD patients under the threshold levels of 8.5 pg/mL and 5.5 pg/mL for serum IL-6 were in 70% and 66% in clinical and endoscopic remission, respectively. Serum sIL-2R, CRP levels and FC values of UC patients were associated in univariate analysis with clinical and endoscopic remission. In multivariate analysis CRP and FC values were associated with clinical remission and serum sIL-2R as well as FC levels with endoscopic remission. UC patients under the threshold levels of 759 IU/mL and 646 IU/mL for serum sIL-2R were in 76% and 76% in clinical and endoscopic remission, respectively. Beside CRP and FC, serum IL-6 levels in CD patients and sIL-2R levels in UC patients can be a further useful non-invasive biomarker to identify the disease activity status.

Novel mass spectrometry‐based comprehensive lipidomic analysis of plasma from patients with inflammatory bowel disease

Lipids play important roles in inflammation and may be involved in the pathophysiology of inflammatory bowel disease (IBD). Here, we evaluated the characteristics of the plasma lipid profile in patients with IBD. Plasma samples were collected from 20 patients with Crohn's disease (CD), 20 patients with ulcerative colitis (UC), and 10 healthy volunteers (HVs) after overnight fasting. The subjects were men between 20 and 49years of age with no history of hyperlipidemia. A total of 698 molecular species in 22 lipid classes were analyzed by ultra-performance liquid chromatography-electrospray ionization-tandem mass spectrometry. Lipid classes of lysophosphatidic acid, lysophosphatidylserine (LPS), phosphatidylserine (PS), and shingosine-1-phosphate (S1P) were significantly increased in UC patients compared with the HV. The LPS, PS, and S1P levels were significantly increased, while those of lysophosphatidylinositol and phosphatidylcholine were significantly decreased in CD patients compared with HV. Among PS species, the levels of PSacyl (PSa) 40:3, PSa 38:3, and PSa 42:4 were significantly higher in CD patients, both active and remissive stage, than in HV. The LPS 18:0 level was significantly higher in CD and UC patients compared with HV. PSa 40:3 and PSa 38:3 levels positively correlated with the Crohn's Disease Activity Index, erythrocyte sedimentation rate, and platelet count and negatively correlated with hemoglobin, hematocrit, and albumin levels in CD patients. The lipid profile in IBD patients exhibits significant alterations, and PS levels are associated with clinical disease activity in CD patients.

Low serum bilirubin, albumin, and uric acid levels in patients with Crohn's disease.

Serum concentrations of bilirubin, albumin, and uric acid (UA) play important roles in controlling oxidative stress. Until now, there are few researches related to the relationship between oxidative stress and Crohn's disease (CD); furthermore, no such study has been reported from China. Our aim was to evaluate serum bilirubin, albumin, and UA levels in CD patients and relate them to disease activity.Seventy-one patients diagnosed with CD and 125 sex- and age-matched healthy individuals were retrospectively analyzed during the same period. Clinical characteristics and laboratory parameters were analyzed in CD patients and healthy control groups.Serum levels of bilirubin, albumin, and UA in patients with CD were significantly lower than those in the healthy control group. Correlation analysis demonstrated that serum concentrations of total bilirubin, direct bilirubin, indirect bilirubin, albumin, and UA were negatively related to disease activity in patients with CD (r = −0.620, P < .001; r = −0.304, P < .05; r = −0.623, P < .001; r = −0.408, P < .01; and r = −0.296, P < .05; respectively).Serum bilirubin, albumin and UA levels were significantly lower in CD patients, suggesting potential correlations between serum bilirubin, albumin, and UA levels and disease activity in CD patients. In addition, the noninvasive biochemical index may be potential markers for assessing the disease activity of patients with CD.

Serum Serotonin/Tryptophan Ratio Correlates with Inflammation in Crohn's Disease Patients

Inflammatory bowel diseases (IBD), which primarily consist of ulcerative colitis and Crohn's disease (CD) are characterized by persistent inflammation of the gastrointestinal (GI) tract. The current standard for diagnosis of IBD utilizes invasive methods including endoscopy and tissue biopsy, with blood tests being less specific for IBD. Substantial evidence has implicated involvement of the neurohormone serotonin (5‐hydroxytryptamine, 5‐HT) in the pathophysiology of IBD. Both mRNA and protein levels of serotonin transporter (SERT) have been consistently shown to be decreased in the intestinal mucosa of IBD patients. However, there is disagreement between previous studies on whether mucosal 5‐HT is increased or decreased in IBD. Furthermore, serum levels, rather than tissue levels, of 5‐HT in patients with IBD have not yet been examined. The current study examined 5‐HT, tryptophan (TRP), and kynurenine (KYN) levels in CD patients to test the hypothesis that 5‐HT serum levels correlate with inflammation.MethodsSerum samples were obtained from a German cohort of 45 CD patients and were categorized into one of three groups based upon their CD activity index (CDAI) and disease history. 15 patients were in remission (CDAI &lt; 150), 15 patients had active disease (CDAI &gt; 450), and 15 patients had chronic disease characterized by persistence of clinical symptoms and no lasting remission after adequate drug therapy. Age, gender, and race were not significantly different between the three groups. LC‐MS was used to measure 5‐HT, TRP, and KYN levels in the serum samples and Luminex Multiplex ELISA was used to measure cytokine levels. Results are shown as 95% confidence intervals of the mean.ResultsTRP and its derivatives 5‐HT and KYN were measured in CD patient serum: 5‐HT (0.65 – 0.93 μM), TRP (50.93 – 59.67 μM), and KYN (1.94 – 2.55 μM). To normalize for differences in body TRP levels, data were calculated as a ratio between 5‐HT and TRP (5‐HT/TRP) and between KYN and TRP (KYN/TRP). There were no significant differences in 5‐HT, TRP, and KYN levels between remission, active, and chronic groups. However, the 5‐HT/TRP ratio was significantly elevated in CD patients with active disease (0.014 – 0.034, p &lt; 0.01) as compared either to patients in remission (0.009 – 0.017) or with chronic disease (0.009 – 0.018) with no difference in KYN/TRP ratio. Further, the 5‐HT/TRP ratio positively correlated with serum C‐reactive protein levels (r = 0.47, p &lt; 0.01), serum IL‐1β levels (r = 0.52, p &lt; 0.01), and serum TNF‐α (r = 0.36, p &lt; 0.05). In conclusion, we have shown that the serum 5‐HT/TRP ratio can discriminate between active disease and chronic disease or remission among CD patients. In addition, the 5‐HT/TRP ratio correlates with established serum markers of inflammation in CD patients, emphasizing the important role that 5‐HT plays in the pathophysiology of IBD.Support or Funding InformationNIDDK NIH R01 DK098170, NIDDK NIH F30 DK113703This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Significance of measurement of serum trough level and anti-drug antibody of adalimumab as personalised pharmacokinetics in patients with Crohn's disease: asubanalysis of the DIAMOND trial.

SummaryBackgroundSignificance of monitoring adalimumab trough levels and anti‐adalimumab antibodies (AAA) for disease outcome in Crohn's disease (CD) patients remained unclear.AimTo evaluate the association of adalimumab trough levels and AAA at week 26 with clinical remission at week 52, the effect of azathiopurine on AAA and factors influencing trough levels in CD patients in the DIAMOND trial.MethodsWe performed this study using adalimumab trough levels, AAA at week 26 and 6‐thioguanine nucleotide (TGN) in red blood cells at week 12. A multiple regression model and receiver operating analysis was performed to identify factors influencing adalimumab trough levels and AAA, and adalimumab thresholds for predicting disease activity.ResultsThere was a significant difference of adalimumab trough level at week 26 between patients with disease remission and without at week 52 (7.7 ± 3.3 μg/mL vs 5.4 ± 4.3 μg/mL: P <.001). Adalimumab trough level of 5.0 μg/mL yielded optimal sensitivity and specificity for remission prediction (80.2% and 55.6%, respectively). AAA development at week 26 significantly affected remission at week 52 (P = .021), which was strongly associated with adalimumab trough levels. Female gender and increasing body weight were independently associated with low adalimumab trough levels, and female gender was associated with AAA development. A cut‐off 6TGN level of >222.5 p mol/8 ×108 RBCs yielded sensitivity (100%) and specificity (60.6%) for AAA negativity.ConclusionAdalimumab trough levels and AAA occurrence were significantly associated with clinical remission. Higher 6TGN affected AAA negativity. The combination therapy is beneficial in some relevant aspects for CD patients. (UMIN Registration No. 000005146)

Status of serum vitamin B12 and folate in patients with inflammatory bowel disease in China.

Background/AimsInflammatory bowel disease (IBD) primarily involves the intestinal tract and can affect vitamin absorption. This study was designed to assess the prevalence of vitamin B12 and folate deficiencies in patients with IBD, and to identify the risk factors associated with abnormal serum vitamin B12 and folate levels.MethodsWe evaluated the medical records of 195 patients with Crohn's disease (CD) and 62 patients with ulcerative colitis (UC), and selected 118 healthy subjects for the control group.ResultsThere were more CD patients with vitamin B12 deficiency than UC patients (14.9% vs. 3.2%, P=0.014) and controls (14.9% vs. 4.2%, P=0.003). The prevalence of folate deficiency was higher in CD patients than in controls (13.3% vs. 3.4%, P=0.004). There were no significant differences in the serum vitamin B12 and folate statuses of the UC and control groups. Patients with prior ileal or ileocolic resection showed a higher prevalence of abnormal vitamin B12 levels than those without prior resection (n=6/16, n=23/179; P=0.018). A disease duration within 5 years was a risk factor of abnormal folate levels in CD patients.ConclusionsThis study showed that vitamin B12 and folate deficiencies were more common in patients with CD than in UC patients and controls. Prior ileal or ileocolonic resection was a risk factor of serum vitamin B12 abnormalities, and a disease duration within 5 years was a risk factor of low serum folate levels in CD patients.

Bone Metabolism and the c.-223C>T Polymorphism in the 5'UTR Region of the Osteoprotegerin Gene in Patients with Inflammatory Bowel Disease.

Osteoporosis is more frequent in inflammatory bowel disease (IBD) patients. A reduction in bone mineral mass in these individuals is caused not only by inflammatory processes in the bowel, because osteoporosis occurs already in very young IBD patients and in newly diagnosed individuals who have not yet undergone any pharmacological treatment. One of individual determinants of the bone turnover parameters is osteoprotegerin (OPG) encoded by the TNFRSF11B gene. The c.-223C > T polymorphism in this gene has been extensively studied in post-menopausal osteoporosis patients. However, no such studies exist for osteoporosis related to IBD. The aim of our study was to determine whether the c.-223C > T (rs2073617) polymorphism in the 5′UTR region of the gene encoding osteoprotegerin is a functional polymorphism which may change the gene expression and resulting OPG levels, and so be associated with osteopenia and osteoporosis, and impaired bone metabolism in Crohn’s disease and ulcerative colitis patients. Our study included 198 IBD patients and 41 healthy controls. Lumbar spine and femoral neck bone mineral density, T-score, Z-score as well as OPG, RANKL, vitamin D, calcium and interleukin 4 and 10 concentrations were determined for all study subjects. Genotyping of the TNFRSF11B polymorphic site was performed by restriction fragment length polymorphism technique. Statistical analyses were conducted using Statistica software. Odds ratios, 95 % confidence intervals, and P values were calculated using the HWE calculator. Our results did not allow determining an unequivocal association between the polymorphic variants of the TNFRSF11B 5′UTR region and a susceptibility to osteoporosis in IBD patients. We have shown, however, that the c.-223T allele was twice as more frequent in Crohn’s disease (CD) patients than among controls (OR = 1.99, P value = 0.009). Interestingly, average osteoprotegerin levels in CD patients did not significantly differ from those in controls, whereas in ulcerative colitis patients, OPG levels were significantly lower. We have concluded that low OPG levels may be associated with osteoporosis in ulcerative colitis, but it is not correlated with the c.-223C > T polymorphism in the TNFRSF11B gene. In CD patients, in turn, we observed increased RANKL levels. Our observations confirm different pathogeneses of Crohn’s disease and ulcerative colitis as well as different molecular backgrounds of osteoporosis associated with these two diseases.

Chemerin, visfatin, and vaspin serum levels in relation to bone mineral density in patients with inflammatory bowel disease.

There is evidence that fat mass is correlated with bone mineral density (BMD) in inflammatory bowel disease (IBD), but data on the role of adipokines on this association are limited. The aim of this study was to investigate the serum levels of chemerin, visfatin, and vaspin, hormones that act as adipokines, in relation to BMD in patients with ulcerative colitis (UC) and Crohn's disease (CD). Serum from 120 IBD patients (68 CD, 52 UC) and 98 matched healthy controls (HC) was collected. Chemerin, visfatin, and vaspin levels were assessed using an enzyme-linked immunosorbent assay. BMD was determined for the lumbar spine and the proximal femur using dual-energy X-ray absorptiometry. Full-body composition scans were analyzed using enCORE software based on the absorptiometry system. Serum chemerin was higher in IBD patients than HC [CD 13.67.1±5.8, UC 13.9±4.3 vs. HC 7.8±2.6 ng/ml, odds ratio (OR): 0.95, 95% confidence interval (CI) 0.93-0.98, P<0.0001]. Serum visfatin levels in CD patients were significantly higher than those in UC patients (9.3±14.01 vs. 6.5±7.2 ng/ml, OR: 0.86, 95% CI 0.80-0.92, P=0.039). In multivariate logistic regression analysis, a significant independent association of osteoporosis (T-score ≤2.5 SD) with age (OR: 1.04, 95% CI 1.01-1.08, P=0.02), visfatin (OR: 0.78, 95% CI 0.63-0.97, P=0.02), and chemerin levels (OR: 0.83, 95% CI 0.70-0.98, P=0.03), but not with BMI or body composition, was found. Serum visfatin and chemerin levels are associated with the development of osteoporosis in IBD. These results suggest a role of visfatin and chemerin in the pathophysiology of osteoporosis in IBD.

Effects of vitamin D supplementation on intestinal permeability, cathelicidin and disease markers in Crohn's disease: Results from a randomised double-blind placebo-controlled study.

Vitamin D (vitD) supplementation may prolong remission in Crohn's disease (CD); however, the clinical efficacy and mechanisms are unclear. To determine changes in intestinal permeability (IP), antimicrobial peptide (AMP) concentrations and disease markers in CD, in response to vitD supplementation. In a double-blind randomised placebo-controlled study, we assigned 27 CD patients in remission to 2000 IU/day vitD or placebo for 3 mos. We determined IP, plasma cathelicidin (LL-37 in ng/mL), human-beta-defensin-2 (hBD2 in pg/mL), disease activity (Crohn's Disease Activity Index (CDAI)), C-reactive protein (CRP in mg/L), fecal calprotectin (µg/g), Quality of Life (QoL) and serum 25-hydroxyvitamin D (25(OH)D in nmol/L) at 0 and 3 mos. At 3 mos., 25(OH)D concentrations were significantly higher in those whom were treated (p < 0.001). Intra-group analysis showed increased LL-37 concentrations (p = 0.050) and maintenance of IP measures in the treated group. In contrast, in the placebo group, the small bowel (p = 0.018) and gastro-duodenal permeability (p = 0.030) increased from baseline. At 3 mos., patients with 25(OH)D ≥ 75 nmol/L had significantly lower CRP (p = 0.019), higher QoL (p = 0.037), higher LL-37 concentrations (p < 0.001) and non-significantly lower CDAI scores (p = 0.082), compared to those with levels <75 nmol/L. Short-term treatment with 2000 IU/day vitD significantly increased 25(OH)D levels in CD patients in remission and it was associated with increased LL-37 concentrations and maintenance of IP. Achieving 25(OH)D ≥ 75 nmol/l was accompanied by higher circulating LL-37, higher QoL scores and reduced CRP. Registered at ClinicalTrials.gov (NCT01792388).

P-106 Evaluation of the Association Between Fecal Lactoferrin Status and Clinical, Serologic and Endoscopic Outcomes

In the SONIC trial, patients with Crohn's disease (CD) treated with infliximab (IFX) alone or in combination with azathioprine achieved higher rates of mucosal healing (MH) at wk 26 than patients treated with azathioprine alone. In a post-hoc analysis of SONIC, clinical symptoms as measured by the Crohn's Disease Activity Index (CDAI) were inaccurate to detect MH1. Recently, fecal biomarkers, such as lactoferrin (LF) and calprotectin, are being increasingly used in monitoring the activity of CD. This post-hoc analysis of SONIC reports on the association of fecal LF to clinical, serologic and endoscopic outcomes, and serum IFX levels in CD patients enrolled in centers in the United States who had evaluable measurements of LF, CDAI, serum CRP, and ileocolonoscopy both at baseline and wk 26, and evaluable serum IFX levels at wk 30. LF was measured by an ELISA assay and expressed as positive (pooled +2, +3, and +4 scores) or negative (pooled negative and +1 scores). MH was defined as absence of any ulceration. Corticosteroid-Free Clinical Remission (CFCR) was defined as clinical remission (CDAI <150) and no treatment with corticosteroids for >6 weeks. Post-hoc analyses were performed using the Fisher's exact test and the Kruskal-Wallis test where appropriate. Among the 109 patients (median age 43 yrs, 54% male), 33.9% were LF+ and 19.3% were LF- at both baseline and wk26; 36.7% patients changed from LF+ to LF- from baseline at wk 26 whereas 10.1% changed from LF- to LF+ (P < 0.0001). At baseline, LF was significantly associated with CDAI (P = 0.024) and serum CRP (P = 0.01) (Table 1); in addition, LF showed a trend of association with normal levels of CRP (65.6% versus 36.4% with CRP <0.8 mg/dL for LF—and LF+, respectively, P = 0.064, Table 1). At wk 26, there was no association between LF and CDAI (P = 0.09), serum CRP (P = 0.18), CRP normalization (0.512), CFCR (P = 0.245) (Table 1). The association between LF and MH (ulceration) was evaluated in 168 patients at baseline and 110 patients at wk 26 who had both LF and MH data at these measurement points. At baseline, ulcerations were not found in 43.1% (25/58) LF- patients versus 21.8% (24/110) LF+ patients (sensitivity 0.72, specificity 0.51, P = 0.004). In contrast, at wk26, 44.3% (27/61) of LF- patients versus 30.6% (15/49) LF+ patients had MH (sensitivity 0.50, specificity 0.64, P = 0.143). In this post-hoc analysis of the SONIC trial, LF was collected as a categorical variable. The association between LF and other clinical parameters may be better estimated if LF was on a continuous scale. Semi-quantitative LF does not seem to be a good biomarker in CD. Although it was useful to identify active clinical and biological disease at baseline, it could not confirm clinical, biological, or endoscopic remission at wk26. 1. L Peyrin-Biroulet, WJ Sandborn, et al. Clinical disease activity, C-reactive protein normalization and mucosal healing in Crohn's disease in the SONIC trial. Gut. 2013;63:88–95.

Vitamin D deficiency in Crohn's disease and healthy controls: A prospective case–control study in the Netherlands

Background and aimsVitamin D deficiency has been observed in a wide range of medical conditions including Crohn's disease (CD). We aimed to assess whether CD patients have lower vitamin D levels than healthy controls, and to determine risk factors for vitamin D deficiency. Methods25(OH)D was measured by chemiluminescent immunoassay in serum obtained from 101 CD patients and 41 controls. Demographics, sunlight exposure, dietary vitamin D intake, comorbidities and medication were recorded using validated questionnaires. In CD patients the Harvey–Bradshaw index, Montreal classification and surgical resections were also evaluated. 25(OH)D levels of >75nmol/L, between 50 and 75nmol/L and <50nmol/L were considered as normal, suboptimal and deficient, respectively. ResultsVitamin D levels were rather low but comparable among CD patients and controls (mean 25(OH)D 51.6nmol/L(±26.6) in CD, and 60.8nmol/L(±27.6) in controls. Multivariate regression analysis revealed BMI, sun protection behaviour, non-Caucasian ethnicity, no use of tanning beds, and no holidays in the last year as significantly associated with serum 25(OH)D levels in CD patients (R=0.62). In the control group no statistically significant factors were identified that had an impact on 25(OH)D serum levels. ConclusionsVitamin D deficiency is common in CD patients, but also in healthy controls. Appropriate vitamin D screening should be advised in patients with CD. Moreover, the positive effect of sunlight on the vitamin D status should be discussed with CD patients, but this should be balanced against the potential risk of developing melanomas, especially in patients using thiopurines.

Serum concentrations of insulin-like growth factor-binding protein 5 in Crohn’s disease

To investigate serum insulin-like growth factor-binding protein 5 (IGFBP-5) levels and intestinal IGFBP-5 expression in patients with Crohn's disease (CD). We analyzed the serum concentrations and intestinal expression of IGFBP-5 in 42 patients with CD, of whom 26 had endoscopically or radiologically proven stricture formation. Nine of the 42 patients had active disease, with a Crohn's disease activity index > 150. Serum IGFBP-5 levels were analyzed in 20 healthy controls matched by sex and age to the CD patients. Serum IGFBP-5 was measured using an enzyme-linked immunosorbent assay. Intestinal tissue was obtained from patients through endoscopic biopsies. IGFBP-5 expression was detected using immunohistochemistry and was scored semiquantitatively. The median serum IGFBP-5 concentrations of CD patients were significantly lower compared with healthy controls [median 7.2 (IQR: 5.5-11.3) ng/mL vs 11.3 (8.0-44.6) ng/mL, P < 0.001]. There was no significant difference between median serum IGFBP-5 levels in CD patients with or without stricture formation [6.9 (5.5-11.3) ng/mL vs 7.8 (5.3-10.1) ng/mL, P = 0.815]. The serum IGFBP-5 levels were not significantly different between patients with active disease and inactive disease [7.2 (6.5-7.6) ng/mL vs 7.2 (5.5-11.3) ng/mL, P = 0.890]. However, a significant correlation was observed between serum IGFBP-5 levels and platelet count (PLT) (r = 0.319, P = 0.0395). No significant correlation was found between tissue IGFBP-5 immunohistochemical staining intensity scores and serum IGFBP-5 levels. No significant difference was found when comparing the serum IGFBP-5 levels among the patients with different tissue IGFBP-5 staining scores (absent/very weak, weak, moderate or strong). There was a significant correlation between tissue IGFBP-5 staining scores and white blood cell count (r = 0.391, P = 0.01) and PLT (r = 0.356, P = 0.021). Our results indicate that serum IGFBP-5 concentrations were lower in CD patients compared to healthy controls regardless of disease activity or the presence of stricture formation.

Enzymes involved in l-carnitine biosynthesis are expressed by small intestinal enterocytes in mice: Implications for gut health

BackgroundCarnitine is essential for mitochondrial β-oxidation of long-chain fatty acids. Deficiency of carnitine leads to severe gut atrophy, ulceration and inflammation in animal models of carnitine deficiency. Genetic studies in large populations have linked mutations in the carnitine transporters OCTN1 and OCTN2 with Crohn's disease (CD), while other studies at the same time have failed to show a similar association and report normal serum carnitine levels in CD patients. MethodsIn this report, we have studied the expression of carnitine-synthesizing enzymes in intestinal epithelial cells to determine the capability of these cells to synthesize carnitine de novo. We studied expression of five enzymes involved in carnitine biosynthesis, namely 6-N-trimethyllysine dioxygenase (TMLD), 4-trimethylaminobutyraldehyde dehydrogenase (TMABADH), serine hydroxymethyltransferase 1 and 2 (SHMT1 and 2) and γ-butyrobetaine hydroxylase (BBH) by real-time PCR in mice (C3H strain). We also measured activity of γ-BBH in the intestine using an ex vivo assay and localized its expression by in situ hybridization. ResultsOur investigations show that mouse intestinal epithelium expresses all five enzymes required for de novo carnitine biosynthesis; the expression is localized mainly in villous surface epithelial cells throughout the intestine. The final rate-limiting enzyme γ-BBH is highly active in the small intestine; its activity was 9.7±3.5pmol/mg/min, compared to 22.7±7.3pmol/mg/min in the liver. ConclusionsWe conclude that mouse gut epithelium is able to synthesize carnitine de novo. This capacity to synthesize carnitine in the intestine may play an important role in gut health and can help explain lack of clinical carnitine deficiency signs in subjects with mutations with OCTN transporters.

PTU-106 A prospective single centre evaluation of the intra-individual variability of faecal calprotectin in quiescent Crohn's disease patients

IntroductionFaecal calprotectin (FC) has become an established non-invasive, sensitive marker of intestinal inflammation in Crohn's disease (CD). A single measurement of FC is used in clinical practice to aid decision...

The usefulness of factor XIII levels in Crohn's disease

Background and AimsThe assessment of inflammatory activity in Crohn's disease (CD) is challenging, and no specific laboratory marker is currently available. Several studies have reported decreased serum factor XIII levels in CD patients as a function of disease activity. We aimed to determine whether the factor XIII level could be a marker for the evolution of CD. MethodsIn this prospective, single-centre trial, 129 patients were included and categorised into two groups: functional bowel disorders (FBDs, n=42) and CD (n=86). The CD group was divided into two subgroups depending on disease activity, as defined by the Crohn's Disease Activity Index score: active disease (CDa, n=41) and disease remission (CDb, n=45). The factor XIII levels were evaluated for each patient. Serial factor XIII levels were evaluated in the patients within the CDa subgroup. ResultsThe factor XIII levels were significantly different between the FBD (117.69%) and CD (101.89%) groups (p=0.009) but there was no significant difference between the CDa and CDb subgroups (99.04% vs 104.65%, p>0.05), and the levels did not vary during follow-up for the patients in the CDa subgroup. By multivariate analysis, factor XIII levels did not correlate with the time course of disease evolution, CRP, serum fibrin levels, platelet count, disease distribution within the bowel, or the presence of a fistulising form of CD. ConclusionsOur results confirm that factor XIII levels are decreased in CD patients but cannot be recommended as a marker for the disease activity.

Serial Changes of Cytokines in Crohn's Disease Treated with Infliximab

Infliximab (IFX) has been reported to be useful as induction therapy and/or maintenance therapy for Crohn's disease (CD). However, the effects of IFX on serial changes in cytokine levels have not been well characterized. We examined cytokine levels in CD patients before and after administration of IFX. A total of 37 patients with active CD were enrolled (24 men, 13 women; mean age, 31.9 years). Patients were given IFX 5mg/kg intravenously. Serum levels of 17 cytokines were simultaneously determined using a Bio-Plex suspension array system before treatment and, 4 and 8 weeks after IFX treatment. Patients were divided into two groups according to the disease duration: the 'early treatment' group and the 'in progress' treatment group. Serum IL-6, IL-7, IL-8 and MIP-1ß levels were significantly decreased in CD patients after IFX treatment compared to before treatment (p<0.05). In particular, serum levels of IL-8, IL-12 and MIP-1ß decreased significantly after IFX treatment in the 'in progress' treatment group. The changes in CD activity score (CDAI) after treatment were positively correlated with the changes of serum MIP-1ß. IFX treatment reduces serum levels of IL-6, IL-7, IL-8, IL-12 and MIP-1ß, which appears to be mediated by the inhibition of inflammation in CD.

Deficit of Gammadelta T Lymphocytes in the Peripheral Blood of Patients with Crohn’s Disease

Gammadelta T lymphocytes are an important component of innate immunity. Previous studies have shown their role in the development of Crohn's-like colitis in mice. The aim of this study was to measure the γδ T lymphocyte levels in Crohn's disease (CD) patients. A prospective study of 40 patients with CD compared with 40 healthy subjects (control group) matched by age and sex was undertaken. Lennard-Jones criteria were used for the diagnosis of CD. Disease activity was measured with the Crohn's disease activity index (CDAI). New patients, patients in remission, and patients with active disease were evaluated. Lymphocytic populations of CD3+, CD4+, CD8+, CD56+, CD19+, and αβ and γδ subsets were measured in the peripheral blood of all participants. The levels of CD3+, CD4+, CD8+, and CD19+ lymphocytes were decreased in CD patients compared with the control group (P = 0.002, 0.049, 0.003, and 0.023, respectively). Although both γδ and αβ T lymphocytes were lower in patients with CD, γδ T subsets showed the lowest levels in CD patients (mean 0.0259 × 10(9)/l) versus healthy controls (mean 0.0769 × 10(9)/l), P < 0.001. In particular, γδ CD8+ T subsets (mean 0.0068 × 10(9)/l) had the largest difference compared to the control group (mean 0.0199 × 10(9)/l), P = 0.008. There is a decrease in the global lymphocyte population in the peripheral blood of patients with CD compared to healthy controls. This decrease is more evident in γδ T lymphocytes, especially γδ CD8+ T subsets. Our conclusion is that these results support the theory that a complex alteration of immune responses that affects the total numbers and function of γδ T cells is present in CD.

The functional −374T/A polymorphism of the receptor for advanced glycation end products may modulate Crohn's disease

The receptor for advanced glycation end products (RAGE) is involved in innate immune mechanisms. Polymorphisms of the RAGE gene have been described as a factor amplifying inflammation in susceptible patients, but the association with Crohn's disease (CD) is not known. The coding RAGE polymorphism G82S (rs2070600) and two promoter polymorphisms, -374T/A (rs1800624) and -429T/C (rs1800625), were studied in two samples from Germany and the United States consisting of 421 and 317 CD patients and 549 and 218 controls, respectively. To test the functional relevance, additional data on serum soluble RAGE (sRAGE), tissue RNA, and protein levels were collected and immunohistochemical stainings of bowel tissue of CD patients and healthy controls as well as models of experimental (dextran sodium sulfate-induced) colitis in RAGE knockout and wild-type mice were performed. The -374T/A RAGE promotor single nucleotide polymorphism (SNP) was negatively associated with CD (odds ratio = 0.708, 95% confidence interval = 0.535-0.938, P = 0.016) and with stenosis (OR = 0.627, P = 0.04) in the German sample. Transmission disequilibrium testing confirmed an undertransmission of the -374A allele. Serum sRAGE levels were higher in patients in complete remission of the -374AA/TA group (1,975 ± 299 pg/ml; -374TT group: 1,310 ± 153 pg/ml SE, P < 0.05) and showed a trend toward decreased levels in CD patients with active disease compared with CD patients in remission. Further in vitro and in vivo studies indicated that an increase of sRAGE ameliorates inflammation. The -429T/C and the G82S polymorphism were not associated with CD. The -374T/A RAGE polymorphism leading to facilitated RAGE gene transcription may to some degree protect from developing a stricturing subphenotype of CD, most likely by increasing levels of sRAGE, which neutralizes proinflammatory mediators.