Prolactin granulogenesis is associated with increased secretogranin expression and aggregation in the Golgi apparatus of GH4C1 cells.

The GH4C1 pituitary tumor cell line (GH cells) serves as a model system to study the role of the granins in the packaging of PRL into secretory granules. The number of secretory granules containing PRL and two members of the granin family, chromogranin-B (CgB) and secretogranin-II (SgII), can be hormonally manipulated. In the present study we have investigated whether 1) granulogenesis in GH cells is preceded by condensation of the granins and PRL in the Golgi; 2) granulogenesis is preceded by an increase in granin expression in GH cells; and 3) PRL and the granins aggregate in vitro under high calcium, low pH conditions. GH cells were treated for up to 3 days with 17 beta-estradiol (1 nM), insulin (300 nM), and epidermal growth factor (10 nM) and were fixed in 4% paraformaldehyde for immunocytochemistry or harvested for RNA isolation and Northern blot analysis. After 1 day of hormone treatment, there was a significant increase in staining for PRL and the granins in the Golgi apparatus, which was identified using an antibody to MG-160. After 3 days of hormone treatment, PRL and granin staining was also found in a perinuclear region that was not stained with anti-MG-160 antibody, most likely representing secretory granules. An increase in PRL and granin expression contributed to increased Golgi staining, as the steady state levels of CgB, SgII, and PRL mRNA increased 186 +/- 14%, 203 +/- 7%, and 337 +/- 5% above control levels, respectively, within 6 h after hormone treatment. An in vitro aggregation system was used to determine whether PRL and the granins coprecipitate under high calcium, low pH conditions, which are thought to be characteristic of the trans-Golgi and secretory granules. Aggregation of the granins CgB and SgII was negligible during overnight dialysis against a buffer containing 150 mM NaCl and 10 mM 2[N-morpholino]ethanesulfonic acid-NaOH (pH 5.5) in the absence of calcium. There was significant aggregation of PRL under these conditions. When dialysis was performed in the presence of 10 mM CaCl2, PRL, CgB, and SgII coaggregated. This study indicates that increased expression and aggregation of the granins is associated with PRL granulogenesis in hormone-treated GH cells. However, the role of the granins may not be obligatory, as some cells can store PRL in the absence of detectable levels of CgB and SgII, and PRL has the capacity to self-aggregate.     

Prolactin and secretogranin-II, a marker for the regulated pathway, are secreted in parallel by pituitary GH4C1 cells.

The granins are a family of tyrosine-sulfated secretory proteins. Two members of this family, chromogranin-B (CgB) and secretogranin-II (SgII), are found in GH4C1 cells, a pituitary cell line that secretes PRL and GH. We have compared the spontaneous and regulated secretion of CgB and SgII with that of PRL in GH4C1 cells and have assessed the importance of granin sulfation on granin and PRL processing and secretion. CgB and SgII were identified by metabolic labeling with [35S]SO4, which was predominantly incorporated into two bands of 105,000 (CgB) and 84,000 (SgII) mol wt. The secretion of [35S]SgII and [35S]PRL from GH4C1 cells simultaneously labeled with 35S-labeled SO4 and methionine showed similar kinetics over 60 min, suggesting that the two proteins are similarly processed. CgB, SgII, and PRL were released in parallel after 10-min treatment with secretagogues (high K+ and BAY K8644, 8-bromo-cAMP, a phorbol ester, and TRH). Hypertonicity and substitution of chloride with isethionate, which inhibit stimulated PRL release, reduced the amount of CgB and SgII released in response to secretagogues, but not basally. Cells were labeled with [35S]SO4 with or without 10 mM chlorate, which inhibits sulfation by more than 90%, and media and cells were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, autoradiography, and immunoblotting using an antibody directed against the N-terminus of SgII. Chlorate reduced [35S]SO4 labeling of CgB and SgII, but had little effect on immunoreactive SgII in cells or media. Inhibiting sulfation with chlorate did not change the amount of PRL or GH synthesized and secreted by GH4C1 cells, basally or in response to secretagogues, or the induction of PRL storage by insulin, estrogen, and epidermal growth factor. The results show that granins are released from GH4C1 cells in parallel with GH and PRL under basal and stimulated conditions, and that sulfation is not essential for normal packaging and processing of these secretory proteins. The data suggest a model in which PRL, CgB, and SgII are sorted to the regulated pathway and released from this pathway basally as well as under stimulated conditions.     

Fecal calprotectin and lactoferrin as predictors of relapse in patients with quiescent ulcerative colitis during maintenance therapy

Purpose

This prospective study was to evaluate the significance of fecal calprotectin and lactoferrin for the prediction of ulcerative colitis (UC) relapse.

Methods

Eighty UC patients in remission for ≥3 months on mesalamine as maintenance therapy were included. At entry, stool samples were collected for the measurement of calprotectin and lactoferrin. All patients were followed up for the following 12 months. To identify predictive factors for relapse, time-dependent analyses using the Kaplan-Meier graphs and Cox's proportional hazard model were applied.

Results

During the 12 months, 21 patients relapsed. Mean calprotectin and lactoferrin levels were significantly higher in patients with relapse than those in remission (calprotectin—173.7 vs 135.5 μg/g, P?=?0.02; lactoferrin—165.1 vs 130.7 μg/g, P?=?0.03). A cutoff value of 170 μg/g for calprotectin had a sensitivity of 76 % and a specificity of 76 % to predict relapse, while a cutoff value of 140 μg/g for lactoferrin had a sensitivity of 67 % and a specificity of 68 %. In a multivariate analysis, calprotectin (≥170 μg/g) was a predictor of relapse (hazard ratio, 7.23; P?=?0.002). None of the following parameters were significantly associated with relapse: age, gender, duration of UC, number of UC episode, severity of the previous episode, extent of UC, extraintestinal manifestation, and lactoferrin level.

Conclusions

Fecal calprotectin showed a higher sensitivity and specificity than fecal lactoferrin for predicting UC relapse. Fecal calprotectin level appeared to be a significant predictor of relapse in patients with quiescent UC on mesalamine as maintenance therapy.     

Surrogate markers of intestinal inflammation are predictive of relapse in patients with inflammatory bowel disease

BACKGROUND & AIMS: Prediction of relapse of inflammatory bowel disease has important implications for therapeutic strategies. We assessed whether measurement of intestinal permeability and inflammation could predict relapse of inflammatory bowel disease (IBD). METHODS: Forty-three patients with Crohn's disease (CD) and 37 with ulcerative colitis (UC) in clinical remission provided a stool sample to be assayed for calprotectin (a neutrophil-specific marker), and patients with CD additionally underwent a small intestinal permeability test. Relapse was defined using clinical disease activity indices. RESULTS: Twenty-five (58%) patients with CD and 19 (51%) with UC had a relapse over the 12-month period. Median calprotectin levels in the relapse groups (122 mg/L for CD, 123 mg/L for UC; normal <10 mg/L) differed significantly (P<0.0001) from those of the nonrelapse groups (41.5 mg/L for CD, 29.0 mg/L for UC). At 50 mg/L, the sensitivity and specificity of calprotectin for predicting relapse in all patients with IBD were 90% and 83%, respectively. Permeability in the CD patients who relapsed (median, 0.075; normal <0.04) differed significantly (P = 0. 004) from that in the nonrelapse group (median, 0.038). At the level of 0.05, the sensitivity and specificity of permeability in predicting relapse were 84% and 61%, respectively. CONCLUSIONS: Fecal calprotectin predicts clinical relapse of disease activity in patients with CD and UC, whereas small intestinal permeability is a useful predictor of relapse in patients with small intestinal CD.     

Characterisation of N-terminal chromogranin A and chromogranin B in mammals by region-specific radioimmunoassays and chromatographic separation methods

Chromogranin A (CgA) and chromogranin B (CgB) are acidic proteins stored in and released from hormone granules in endocrine and neuroendocrine tissue. The chromogranins are postulated to serve as pro-hormones to generate biologically active peptides, which may influence hormonal release and vascular functions or have antibacterial functions. Although N-terminal and C-terminal regions show some species amino acid homology, the chromogranins as a whole display considerable interspecies differences, which prevents their use in comparative studies of biological functions. We present four new radioimmunoassays for the measurement of defined N-terminal regions of CgA and CgB. A new radioimmunoassay for measurement of intact bovine CgA has also been developed. With these assays and two previously published ones, we have compared the cross-reactivity of chromogranins from man, cattle, sheep, goat, pig and horse and compared adrenomedullar content and serum levels of CgA from these species. We have also studied the influence of peptide concentrations and the ionic strength of the mobile phase on molecular weight estimations. Assays with antibodies directed against the N-terminal parts of CgA and CgB showed sufficient interspecies cross-reactivity to allow comparative quantification of the circulating levels in man, cattle, sheep, goat, pig and horse. Assays measuring the intact human or bovine CgA were not suitable for comparative purposes in samples from sheep, goat, pig and horse. Molecular interactions between vasostatin immunoreactive material and intact bovine CgA were demonstrated in gel permeation studies, suggesting that conclusions about the degree of N-terminal processing from elution profiles should be made with caution. Reliable interspecies comparison of chromogranins is difficult, but measurements with region-specific assays may be helpful to study concentrations of chromogranins and chromogranin-related peptides.     

Fecal Excretion of Calprotectin in Colorectal Cancer Relationship to Tumor Characteristics

Background: The aim of this study was to evaluate fecal calprotectin in patients treated for colorectal cancer. Furthermore, the changes in fecal calprotectin concentration from before to after surgery were investigated. Methods: In 155 patients with newly diagnosed colorectal cancer, two spot samples were taken from the same feces on two consecutive days. Results: Three ways of evaluating calprotectin excretion were compared, (1st spot 1st stool; maximum of 1st spot 1st stool and 2nd spot 1st stool; maximum of 1st spot 1st stool and 1st spot 2nd stool) and gave similar results with median fecal calprotectin values 47 mg/l, 52 mg/l and 54 mg/l, respectively. Median calprotectin concentration did not differ significantly between different tumor stages, although the levels were slightly lower in Dukes stage A tumor than in the rest of the stages. Neither were there any differences in the concentrations related to the localization, size or the histological grading of the carcinoma. As the currently used cut-off level for fecal calprotectin is 10 mg/l, 87% of all patients had elevated fecal calprotectin. Seventy-nine percent of the patients had levels above 15 mg/l and 74% had levels above 20 mg/l (1st spot 1st stool). In patients who delivered fecal samples after the operation the calprotectin value fell significantly from a preoperative median value of 45 mg/l to 14 mg/l after the resection. Conclusions: The majority of patients with colorectal cancer have increased fecal concentration of calprotectin. One single fecal spot seems to be sufficient for determination of the calprotectin level. Measurement of fecal calprotectin may possibly become of value as a marker for colorectal cancer, although calprotectin, similar to fecal occult blood (FOB) tests, is a non-specific test for colorectal pathology, also being elevated in inflammatory bowel diseases. Further investigation of its specificity is therefore needed.     

Clinical significance of faecal calprotectin levels in patients with ulcerative colitis]

AIMS: To assess the clinical significance of faecal calprotectin levels (a neutrophil protein) in patients with ulcerative colitis (UC). METHODS: 25 patients with UC provided stool samples for calprotectin assay and the amount of calprotectin was related to UC disease activity index in each patient. Of 25 patients 4 with prednisolone refractory UC received 10 granulocyte and monocyte adsorption apheresis (GMCAP) sessions of 60 minutes duration, flow rate 30 mL per minute for 10 consecutive weeks. RESULTS: Calprotectin level in consecutive faecal samples from three patients was stable. However, increased calprotectin levels were significantly (p < 0.005) associated with Matts's endoscopic index, reflecting the level of colorectal inflammation. The 4 patients who received GMCAP therapy had a clinical activity index < 2 at week 7, the calprotectin level declined with improving Matts' index. CONCLUSIONS: Assay of faecal calprotectin holds promise as a sensitive biomarker to identify colorectal inflammation.     

High fecal calprotectin levels are associated with SARS-CoV-2 intestinal shedding in COVID-19 patients: A proof-of-concept study

BACKGROUNDOne third of coronavirus disease 2019 (COVID-19) patients have gastrointestinal symptoms. Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) RNA has been detected in stool samples of approximately 50% of COVID-19 individuals. Fecal calprotectin is a marker of gastrointestinal inflammation in the general population.AIMTo investigate if fecal calprotectin correlates with SARS-CoV-2 intestinal shedding in COVID-19 patients with pneumonia.METHODSPatients with SARS-CoV-2 pneumonia admitted to the Infectious Disease Unit (University Hospital of Trieste, Italy) from September to November 2020 were consecutively enrolled in the study. Fecal samples were collected and analyzed for quantification of fecal calprotectin (normal value < 50 mg/kg) and SARS-CoV-2 RNA presence by polymerase chain reaction (PCR). Inter-group differences were determined between patients with and without diarrhea and patients with and without detection of fecal SARS-CoV-2.RESULTSWe enrolled 51 adults (40 males) with SARS-CoV-2 pneumonia. Ten patients (20%) presented with diarrhea. Real-time-PCR of SARS-CoV-2 in stools was positive in 39 patients (76%), in all patients with diarrhea (100%) and in more than two thirds (29/41, 71%) of patients without diarrhea. Obesity was one of the most common comorbidities (13 patients, 25%); all obese patients (100%) (P = 0.021) tested positive for fecal SARS-CoV-2. Median fecal calprotectin levels were 60 mg/kg [interquartile range (IQR) 21; 108]; higher fecal calprotectin levels were found in the group with SARS-CoV-2 in stools (74 mg/kg, IQR 29; 132.5) compared to the group without SARS-CoV-2 (39 mg/kg, IQR 14; 71) (P < 0.001).CONCLUSIONHigh fecal calprotectin levels among COVID-19 patients correlate with SARS-CoV-2 detection in stools supporting the hypothesis that this virus can lead to bowel inflammation and potentially to the ‘leaky gut’ syndrome.     

Direct estradiol down-regulation of secretogranin II and chromogranin A mRNA levels in rat pituitary cells.

Estradiol (E2) has been previously shown to negatively regulate, in vivo, the secretogranin (SgII) and chromogranin A (CgA) mRNA levels in the rat pituitary. Using cultured pituitary cell aggregates, experiments were undertaken to discriminate between direct or indirect effects of E2 on SgII and CgA levels. SgII, CgA and gonadotropin alpha- and beta-subunit mRNA levels were determined by Northern blotting. SgII and CgA protein levels were quantitated by Western blotting, and by immunoprecipitation of radioactive SgII after [35S]methionine labeling. Increasing concentrations of E2 (from 10(-12) M to 10(-8) M) in the culture medium promoted a decrease of SgII and CgA mRNA levels to 30% and 50% of the control, respectively, after 72.h treatment. By contrast, none of the gonadotropin subunit mRNAs exhibited changes in concentration. A 24 h treatment with 10(-8) M E2 was sufficient to promote such a decrease in SgII and CgA mRNAs. Quantitation of the proteins after Western blotting revealed that 10(-8) M E2 lowered by 30% in CgA content of aggregates (P < 0.05 vs. control) while SgII content remained unaffected. Moreover, quantitation of the newly synthesized SgII by immunoprecipitation of the 35S-labeled SgII gave evidence for a lack of E2 effect. These data demonstrate: (1) a direct effect of E2 on the pituitary cells to down-regulate SgII and CgA mRNA steady-state levels; (2) though contained within the same secretory granules, a distinct pathway for negative E2 regulation of the gonadotropins and both granins; and (3) a differential effect of E2 on cell SgII and CgA contents as was previously demonstrated in vivo.     

Relationship Between Fecal Calprotectin, Intestinal Inflammation, and Peripheral Blood Neutrophils in Patients with Active Ulcerative Colitis

Active ulcerative colitis (UC) is associated with elevated granulocytes and monocytes/macrophages (GM) which show activation behavior and increased survival time. Further, fecal calprotectin (a stable neutrophil protein) level parallels intestinal inflammation and can predict UC relapse. Since GM are major sources of inflammatory cytokines and chemokines, they are suspected to have roles in the initiation and perpetuation of UC. Our objective was to investigated relationships between peripheral blood (PB) neutrophils, calprotectin, and UC disease activity. Full PB and calprotectin were determined in 69 healthy controls and 31 patients with UC, then 7 randomly selected patients received GM adsorptive apheresis (GMA) with Adacolumn, 10 sessions of 60-min duration each. Patients with UC had higher neutrophil counts (P < 0.001), but lower lymphocyte counts (P < 0.001) compared with controls. Further, fecal calprotectin levels showed a correlation with UC clinical activity index (CAI; P < 0.001) and mucosal inflammation (P < 0.001). Following GMA, there were falls in neutrophils (P < 0.02), CAI (P < 0.02) and calprotectin (P < 0.02). In conclusion, GM appear to contribute to intestinal inflammation and UC activity and reduction of these cells by GMA should benefit patients with active UC. Further, the correlations among calprotectin, UC activities, and PB neutrophils should serve as the basis for preemptive actions to control this disease.     

Validation of a point-of-care desk top device to quantitate fecal calprotectin and distinguish inflammatory bowel disease from irritable bowel syndrome

Background and aimsThe neutrophil protein calprotectin has been investigated as a surrogate marker for intestinal inflammation. This study was designed to contrast fecal calprotectin levels in patients with inflammatory and non-inflammatory intestinal diseases and to compare the results obtained from the standard ELISA-based method with those obtained from a novel desk-top device.MethodsSoluble proteins were extracted from stool samples of 50 participating patients, including those diagnosed with Ulcerative Colitis, Crohn's Disease or IBS, and volunteers with no known intestinal problems. Calprotectin was assessed in the extracted material using the “desk top” Bühlmann Quantum Blue Reader® or by standard ELISA techniques.ResultsThe mean concentration of calprotectin in the IBD patients group was significantly higher than the mean concentration found in IBS patients and healthy controls (p = 0.01). Calprotectin concentrations in IBS patients and controls were indistinguishable. IBD patients that had undergone recent surgery displayed scores similar to controls and IBS patients. Excluding these patients yielded a specificity of 100% for results from both CD and UC patients and an accuracy rate of 1 for CD and 0.89 for UC patients in ROC analysis. Quantum Blue Reader® calprotectin levels were available within 30 min and correlated well with results derived from standard ELISA assays, which took over 8 h to complete.ConclusionOur results confirm the effective use of fecal calprotectin levels in differentiating non-inflammatory from active inflammatory intestinal diseases. The desk top Bühlmann Quantum Blue Reader® exhibits a fast, non-invasive, and reliable way of identifying an inflammatory intestinal disease.     

钙卫蛋白、乳铁蛋白在溃疡性结肠炎患者中表达的临床研究

背景:溃疡性结肠炎(UC)是慢性反复发作的肠道炎性疾病,评估疾病活动度对其疗效判断非常重要,实验室标记物尤其是粪便标记物可很好地反映疾病活动度。目的:探讨钙卫蛋白(Cal)、乳铁蛋白(Lf)在UC患者结肠黏膜和粪便中表达的临床意义。方法:选取1998年1月～2008年1月安徽医科大学第一附属医院收治的具备完整结肠镜或手术病理检查结果的UC患者120例。以临床活动度指数(CAI)评分进行疾病分期,以免疫组化SP法检测结肠黏膜Cal、Lf表达,以ELISA法检测粪便Cal、Lf含量。结果:活动期UC患者结肠黏膜中均表达Cal、Lf,缓解期无或仅弱表达。活动期UC患者粪便Cal、Lf含量显著高于缓解期(P0.01)。UC患者结肠黏膜Cal和Lf表达以及粪便Cal和Lf含量均与CAI评分呈正相关(P=0.000)。粪便Cal和Lf含量与结肠黏膜Cal和Lf表达亦呈正相关(r=0.588,P=0.000;r=0.519,P=0.000),且判断UC活动性的敏感性和特异性均较高。结论:UC患者结肠黏膜Cal、Lf表达可反映临床严重度,粪便Cal、Lf含量与UC活动性显著相关。     

Does fecal calprotectin predict relapse in patients with Crohn's disease and ulcerative colitis?

Background and aimsAn evaluation is made of the utility of fecal calprotectin in predicting relapse in patients with inflammatory bowel disease (IBD). The possible differences in its predictive capacity in Crohn's disease (CD) versus ulcerative colitis (UC), and the different phenotypes, are also examined.MethodsThis is a prospective study with 135 patients diagnosed with IBD in clinical remission for at least 3 months. The patients submitted a stool sample within 24 hours after the baseline visit, for the measurement of fecal calprotectin. All patients were followed-up on for one year.ResultsSixty-six patients had CD and 69 UC. Thirty-nine (30%) suffered from relapse. The fecal calprotectin concentration was higher among the patients with relapse than in those that remained in remission: 444 µg/g (95% CI 34–983) versus 112 µg/g (95% CI 22–996); p < 0.01. Patients with CD and calprotectin > 200 µg/g relapsed 4 times more often than those with lower marker concentrations. In UC, calprotectin > 120 µg/g was associated with a 6-fold increase in the probability of disease activity outbreak. The predictive value was similar in UC and CD with colon involvement and inflammatory pattern. In this group, calprotectin > 120 µg/g predicted relapse risk with a sensitivity of 80% and a specificity of 60%. Relapse predictive capacity was lower in patients with ileal disease.ConclusionsFecal calprotectin may be a useful marker for predicting relapse in patients with IBD. Its predictive value is greater in UC and CD with colon involvement and inflammatory pattern, compared with ileal CD.     

Differential expression and processing of chromogranin A and secretogranin II in relation to the secretory status of endocrine cells

Chromogranin A (CgA) and secretogranin II (SgII) are neuroendocrine secretory proteins that participate in regulation of the secretory pathway and also serve as precursors of biologically active peptides. To investigate whether there is a relationship between the expression, distribution, and processing of CgA and SgII and the degree of secretory activity, we employed two melanotrope subpopulations of the pituitary intermediate lobe that exhibit opposite secretory phenotypes. Thus, although one of the melanotrope subtypes shows high secretory activity, the other exhibits characteristics of a hormone storage phenotype. Our data show that SgII expression levels were higher in secretory melanotropes, whereas CgA expression showed similar rates in both cell subsets. The use of various antibodies revealed the presence of the unprocessed proteins as well as three CgA-derived peptides (67, 45, and 30 kDa) and six SgII-derived peptides (81, 66, 55, 37, 32, and 30 kDa) in both subpopulations. However, the smallest molecular forms of both granins predominated in secretory melanotropes, whereas the largest SgII- and CgA-immunoreactive peptides were more abundant in storage melanotropes, which is suggestive of a more extensive processing of granins in the secretory subset. Confocal microscopy studies showed that CgA immunoreactivity was higher in storage cells, but SgII immunoreactivity was higher in secretory melanotropes. Taken together, our results indicate that SgII and CgA are differentially regulated in melanotrope subpopulations. Thus, SgII expression is strongly related to the secretory activity of melanotrope cells, whereas CgA expression may not be related to secretory rate, but, rather, to hormone storage in this endocrine cell type.     

TWEAK is not elevated in patients with newly diagnosed inflammatory bowel disease

Objective: Tumor necrosis factor-like weak inducer of apoptosis (TWEAK) may be involved in the pathogenesis of inflammatory bowel disease. The aim was to investigate if TWEAK may reflect disease activity in inflammatory bowel disease.

Materials and methods: In this cohort study, 139 consecutive patients with newly diagnosed and previously untreated inflammatory bowel disease – 95 with ulcerative colitis (UC) and 44 with Crohn’s disease (CD) – underwent colonoscopy. Disease activity was assessed by the Mayo score and the Mayo endoscopic score (MES) for UC, or the Simple Endoscopic Score (SES) for CD. Serum C-reactive protein (CRP) and fecal calprotectin were measured in IBD patients, as were plasma TWEAK levels in patients and 85 healthy subjects. Associations between TWEAK levels and disease activity markers were explored.

Results: In the total IBD group, the median (interquartile range) TWEAK level was 430?pg/ml (109–6570), in UC 502?pg/ml (109–4547) and in CD patients 352?pg/ml (101–9179), respectively. Healthy subjects had a median (IQR) TWEAK of 307?pg/ml (63–3492). There were no significant differences in TWEAK levels between the total IBD group and healthy control subjects, nor between UC and CD, or between UC/CD and healthy subjects. Furthermore, we found no significant associations between Mayo scores, MES-UC, SES-CD, CRP, and fecal calprotectin with plasma TWEAK levels.

Conclusions: Plasma TWEAK levels do not reflect disease activity or the grade of inflammation in patients with newly diagnosed inflammatory bowel disease. NCT01551563.     

The utility of fecal calprotectin in predicting the need for escalation of therapy in inflammatory bowel disease

Background and aims: Fecal calprotectin is an important biomarker used in the evaluation of inflammatory bowel disease. It has proven to be an effective tool in initial screening as well monitoring response to therapy. The aim of this study is to examine the utility of fecal calprotectin both as a predictor for the escalation of therapy in established inflammatory bowel disease and as a predictor of de novo diagnosis.

Methods: Patients with signs and symptoms concerning for inflammatory bowel disease presenting to outpatient clinics were recruited to provide fecal calprotectin stool samples prior to endoscopic evaluation. Patients were followed up for at least one year and monitored clinically for any change in symptomatology, escalation of therapy or development of IBD, confirmed endoscopically.

Results: A total of 126 patients, of whom 72 were known to have underlying inflammatory bowel disease, were included in the final analysis. Among the patients with elevated fecal calprotectin levels and known inflammatory bowel disease, 66% (33/50) went on to have escalation of therapy within 12 months compared to 18% (4/22) if the fecal calprotectin levels were in the normal range (p?<?.0001). For the remaining patients who at baseline did not have inflammatory bowel disease and a normal endoscopic evaluation, elevated fecal calprotectin resulted in no cases (0/17) of a new diagnosis in the next 12 months.

Conclusions: Fecal calprotectin is a useful test for predicting escalation of therapy in established inflammatory bowel disease.     

粪便生物标记物检测对溃疡性结肠炎的诊断价值研究

目的研究两种粪便生物标记物乳铁蛋白、钙卫蛋白诊断溃疡性结肠炎(ulcerative colitis,UC)活动性的价值。方法选择确诊的UC患者72例作为研究组,60例经结肠镜检查均正常的患者作为对照组,留取结肠镜检查3 d内的粪便样本5~10 g,应用ELISA方法进行粪便乳铁蛋白、钙卫蛋白检测。结果粪便乳铁蛋白、钙卫蛋白水平在缓解组和对照组之间比较差异均无统计学意义(P0.05);活动组分别与缓解组和对照组比较差异均有显著统计学意义(P0.01);活动期各组之间相互比较差异均有统计学意义(P0.01)。粪便乳铁蛋白、钙卫蛋白水平与UC内镜分级标准呈正相关(r=0.873;0.891,P0.01)。结论粪便乳铁蛋白、钙卫蛋白检测能够较准确地诊断UC活动期和缓解期,对临床治疗有指导作用。     

Mutual relationships between chromogranins A and B and gastrin in individual gastrin cells.

The chromogranins A and B (CgA and CgB, respectively), originally detected in the adrenal medulla, are present in various endocrine organs. Remarkably, their immunoreactivities vary among different endocrine cell types and also within a given endocrine cell population. With densitometric techniques at the cellular level, individual gastrin cells (n = 318) from guinea pig antral mucosa were studied to measure their content of immunoreactive CgA, CgB, and gastrin. The composition of these secretory proteins in individual gastrin cells varied considerably but with predictable components. Statistical evaluation of the data showed that immunoreactivities for gastrin and CgA correlated negatively in these cells; CgA and CgB immunoreactivities also correlated inversely. On the other hand, immunoreactivities for gastrin and CgB exhibited a high positive correlation. The mutual relationships between gastrin, CgA, and CgB suggest that under physiological conditions biosynthetic pathways of these secretory constituents are linked to each other in individual gastrin cells.     

Accuracies of fecal calprotectin,lactoferrin, M2-pyruvate kinase,neopterin and zonulin to predict the response to infliximab in ulcerative colitis

BackgroundFecal markers might predict the response to anti-TNFα in ulcerative colitis (UC).AimsTo compare the performance of fecal calprotectin (fCal), lactoferrin (fLact), M2-PK (fM2-PK), neopterin (fNeo), and zonulin (fZon) to predict the response to therapy in active UC patients.MethodsDisease activity from 31 consecutive patients with an active UC, treated with infliximab (IFX) was assessed by the Mayo score at baseline and at week 14 and by the partial Mayo score at W52 and stool samples collected for fecal marker measurements at W0, W2, and W14.ResultsAt W14, 19 patients (61%) were responders to IFX induction. The median levels of fCal, fLact and fM2-PK drop dramatically from baseline to W14 in clinical responders. At W2, fM2-PK, fLact and fCal levels predicted accurately the response to IFX induction. At W14, fLact, fCal, and fM2-PK were individually reliable markers to predict sustained response at W52. The performances of fNeo and fZon were weaker in this setting.ConclusionsThe performance of fM2-PK at W2 to predict response to induction therapy with IFX was superior to that of fLact and fCal, whereas monitoring fLact was the best tool to predict adequately the course of the disease at one year under maintenance IFX in UC.