Median Calprotectin Levels Research Articles

Serum Calprotectin Levels in Vitiligo Patients and Disease Relation.

Vitiligo is characterized as melanocyte loss in skin and mucous membranes, the pathogenesis of which has not yet been clarified. Calprotectin is a protein secreted from neutrophils, monocytes, and dendritic cells which has an effect on cytokine receptor regulation and the production of reactive oxygen radicals. It has been the subject of research in various inflammatory and autoimmune diseases, yet not investigated in vitiligo. The aim of our study was to investigate the role of calprotectin in the etiopathogenesis of vitiligo and its relationship with clinical subtypes and disease scores. Forty-four vitiligo patients with lack of autoimmune disease and 36 age- and sex-matched healthy controls were involved in the study. Serum calprotectin levels were measured by ELISA. The results were compared with the control group, and the relationship between patients' demographic characteristics, skin phototypes, disease type, disease scores (Vitiligo Area Scoring Index and Vitiligo Disease Activity Score), disease duration, and age at onset were evaluated. The median serum calprotectin level was 454.08 pg/ml (41.19-873.41) in the patient group, and the median serum calprotectin level was 223.17 pg/ml (44.88-1044.43) in the control group. Serum calprotectin level was significantly higher in the patient group than in the control group (P = 0.016). No correlation was found between serum calprotectin level and disease scores, disease duration, age, or age of onset of disease (P > 0.05). In our study, serum calprotectin levels in the patient group were found to be significantly higher than in the control group. Our findings and the existing literature on calprotectin suggest its potential involvement in the pathogenesis of vitiligo, independent of disease progression and patient characteristics.

P553 The safety and efficacy of adalimumab 80mg weekly as maintenance therapy in pediatric Crohn’s Disease

Abstract Background Intensification of adalimumab (ADL) dosing to weekly 40 mg injections, in response to low drug levels, has been shown to provide beneficial outcomes in pediatric patients with Crohn’s disease (CD). Our study aimed to evaluate the safety and efficacy of weekly 80 mg ADL administration in children with CD. Methods In this retrospective cohort study conducted across five Israeli centers, we reviewed the medical records of pediatric CD patients who received a high dose of ADL 80 mg weekly injections between 2016 and 2023. Collected data included demographic characteristics, disease features, laboratory studies, and treatment outcomes. Results Thirty-two children with CD were included: mean age 15.8 (±1.7) years at intensification, 21 male (66%), 18 (56%) with L3 phenotype. The median time to ADL 80 mg intensification from ADL induction was 48.4 weeks (IQR 23.1-122.5). The mean weighted Pediatric Crohn's Disease Activity Index (wPCDAI) was 28.5 (±16.9) at the time of intensification and the median calprotectin and C-reactive protein levels were 937 μg/g (IQR 540-1410) and 1.3 mg/dL (IQR 0.6-5.2), respectively. Clinically active disease was the main reason for ADL intensification (30, 94%). Baseline ADL levels were available in 30 patients (94%) with a median of 3.8 μg/mL (IQR 2.4-7.2). Among these, 23 children (77%) failed to achieve a target of ≥ 7.5 μg/mL. The median follow-up duration from the intensification dose was 91.2 weeks (IQR 53.6-149.1). Corticosteroids were required in 5/32 (15.6%) children, with a median time of 32.3 weeks (IQR 10.7-51.3) from intensification. There was no statistically significant difference in steroid utilization rates between children who achieved a target baseline drug level of ≥ 7.5 μg/mL and those who did not (p=0.934). Thirteen individuals (41%) discontinued ADL treatment, within a median of 24.7 weeks (IQR 11.7-57.1) from intensification. CD-related exacerbation, hospitalization, and surgery rates were 9 (28%), 4 (13%), and 2 (6%), respectively. No statistically significant differences were found in exacerbation (p=0.406), hospitalization (p=0.322), or surgery rates (p=0.427) between children who achieved a baseline ADL trough concentration level of ≥ 7.5 μg/mL and those who did not. Overall, ADL intensification was safe, but 3 (9%) patients developed new-onset psoriasis. Conclusion Our findings support the safety and efficacy of administering ADL at a weekly dosage of 80 mg as maintenance therapy in pediatric patients with moderate to severe CD.

Serum Calprotectin Levels in Different Subtypes of Juvenile Idiopathic Arthritis (JIA) and Its Correlation with Quantitative CRP and JADAS-27.

To study the serum calprotectin levels in children with juvenile idiopathic arthritis (JIA) and correlate it with C-reactive protein (CRP) and the Juvenile Arthritis Disease Activity Score-27 (JADAS-27). This was a cross-sectional observational study done between November 2017 and March 2019. Fifty treatment-naive children, aged 1 to 18 y with the diagnosis of JIA as per the International League of Associations of Rheumatology (ILAR) criteria were enrolled. Assessment of disease activity was done according to the Juvenile Arthritis Disease Activity Score (JADAS-27). Determination of serum calprotectin and CRP levels was done by immunoassay. The correlation between calprotectin levels with CRP and JADAS-27 was calculated. Of the 50 patients with JIA included in the study, there were 18 female and 32 male children. The median age of presentation to the hospital was 9 y (IQR 5.82-13). The median JADAS-27 was 14 (IQR 6, 20.25). The median serum calprotectin level was 45,375 ng/mL (IQR 30,725, 52,270; range 8,560-63,160 ng/mL). The median CRP was 35.4mg/L (IQR 3.48, 80.3; range 0.02 and 107.4mg/L). The levels of calprotectin in different JIA subtypes were not statistically different using Kruskal-Wallis test. The study also demonstrated a positive correlation between serum calprotectin with CRP and the JADAS-27 (r = 0.418). The calprotectin levels in JIA were significantly higher than those reported in the literature irrespective of the subtype. Serum calprotectin positively correlated with CRP and JADAS-27 in children with JIA.

Relationship between the Levels of Calprotectin and Soluble Receptor for Advanced Glycation End Products with Abdominal Aortic Aneurysm Diameter: A Preliminary Clinical Trial.

An abdominal aortic aneurysm (AAA) is a dilatation of the abdominal aorta greater than 50% of the diameter of a healthy aorta. Previous experimental studies confirm the effect of calprotectin (CAL) on the onset of arterial pathology. It has been suggested that low levels of soluble receptors for advanced glycation end products (RAGEs) increase levels of cytokines that lead to the inhibition of matrix metalloproteinases (MMPs), affecting AAA formation. This study aimed to analyze the correlation of levels of RAGE and CAL with AAA diameter. A group of 32 patients aged 50–75 with diagnosed AAA was enrolled in the study. This group of patients was further divided into three subgroups based on AAA diameter: (1) <4.5 cm, (2) 4.5–5.5 cm, (3) >5.5 cm. Peripheral blood was drawn from all participants on admission to measure the serum CAL and RAGE levels. An enumeration survey was performed three months after AAA surgical treatment. CAL and RAGE plasma levels were measured with the enzyme-linked immunosorbent assay (ELISA). The median CAL levels were 2273.0 ng/mL before and 1217.0 ng/mL after treatment. There was a statistically significant decrease in CAL levels following the surgical treatment (p = 0.003). The correlation analysis between CAL levels and RAGE levels before and after surgical treatment showed no statistically significant correlations. In addition, there were no statistically significant correlations between CAL and RAGE levels with AAA size. In conclusion, CAL levels appear to be a significant marker in patients with AAA. There is an almost twofold decrease in CAL levels after AAA excision.

CORRELATION OF FECAL, PLASMA, SERUM, AND SALIVARY CALPROTECTIN TO ENDOSCOPIC AND HISTOLOGIC OUTCOMES IN PATIENTS WITH CROHN’S DISEASE

Abstract BACKGROUND Monitoring disease activity in Crohn’s disease is important as achieving endoscopic remission significantly improves clinical outcomes. Fecal calprotectin is a common non-invasive biomarker for monitoring disease activity. However, fecal calprotectin does not always correlate with other disease activity assessments and stool samples can be difficult to collect and inconvenient to return. Thus, we aimed to compare the association between calprotectin levels in stool, saliva, and blood, and assess to their correlation with endoscopic and histologic outcomes among patients with Crohn’s disease. METHODS Nineteen patients with Crohn’s disease were included in the data analysis. Patients collected stool, blood and saliva samples within 2 days of colonoscopy. Samples were processed and stored at -80C, then assessed for calprotectin via ELISA. The Simple Endoscopic Score – Crohn’s Disease (SES-CD) was scored at the time of colonoscopy. The biopsy sample with the most severe disease activity was assessed by a single pathologist for the Robarts Histological Index (RHI). RESULTS All participants had endoscopic data and blood samples (n=19), 18 had saliva samples, and 18 had stool samples. Participants had a mean age of 32.4 (SD: 7.4). Seventy percent (n=14) were female and 45% were on biologics. The median calprotectin levels were 65.4 ug/g (IQR: 8.5, 385.0) for fecal calprotectin, 23.9 ng/ml (IQR: 15.9, 40.8) for plasma calprotectin, 977.7 ng/ml (IQR: 733.3, 1207.5) for serum calprotectin, and 3761.6 ng/ml (IQR: 874.0, 8267.2) for salivary calprotectin. Correlations (as represented by Pearson’s correlation coefficients) with endoscopic disease activity (SES-CD) were 0.83 for fecal calprotectin, 0.28 for plasma calprotectin, 0.24 for serum calprotectin, and 0.02 for salivary calprotectin. Correlations with histologic disease activity (RHI) were 0.80 for fecal calprotectin, 0.56 for plasma calprotectin, 0.16 for serum calprotectin, and -0.03 for salivary calprotectin. CONCLUSION We present preliminary findings of the relationship between calprotectin from fecal, blood, and saliva samples with endoscopic and histologic outcomes in a small sample of patients with Crohn’s disease. Larger sample sizes are needed to further examine the possible role of blood-based calprotectin assessments. In the current study, only two participants had very high SES-CD scores which influenced our ability to understand the relationship of calprotectin to endoscopic disease activity in this subset. Of interest, plasma calprotectin had a higher correlation with histology than serum, which aligns with previous studies.

Correlation of calprotectin serum levels with degrees of endometriosis: A cross-sectional study.

BackgroundEndometriosis is closely associated with delayed diagnosis due to the lack of a definitive and sensitive noninvasive approach. The use of calprotectin in inflammatory process has been demonstrated in various inflammatory diseases. Calprotectin has a significant correlation with high-sensitivity C-reactive protein (hs-CRP) and could be used as an inflammatory marker. No study thus far has evaluated the correlation between calprotectin and endometriosis.ObjectiveTo determine the correlation of calprotectin with the degree of endometriosis in order to help clinicians in establishing better early detection and management. Materials and Methods In this cross-sectional study, 46 women referred to the Cipto Mangunkusumo, Fatmawati, and Persahabatan Hospitals in Jakarta, Indonesia between July 2017 and April 2018 were enrolled, and their blood serum were taken a day before surgery. Calprotectin serum level was treated using the PhicalⓇ ELISA method. After the diagnosis of endometriosis was confirmed through pathological examination, the final diagnosis of endometriosis could be established. The degree of endometriosis was classified according to the revised American Society for Reproductive Medicine (ASRM) classification.ResultsThe prevalence of minimal, mild, moderate, and severe degrees of endometriosis were 15.2, 39.1, 34.8, and 10.9%, respectively. The median serum calprotectin levels for minimal, mild, moderate, and severe endometriosis were 138.98, 121.49, 124.16, and 122.82 mg/mL, respectively. No correlation was observed between calprotectin and the degrees of endometriosis (r = –0.16, p = 0.278).ConclusionThere is no correlation between calprotectin serum levels and the degrees of endometriosis.

Seven Weeks of High-Dose Vitamin D Treatment Reduces the Need for Infliximab Dose-Escalation and Decreases Inflammatory Markers in Crohn's Disease during One-Year Follow-Up.

Background: Seven weeks of high-dose vitamin D treatment decreases intestinal IL17A and IFN-γ mRNA expression in active Crohn’s disease (CD). In this follow-up study, we investigated whether seven-week vitamin D treatment affected the infliximab response in the following 45 weeks compared to placebo. Methods: CD patients (n = 40) were initially randomised into four groups: infliximab + vitamin-D; infliximab + placebo-vitamin-D; placebo-infliximab + vitamin-D; and placebo-infliximab + placebo-vitamin-D. Infliximab (5 mg/kg) or placebo-infliximab was administered at weeks 0, 2 and 6. Vitamin D (5 mg bolus followed by 0.5 mg/day for 7 weeks) or placebo-vitamin D was handed out. After the 7-week vitamin D period, all patients received infliximab during follow-up. Results are reported for Group D+ (infliximab + vitamin-D and placebo-infliximab + vitamin-D) and Group D- (infliximab + placebo-vitamin-D and placebo-infliximab + placebo-vitamin-D). Results: Group D- patients had greater needs for infliximab dose escalation during follow-up compared to group D+ (p = 0.05). Group D+ had lower median calprotectin levels week 15 (p = 0.02) and week 23 (p = 0.04) compared to group D-. Throughout follow-up, group D+ had 2.2 times (95% CI: 1.1–4.3) (p = 0.02) lower median CRP levels compared with group D-. Conclusions: Seven weeks high-dose vitamin D treatment reduces the need for later infliximab dose-escalation and reduces inflammatory markers. EudraCT no. 2013-000971-34.

Faecal Inflammatory Biomarkers and Gastrointestinal Symptoms after Bariatric Surgery: A Longitudinal Study

Background: Bariatric surgery induces various gastrointestinal (GI) modifications. We performed the first study longitudinally assessing the effect of bariatric surgery on faecal inflammatory biomarker levels and its relation with GI complaints. Method: Faecal calprotectin, lactoferrin, and calgranulin-C levels were determined in 41 patients (34 Roux-en-Y [RYGB], 7 sleeves) before and at 6–16 weeks, 6 months, and 1 year after surgery. Changes in biomarker levels and percentage of patients above reference value were determined. Gastrointestinal symptom rating scale (GSRS) was used to assess GI complaints at corresponding time points. The postoperative relation between GSRS score and biomarker levels above reference value was investigated. Results: After RYGB, median calprotectin levels are significantly higher (>188, 104–415 μg/g) than before surgery (40, 19–78 μg/g; p < 0.001), and over 90% of patients have levels above reference value 1 year after surgery. Median lactoferrin was 0.4 (0.2–1.6) μg/g before, and >5.9 (1.8–13.6) μg/g after surgery (p < 0.001). Median calgranulin-C levels remained far below the reference value and were 0.13 (0.05–0.24) μg/g before and <0.23 (0.06–0.33) μg/g after surgery. Similar results were found after sleeve gastrectomy. No difference was seen in GSRS score for patients with calprotectin and lactoferrin levels above reference values. Conclusion: Faecal inflammatory biomarkers calprotectin and lactoferrin, but not calgranulin-C, rise above reference values shortly after bariatric surgery and remain elevated in the majority of patients. The discrepancy between calprotectin and calgranulin-C levels suggests no GI inflammation. Furthermore, patients after RYGB with biomarkers above the population reference value do not seem to have more GI complaints.

Blood calprotectin in children with juvenile idiopathic arthritis: relationship to flare development after discontinuation of treatment.

ObjectivesThe study aim was to prospectively evaluate the relationship between disease flare development in children with juvenile idiopathic arthritis (JIA) after discontinuation of treatment and serum calprotectin levels (MRP8/14).Material and methodsDetermination of blood serum level of calprotectin was performed in 54 patients with inactive JIA from various regions of Ukraine. The inclusion criterion was the existence of an inactive state of the disease in children with JIA for at least 6 months. During 1 week after blood sampling for determination of serum calprotectin (MRP8/14) level the patients were completely discontinued of all therapy. Determination of calprotectin level in blood serum was performed with reagents EK-MRP8/14 Buhlmann (MRP8/14; S100A8/9), Switzerland, using the ELISA method.ResultsThe trial results showed that 3 months after discontinuation of treatment in patients with inactive JIA, the flares developed in 5 out of 54 patients (9.3%). The median calprotectin level before discontinuation of the treatment was 1,700 ng/ml in patients who developed a flare, and 1,500 ng/ml in other studied patients (not statistically significant). At 6 months, the flare had developed in an additional 3 out of 48 (6.3%) of patients, who continued to be followed up, while their median calprotectin serum levels were 1,300 ng/ml and 1,500 ng/ml respectively (not statistically significant). At 12 months, the flares had developed in 13 more out of 45 (28.9%) patients, who continued to be followed up, while the median calprotectin serum level in these patients before discontinuation of treatment was 1,100 ng/ml and 1,650 ng/ml respectively (not statistically significant).ConclusionAfter discontinuation of treatment a flare over the next year of follow-up developed in 38.9% of patients. The study results did not reveal a significant difference in calprotectin level in patients with JIA prior to complete discontinuation of treatment who developed a flare and those without a flare after 3, 6 and 12 months.

The value of calprotectin in synovial fluid for the diagnosis of chronic prosthetic joint infection.

AimsThis study aimed to evaluate calprotectin in synovial fluid for diagnosing chronic prosthetic joint infection (PJI) .MethodsA total of 63 patients who were suspected of PJI were enrolled. The synovial fluid calprotectin was tested by an enzyme-linked immunosorbent assay (ELISA). Laboratory test data, such as ESR, CRP, synovial fluid white blood cells (SF-WBCs), and synovial fluid polymorphonuclear cells (SF-PMNs), were documented. Chi-squared tests were used to compare the sensitivity and specificity of calprotectin and laboratory tests. The area under the curve (AUC) of the receiver operating characteristic (ROC) curve was calculated to determine diagnostic efficacy.ResultsThe median calprotectin level was 776 μg/ml (interquartile range (IQR) 536.5 to 1132) in the PJI group and 54.5 μg/ml (IQR, 38.75 to 78.25) in the aseptic failure (AF) group (p < 0.05). Using a threshold of 173 ug/ml, the sensitivity was 95.2%, with a 97.6% specificity, and the AUC was 0.993. The sensitivity of calprotectin of the antibiotic-treated PJI group was 100% versus 90.9% of the non-antibiotic-treated PJI group. Although 47.6% (ten cases) of the patients in the PJI group received antibiotics before aspiration, the diagnostic efficacy of calprotectin was not affected. The sensitivity and specificity of ESR, CRP, SF-WBCs, and SF-PMNs ranged from 76.2% to 90.5% and 64.3% to 85.7%, respectively.ConclusionCalprotectin in synovial fluid has great diagnostic efficacy for PJI diagnosisand outperformed ESR, CRP, SF-WBCs, and SF-PMNs.Cite this article: Bone Joint Res 2020;9(8):450–457.

COMPARING OF FAECAL CALPROTECTIN LEVELS IN PATIENTS WITH OSTEOARTHRITIS TAKING NSAID TREATMENT AND PATIENTS WITHOUT NSAIDS THERAPY

Faecal calprotectin (FC) level can be increased in several conditions, which are characterised by neutrophilic inflammation. Some medications, particularly NSAIDs, can elevate its level as well. NSAIDs are taken by patients in many chronic conditions, including osteoarthritis (OA). On the other hand, there is growing evidence that osteoarthritis is not only a degenerative disease, but it has a significant inflammatory component. The role of systemic inflammation is well-known in inflammatory joint diseases, but there is some evidence that it can play an essential role in the OA as well. It can suggest that in the OA, the inflammatory changes could be found in the different organs and systems. The aim of this study was to investigate the FC level in patients with osteoarthritis depending on the NSAIDs intake and to compare it to the FC levels in healthy adults. Materials and methods. In this small observational study, we evaluated the FC levels in patients suffering from OA (36 persons), divided them into two groups depending on their NSAIDs intake, and compared it to FC levels in healthy participants (12 persons). We compared the FC levels depending on the selectivity of the NSAIDs taken by our participants, as well. Results. The median calprotectin level in our patients was 72.57 (IQR 20.55-221.57) mg/kg, 95 %CI 26,18-109.01. OA patients had higher levels than the healthy group (p&lt;0.001). OA patients who took NSAIDs had the highest FC levels – 221.57 (IQR 91.56-448.61) – higher, than those who did not take it – 72.57 (IQR 35.26-164.79) (p=0.03) and than healthy participants, who has normal FC levels (p&lt;0.001); the FC levels of patients who did not take NSAIDs also exceeds healthy subject’s levels (p&lt;0.001). The FC levels in the collective group have a sufficient positive correlation with the duration of NSAIDs intake, VAS score and strong correlation with Lequesne index values. We found that both NSAIDs groups have a significantly greater prevalence of elevated or high FC levels than the control group (p&lt;0.001) and that NSAIDs patients significantly more often have high FC levels than those who do not take NSAIDs (p=0.035). When comparing FC levels in patients depending on the type of NSAIDs they take, we found that those who take non-selective NSAIDs has significantly higher FC levels than those who do not take NSAIDs – 264.1 (IQR 89.72-464.67) to 25.65 (IQR 19.5-75.33) (p=0.0003). The FC levels of who take selective NSAIDs – 98.53 (91.56-105.5) – did not differ significantly to non-selective NSAIDs taker’s group values and non-selective NSAIDs taker’s values (p&gt;0.05). Conclusions. Patients who suffer from OA have higher FC levels than healthy individuals, and patients with OA who take NSAIDs regularly have higher FC levels than those who do not. The intake of non-selective NSAIDs is associated with higher FC levels, than the intake of high-selective NSAIDs. FC levels of those who take high-selective NSAIDs do not differ statistically from those who do not intake NSAIDs. Further research is needed in this area.

SUN-181 HIGH SERUM CALPROTECTIN LEVELS MAY PREDICT MORTALITY RISK IN HEMODIALYSIS PATIENTS WITH HIGH-PHOSPHATE CONDITIONS

The inflammatory mediator myeloid-related protein 8 (MRP8: S100A8), which forms a heterodimeric complex with a binding partner MRP14 in the bloodstream, plays important roles as an endogenous ligand for toll-like receptor 4, contributing to pathophysiology in various inflammatory diseases. Serum MRP8/14, known as calprotectin (CPT), reportedly became a potential biomarker in inflammatory diseases such as rheumatoid arthritis or ANCA-associated vasculitis, though there is no report in hemodialysis patients. The aim of this study is to investigate the predictive role of serum CPT on mortality in hemodialysis patients. We conducted a multicenter, observational cohort study of 388 Japanese subjects undergoing maintenance hemodialysis in Kumamoto, Japan. Serum CPT levels were measured using an ELISA. The potential associations between serum CPT levels and clinical variables were examined in a cross-sectional study. Multivariable Cox regression was used to estimate the association between serum CPT, high-sensitivity C reactive protein (hs-CRP), white blood cell (WBC) count and mortality, adjusting for possible confounding variables including age, sex, diabetes and others. A median follow-up period was 6.6 years. The mean age of the subjects was 65.3 years; 36.9% were female, and the median vintage was 5.8 years. The median CPT level was 6,108 ng/ml (median in healthy subjects, 2,800) at baseline. The serum CPT levels positively correlated with WBC count (ρ=0.54, P<0.0001) and hs-CRP values (ρ=0.34, P<0.0001). We classified serum CPT and hs-CRP into tertile, and estimated the hazard ratios (HRs) for all-cause mortality in comparison with the lowest tertile. In crude analysis, hs-CRP was an independent predictor of all-cause mortality after adjusting confounding factors (middle vs. low: HR, 2.01; 95% confidence interval [CI], 1.19–3.48 and high vs. low: HR, 2.92; 95% CI, 1.72–5.09). In the analysis by stratum of phosphate levels, as for hs-CRP, the middle tertile (HR, 1.94; 95%CI, 1.07–3.67) and the highest tertile (HR, 2.92; 95%CI, 1.58–5.63) were each significantly associated with all-cause mortality in the low-phosphate group (cut-off, 6.0 mg/dl), after adjustment for relevant confounding factors. In contrast, elevated serum CPT levels were significantly associated with all-cause mortality in the highest tertile (HR, 14.7; 95%CI, 2.57–277.8) among the high-phosphate group, but not among the low-phosphate group. High serum CPT levels could give a potential predictive marker on mortality in hemodialysis patients with high-phosphate levels, which characteristic differs significantly from that of a conventional inflammatory marker, hs-CRP.

Predominantly Antibiotic-resistant Intestinal Microbiome Persists in Patients With Pouchitis Who Respond to Antibiotic Therapy

Pouchitis that develops in patients with ulcerative colitis after total proctocolectomy and ileal pouch anal anastomosis is usually treated with antibiotics. Some patients have recurrence of flares, or become antibiotic-dependent, and require repeated courses or prolonged periods of antibiotic therapy. We investigated microbial factors associated with response to antibiotic treatment and development of antibiotic dependence in patients with pouchitis. We performed a prospective study of 49 patients who had undergone pouch surgery at a tertiary center. Disease activity was determined based on clinical, endoscopic, and histologic criteria. Pouch phenotype was defined as recurrent-acute pouchitis (n= 6), chronic pouchitis and Crohn's-like disease of the pouch (n= 27), normal pouch from patient with ulcerative colitis (n= 10), and normal pouch from patient with familial adenomatous polyposis (n= 6). Fecal samples (n= 234) were collected over time during or in the absence of antibiotic treatment (ciprofloxacin and/or metronidazole). Thirty-three patients were treated with antibiotics, for a median of 425 days of cumulative antibiotic therapy, during follow-up. Calprotectin was measured and fecal DNA was sequenced using shotgun metagenomics and analyzed with specifically designed bioinformatic pipelines. Bacterial strains were isolated from fecal samples. We assessed their ciprofloxacin resistance and ability to induce secretion of inflammatory cytokines by HT-29 intestinal epithelial cells. Most antibiotic-treated patients (79%) had a clinical response to each course of antibiotics; however, 89% of those who completed a 4-week course relapsed within 3 months. Median calprotectin levels decreased by 40% in response to antibiotics. Antibiotic treatment reduced disease-associated bacteria such as Clostridium perfringens, Ruminococcus gnavus, and Klebsiella pneumoniae, but also beneficial species, such as Faecalibacterium prausnitzii. The microbiomes of antibiotic-responsive patients were dominated by facultative anaerobic genera (Escherichia, Enterococcus, and Streptococcus), with multiple ciprofloxacin-resistance mutations in drug target genes and confirmed drug resistance. However, these strains had lower potential for virulence and did not induce secretion of inflammatory cytokines by epithelial cells. After antibiotic cessation, patients had an abrupt shift in microbiome composition, with blooms of oral and disease-associated bacteria. In addition, antibiotic treatment enriched for strains that acquired multidrug resistance loci, encoding enzymes that confer resistance to nonrelated antibiotics, including extended-spectrum beta-lactamases. The efficacy of antibiotic treatment of pouchitis might be attributed to the establishment of an antibiotic-resistant microbiome with low inflammatory potential. This microbiome might provide resistance against colonization by bacteria that promote inflammation. To avoid progression to antibiotic-dependent disease and its consequences, strategies such as short-term alternating antibiotics and nutrition- and microbiome-based interventions should be considered.

Usefulness of fecal calprotectin as a biomarker of microscopic colitis in a cohort of patients with chronic watery diarrhoea of functional characteristics

BackgroundInformation on the use of fecal markers in microscopic colitis screening is limited. AimTo evaluate the risk variables associated with a diagnosis of microscopic colitis including fecal calprotectin. MethodsPatients submitted for a colonoscopy due to chronic watery diarrhea fulfilling criteria of functional disease were evaluated. Colonic mucosa was normal but mild erythema and edema was allowed. Fecal calprotectin was analyzed. A logistic regression was used to evaluate variables associated with both raised fecal calprotectin and a diagnosis of microscopic colitis. Results94 patients were included, 30 were diagnosed with microscopic colitis and 64 made up the control group. Median calprotectin levels were 175 (IQR, 59–325) for the microscopic colitis and 28 (IQR, 16–111) for the control group (p < 0.001). The optimal cut-off for fecal calprotectin was >100 μg/g (AUC, 0.73), with 67% sensitivity and 75% specificity. The number of drugs used ≥3 (OR, 3.9; CI, 1.4–10.4) and microscopic colitis diagnosis (OR, 6; CI, 2.2–16.3) were associated with raised calprotectin levels. Age >60 years (OR, 3.8; CI, 1.4–10.1) and calprotectin levels (OR, 5.3; CI, 2–14.1) were associated with a risk of microscopic colitis. ConclusionsElevated fecal calprotectin concentrations are often seen in microscopic colitis, and may be helpful in the diagnosis of women over 60 with chronic watery diarrhea.

Levels of wound calprotectin and other inflammatory biomarkers aid in deciding which patients with a diabetic foot ulcer need antibiotic therapy (INDUCE study).

AimsDeciding if a diabetic foot ulcer is infected in a community setting is challenging without validated point‐of‐care tests. Four inflammatory biomarkers were investigated to develop a composite algorithm for mildly infected diabetic foot ulcers: venous white cell count, C‐reactive protein (CRP) and procalcitonin, and a novel wound exudate calprotectin assay. Calprotectin is a marker of neutrophilic inflammation.MethodsIn a prospective study, people with uninfected or mildly infected diabetic foot ulcers who had not received oral antibiotics in the preceding 2 weeks were recruited from community podiatry clinics for measurement of inflammatory biomarkers. Antibiotic prescribing decisions were based on clinicians’ baseline assessments and participants were reviewed 1 week later; ulcer infection was defined by clinicians’ overall impression from their two assessments.ResultsSome 363 potential participants were screened, of whom 67 were recruited, 29 with mildly infected diabetic foot ulcers and 38 with no infection. One participant withdrew early in each group. Ulcer area was 1.32 cm2 [interquartile range (IQR) 0.32–3.61 cm2] in infected ulcers and 0.22 cm2 (IQR 0.09–1.46 cm2) in uninfected ulcers. Baseline CRP for mild infection was 9.00 mg/ml and 6.00 mg/ml for uninfected ulcers; most procalcitonin levels were undetectable. Median calprotectin level in infected diabetic foot ulcers was 1437 ng/ml and 879 ng/ml in uninfected diabetic foot ulcers. Area under the receiver operating characteristic curve for a composite algorithm incorporating calprotectin, CRP, white cell count and ulcer area was 0.68 (95% confidence intervals 0.52–0.82), sensitivity 0.64, specificity 0.81.ConclusionsA composite algorithm including CRP, calprotectin, white cell count and ulcer area may help to distinguish uninfected from mildly infected diabetic foot ulcers. Venous procalcitonin is unhelpful for mild diabetic foot ulcer infection.

Faecal biomarkers for screening small bowel inflammation in patients with Crohn's disease: a prospective study.

Background:The value of faecal biomarkers for screening small bowel inflammation in patients with Crohn’s disease (CD) remains to be elucidated. This prospective study was to evaluate the utility of faecal biomarkers for detecting small intestinal inflammation.Methods:A total of 122 consecutive patients with a diagnosis of CD in the small intestine were screened for eligibility. Computed tomography enterography (CTE) was undertaken to evaluate small bowel inflammation followed by colonoscopy to confirm no large bowel involvement. Seventy eligible patients with inflammation confined to the small intestine were included. Faecal samples were collected for assaying calprotectin, lactoferrin and haemoglobin. For assessing the degree of small bowel inflammation, a semi-quantitative scoring system (CTE0, normal; CTE1, mild; CTE2, moderate; CTE3, severe) was applied.Results:The median calprotectin, lactoferrin and haemoglobin levels were significantly higher in patients with small bowel inflammation, CTE scores 1–3 (n = 42) versus 0 (n = 28): calprotectin, 330 versus 40 ng/ml, p < 0.0001; lactoferrin, 14 versus 3 ng/ml, p < 0.0001; haemoglobin, 29.5 versus 6.5 ng/ml, p = 0.005. There was a strong positive relationship between the faecal biomarkers and CTE score: calprotectin, p < 0.0001; lactoferrin, p < 0.0001; haemoglobin, p = 0.0004. A cutoff value of 140 ng/ml for calprotectin had a sensitivity of 69% and a specificity of 82% with an area under the receiver operating characteristic curve (AUC) of 0.82 to detect small bowel inflammation (CTE scores 1–3), while lactoferrin 6 ng/ml had a sensitivity of 69% and a specificity of 79% with an AUC of 0.83, and haemoglobin 9 ng/ml showed a sensitivity of 71% and a specificity of 39% with an AUC of 0.70.Conclusions:Faecal calprotectin, lactoferrin, and to a lesser degree haemoglobin are relevant biomarkers for screening small bowel inflammation in CD patients without large bowel involvement. Further well-designed large-scale studies in this clinical setting should strengthen our findings.

Synovial calprotectin

Recently, several synovial biomarkers have been introduced into the algorithm for the diagnosis of a prosthetic joint infection (PJI). Alpha defensin is a promising biomarker, with a high sensitivity and specificity, but it is expensive. Calprotectin is a protein that is present in the cytoplasm of neutrophils, is released upon neutrophil activation and exhibits anti-microbial activity. Our aim, in this study, was to determine the diagnostic potential of synovial calprotectin in the diagnosis of a PJI. In this pilot study, we prospectively collected synovial fluid from the hip, knee, shoulder and elbow of 19 patients with a proven PJI and from a control group of 42 patients who underwent revision surgery without a PJI. PJI was diagnosed according to the current diagnostic criteria of the Musculoskeletal Infection Society. Synovial fluid was centrifuged and the supernatant was used to measure the level of calprotectin after applying a lateral flow immunoassay. The median synovial calprotectin level was 991 mg/L (interquartile range (IQR) 154 to 1787) in those with a PJI and 11 mg/L (IQR 3 to 29) in the control group (p < 0.0001). Using a cut-off value of 50 mg/L, this level showed an excellent diagnostic accuracy, with an area under the curve of 0.94. The overall sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) was 89%, 90%, 81% and 95% respectively. The NPV was 97% in the nine patients with a chronic PJI. Synovial calprotectin may be a valuable biomarker in the diagnosis of a PJI, especially in the exclusion of an infection. With a lateral flow immunoassay, a relatively rapid quantitative diagnosis can be made. The measurement is cheap and is easy to use. Cite this article: Bone Joint J 2017;99-B:660-5.

Effectiveness and safety of infliximab biosimilar CT-P13 in treating ulcerative colitis: a real‑life experience in IBD primary centers

The aim of this study was to assess the efficacy and safety of infliximab biosimilar (IFX) IFX CT-P13 in inducing and maintaining remission in ulcerative colitis (UC) outpatients in Italian primary gastroenterology centers. Patients were prospectively assessed at entry, after 8, 12, 24, 36, and therefore 52 weeks. Clinical activity was rated as per the Mayo Score. The primary endpoint was reaching of clinical remission (Mayo Score ≤2). Several secondary endpoints were clinical response to treatment, reaching of mucosal healing (MH), safety of the drug. Twenty-nine patients (16 males and 13 females, mean age 45 years, range 35-42 years) were enrolled. Eleven (37.9%) patients had previous exposure to other anti-TNF-α. Clinical remission was present in 78.5% at week 24, and in 100% at 12-month follow-up. Subgroup analysis did not reveal significant differences in clinical remission between IFX-naïve patients and patients switching from originator to IFX biosimilar. A clinical response was observed in 92.3% at week 8, in 50.0% at week 16, in 100% at week 36 and in 100% at 12-month follow-up. MH occurred in 85.7% at week 24, and in 100% at 12-month follow-up Reduction of steroids was achieved in 92.3% at week 8, and in 100% during follow-up. One patient underwent proctocolectomy 3 weeks after starting IFX CT-P13. The median C-reactive protein and calprotectin levels during follow-up were significantly reduced during follow-up. No adverse events were observed during follow-up. IFX CT-P13 seems to be very effective and safe in real-life experience at primary IBD centers.

Elevated calprotectin in patients with atrial fibrillation with and without heart failure

Calprotectin is an inflammatory marker, which has been found elevated in patients suffering from cardiac conditions, e.g. myocardial infarction, unstable angina and chronic heart failure. Inflammation has further been linked to atrial fibrillation (AF). However, the association between calprotectin and AF is unknown. We aimed to compare calprotectin levels in patients suffering from AF with healthy adults. In addition, AF patients with and without heart failure were compared. Calprotectin was measured in patients undergoing elective direct current cardioversion for AF. Calprotectin was determined before, 4 hours and 3 months after cardioversion. Healthy blood donors were used to verify the reference interval for calprotectin. In total, 104 prospectively enrolled patients were included. The median serum calprotectin level for AF patients was 1.6 μg/mL before cardioversion. Calprotectin levels increased significantly 4 h (1.9 μg/mL) and 3 months (2.2 μg/mL) after cardioversion. Blood donors’ median serum calprotectin (1.3 μg/mL) was significantly lower than AF patients. AF patients with heart failure had significantly higher calprotectin at baseline compared with AF patients without a history of heart failure (2.0 μg/mL vs. 1.5 μg/mL). The difference was not significant at 4 h (2.0 μg/mL vs. 1.7 μg/mL) or 3 months (2.5 μg/mL vs. 2.2 μg/mL). In conclusion, the calprotectin levels in patients with AF were significantly higher than healthy blood donors and were further increased after cardioversion. AF patients with heart failure had significantly higher levels of calprotectin than AF patients without heart failure.

Calprotectin as a Biomarker for Melioidosis Disease Progression and Management.

Melioidosis is a neglected tropical disease that is caused by the bacterium Burkholderia pseudomallei and is underreported in many countries where the disease is endemic. A long and costly administration of antibiotics is needed to clear infections, and there is an unmet need for biomarkers to guide antibiotic treatment and increase the number of patients that complete therapy. We identified calprotectin as a lead biomarker of B. pseudomallei infections and examined correlations between this serum protein and the antibiotic treatment outcomes of patients with melioidosis. Serum levels of calprotectin and C-reactive protein were significantly higher in patients with melioidosis and nonmelioidosis sepsis than in healthy controls. Median calprotectin levels were higher in patients with melioidosis than in those with nonmelioidosis sepsis, whereas C-reactive protein levels were similar in both groups. Notably, intensive intravenous antibiotic treatment of patients with melioidosis resulted in lower levels of calprotectin and C-reactive protein (P < 0.0001), coinciding with recovery. The median percent reduction of calprotectin and C-reactive protein was 71% for both biomarkers after antibacterial therapy. In contrast, we found no significant differences in calreticulin levels between the two melioidosis treatment phases. Thus, reductions in serum calprotectin levels were linked to therapeutic responses to antibiotics. Our results suggest that calprotectin may be a sensitive indicator of melioidosis disease activity and illustrate the potential utility of this biomarker in guiding the duration of antibiotic therapy.