Inflammatory Bowel Syndrome Research Articles

Human Catestatin Alters Gut Microbiota Composition in Mice.

The mammalian intestinal tract is heavily colonized with a dense, complex, and diversified microbial populations. In healthy individuals, an array of epithelial antimicrobial agents is secreted in the gut to aid intestinal homeostasis. Enterochromaffin cells (EC) in the intestinal epithelium are a major source of chromogranin A (CgA), which is a pro-hormone and can be cleaved into many bioactive peptides that include catestatin (CST). This study was carried out to evaluate the possible impact of CST on gut microbiota in vivo using a mouse model. The CST (Human CgA352−372) or normal saline was intrarectally administered in C57BL/6 male mice for 6 days and then sacrificed. Feces and colonic mucosa tissue samples were collected, DNA was extracted, the V4 region of bacterial 16S rRNA gene was amplified and subjected to MiSeq Illumina sequencing. The α-diversity was calculated using Chao 1 and β-diversity was determined using QIIME. Differences at the genus level were determined using partial least square discriminant analysis (PLS-DA). Phylogenetic investigation of communities by reconstruction of unobserved states (PICRUSt) was used to predict functional capacity of bacterial community. CST treatment did not modify bacterial richness in fecal and colonic mucosa-associated microbiota; however, treatment significantly modified bacterial community composition between the groups. Also, CST-treated mice had a significantly lower relative abundance of Firmicutes and higher abundance of Bacteroidetes, observed only in fecal samples. However, at lower phylogenetic levels, PLS-DA analysis revealed that some bacterial taxa were significantly associated with the CST-treated mice in both fecal and colonic mucosa samples. In addition, differences in predicted microbial functional pathways in both fecal and colonic mucosa samples were detected. The results support the hypothesis that CST treatment modulates gut microbiota composition under non-pathophysiological conditions, however, the result of this study needs to be further validated in a larger experiment. The data may open new avenues for the development of a potential new line of antimicrobial peptides and their use as therapeutic agents to treat several inflammatory conditions of the gastrointestinal tract, such as inflammatory bowel disease (IBD), inflammatory bowel syndrome (IBS), or other health conditions.

Hypogonadotropic Hypogonadism in Males with Glycogen Storage Disease Type 1.

Glycogen storage disease type 1 is an autosomal recessive disorder with an incidence of 1 in 100,000. Long-term complications include chronic blood glucose lability, lactic academia, short stature, osteoporosis, delayed puberty, gout, progressive renal insufficiency, systemic or pulmonary hypertension, hepatic adenomas at risk for malignant transformation, anemia, vitamin D deficiency, hyperuricemic nephrocalcinosis, inflammatory bowel syndrome (type 1b), hypertriglyceridemia, and irregular menstrual cycles. We describe hypogonadotropic hypogonadism as a novel complication in glycogen storage disease (GSD) type 1. Case Studies and Methods: Four unrelated patients with GSD 1a (N = 1) and 1b (N = 3) were found to have hypogonadotropic hypogonadism diagnosed at different ages. Institutional Research Ethics Board approval was obtained as appropriate. Participant consent was obtained. A retrospective chart review was performed and clinical symptoms and results of investigations summarized as a case series. All patients were confirmed biochemically to have low luteinizing hormone (LH) and follicular stimulating hormone (FSH), and correspondingly low total testosterone. Clinical symptoms of hypogonadism varied widely. Investigations for other causes of hypogonadotropic hypogonadism were unremarkable. In addition, all patients were found to have disproportionately low bone mineral density at the lumbar spine compared to the hip. Common to all patients was erratic metabolic control, including recurrent hypoglycemia and elevated lactate levels. Recurrent elevations in cortisol in response to hypoglycemia may be the underlying pathology leading to suppression of gonadotropin-releasing hormone (GnRH) release. Incorporating clinical and/or biochemical screening of the hypothalamic-pituitary-gonadal axis may be important in the management of this disease. Testosterone therapy however needs to be carefully considered because of the risk of hepatic adenomas.

Prevalence and Impact of Inflammatory Bowel Disease-Irritable Bowel Syndrome on Patient-reported Outcomes in CCFA Partners

Prevalence and Impact of Inflammatory Bowel Disease-Irritable Bowel Syndrome on Patient-reported Outcomes in CCFA Partners

Protective Effects of Lignite Extract Supplement on Intestinal Barrier Function in Glyphosate-Mediated Tight Junction Injury

Background: Disruption of tight junctions is associated with various inflammatory diseases, including inflammatory bowel syndrome. Tight junctions form a protective barrier throughout the intestinal tract that is central to health. Glyphosate is known to inhibit tight junction function. The aim of this study was to assess the effect of a soil-derived, lignite extract dietary supplement in protecting against glyphosate’s effect on the tight junctions in cell culture. Methods: Small bowel (IEC-6) and colon epithelium (Caco-2) cells were incubated until a stable Transepithelial Electrical Resistance (TEER) was measured. A control and the lignite extract supplement (20% concentration) were placed on the cells and left overnight. These cells were then treated with and without glyphosate at 10 mg/ml. TEER was measured at 30 min. The expression of the tight junctions was determined by immunofluorescent microscopy of the ZO-1 tight junction element before and after overnight incubation, and following glyphosate exposure. Results: TEER increased with the lignite extract on IEC-6 (95%) and Caco-2 (35%) compared to the control. Glyphosate dramatically decreased the TEER in both IEC-6 (80%) and Caco-2 (76%) cells. The lignite extract blocked the glyphosatemediated decrease in TEER. Conclusion: The lignite extract supplement blocked a glyphosate-dependent decrease in TEER in both cell lines. If found efficacious in clinical trials, RESTORE could prove helpful in combating intestinal sensitivities caused by glyphosatemediated tight junction disruption.

AB294. SPR-21 Sex differences in bladder dysfunction in response to enteric neuronal NFκB overactivation and experimentally induced colitis in mice

ObjectiveIn experimental models, animals with induced colitis have increased urinary frequency and abnormalities in bladder contraction to cholinergic and electric field stimulation (EFS). Nuclear factor kappa B (NFκB) is a major mediator of the inflammatory response in inflammatory bowel disease and syndrome. The aim of the present study was to determine the effect of increased NFκB signalling in enteric neurons on bladder contraction with and without experimentally induced colitis.MethodsCalretinin-Cre-ERT2-IKK2CA/tdTomato transgenic (TG) mice were generated and compared to littermate wild-type (WT) controls. At two months of age, the mice were treated for three days with tamoxifen causing continuous NFκB activation in enteric neuronal cells. One month later, the mice were treated with vehicle or 2.5% dextran sulfate sodium (DSS) for 7 days to induce colitis. Full thickness urinary bladder strips were stimulated with potassium and then EFS to induce nerve mediated contraction. The effect of atropine to inhibit muscarinic receptors and alpha, beta methylene ATP (ATP) to desensitize purinergic receptors was determined.ResultsEven though the bladder contractile response to high potassium stimulation was similar between sexes in both WT and TG mice, the EFS induced maximal contraction was greater in WT males than in WT females. Bladder strips from WT female mice had a significantly greater atropine resistant (purinergic) component of EFS induced contraction, compared to WT male mice, but this was not found in TG mice. The cholinergic component of EFS induced contraction was not different between male and female mice (WT or TG). Acute colitis significantly reduced KCl stimulated and nerve mediated contraction in bladder strips from WT female and male mice, and from TG female mice. Constitutive activation of neuronal NFκB preserved the nerve mediated and KCL induced contractile response in male mice. Acute colitis increased the purinergic component of nerve mediated contractions in male WT mice only.ConclusionsExperimentally induced colitis causes decreased bladder contraction. Neuronal NFκB signalling rescued the decreased bladder contraction due to colitis, perhaps through purinergic regulation in males. Neurogenic inflammation in the bladder may cause a larger degree of bladder dysfunction in female than in male mice perhaps because females have a greater purinergic component of nerve mediated bladder contraction that does not further increase due to colitisFunding Source(s)RO1DK075684; P2ODK097819

Radiation exposure in patients with inflammatory bowel disease and irritable bowel syndrome in the years 2001–2011

Objectives: To compare cumulative ionizing radiation in patients with inflammatory bowel disease (IBD) and irritable bowel syndrome (IBS) for the years 2001–2011. To study how radiation exposure change over time in patients with newly diagnosed IBD and factors associated with radiation exposure.Material and methods: All radiological investigations performed between 1 January 2001 and 31 December 2011 were retrospectively recorded in patients with Crohn’s disease (CD) (n = 103), ulcerative colitis (UC) (n = 304) and IBS (n = 149). Analyses were done with Mann–Whitney and Chi-Square test.Results: The median total cumulative radiation exposure in mSv for CD (20.0, inter quartile range (IQR) 34.8), UC (7.01, IQR 23.8), IBS (2.71, IQR 9.15) and the proportion of patients who had been exposed for more than 50 mSv during the study period (CD 19%, UC 11%, IBS 3%) were significantly higher in the patients with CD compared to patients with UC (p < .001) and IBS (p < .001), respectively. In turn, patients with UC had significantly higher doses than patients with IBS (p = .005). Risk factors for radiation exposure were female gender (CD), early onset (UC), ileocolonic location (CD), previous surgery (CD and UC), depression (IBS) and widespread pain (IBS). In newly diagnosed CD, there was a significant decline in median cumulative radiation dose in mSv (17.2 vs. 12.0; p = .048) during the study period.Conclusions: Patients with CD are at greatest risk for high cumulative radiation exposure, but there is a decline in exposure during the late 2000s. Non-colectomized patients with UC and patients with IBS have a relatively low risk of cumulative radiation exposure.

Diuretics and Malnutrition-Induced Wernicke’s Encephalopathy in the Critically Ill Patient

SESSION TITLE: Student/Resident Case Report Poster - Critical Care V SESSION TYPE: Student/Resident Case Report Poster PRESENTED ON: Tuesday, October 25, 2016 at 01:30 PM - 02:30 PM INTRODUCTION: Wernicke Encephalopathy has been traditionally associated with alcohol abuse. Among non alcoholic causes are dialysis, diuretic use, malnutrition, parenteral nutrition, bariatric surgery, cancer and AIDS. Wernicke encephalopathy is a heterogeneous entity not only limited to the described triad of ophthalmoplegia, encephalopathy and ataxia. CASE PRESENTATION: 43-year-old male with history of aspergillosis was admitted with pneumonia. Due to increased work of breathing he required endotraqueal intubation. The patient looked cachectic, HR: 100 bpm, BP:130/70, and on lung auscultation bilateral rales were noted. He failed weaning trials for two weeks requiring tracheostomy placement. He also developed renal failure requiring dialysis and high doses of diuretics. His mental status after weaning sedation remained abnormal and multiple attempts to switch to trach collar were also unsuccessful. On his neurological exam completely off sedation, he could not follow commands or communicate. He also had a fixed look and unable to track with eyes, concerning for ophthalmoplegia. CT Head showed no abnormalities and an EEG resulted negative for seizures. Repeated blood cultures, TSH and cortisol levels were normal. The etiology of his abnormal mental status was unclear but given his poor nutritional status, diuretic use, critically ill state and what seemed to be an unspecific encephalopathy with ophthalmoplegia, thiamine deficiency was entertained. High doses of IV thiamine was started with dramatic improvement. He progressively started following commands and communicating and within 48 hours he was able to be completely weaned to trach collar. DISCUSSION: Wernicke encephalopathy can present with different sign and symptoms that may be overlooked by a physician. In early stages there is gait and trunk incoordination, ocular abnormalities or mental status changes, and infrequently hemodynamic instability hypothermia, behavioral changes and seizures. Late stages present with hyperthermia, spastic paralysis or increase muscle tone, choreic dyskinesia and coma.1 Wernicke can present in non-alcoholic patient in the setting of malnutrition, parenteral nutrition, cancer, AIDS, illness, Inflammatory bowel syndrome, diuretic use, peritoneal dialysis, hemodialysis and bariatric surgery.2 CONCLUSIONS: Diagnosing thiamine deficiency and Wernicke encephalopathy can be challenging in non-alcoholic. Wernicke can be treated with intravenous thiamine and its development can be prevented in at risk patients with proper supplementation. No agreement on dosage has yet been stablished but it is clear that high doses of thiamine are effective in the treatment of Wernicke encephalopathy. 2,3. Reference #1: Sechi GP, et al. Wernicke’s encephalopathy: new clinical settings and recent advances in diagnosis and management. Lancet Neurol 2007; 6: 442-55 Reference #2: Hutcheon DA. Malnutrition-induced Wernicke’s encephalopathy following a water-only fasting diet. Nutr Clin Pract. 2015;30:92-99. DISCLOSURE: The following authors have nothing to disclose: Konstantinos Sdrimas, Supakanya Wongrakpanich, Jean Bustamante Alvarez No Product/Research Disclosure Information

Hypogonadotropic Hypogonadism in Males with Glycogen Storage Disease Type 1

Background: Glycogen storage disease type 1 is an autosomal recessive disorder with an incidence of 1 in 100,000. Long-term complications include chronic blood glucose lability, lactic academia, short stature, osteoporosis, delayed puberty, gout, progressive renal insufficiency, systemic or pulmonary hypertension, hepatic adenomas at risk for malignant transformation, anemia, vitamin D deficiency, hyperuricemic nephrocalcinosis, inflammatory bowel syndrome (type 1b), hypertriglyceridemia, and irregular menstrual cycles. We describe hypogonadotropic hypogonadism as a novel complication in glycogen storage disease (GSD) type 1.

Expression, localization and possible functions of aquaporins 3 and 8 in rat digestive system

Although aquaporins (AQPs) play important roles in transcellular water movement, their precise quantification and localization remains controversial. We investigated expression levels and localizations of AQP3 and AQP8 and their possible functions in the rat digestive system using real-time polymerase chain reactions, western blot analysis and immunohistochemistry. We investigated the expression levels and localizations of AQP3 and AQP8 in esophagus, forestomach, glandular stomach, duodenum, jejunum, ileum, proximal and distal colon, and liver. AQP3 was expressed in the basolateral membranes of stratified epithelia (esophagus and forestomach) and simple columnar epithelia (glandular stomach, ileum, and proximal and distal colon). Expression was particularly abundant in the esophagus, and proximal and distal colon. AQP8 was found in the subapical compartment of columnar epithelial cells of the jejunum, ileum, proximal colon and liver; the most intense staining occurred in the jejunum. Our results suggest that AQP3 and AQP8 play significant roles in intestinal function and/or fluid homeostasis and may be an important subject for future investigation of disorders that involve disruption of intestinal fluid homeostasis, such as inflammatory bowel disease and irritable bowel syndrome.

P-021 YI Prevalence and Impact of Inflammatory Bowel Disease-Irritable Bowel Syndrome (IBD-IBS) on Patient Reported Outcomes in CCFA Partners

P-021 YI Prevalence and Impact of Inflammatory Bowel Disease-Irritable Bowel Syndrome (IBD-IBS) on Patient Reported Outcomes in CCFA Partners

Key Questions for Translation of FFA Receptors: From Pharmacology to Medicines.

The identification of fatty acids as ligands for the G-protein coupled free fatty acid (FFA) receptor family over 10 years ago led to intensive chemistry efforts to find small-molecule ligands for this class of receptors. Identification of potent, selective modulators of the FFA receptors and their utility in medicine has proven challenging, in part due to their complex pharmacology. Nevertheless, ligands have been identified that are sufficient for exploring the therapeutic potential of this class of receptors in rodents and, in the case of FFA1, FFA2, FFA4, and GPR84, also in humans. Expression profiling, the phenotyping of FFA receptor knockout mice, and the results of studies exploring the effects of these ligands in rodents have uncovered a number of indications where engagement of one or a combination of FFA receptors might provide some clinical benefit in areas including diabetes, inflammatory bowel syndrome, Alzheimer's, pain, and cancer. In this chapter, we will review the clinical potential of modulating FFA receptors based on preclinical and in some cases clinical studies with synthetic ligands. In particular, key aspects and challenges associated with small-molecule ligand identification and FFA receptor pharmacology will be addressed with a view of the hurdles that need to be overcome to fully understand the potential of the receptors as therapeutic targets.

Emerging Vascular Risk Factors in Women: Any Differences from Men?

Gender differences have been reported for traditional vascular risk factors such as smoking, obesity, diabetes, hypertension, dyslipidemia, age and family history of premature coronary heart disease. The prevalence, severity, associations and response to treatment of several emerging cardiovascular disease (CVD) risk factors may also differ between men and women. Such CVD risk factors include certain inflammatory and hemostatic markers, endothelial dysfunction, homocysteine, lipid disorders, microalbuminuria/proteinuria, coronary artery calcium score, arterial stiffness, periodontitis, inflammatory bowel syndrome, obstructive sleep apnea, impaired glucose metabolism, metabolic syndrome and non-alcoholic fatty liver disease. Further larger prospective studies are needed to establish these relationships. Hormone replacement therapy may also affect vascular risk. These data should be taken into consideration when assessing and treating CVD risk in women.

OC-107 Gas chromatography sensor platform for diagnosing inflammatory bowel disease and irritable bowel syndrome using faecal samples: Abstract OC-107 Table 1

Introduction Inflammatory bowel disease (IBD) is a chronic gastrointestinal disease caused by an aberrant immune response in the gut. Irritable bowel syndrome (IBS) is a common disease that may be confused with IBD. Faecal calprotectin concentration is the current recommended biomarker to rule out IBD, but there are no tests to positively diagnose IBS. Consequently, expensive and invasive methods involving colonoscopy and biopsy are still used by many clinicians to rule out other pathologies. Here, we propose a platform for accurately diagnose IBD and IBS based on faecal samples. Method The platform is composed of a gas chromatography coupled to a metal oxide gas sensor (OdoReader) and a computer algorithm. The OdoReader separates the volatile organic compounds (VOCs) present in biological samples, while the computer algorithm identifies resistance patterns associated with specific medical conditions and builds mathematical models to classify unknown samples. This platform was applied to faecal samples collected from 98 patients attending the gastroenterology clinic: 45 patients with active IBD; 24 patients with diarrhoea-predominant IBS; and 29 healthy donors (Control). The following comparisons were performed: IBD vs. IBS; IBD vs. Control; IBS vs. Control; IBD vs. non-IBD (IBS + Control); IBS vs. non-IBS (IBD + Control); Control vs. non-Control (IBD + IBS). The mathematical models developed were validated through rigorous validation schemes namely 10-Fold Cross Validation, Double Cross Validation and their Monte Carlo variations. Results Conclusion This is the first description of an investigation for the positive diagnosis of IBS. The platform presented here classified samples with accuracies from 85% to 94% using rigorous validation schemes. These schemes provide an estimate of out-of-sample predictive accuracy for similar populations. Therefore, the results reported here indicate a successful differentiation of biological samples from patients with IBD, IBS and healthy donors. These results also indicate a better performance than calprotectin alone for diagnosing IBD, which has been reported as 85% before the application of validation schemes. This platform could be used at point of care for non-invasive diagnosis of IBS and IBD. Disclosure of interest None Declared.

Migraine associated with gastrointestinal disorders: review of the literature and clinical implications.

Recent studies suggest that migraine may be associated with gastrointestinal (GI) disorders, including irritable bowel syndrome (IBS), inflammatory bowel syndrome, and celiac disease. Here, an overview of the associations between migraine and GI disorders is presented, as well as possible mechanistic links and clinical implications. People who regularly experience GI symptoms have a higher prevalence of headaches, with a stronger association with increasing headache frequency. Children with a mother with a history of migraine are more likely to have infantile colic. Children with migraine are more likely to have experienced infantile colic compared to controls. Several studies demonstrated significant associations between migraine and celiac disease, inflammatory bowel disease, and IBS. Possible underlying mechanisms of migraine and GI diseases could be increased gut permeability and inflammation. Therefore, it would be worthwhile to investigate these mechanisms further in migraine patients. These mechanisms also give a rationale to investigate the effects of the use of pre- and probiotics in migraine patients.

Pregnancy Outcomes in Women with Inflammatory Bowel Syndrome Following Exposure to Thiopurines and Anti-Tumor Necrosis Factor Drugs: A Meta-Analysis and Systematic Review.

Pregnancy Outcomes in Women with Inflammatory Bowel Syndrome Following Exposure to Thiopurines and Anti-Tumor Necrosis Factor Drugs: A Meta-Analysis and Systematic Review.

1H NMR-based urinary metabolic profiling reveals changes in nicotinamide pathway intermediates due to postnatal stress model in rat.

The maternal separation protocol in rodents is a widely recognized model of early life stress allowing acute and chronic physiological consequences to be studied. An (1)H NMR-based metabolomic approach was applied to urines to evaluate the systemic metabolic consequences of maternal separation stress in female rats after the beginning of weaning and 4 weeks later when the rats were reaching adulthood. Furthermore, because maternal separation is considered as a model mimicking the inflammatory bowel syndrome, the lactulose/mannitol test was used to evaluate the influence of postnatal maternal separation on gut permeability and mucosal barrier function by (1)H NMR spectroscopy analysis of urine. The results showed no statistical differences in gut permeability due to maternal separation. The application of ANOVA simultaneous component analysis allowed the contributions of physiological adaptations to the animal's development to be separated from the metabolic consequences due to postnatal stress. Systemic metabolic differences in the maternally separated pups were mainly due to the tryptophan/NAD pathway intermediate levels and to the methyladenosine level. Urinary NMR-based metabolic profiling allowed us to disentangle the metabolic adaptive response of the rats to postnatal stress during the animal's growth, highlighting the metabolic changes induced by weaning, gut closure, and maturity.

Oral Serum-derived Bovine Immunoglobulin/Protein Isolate (SBI) in a Large, Single Center Experience Improves Symptomatology in Patients With Inflammatory Bowel Disease and Irritable Bowel Syndrome

Oral Serum-derived Bovine Immunoglobulin/Protein Isolate (SBI) in a Large, Single Center Experience Improves Symptomatology in Patients With Inflammatory Bowel Disease and Irritable Bowel Syndrome

The Interplay between the Microbiota and Enterohemorrhagic Escherichia coli

The gastrointestinal tract of mammals is home to a plethora of microbial species that comprise the microbiota. The role of the microbiota in human health is at the forefront of science in recent years, because it is now appreciated that this intricate microbe-host association shapes the host's immune response and physiology. Many diseases are associated with changes in the microbiota, called dysbiosis. Dysbiosis is associated with obesity, metabolic syndromes, inflammatory bowel-disease, inflammatory bowel syndrome, cancer, diabetes, allergies, and autism. The microbiota is largely regarded as a barrier to enteric infections, such as with enterohemorrhagic Escherichia coli (EHEC). However, the interactions between pathogens and the microbiota are largely unknown, as is how these interactions influence the outcome of enteric disease. The microbial composition of the gastrointestinal tract shapes the landscape in which EHEC survives within the host. This organism competes for nutrients derived from the host diet, liberates additional resources from dietary and host sources, and produces signaling molecules sensed by EHEC to direct gene expression. To successfully colonize the recto-anal junction of a ruminant, the EHEC reservoir, or the colon of a human, an accidental host, EHEC must alter its physiology to survive within the host digestive tract. In this article, we explore the classes of molecules produced or modified by the microbiota that appear to be instrumental in governing virulence gene expression of EHEC. We also explore how interaction with different microbiotas influences EHEC infectivity and host interaction.

Simultaneous automated screening and confirmatory testing for vasculitis-specific ANCA.

Anti-neutrophil cytoplasmic antibodies (ANCA) are the serological hallmark of small vessel vasculitis, so called ANCA-associated vasculitis. The international consensus requires testing by indirect immunofluorescence (IIF) on human ethanol-fixed neutrophils (ethN) as screening followed by confirmation with enzyme-linked immunosorbent assays (ELISAs). This study evaluates the combination of cell- and microbead-based digital IIF analysis of ANCA in one reaction environment by the novel multiplexing CytoBead technology for simultaneous screening and confirmatory ANCA testing. Sera of 592 individuals including 118 patients with ANCA-associated vasculitis, 133 with rheumatoid arthritis, 49 with infectious diseases, 77 with inflammatory bowel syndrome, 20 with autoimmune liver diseases, 70 with primary sclerosing cholangitis and 125 blood donors were tested for cytoplasmic ANCA (C-ANCA) and perinuclear ANCA (P-ANCA) by classical IIF and ANCA to proteinase 3 (PR3) and myeloperoxidase (MPO) by ELISA. These findings were compared to respective ANCA results determined by automated multiplex CytoBead technology using ethN and antigen-coated microbeads for microbead immunoassays. There was a good agreement for PR3- and MPO-ANCA and a very good one for P-ANCA and C-ANCA by classical and multiplex analysis (Cohen's kappa [κ] = 0.775, 0.720, 0.876, 0.820, respectively). The differences between classical testing and CytoBead analysis were not significant for PR3-ANCA, P-ANCA, and C-ANCA (p<0.05, respectively). The prevalence of confirmed positive ANCA findings by classical testing (IIF and ELISA) compared with multiplex CytoBead analysis (IIF and microbead immunoassay positive) resulted in a very good agreement (κ = 0.831) with no significant difference of both methods (p = 0.735). Automated endpoint-ANCA titer detection in one dilution demonstrated a very good agreement with classical analysis requiring dilution of samples (κ = 0.985). Multiplexing by CytoBead technology can be employed for simultaneous screening and quantitative confirmation of ANCA. This novel technique provides fast and cost-effective ANCA analysis by automated digital IIF for the first time.

Insights into desmosome biology from inherited human skin disease and cardiocutaneous syndromes.

The importance of desmosomes in tissue homeostasis is highlighted by natural and engineered mutations in desmosomal genes, which compromise the skin or heart and in some instances both. Desmosomal gene mutations account for 45-50% of cases of arrhythmogenic right ventricular cardiomyopathy, and are mutated in an array of other disorders such as striate palmoplantar keratoderma, hypotrichosis with or without skin vesicles and lethal acantholytic epidermolysis bullosa. Recently, we reported loss-of-function mutations in the human ADAM17 gene, encoding for the 'sheddase' ADAM17, a transmembrane protein which cleaves extracellular domains of substrate proteins including TNF-α, growth factors and desmoglein (DSG) 2. Patients present with cardiomyopathy and an inflammatory skin and bowel syndrome with defective DSG processing. In contrast, the dominantly inherited tylosis with oesophageal cancer appears to result from gain-of-function in ADAM17 due to increased processing via iRHOM2. This review discusses the heterogeneity of mutations in desmosomes and their regulatory proteins.