Evaluation of an automated algorithm for interpretation of lupus anticoagulant testing

Introduction: Lupus anticoagulant (LAC) testing is a multistep procedure including screening, mixing, and confirmation tests. STA Coag Expert is a software module for STA R Max and STA Compact Max analyzers which includes an on-demand LAC algorithm, based on ISTH guidelines, for automatic interpretation, calculation, and launch of assays in LAC interpretation (Stago coag algorithm). Materials and methods: One hundred ninety four patient samples were analyzed in parallel and interpreted manually and automatically by LAC algorithms. LAC algorithms use identical flowcharts and cutoff values as in daily practice. Differently, it only uses index of circulating anticoagulant (ICA), whereas in routine also normalized ratios were assessed for interpretation of mixing tests. Interpretation of dRVVT and aPTT pathways and final conclusions were compared between both approaches. Results: Compared to routine interpretation, LAC algorithm showed a sensitivity of 94% and a specificity of 100% for LAC detection, when discrepancies due to measured clotting times between both analyzers were excluded. Three false negatives were due to different interpretation of dRVVT mixing test. Discrepancies in interpretation of the aPTT mixing test (n=11) did not result in discrepant final LAC result, all having negative confirmation tests. No false positives were observed. With LAC algorithm, hands-on time reduced from 200 to 80minutes. Conclusion: The LAC algorithm of the STA Coag Expert shows good comparability to the manual interpretation of LAC and may be used to assist laboratories in automatic launching of additional tests and in interpretation of LAC according to ISTH guidelines. This way the STA Coag Expert LAC algorithm may improve interlaboratory and STA comparability of LAC results

Evaluation of the primary biliary cholangitis-related serologic profile in a large cohort of Belgian systemic sclerosis patients

Background: Systemic sclerosis (SSc) and primary biliary cholangitis (PBC) are autoimmune diseases that may occur concomitantly and are both strongly associated with disease-specific autoantibodies. This study investigated the prevalence and fine specificity of PBC-specific serology (PBC-Ab) and associations with the SSc-subtypes and SSc-specific antibodies as well as the association with cholestatic liver enzymes. Furthermore, three different techniques for the detection of PBC-Ab were compared. Methods: Serum of 184 Belgian SSc patients with a known SSc-antibody profile, was analyzed for PBC-Ab (antimitochondrial antibodies [AMA], anti-Gp210, anti-Sp100 and anti-PML) using indirect immunofluorescence (IIF) analysis on human epithelioma-2000 (HEp-2000) cells (ANA-IIF, Immunoconcepts) and liver-kidney-stomach tissue sections (IIF-LKS) (Menarini), and a line immunoblot (LB) (Eurolmmun). Alkaline phosphatase/gamma-glutamyl transferase (ALP/GGT) were evaluated at time of first sampling (t0) and after 3 years of follow-up (t3). Results: PBC-Ab were present in 13% of patients and significantly correlated with centromere antibodies (anti-CENP-B), but not correlated with the limited cutaneous SSc subgroup (lcSSc). The most frequent reactivities were AMA (11%, with 9% AMA-M2) and Sp-100 antibodies (5%), showing a major overlap. There was no relevant association between the presence of PBC-Ab and ALP or GGT elevation at t0 nor at t3. Detection of AMA with IIF-LKS is comparable to LB. ANA-IIF screening was less sensitive compared to LB. Conclusions: A wide range of PBC-Ab is detectable in SSc in the absence of cholestatic liver enzyme elevations, even after 3 years of follow-up. However, as these antibodies may precede PBC-disease up to 10 years further prospective follow-up of our cohort will be necessary

Active von Willebrand Factor in patients with a bleeding diathesis

Introduction: Increased levels of circulating von Willebrand Factor (VWF) in its active, platelet-binding conformation have been implicated in the pathogenesis of several thrombotic conditions as well as bleeding conditions characterized by severe thrombocytopenia. However, it is unclear whether the proportion of activated VWF in the circulation also plays a role in patients with mild to moderate bleeding without thrombocytopenia. Methods: Citrated plasma samples were collected from 145 patients with a bleeding diathesis with unknown cause. Active VWF levels were measured with an in-house developed ELISA assay. In addition, VWF antigen (VWF:Ag), VWF ristocetin cofactor activity (VWF:RCo) and (flow-cytometric) platelet-VWF binding (Plt:VWF) were determined. Results: Active VWF levels were on average mildly, but not significantly, lowered in patients with a bleeding diathesis compared to the reference interval (especially in individuals with non-O blood groups). Active VWF was not significantly different for subjects with (median 107.4%, IQR 18.3) versus without (median 111.1%, IQR 32.3%) an increased bleeding score, nor between subjects with suspected VWD (median 104%, IQR 20.6%) versus other suspected causes of bleeding diathesis (median 111.7%, IQR 33.3%). Conclusion: In this clinically heterogeneous population of patients with a mild bleeding phenotype, quantification of active VWF levels does not have added diagnostic value to VWF:Ag and VWF activity assays in the diagnosis of unexplained bleeding disorders

Intestinal barrier dysfunction in inflammatory bowel diseases

The etiology of human inflammatory bowel diseases (IBDs) is believed to involve inappropriate host responses to the complex commensal microbial flora in the gut, although an altered commensal flora is not completely excluded. A multifunctional cellular and secreted barrier separates the microbial flora from host tissues. Altered function of this barrier remains a major largely unexplored pathway to IBD. Although there is evidence of barrier dysfunction in IBD, it remains unclear whether this is a primary contributor to disease or a consequence of mucosal inflammation. Recent evidence from animal models demonstrating that genetic defects restricted to the epithelium can initiate intestinal inflammation in the presence of normal underlying immunity has refocused attention on epithelial dysfunction in IBD. We review the components of the secreted and cellular barrier, their regulation, including interactions with underlying innate and adaptive immunity, evidence from animal models of the barrier's role in preventing intestinal inflammation, and evidence of barrier dysfunction in both Crohn's disease and ulcerative colitis. (Inflamm Bowel Dis 2008

Surveillance of endoscopes : comparison of different sampling techniques

OBJECTIVE: To compare different techniques of endoscope sampling to assess residual bacterial contamination. DESIGN: Diagnostic study. SETTING: The endoscopy unit of an 1,100-bed university hospital performing similar to 13,000 endoscopic procedures annually. METHODS: In total, 4 sampling techniques, combining flushing fluid with or without a commercial endoscope brush, were compared in an endoscope model. Based on these results, sterile physiological saline flushing with or without PULL THRU brush was selected for evaluation on 40 flexible endoscopes by adenosine triphosphate (ATP) measurement and bacterial culture. Acceptance criteria from the French National guideline (<25 colony-forming units [CFU] per endoscope and absence of indicator microorganisms) were used as part of the evaluation. RESULTS: On biofilm-coated PTFE tubes, physiological saline in combination with a PULL THRU brush generated higher mean ATP values (2,579 relative light units [RLU]) compared with saline alone (1,436 RLU; P=.047). In the endoscope samples, culture yield using saline plus the PULL THRU (mean, 43 CFU; range, 1-400 CFU) was significantly higher than that of saline alone (mean, 17 CFU; range, 0-500 CFU; P<.001). In samples obtained using the saline+PULL THRU brush method, ATP values of samples classified as unacceptable were significantly higher than those of samples classified as acceptable (P=.001). CONCLUSION: Physiological saline flushing combined with PULL THRU brush to sample endoscopes generated higher ATP values and increased the yield of microbial surveillance culture. Consequently, the acceptance rate of endoscopes based on a defined CFU limit was significantly lower when the saline+PULL THRU method was used instead of saline alone

Transcriptional Activation of the Adenoviral Genome Is Mediated by Capsid Protein VI

Gene expression of DNA viruses requires nuclear import of the viral genome. Human Adenoviruses (Ads), like most DNA viruses, encode factors within early transcription units promoting their own gene expression and counteracting cellular antiviral defense mechanisms. The cellular transcriptional repressor Daxx prevents viral gene expression through the assembly of repressive chromatin remodeling complexes targeting incoming viral genomes. However, it has remained unclear how initial transcriptional activation of the adenoviral genome is achieved. Here we show that Daxx mediated repression of the immediate early Ad E1A promoter is efficiently counteracted by the capsid protein VI. This requires a conserved PPxY motif in protein VI. Capsid proteins from other DNA viruses were also shown to activate the Ad E1A promoter independent of Ad gene expression and support virus replication. Our results show how Ad entry is connected to transcriptional activation of their genome in the nucleus. Our data further suggest a common principle for genome activation of DNA viruses by counteracting Daxx related repressive mechanisms through virion proteins

Terrestrial Very-Long-Baseline Atom Interferometry:Workshop Summary

This document presents a summary of the 2023 Terrestrial Very-Long-Baseline Atom Interferometry Workshop hosted by CERN. The workshop brought together experts from around the world to discuss the exciting developments in large-scale atom interferometer (AI) prototypes and their potential for detecting ultralight dark matter and gravitational waves. The primary objective of the workshop was to lay the groundwork for an international TVLBAI proto-collaboration. This collaboration aims to unite researchers from different institutions to strategize and secure funding for terrestrial large-scale AI projects. The ultimate goal is to create a roadmap detailing the design and technology choices for one or more km-scale detectors, which will be operational in the mid-2030s. The key sections of this report present the physics case and technical challenges, together with a comprehensive overview of the discussions at the workshop together with the main conclusions

ARIA digital anamorphosis : Digital transformation of health and care in airway diseases from research to practice

Digital anamorphosis is used to define a distorted image of health and care that may be viewed correctly using digital tools and strategies. MASK digital anamorphosis represents the process used by MASK to develop the digital transformation of health and care in rhinitis. It strengthens the ARIA change management strategy in the prevention and management of airway disease. The MASK strategy is based on validated digital tools. Using the MASK digital tool and the CARAT online enhanced clinical framework, solutions for practical steps of digital enhancement of care are proposed.Peer reviewe

International genome-wide meta-analysis identifies new primary biliary cirrhosis risk loci and targetable pathogenic pathways.

Primary biliary cirrhosis (PBC) is a classical autoimmune liver disease for which effective immunomodulatory therapy is lacking. Here we perform meta-analyses of discovery data sets from genome-wide association studies of European subjects (n=2,764 cases and 10,475 controls) followed by validation genotyping in an independent cohort (n=3,716 cases and 4,261 controls). We discover and validate six previously unknown risk loci for PBC (Pcombined<5 × 10(-8)) and used pathway analysis to identify JAK-STAT/IL12/IL27 signalling and cytokine-cytokine pathways, for which relevant therapies exist