bluuebunny/crossref_metadata_2025 huggingface dataset:
Evolution of Methods for Measurement of HDL-Cholesterol: From Ultracentrifugation to Homogeneous Assays
Authors: Warnick G Russell , Nauck Matthias , Rifai Nader
Published: January 2020

Background: Adoption of automated homogeneous assays for HDL-cholesterol (HDL-C) is increasing, driven by the need of clinical laboratories to cope with increasing workloads while containing costs. However, performance characteristics of homogeneous assays often differ in important aspects from those of the earlier precipitation methods. This review provides an overview of the new generation of homogeneous assays for HDL-C within the historical context of the evolution of methods and the efforts to standardize measurements of the lipoproteins.

Approach: This is a narrative review based on method evaluations conducted in the laboratories of the authors as well as on relevant publications, especially comparative evaluation studies, from the literature. Publications considered here have been collected by the authors over the past 30 years of involvement as methods for HDL-C made the transition from their early use in lipid research laboratories to clinical laboratories and the recent emergence of homogeneous assays.

Content: The presentation includes descriptions of methodologies, including homogeneous, precipitation, electrophoresis, and ultracentrifugation assays. Reference methods and recommended approaches for assessing accuracy are described. Accuracy and imprecision are summarized in the context of the National Cholesterol Education Program (NCEP) standards for analytical performance. The effects of interfering substances and preanalytical sources of variation are presented.

Summary: Homogeneous assays have been shown to be reasonably well suited for use in routine clinical laboratories, generally meeting the NCEP criteria for precision, accuracy, and total error. However, discrepant results compared with the reference methods have been observed with some of the assays, and the sources of discrepancies are not well characterized. Some homogeneous reagents have not been thoroughly evaluated. At least three of the reagents have experienced successive adjustments in formulation; hence, the reagents may not yet be fully optimized. For these reasons, the homogeneous assays cannot be confidently recommended for use in long-term clinical trials and other research applications without thorough validation.
Effects of Blood-Processing Protocols on Fetal and Total DNA Quantification in Maternal Plasma
Authors: Chiu Rossa W K , Poon Leo L M , Lau Tze K , Leung Tse N , Wong Eric M C , Lo Y M Dennis
Published: January 2020

Background: Recently, apoptotic cells have been found in plasma obtained by centrifugation of blood from pregnant women, raising the question of what constitutes plasma and whether plasma is truly cell free. We compared the effects of different blood-processing protocols on the quantification, DNA composition, and day-to-day fluctuation of fetal and total DNA in maternal plasma.

Methods: Blood samples were collected from healthy pregnant women. The blood sample from each individual was simultaneously processed by different means, including the following: Percoll separation, centrifugation, microcentrifugation, and filtration. The resulting plasma aliquots were subjected to real-time quantitative amplification of the β-globin (for total DNA) and SRY (for fetal DNA) genes. The differences in the β-globin and SRY DNA concentrations and the degree of variation between the various plasma aliquots were assessed statistically.

Results: Different protocols of blood processing significantly affected the quantification and the day-to-day fluctuation of total (P <0.001), but not fetal (quantification, P = 0.336; fluctuation, P = 0.206), DNA in maternal plasma. The quantitative difference could be attributed to the fact that efficacies of different protocols for generating cell-free plasma vary. Processing blood samples by centrifugation followed by filtration or microcentrifugation is effective in producing cell-free plasma.

Conclusions: Standardization in plasma-processing protocols is needed for maternal plasma DNA analysis, especially for quantification of total DNA in maternal plasma. Such preanalytic factors may also affect other applications of plasma DNA analysis.
Association of Increased Ferritin with Premature Coronary Stenosis in Men
Authors: Haidari Mehran , Javadi Ebrahim , Sanati Arashmidos , Hajilooi Mehrdad , Ghanbili Jafar
Published: January 2020

Background: Body iron status has been implicated in atherosclerotic cardiovascular disease. The main hypothesis is that high iron status is associated with increased oxidation of LDL. We investigated the potential role of ferritin as an additional risk factor promoting atherosclerosis among a young population with coronary artery disease (CAD).

Methods: Four hundred consecutive patients (218 males, 182 females) referred for diagnostic coronary angiography were examined, and risk factors for CAD, lipids, C-reactive protein (CRP), and ferritin concentrations were recorded for all participants.

Results: Ferritin was higher in the male patients with CAD (121 μg/L; range, 56–258 μg/L) than in the men without significant CAD (73 μg/L; range, 32–138 μg/L; P <0.002). Multiple logistic regression analysis, after adjustment for the established coronary risk factors, showed ferritin as an independent discriminating risk factor for CAD (P <0.01). Men in the highest quartile of ferritin had an odds ratio (OR) of 1.62 [95% confidence interval (95% CI), 1.12–2.42; P <0.01] compared with men in the lowest quartile of ferritin. The association between ferritin and CAD was more pronounced in male patients ≤50 years (OR = 2.65; 95% CI, 1.35–5.51; P <0.003). Ferritin was significantly higher in diabetic male patients in comparison with nondiabetic male patients [168 μg/L (range, 74–406 μg/L) vs 106 μg/L (range, 44–221 μg/L), respectively; P <0.002]. No association was observed between ferritin and CAD among the female patients.

Conclusion: Our data suggest that increased ferritin might be an independent predictor of premature CAD in male Iranian patients.
Nitrogen Metabolism and Bone Metabolism Markers in Healthy Adults during 16 Weeks of Bed Rest
Authors: Scheld Kerstin , Zittermann Armin , Heer Martina , Herzog Birgit , Mika Claudia , Drummer Christian , Stehle Peter
Published: January 2020

Background: The associations between nitrogen metabolism and bone turnover during bed rest are still not completely understood.

Methods: We measured nitrogen balance (nitrogen intake minus urinary nitrogen excretion) and biochemical metabolic markers of calcium and bone turnover in six males before head-down tilt bed rest (baseline), during 2, 10, and 14 weeks of immobilization, and after reambulation.

Results: The changes in nitrogen balance were highest between baseline and week 2 (net change, −5.05 ± 1.30 g/day; 3.6 ± 0.6 g/day at baseline vs −1.45 ± 1.3 g/day at week 2; P<0.05). In parallel, serum intact osteocalcin (a marker of bone formation) was already reduced and renal calcium and phosphorus excretions were increased at week 2 (P <0.05). Fasting serum calcium and phosphorus values and renal excretion of N-telopeptide (a bone resorption marker) were enhanced at weeks 10 and 14 (P <0.05–0.001), whereas serum concentrations of parathyroid hormone, calcitriol, and type I collagen propeptide (a marker of bone collagen formation) were decreased at week 14 (P <0.05–0.01). Significant associations were present between changes of serum intact osteocalcin and 24-h calcium excretion (P <0.001), nitrogen balance and 24-h phosphorus excretion (P <0.001), nitrogen balance and renal N-telopeptide excretion (P <0.05), and between serum osteocalcin and nitrogen balance (P <0.025).

Conclusions: Bone formation decreases rapidly during immobilization in parallel with a higher renal excretion of intestinally absorbed calcium. These changes appear in association with the onset of a negative nitrogen balance, but decreased bone collagen synthesis and enhanced collagen breakdown occur after a time lag of several weeks.
Telomerase Detection in Body Fluids
Authors: Hess Jennifer L , Highsmith W Edward
Published: January 2020

Background: Telomerase is a ribonucleoprotein that maintains chromosomal telomere length. Telomerase is not active in nonmalignant somatic cells, but is activated in most human cancers. Telomerase activity in easily obtainable body fluids that bathe tumors may be a useful cancer marker, especially when used in conjunction with conventional cytology.

Approach: Results from studies that assayed telomerase activity in easily obtainable body fluids are reviewed.

Content: The telomerase repeat amplification protocol (TRAP) assay has been used to measure telomerase activity in body fluids, including ascites, pleural effusions, pelvic washes, bronchial washings, bronchial lavage, urine, bladder washings, oral rinses, and plasma. Telomerase activity has sensitivities of 60–90% as a tumor marker with clinical specificities for cancer of ∼90%. Telomerase activity is more sensitive than conventional cytology, the sensitivity of which was 40–65% in various studies.

Summary: Telomerase activity in body fluids, as measured by the TRAP assay, is a sensitive potential tumor marker that might help increase the cancer detection rate and the cancer treatment success rate when combined with conventional cytology.
Stability of Endogenous and Added RNA in Blood Specimens, Serum, and Plasma
Authors: Tsui Nancy BY , Ng Enders KO , Lo YM Dennis
Published: January 2020

Background: Circulating RNA in plasma/serum is an emerging field for noninvasive molecular diagnosis. Because RNA is widely thought to be labile in the circulation, we investigated the stability and various preanalytical factors that may affect RNA concentrations in blood specimens.

Methods: Blood samples were collected from 65 healthy volunteers. The effects of two preanalytical variables were studied: (a) time delay in processing of EDTA blood and clotted blood after venesection, and (b) freezing and thawing of plasma and serum. The lability of free added RNA in plasma was also investigated. Plasma/serum RNA was measured by a real-time quantitative reverse transcription-PCR assay for glyceraldehyde 3-phosphate dehydrogenase mRNA, whereas DNA was measured by a real-time quantitative PCR assay for the β-globin gene.

Results: No significant difference was found for plasma RNA concentrations obtained from uncentrifuged EDTA blood that had been left at 4 °C for 0, 6, and 24 h (P =0.182). On the other hand, the serum RNA concentrations increased significantly over 24 h when uncentrifuged clotted blood was stored at 4 °C (P <0.05). In comparison, >99% of the free added RNA could no longer be amplified after incubation in plasma for 15 s. Never-frozen plasma, freeze-thawed plasma, and thawed plasma left at room temperature for 1 h showed no significant differences in RNA concentration (P =0.465). No significant difference was observed for freeze-thawed serum (P = 0.430).

Conclusions: Plasma RNA is stable in uncentrifuged EDTA blood stored at 4 °C, but to obtain a stable serum RNA concentration, uncentrifuged clotted blood should be stored at 4 °C and processed within 6 h. A single freeze/thaw cycle produces no significant effect on the RNA concentration of plasma or serum.
Comprehensive Scanning of the Entire Mitochondrial Genome for Mutations
Authors: Wong Lee-Jun C , Liang Min-Hui , Kwon Haeyoung , Park John , Bai Ren-Kui , Tan Duan-Jun
Published: January 2020

Background: Definitive molecular diagnosis of mitochondrial disorders has been greatly hindered by the tremendous clinical and genetic heterogeneity, the heteroplasmic condition of pathogenic mutations, and the presence of numerous homoplasmic mitochondrial DNA (mtDNA) variations with unknown significance. We used temporal temperature gradient gel electrophoresis (TTGE) to detect heteroplasmic mutations from homoplasmic variations in the whole mitochondrial genome.

Methods: We screened 179 unrelated patients by TTGE with use of 32 overlapping primer pairs. Mutations were identified by direct sequencing of the PCR products and confirmed by PCR with allele-specific oligonucleotide or restriction fragment length polymorphism analysis.

Results: We detected 71 heteroplasmic and 647 homoplasmic banding patterns. Sequencing of the heteroplasmic fragments identified 68 distinct novel mutations and 132 reported sequence variations and mutations; most of them occurred only once. The deleterious nature of some of the novel mutations was established by analyzing the asymptomatic family members and the biochemical and molecular characteristics of the mutation. When the number of mutations was normalized to the size of the region, the occurrence of mutations was 2.4 times more frequent in the tRNA genes than in the mRNA (protein coding) regions.

Conclusions: Screening by TTGE detects low proportions of mutant mtDNA and distinguishes heteroplasmic from homoplasmic variations. Results from comprehensive molecular analysis should be followed up with clinical correlation to establish a guideline for complete mutational analysis of the entire mitochondrial genome and to facilitate the diagnosis of mitochondrial disorders.
Simple Method for Quantification of Bence Jones Proteins
Authors: Salomo Morten , Gimsing Peter , Nielsen Lars B
Published: January 2020

Background: Quantification of free monoclonal light chains in urine [Bence Jones proteins (BJPs)] is used to diagnose multiple myeloma and to evaluate response to treatment. We have developed and evaluated an optimized approach for quantification of BJPs.

Methods: High-resolution gel electrophoresis of unconcentrated urine and albumin calibrators was carried out on Sebia’s Hydrasys instrument with Hydragel HR agarose gels. After staining with acid violet, the gels were scanned densitometrically. The staining intensities of BJP bands relative to the staining intensities of albumin solutions were used to determine the BJP concentrations. Results for patient samples were compared with conventional agarose gel electrophoresis on concentrated samples.

Results: The relationships between staining intensity and the protein concentrations of albumin and BJPs were linear up to protein concentrations of ∼2000 mg/L. The detection limit was ∼20 mg/L. The interassay imprecision (CV) was ∼8% (n = 23, duplicate analysis), and the results (y) showed a close positive relationship to the comparison method: slope = 0.82 (confidence interval, 0.75-0.88); y-intercept = 34 (−14 to 81) mg/L; n = 29; r2 = 0.96.

Conclusions: Agarose gel electrophoresis of unconcentrated urine samples together with a series of albumin calibrators followed by acid violet staining and densitometric scanning is sufficiently reproducible and sensitive to quantify clinically relevant BJPs.
Drug Testing in Blood: Validated Negative-Ion Chemical Ionization Gas Chromatographic–Mass Spectrometric Assay for Determination of Amphetamine and Methamphetamine Enantiomers and Its Application to Toxicology Cases
Authors: Peters Frank T , Kraemer Thomas , Maurer Hans H
Published: January 2020

Background: Enantioselective analysis of amphetamine (AM) or methamphetamine (MA) in urine is already a well-established tool for differentiation of illicit from therapeutic ingestion of AM or MA derivatives. However, because of the increasing importance of plasma or serum in analytical toxicology, a method for enantioselective analysis of AM and MA in these matrices is needed.

Methods: AM and/or MA were extracted from 0.2 mL of blood plasma or serum by mixed-mode solid-phase extraction. After derivatization with S-(−)-heptafluorobutyrylprolyl chloride, the resulting diastereomers were separated by gas chromatography on a HP-5MS column during a 15-min program and detected by mass spectrometry in the negative-ion chemical ionization mode (NICI-GC-MS). The method was fully validated and applied to >50 samples from authentic toxicology cases.

Results: The derivatized AM and MA enantiomers were well separated and sensitively detected. The method was linear from 5 to 250 μg/L per enantiomer with analytical recoveries, accuracy, and within- and between-run precision well within required limits. Extraction yields were 88.9–98.6%. Implications of concentrations and enantiomeric composition of AM and MA in the authentic samples were considered.

Conclusions: This sensitive, reliable, rapid NICI-GC-MS assay is suitable for enantioselective determination of AM and MA in blood plasma or serum samples.
Persistent Müllerian duct syndrome due to anti-Müllerian hormone receptor 2 microdeletions: a diagnostic challenge
Authors: Tosca L , Giltay J C , Bouvattier C , Klijn A J , Bouligand J , Lambert A S , Lecerf L , Josso N , Tachdjian G , Picard J Y
Published: January 2020

The persistent Müllerian duct syndrome (PMDS) is defined by the persistence of Müllerian derivatives in an otherwise normally virilized 46,XY male. It is usually caused by mutations in either the anti-Müllerian hormone (AMH) or AMH receptor type 2 (AMHR2) genes. We report the first cases of PMDS resulting from a microdeletion of the chromosomal region 12q13.13, the locus of the gene for AMHR2. One case involved a homozygous microdeletion of five exons of the AMHR2 gene. In the second case, the whole AMHR2 gene was deleted from the maternally inherited chromosome. The patient’s paternal allele carried a stop mutation, which was initially thought to be homozygous by Sanger sequencing. Diagnostic methods are discussed, with an emphasis on comparative genomic hybridization and targeted massive parallel sequencing.
Interrogating the Gut-Brain Axis in the Context of Inflammatory Bowel Disease: A Translational Approach
Authors: Collins Stephen M
Published: January 2020

This review examines preclinical and clinical studies relevant to our understanding of how the bidirectional gut-brain axis influences the natural history of inflammatory bowel disease. Preclinical studies provide proof of concept that preexisting behavioral illness, such as depression, results in increased susceptibility to inflammatory stimuli and that commonly used classes of antidepressants protect against this vulnerability. However, clinical studies suggesting behavioral illness as a risk factor for IBD and a protective role for antidepressants have relied primarily on symptom-reporting rather than objective measurements of inflammation. In terms of gut-to-brain signaling, there is emerging evidence from preclinical and clinical observation that intestinal inflammation alters brain functions, including the induction of mood disorders, alteration of circadian rhythm both centrally and peripherally, and changes in appetitive behaviors. Furthermore, preclinical studies suggest that effective treatment of intestinal inflammation improves associated behavioral impairment. Taken together, the findings of this review encourage a holistic approach to the management of patients with IBD, accommodating lifestyle issues that include the avoidance of sleep deprivation, optimized nutrition, and the monitoring and appropriate management of behavioral disorders. The review also acknowledges the need for better-designed clinical studies evaluating the impact of behavioral disorders and their treatments on the natural history of IBD, utilizing hard end points to assess changes in the inflammatory process as opposed to reliance on symptom-based assessments. The findings of the review also encourage a better understanding of changes in brain function and circadian rhythm induced by intestinal inflammation.
Temporal Gut Microbial Changes Predict RecurrentClostridiodes DifficileInfection in Patients With and Without Ulcerative Colitis
Authors: Lee Allen A , Rao Krishna , Limsrivilai Julajak , Gillilland Merritt , Malamet Benjamin , Briggs Emily , Young Vincent B , Higgins Peter D R
Published: January 2020

Background

Ulcerative colitis (UC) carries an increased risk of primary and recurrent Clostridiodes difficile infection (rCDI), and CDI is associated with UC flares. We hypothesized that specific fecal microbial changes associate with UC flare and rCDI.

Methods

We conducted a prospective observational cohort study of 57 patients with UC and CDI, CDI only, and UC only. Stool samples were collected at baseline, at the end of antibiotic therapy, and after reconstitution for 16S rRNA sequencing. The primary outcomes were recurrent UC flare and rCDI. Logistic regression and Lasso models were constructed for analysis.

Results

There were 21 (45.7%) patients with rCDI, whereas 11 (34.4%) developed UC flare. Patients with rCDI demonstrated significant interindividual (P = 0.008) and intraindividual differences (P = 0.004) in community structure by Jensen-Shannon distance (JSD) compared with non-rCDI. Two cross-validated Lasso regression models predicted risk of rCDI: a baseline model with female gender, hospitalization for UC in the past year, increased Ruminococcaceae and Verrucomicrobia, and decreased Eubacteriaceae, Enterobacteriaceae, Lachnospiraceae, and Veillonellaceae (AuROC, 0.94); and a model 14 days after completion of antibiotics with female gender, increased Shannon diversity, Ruminococcaceae and Enterobacteriaceae, and decreased community richness and Faecalibacterium (AuROC, 0.9). Adding JSD between baseline and post-treatment samples to the latter model improved fit (AuROC, 0.94). A baseline model including UC hospitalization in the past year and increased Bacteroidetes was associated with increased risk for UC flare (AuROC, 0.88).

Conclusion

Fecal microbial features at baseline and after therapy predict rCDI risk in patients with and without UC. These results may help risk stratify patients to guide management.
P101 SARCOPENIA IS ASSOCIATED WITH INCREASED RISK OF INFECTION IN IBD PATIENTS OLDER THAN 50 YEARS STARTING BIOLOGIC MEDICATIONS
Authors: Campbell James , Teigen Levi , Feussom Ghislaine , Price Kathleen , Cogswell Rebecca , Colombel Jean-Frederic , Shmidt Eugenia
Published: January 2020

Background

The relationship between sarcopenia and clinical outcomes outside of the post-operative setting has not been well-studied in IBD patients. We aimed to characterize the prevalence of sarcopenia and its association with adverse events and clinical response in IBD patients starting on new biologic medications.

Methods

We identified a retrospective cohort of adult (≥18 years old) IBD patients at the University of Minnesota with an abdominal CT or MRI within 6 months prior to or one month after starting a new biologic medication. Baseline demographics, disease severity (based on Physician’s Global Assessment from GI encounters), laboratory values and clinical outcomes for one year after the start of medications were obtained from chart review. The primary endpoint was incidence of adverse events (defined as infection, need for surgery, or hospitalization) within one year after medication start. The secondary outcome was clinical response as assessed based on clinical, endoscopic, and radiographic follow-up. Skeletal muscle index (SMI) measures were obtained from CT/MRI images at the L3 vertebral level. Published cut-offs (SMI of 38.5 cm2/m2 for women and 52.4 cm2/m2 for men) were used to determine the presence of sarcopenia. Associations between sarcopenia and outcomes were analyzed using logistic regression and age stratification was performed.

Results

Ninety-four patients (74 CD, 20 UC, 47% male, mean age 38 years) were included in the analysis. The majority of patients had moderate disease severity or worse (83%) at the time of medication start, and 75% were started on anti-TNF medications. Overall prevalence of sarcopenia was 57% (57% CD, 60% UC). In the entire cohort, sarcopenia was not associated with increased rates of adverse events (OR = 0.516, 95% CI 0.217–1.225; p= 0.1334). However, in patients ≥ 50 years of age (n=23), those with sarcopenia had higher risk of infection at 1 year compared to those without sarcopenia (85% vs. 20%; OR = 21.99, 95% CI 3.086–262.515; p = 0.0051). The majority of infections were those that required antibiotics or antivirals, but not hospitalization. Examples included sinusitis, upper respiratory infections, periodontitis, urinary tract infection, herpes simplex, and two cases of C. difficile. Furthermore, when controlling for disease duration and concomitant steroid use, the association between sarcopenia and infection remained significant. In this age subgroup, there was no association between sarcopenia and clinical response to therapy.

Conclusions

In our cohort, sarcopenia was present in the majority of adult IBD patients starting new biologic medications. In patients ≥ 50 years old, sarcopenia was associated with an increased risk of infections. Further work is needed to validate these findings and to understand which patients may benefit from sarcopenia assessment prior to biologic start.
18 A HUMAN-DERIVED 5-MER PEPTIDE (MTADV), WHICH RESTRICTIVELY ALLEVIATES THE PRO-INFLAMMATORY ACTIVITY OF SERUM AMYLOID A (SAA), SUBSTANTIALLY AMELIORATES IBD PATHOLOGY: NEW POTENTIAL DRUG (MTADV) AND THERAPEUTIC TARGET CANDIDATE (SAA) FOR IBD
Authors: Naor David , Hemed-Shaked Maayan , Cowman Mary , Kim Jin , Armengol Jordi , Alemany Jorge , Kanduc Darja
Published: January 2020

A human anti-inflammatory 5-MER peptide MTADV (methionine-threonine-alanine-aspartic acid-valine) derived from a sequence of a pro-inflammatory CD44 variant was first described by us in Nedvetzki et al., J. Clin. Invest. 111,1211-1220, 2003. A NCBI BLAST RefSeq protein sequence database, that contains at present 81,347 human protein sequences, reveals that only one additional protein contains this unique MTADV sequence of alternating hydrophobic and hydrophilic amino acids. This human peptide displays an efficient anti-inflammatory effect in Inflammatory Bowel Disease (IBD), Rheumatoid Arthritis and Multiple sclerosis mouse models, all share amyloid proteins in the relevant target organs. We show here that the 5-MER peptide, administered after the onset of disease,either by SC injections or oral delivery, inhibited the pathological activity of two IBD models (TNBS and DSS), as indicated by histopathology analysis (Ameho’s score) or Bioluminescence Imaging, sensing the release of Reactive Oxdative Species (ROS), which reflect colon inflammation (Fig 1). The 5-MER MTADV peptide, while inhibiting chronic inflammation, neither interferes with normal immune responses nor induce neutralizing antibodies. A pull-down experiment with mass spectrometric analysis revealed that the 5-MER peptide specifically binds to serum amyloid A (SAA), a stress pro-inflammatory protein, which generates aggregated amyloid deposits in the IBD colon (de Villiers at al Cytokine 12:1337-1347,2000). Our studies further provide in vitro evidence, strengthened by in vivo experiments, that SAA is a significant target of this pentamer. To this end, the 5-MER peptide (but not the corresponding scrambled peptide) inhibits the release of the pro-inflammatory cytokines IL-6 and IL-1β from SAA-activated fibroblasts. Furthermore, the 5-MER peptide was found to retard the early stages of amyloid-type aggregation of SAA in solution (Fig 2). Adopting the β-sheet conformation, MTADV would display opposing hydrophobic and hydrophilic faces that could interact with the β-sheet-forming amyloidogenic sequences in SAA. This suggests that the mechanism of action for the 5-MER peptide in vivo may depend on its ability to slow the aggregation of SAA, thus reduce its contribution to chronic inflammation. Finally, using bioinformatics and qRT-PCR, we have found the pentamer up-regulates set of genes involved in resistance to chronic inflammations. Hence, our study provides both a new potential drug (MTADV) and a new therapeutic target candidate (SAA) for IBD.
P154 THE ANTI-INFLAMMATORY EFFECT OF A PLANT-BASED DIET IN DSS-INDUCED COLITIS
Authors: Basson Abigail , Kulpins Danielle , LaSalla Alexandria , Di Martino Luca , Rodriguez-Palacios Alexander , Cominelli Fabio
Published: January 2020

Background

Inflammatory Bowel Disease (IBD) results from an imbalanced immune response toward the gut microbiota. Disruptions in the gut microbiota are proposed as critical elements that could foster pro-inflammatory imbalances and cause active flares IBD. Diet has been shown to have a pro-inflammatory effect in Crohn’s disease (CD) but not much is known on the anti-inflammatory effect of a plant-based diet. Gut microbial modulation via diet is a needed strategy for the therapeutic management of CD; however, no specific recommendations exist for CD patients. We hypothesized that plant-based diet may have a protective effect on the severity of acute dextran sodium sulfate (DSS)-induced colitis.

Methods

20-wk old age-sex matched C57BL/6 mice were fed a plant-based diet or standard chow for 7 days. Mice were then administered 3% DSS for 7 days and sacrificed 2 days later. Mice were individually caged and maintained on Aspen bedding. A fecal homogenization protocol was performed prior to the start of the experiment. We monitored body weight, fecal myeloperoxidase activity, water and food intake in all mice. Epithelial permeability (FITC-dextran) and endoscopic assessment were performed before and after DSS treatment. Post mortem blood, mesenteric lymph node, colon and liver were collected. Colon morphology was evaluated using 3D-stereomicroscopy. Histological assessment was performed in a blinded manner. Analysis of samples by Illumina 16S protocols, real-time quantitative PCR (qPCR), flow cytometry.

Results

Mice fed the plant-based diet had decreased DSS-induced weight loss, myeloperoxidase activity and mortality compared to chow-fed mice (P<0.05). Plant-fed mice also had significantly lower colonoscopy and histological scores, with colonoscopic images showing decreased transparency and increased presence of erosion and strictures in chow-fed mice. Compared to the chow-fed mice, the concentration of FITC-D in the plasma of plant-fed mice was significantly lower (Mann-Whitney P=0.004, power 0.98). Bacterial quantification qPCR CT-values (DNA copy abundance) revealed differences in the microbiota composition with increased abundance for Lactobacilli in the context of a generic/universal 16S bacterial primer and the host gut cell DNA copies using β-actin in the plant diet mice. Illumina 16S rRNA gene sequencing of fecal genomic DNA samples is in progress.

Conclusion

Our results show that a plant-based diet has a protective effect on DSS-induced colitis. Further studies are needed to investigate the potential underlying mechanisms. These findings may have great potential for therapeutic dietary interventions in individuals with IBD.
P067 IMPACT OF MIRIKIZUMAB TREATMENT ON HEALTH-RELATED QUALITY OF LIFE IN PATIENTS WITH ULCERATIVE COLITIS: A PHASE 2 STUDY ANALYSIS USING THE SF-36 V2 STANDARD
Authors: Lichtenstein Gary , Feagan Brian , Tuttle Jay , Morris Nathan , Dong Yan , Arora Vipin , Naegeli April , Rubin David
Published: January 2020

Background

Mirikizumab (LY3074828) is a humanized monoclonal antibody directed against the p19 subunit of IL-23, and had demonstrated efficacy in psoriasis, ulcerative colitis (UC), and Crohn’s disease. The effect of miri on health-related quality of life (HRQoL) as measured by the 36-Item Short Form Health Survey v2 Standard (SF-36) was examined in a Phase 2, multicenter, randomized, parallel-arm, double-blind placebo (PBO)-controlled trial (NCT02891226) in patients with moderate to severely active UC.

Methods

Patients were randomized 1:1:1:1 to receive intravenous PBO, miri 50mg or 200mg with possibility of exposure-based (EB) dose increases, or fixed miri 600mg every 4 weeks (Q4W), with efficacy assessment at Week 12. Patients with clinical response to miri at Week 12 were re-randomized 1:1 to a double-blind maintenance treatment of miri 200mg subcutaneously (SC) Q4W or 12 weeks (Q12W) and were treated through Week 52. The SF-36, a 36-item patient-completed questionnaire with recall period past 4-weeks, that measures 8 domains (physical functioning, role-physical, role-emotional, bodily pain, vitality, social functioning, mental health, and general health) and two summary scores, physical component summary (PCS) and mental component summary (MCS), was assessed at baseline and at Weeks 4, 8, 12, 24, 32, and 52. Treatment comparisons were evaluated using mixed effects for repeated measures for mean improvement of PCS, MCS, and domain score changes with treatment, geographic region, and prior biologic UC therapy use as factors.

Results

The PCS was significantly higher in all miri doses at Week 12 (Least Squares Mean [LSM] ± SE; PBO: 45.4±0.8; miri 50mg: 48.2±0.9, p=0.011; 200mg: 48.0±0.8, p=0.022; 600mg: 49.0±0.9, p=0.002; Fig 1A), while the MCS was significantly higher in the miri 200mg and 600mg groups (PBO: 42.5±1.2; miri 50mg: 43.8±1.3; 200mg: 46.1±1.2, p=0.028; 600mg: 48.1±1.3, p<0.001; Fig 1B). Of the 8 SF-36 domains, 7 (physical functioning, role-physical, role-emotional, social functioning, vitality, bodily pain, and mental health) showed statistically significant improvement at Week 12 in at least one miri dose group compared to PBO, while 1 (general health) demonstrated a numerical improvement (Fig 1C). Patients who continued onto randomized maintenance treatment sustained the improved scores through Week 52 (Fig 2). These results paralleled the observed treatment associated benefits on clinical response and remission rates at Weeks 12 and 521.

Conclusion

Mirikizumab treatment results in significant improvements in patient HRQoL demonstrated by significantly improved SF-36 scores after 12 weeks of induction treatment, which were sustained during an additional 40 weeks of maintenance treatment.
P069 ONTAMALIMAB, A FULLY HUMAN MONOCLONAL ANTIBODY AGAINST MUCOSAL ADDRESSIN CELL ADHESION MOLECULE-1, PROVIDES SUSTAINED EXPOSURE FOLLOWING LONG-TERM TREATMENT IN PATIENTS WITH ULCERATIVE COLITIS
Authors: Wang Yi , Marier J F , Kheibarshekan Leila , Kassir Nastya , Martin Patrick
Published: January 2020

Introduction

Ontamalimab is a fully human immunoglobulin G2 monoclonal antibody against mucosal addressin cell adhesion molecule-1 in development for the induction and maintenance of clinical remission in patients with ulcerative colitis (UC). This study aimed to assess the long-term pharmacokinetics (PK) of ontamalimab in patients with UC, and the effects of concomitant medications on PK parameters.

Methods

A 12-week induction study (TURANDOT; NCT01620255) was performed to assess the PK, efficacy and safety of ontamalimab (7.5, 22.5, 75 and 225 mg subcutaneous [s.c.] every four weeks [Q4W]) in patients with UC. Individuals who completed the induction study were eligible for enrollment in an open-label extension (OLE) study (TURANDOT II; NCT01771809) to assess the long-term PK, efficacy and safety of ontamalimab (75 or 225 mg s.c. Q4W up to week 72). Population PK analyses were performed using nonlinear mixed-effects modelling. Exposure-response analyses were performed to assess the relationships between minimum concentration (Cmin,ss) of ontamalimab and clinical response, clinical remission and mucosal healing. The effect of concomitant treatments (used for ≥20% of treatment duration) on PK parameters was also evaluated.

Results

The PK population included 130 (39.8%) women and 197 (60.2%) men, of median age of 40 years. A 1-compartment model with linear elimination adequately described the PK of ontamalimab. Population estimates of apparent clearance (CL/F) and volume of distribution (V/F) were 0.00917 L/h (0.22 L/day) and 7.44 L, respectively. Albumin had a significant effect on the variability of CL/F. Individuals with albumin levels of 30 g/L and 47 g/L are expected to have CL/F values 44% higher and 23% lower, respectively, than a typical patient with an albumin level of 39 g/L. Anti-inflammatory agents affected CL/F, such that CL/F is expected to be 14% higher in patients receiving than not receiving these agents. Other medications including immunosuppressants, steroids and treatments for peptic ulcers and gastroesophageal reflux disease had no effect on CL/F. Weight was the only covariate that significantly affected V/F. The half-life of ontamalimab was 23.4 days. Concentrations of ontamalimab over 72 weeks in the OLE study were consistent with those observed in the 12-week induction study. Ontamalimab Cmin,ss was related to efficacy, such that at week 16 (week 28 in total including induction), patients with higher Cmin,ss values were more likely to have clinical response, clinical remission and mucosal healing than those with lower Cmin,ss.

Conclusion

The exposure to ontamalimab was sustained following prolonged treatment in patients with UC for up to 72 weeks. Higher ontamalimab exposure was associated with a higher probability of clinical response.
P059 SIGNIFICANTLY INCREASED MUCOSAL EOSINOPHILS IN PATIENTS WITH POST-IBD IBS-D SYNDROME MAY HAVE A NEW AVENUE
Authors: Tozlu Mukaddes , Beketaev Ilimbek , Younes Mamoun , Dar Wasim , Ju Changqing , Ertan Atilla
Published: January 2020

Introduction

Inflammatory bowel disease (IBD) patients in remission who present with irritable bowel syndrome with diarrhea (IBS-D) like symptoms pose a diagnostic and therapeutic dilemma that is called post-IBD IBS-D. This syndrome was first reported by Isgar et al. who documented IBS-type symptoms in 33% of their patients with chronic ulcerative colitis (CUC) in remission in 1983. In their meta-analysis of 13 studies incorporating 1,703 patients, Halpin and Ford in 2012 calculated a pooled prevalence of 35% for IBS symptoms among IBD subjects in remission. The post-IBD IBS-D syndrome represents a source of considerable stress, incurs morbidity and impairs quality of life of these patients. The etiology of this syndrome is unknown and may be multifactorial such as environmental, psychological, GI motility disarray, genetic components and possibly by gut microbiome.

Methods

We conducted a retrospective, single-center study of patients with history of IBD and IBS-D like symptoms. The demographic characteristics, type of IBD, colonoscopy, biopsy findings and empirical medical management outcome were analyzed.

Results

We have found significantly increased mucosal eosinophils (>50 HPF in both side of the colon examined) in our patients with post-IBD IBS-D syndrome. In this study, 15 CUC patients and 20 Crohn’s disease patients with this syndrome were investigated and showed no clinical, serological, mucosal or microscopic IBD activity. These patients had non-bloody diarrhea (5–20 x daily), lower abdominal cramps (%90), mild weight loss (4–6 lbs) and fecal incontinence (%25) who were placed on the GI-hypoallergenic diet and budesonide therapy. Sixty seven percent of these 35 patients with post-IBD IBS-D responded well clinically to this management. Increased mucosal eosinophils in these patients with post-IBD IBS-D may have eosinophilic colopathy that may be related to intestinal permeability disarray. The epithelium in these IBD patients who are in remission may have the production of pro-inflammatory cytokines by the eosinophils, especially IL-23 and IL-33. This interesting area investigation is in progress by our research faculty.

Conclusion

In summary, this is an exciting new finding that significantly increased mucosal eosinophils in our patients with post-IBD IBS-D syndrome may have a new avenue, eosinophilic colopathy. Interestingly, these patients have responded to the GI-hypoallergenic diet and budesonide therapy. Obviously, we need more patients in the near future with longer follow-up and we hope that the other investigators may confirm our findings.
P102 ANEMIA IN IBD PATIENTS ASSOCIATED WITH INCREASED HEALTHCARE UTILIZATION
Authors: Blaney Hanna , Mathew Antony , Snellings Robin , Duong Camille , England Julie , Vu Peter , Keum Yo Sup , Hammonds Kendall , Johnson Christopher
Published: January 2020

Introduction

Anemia is a common systemic complication of inflammatory bowel disease (IBD), with reported prevalence up to 74%. Anemia is associated with worse disease outcome and quality of life. The Crohn’s and Colitis Foundation recently published an initiative quantifying the healthcare cost of IBD patients. They found that the presence of anemia was associated with high resource utilization, particularly hospitalization. The purpose of this study was to determine how anemia severity impacts healthcare utilization and if treatment of anemia reduces healthcare utilization.

Methods

An IRB-approved retrospective chart review including patients 18 years of age and older at a tertiary referral center with IBD (Crohn’s Disease (CD) or Ulcerative Colitis (UC)) who had a CBC and an encounter with gastroenterology (GI) from 2014–2018. Demographic, laboratory, treatment, and visit data were extracted from electronic records and records were manually reviewed for verification. Anemia and anemia severity were defined by the World Health Organization classification of anemia.

Results

930 patients with IBD were included in the analysis, with 516 (56%) patients with CD, 409 (44%) with UC, and 5 (0.5%) with unspecified IBD. Of these patients, 502 (54%) had anemia based on lab values, with 194 (39%) having mild anemia, 244 (49%) with moderate anemia, and 64 (13%) with severe anemia. 321 unique patients had at least one admission during this time period. Patients with anemia had more hospitalizations (p=0.0019) and longer hospital stays, averaging 1.9 days longer (p<0.0001), compared to patients with IBD without anemia. Patients with anemia also had statistically more visits to the ED (p=0.0037), GI clinic visits (p<0.0001), and primary care visits (p=0.0036) as compared to patients with IBD without anemia. Furthermore, patients with severe anemia had an increase in number of hospitalizations (p=0.0169), increased length of stay (p=0.0344) and more GI visits (p=0.043) when compared to patients with less severe anemia. Treatment of anemia did not result in a statistically significant reduction in utilization of care as measured through number of hospitalizations, length of stay, or outpatient visits when compared to patients with anemia who were not treated.

Discussion

Our results demonstrate that the presence of anemia is correlated with increased health care utilization in patients with IBD, with increasing anemia severity associated with a higher utilization of care. Our results also suggest that treating anemia does not reduce health care utilization. Anemia has been associated with increased disease activity and could represent a marker of more severe disease, possibly explaining these associations. Future research is needed to determine if treatment of anemia in IBD results in reduced health care visits and costs.
23 EFFECTS OF VEDOLIZUMAB ON EXTRAINTESTINAL MANIFESTATIONS IN INFLAMMATORY BOWEL DISEASE
Authors: Dimopoulos Christina , Hung Kenneth , Proctor Deborah , Ruggiero Eilzabeth , Al-Bawardy Badr
Published: January 2020

Background

Vedolizumab (VDZ) is an α4β7 integrin monoclonal antibody that is effective for both Crohn’s disease (CD) and ulcerative colitis (UC). There is limited data on how vedolizumab’s gut selective mechanism impacts extraintestinal manifestations (EIM) in inflammatory bowel disease (IBD).

Methods

A retrospective study of all IBD patients at our institution who received at least 1 dose of VDZ between 1/1/2014 and 8/1/2019 were included. Primary outcome was rate of new or worsening EIMs after initiation of VDZ. Secondary outcomes were factors associated with new or worsening EIMs after VDZ. Continuous variables were analyzed using an unpaired student’s t-test. Categorical variables were analyzed using a chi-square test or Fisher’s exact test. A multivariable logistic regression model was built to identify factors associated with EIMs. The final model was obtained by sequentially subtracting variables and retaining variables that were independently associated with EIMs while controlling for other factors. We also identified factors associated with new or worsening of EIMs after VDZ therapy using Fisher’s exact testing and univariable logistic regression.

Results

A total of 142 patients were included (53.5% with CD, 41.6% with UC, and 4.9% with indeterminate colitis). The median age was 39 years (range 15–78), and 49.3% were women (Table 1). EIMs were noted in 53.5% (n=76) and included: peripheral arthritis (39.4%), sacroileitis (15.5%), erythema nodosum/pyoderma gangrenosum (9.2%), uveitis (4.9%), and episcleritis (0.7%). Twenty patients had more than one EIM. Significant factors associated with having EIMs included female sex (63.2% with EIM vs 33.3% without, p-value < 0.01), tobacco use (23.7% with EIM vs 6.1% without, p < 0.01), CD (60.5% with EIM vs 45.5% without, p=0.02), and prior biologic agent use (94.7% with EIM vs 74.2% without, p < 0.01). On multivariable analysis, female sex (OR 3.19, 95% CI 1.52–6.72), tobacco use (OR 4.84, 95% CI 1.45–16.23), and prior biologic agent use (OR 4.30, 95% CI 1.30–14.20) were the only significant factors.

Of the 76 patients with EIMs, 71.1% (n=54) had stable or improved EIMs after starting VDZ and 28.9% (n=22) had worsening or new EIMs after VDZ. Patients who were on steroids at the time of VDZ induction were more likely to have stable or improved EIMs compared to who did not receive concomitant steroid therapy (OR for worsened EIMs 0.20, 95% CI 0.06–0.63) (Table 2). Prior biologic use and current combination therapy did not significantly predict worsening or new EIMs.

Conclusion

Female sex, tobacco use, and prior biologic agent use are significantly associated with EIMs of IBD. Post-VDZ, the majority of patients had stable to improved EIMs. Patients on concomitant steroid therapy at the time of VDZ induction were more likely to have improved or stable EIMs compared to patients not on steroids.