bluuebunny/crossref_metadata_2025 huggingface dataset:
Direct Assay for Cobalamin Bound to Transcobalamin (Holo-Transcobalamin) in Serum
Authors: Ulleland Marius , Eilertsen Ingar , Quadros Edward V , Rothenberg Sheldon P , Fedosov Sergey N , Sundrehagen Erling , Örning Lars
Published: January 2020

Background: Only cobalamin carried by transcobalamin (holo-transcobalamin) is available for cellular uptake and hence is physiologically relevant. However, no reliable or accurate methods for quantifying holo-transcobalamin are available. We report a novel holo-transcobalamin assay based on solid-phase capture of transcobalamin.

Methods: A monoclonal antibody specific for human transcobalamin with an affinity constant >1010 L/mol was immobilized on magnetic microspheres to capture and concentrate transcobalamin. The cobalamin bound to transcobalamin was then released and assayed by a competitive binding radioassay. The quantification of holo-transcobalamin was accomplished using calibrators composed of recombinant, human holo-transcobalamin.

Results: The assay was specific for holo-transcobalamin and had a detection limit of 5 pmol/L. Within-run and total imprecision (CV) was 5% and 8–9%, respectively. The working range (CV <20%) was 5–370 pmol/L. Dilutions of serum were linear in the assay range. The recovery of recombinant, human holo-transcobalamin added to serum was 93–108%. A 95% reference interval of 24–157 pmol/L was established for holo-transcobalamin in 105 healthy volunteers 20–80 years of age. For 72 of these sera, holo-haptocorrin and total cobalamin were also determined. Whereas holo-haptocorrin correlated well (r2 = 0.87) with total cobalamin, holo-transcobalamin correlated poorly (r2 = 0.23) with total cobalamin or holo-haptocorrin.

Conclusions: The solid-phase capture assay provides a simple, reliable method for quantitative determination of holo-transcobalamin in serum.
Bioluminescent Method for Detecting Telomerase Activity
Authors: Xu Shun-Qing , He Min , Yu Hong-Ping , Wang Xiao-Yang , Tan Xiang-Lin , Lu Bin , Sun Xi , Zhou Yi-Kai , Yao Qun-Feng , Xu Yong-Jun , Zhang Zhi-Ren
Published: January 2020

Background: Telomerase is a promising biomarker in cancer diagnosis and therapy. The elongation of telomeric repeats catalyzed by telomerase is accompanied by release of six PPi for each TTAGGG repeat (1 pmol PPi/310 pg telomeric repeats). We developed a novel method to measure telomerase activity by use of an enzymatic luminometric PPi assay (ELIPA).

Methods: Extracts of cell lines and tissues were incubated with primer at 30 °C for 30 min. Released PPi was converted to ATP by sulfurylase, and ATP was detected by a luciferase bioluminescence system. The ELIPA results were compared with results obtained with the conventional telomeric repeat amplification (TRAP)-ELISA in 42 lung carcinoma tissues and 27 control tissues without malignancy.

Results: The lower detection limits of ELIPA and TRAP-ELISA were 5 and 10 cells, respectively. The within-run imprecision (CV) of ELIPA was ≤12%. When compared with TRAP-ELISA, the correlation coefficient (r) was 0.79. When we used the cutoff value from ROC analysis to distinguish malignant and nonmalignant tissues, the sensitivity and specificity of ELIPA were 83% and 96%, respectively, whereas the sensitivity and specificity of TRAP-ELISA were 71% and 96%, respectively.

Conclusion: ELIPA is a simple and sensitive homogeneous method to quantify telomerase activity.
Cancer Biomarkers: Easier Said Than Done
Authors: Pritzker Kenneth PH
Published: January 2020

Background: Biological and technical advances have led to greatly increased research and development of cancer biomarkers. This overview lists some of the challenges and barriers to developing novel effective cancer biomarkers and enablers to facilitate cancer biomarker development.

Methods: Current scientific literature regarding development of biomarkers for cancer and other diseases was reviewed.

Results: Challenges to developing cancer biomarkers include better understanding of biological heterogeneity, including host/tumor heterogeneity; analytical factors, such as interferences and analytical sensitivity; clinical pathologic factors, such as current histopathologic standards; and health service and market factors. More standardized biomarker definitions, standardization of cancer biology terminology, and high-quality reference materials (specimen and clinical data repositories) were identified as factors required to support advances in cancer biomarkers.

Conclusions: With the above enablers, novel cancer biomarkers may be useful, both for assessing early and established neoplasia more precisely and for contributing data toward development of novel practical concepts regarding cancer biology.
Measurement of the Noncomplexed Free Fraction of Tissue Inhibitor of Metalloproteinases 1 in Plasma by Immunoassay
Authors: Holten-Andersen Mads N , Christensen Ib Jarle , Nielsen Hans Jørgen , Lilja Hans , Murphy Gillian , Jensen Vibeke , Brünner Nils , Piironen Timo
Published: January 2020

Background: We previously found differences in total concentrations of tissue inhibitor of metalloproteinases 1 (TIMP-1) in plasma from donors and cancer patients. Because TIMP-1 can exist in more than one molecular form, a new immunoassay to specifically detect free TIMP-1 was developed and concentrations were determined in plasma from healthy donors and colorectal cancer (CRC) patients.

Methods: We established and validated an immunoassay for the specific measurement of free TIMP-1 that uses a polyclonal anti-TIMP-1 antibody for capture and a monoclonal anti-TIMP-1 antibody that binds only free TIMP-1 for detection of antigen. Plasma samples from healthy donors and CRC patients were assayed for free TIMP-1. Total TIMP-1 was measured by our previously published assay.

Results: The mean (SD) concentrations of free TIMP-1 were similar in citrate [55.5 (11.5) μg/L] and EDTA plasma [58.9 (13.3) μg/L] from 76 donors (r2 = 0.82). In 154 donors, the ratio of free TIMP-1 [mean (SD), 64.5 (18.0) μg/L] to total TIMP-1 [83.8 (19.8) μg/L] in EDTA plasma was 0.77. Plasma concentrations of free and total TIMP-1 correlated significantly to age (free, r2 = 0.19; total, r2 = 0.27; P <0.0001), increasing 50% over an age span of 45 years. Free and total TIMP-1 were significantly increased in CRC patients (P <0.0001), whereas the ratio of free to total TIMP-1 (mean, 0.58) was significantly lower than in donors.

Conclusions: Most of the TIMP-1 in donor plasma is present in its free form, and free TIMP-1 increases with age. Free and total TIMP-1 are increased in CRC patient plasma, but the ratio of free to total TIMP-1 is significantly lower in these patients than in donors.
Temperature-controlled Primer Limit for Multiplexing of Rapid, Quantitative Reverse Transcription-PCR Assays: Application to Intraoperative Cancer Diagnostics
Authors: Raja Siva , El-Hefnawy Talal , Kelly Lori A , Chestney Melissa L , Luketich James D , Godfrey Tony E
Published: January 2020

Background: Rapid-cycling, real-time PCR instruments bring the opportunity for improved intraoperative detection of metastasis to sentinel lymph nodes. Rapid, standardized, and internally controlled assays need to be developed that are sensitive and accurate.

Methods: We describe rapid, multiplexed, internally controlled, quantitative reverse transcription-PCR (QRT-PCR) assays for tyrosinase and carcinoembryonic antigen mRNAs on the SmartCycler (Cepheid). We used a temperature-controlled primer-limiting approach to eliminate amplification of the endogenous control gene as soon as its signal had reached threshold. Positive-control oligonucleotide mimics were incorporated into all reactions to differentiate failed reactions from true negative samples.

Results: The optimized assays for rapid QRT-PCR yielded results with threshold cycle values that were only 1–2 cycles higher than slower, more conventional protocols. In rapid PCR, the temperature-controlled multiplex assay was quantitative over a dynamic range of at least 15 cycles, compared with only 6 cycles for conventional multiplexing methods. All histologically positive lymph nodes examined were also QRT-PCR positive for the appropriate marker, and the exogenous, internal positive-control mimics produced signals in all negative samples.

Conclusion: Internally controlled, rapid QRT-PCR assays can be performed in an intraoperative time frame and with sufficient sensitivity to detect histologically identified metastases to lymph nodes.
Genetic Covariation between Serum γ-Glutamyltransferase Activity and Cardiovascular Risk Factors
Authors: Whitfield John B , Zhu Gu , Nestler John E , Heath Andrew C , Martin Nicholas G
Published: January 2020

Background: Several studies have shown that variation in serum γ-glutamyltransferase (GGT) in the population is associated with risk of death or development of cardiovascular disease, type 2 diabetes, stroke, or hypertension. This association is only partly explained by associations between GGT and recognized risk factors. Our aim was to estimate the relative importance of genetic and environmental sources of variation in GGT as well as genetic and environmental sources of covariation between GGT and other liver enzymes and markers of cardiovascular risk in adult twin pairs.

Methods: We recruited 1134 men and 2241 women through the Australian Twin Registry. Data were collected through mailed questionnaires, telephone interviews, and by analysis of blood samples. Sources of variation in GGT, alanine aminotransferase (ALT), and aspartate aminotransferase (AST) and of covariation between GGT and cardiovascular risk factors were assessed by maximum-likelihood model-fitting.

Results: Serum GGT, ALT, and AST were affected by additive genetic and nonshared environmental factors, with heritabilities estimated at 0.52, 0.48, and 0.32, respectively. One-half of the genetic variance in GGT was shared with ALT, AST, or both. There were highly significant correlations between GGT and body mass index; serum lipids, lipoproteins, glucose, and insulin; and blood pressure. These correlations were more attributable to genes that affect both GGT and known cardiovascular risk factors than to environmental factors.

Conclusions: Variation in serum enzymes that reflect liver function showed significant genetic effects, and there was evidence that both genetic and environmental factors that affect these enzymes can also affect cardiovascular risk.
Comparison of Two Provocative Tests for Calcitonin in Medullary Thyroid Carcinoma: Omeprazole vs Pentagastrin
Authors: Vitale Giovanni , Ciccarelli Antonio , Caraglia Michele , Galderisi Maurizio , Rossi Riccardo , Del Prete Salvatore , Abbruzzese Alberto , Lupoli Giovanni
Published: January 2020

Background: Provocative tests for calcitonin (CT) are fundamental in the diagnosis and follow-up of C-cell disease and in the detection of hereditary medullary thyroid carcinoma (MTC) carriers with unknown RET mutations. A recent report has proposed omeprazole, which can increase endogenous gastrin (GT), as a new provocative test for MTC.

Methods: We compared the omeprazole test (20 mg twice a day for 4 days) to the pentagastrin test (0.5 μg/kg of body weight) for the diagnosis and management of MTC. Twenty healthy individuals and 20 MTC patients with mildly or moderately increased basal CT serum concentrations underwent the pentagastrin and omeprazole tests.

Results: In MTC patients, the pentagastrin test produced a significantly higher increase in serum CT than did omeprazole. After the pentagastrin injection, several patients reported unpleasant side effects, including substantial tightness in 38 of 40 participants. No adverse effects were observed during the omeprazole test. A significant direct correlation was recorded between CT% (ratio of CT peak to basal value × 100) and GT% (ratio of GT peak to basal value × 100) during the omeprazole test in MTC patients (r = 0.73; P <0.001).

Conclusions: In spite of several adverse effects, pentagastrin remains the best provocative test for the diagnosis of MTC. Omeprazole may be useful when pentagastrin is contraindicated or refused because of the unpleasant side effects, but further validation is needed.
The carnivorous digestive system and bamboo diet of giant pandas may shape their low gut bacterial diversity
Authors: Guo Wei , Chen Yinfeng , Wang Chengdong , Ning Ruihong , Zeng Bo , Tang Jingsi , Li Caiwu , Zhang Mingwang , Li Yan , Ni Qingyong , Ni Xueqin , Zhang Hemin , li Desheng , Zhao Jiangchao , Li Ying
Published: January 2020

Giant pandas have an exclusive diet of bamboo; however, their gut microbiotas are more similar to carnivores than herbivores in terms of bacterial composition and their functional potential. This is inconsistent with observations that typical herbivores possess highly diverse gut microbiotas. It is unclear why the gut bacterial diversity of giant pandas is so low. Herein, the dynamic variations in the gut microbiota of eight giant panda cubs were measured using 16S rRNA gene paired-end sequencing during a dietary switch. Similar data from red panda (an herbivorous carnivore) and carnivorous species were compared with that of giant pandas. In addition, mice were fed a high-bamboo diet (80% bamboo and 20% rat feed) to determine whether a bamboo diet could lower the gut bacterial diversity in a non-carnivorous digestive tract. The diversity of giant panda gut microbiotas decreased significantly after switching from milk and complementary food to bamboo diet. Carnivorous species living on a plant-based diet, including giant and red pandas, possess a lower microbial diversity than other carnivore species. Mouse gut microbiota diversity significantly increased after adding high-fibre bamboo to their diet. Findings suggest that a very restricted diet (bamboo) within a carnivorous digestive system might be critical for shaping a low gut bacterial diversity in giant pandas.
Glucagon-like peptide-1 receptor agonists and the risk of cardiovascular events in diabetes patients surviving an acute myocardial infarction
Authors: Trevisan Marco , Fu Edouard L , Szummer Karolina , Norhammar Anna , Lundman Pia , Wanner Christoph , Sjölander Arvid , Jernberg Tomas , Carrero Juan Jesus
Published: January 2020

Aims

Trial evidence indicates that glucagon-like peptide-1 receptor agonists (GLP-1 RAs) may reduce the risk of cardiovascular (CV) events in patients with diabetes and myocardial infarction (MI). We aimed to expand this observation to routine care settings.

Methods and results

Prospective observational study including all patients with diabetes surviving an MI and registered in the nationwide SWEDEHEART registry during 2010–17. Multivariable Cox regression analyses were used to estimate the association between GLP-1 RAs use and the study outcome, which was a composite of stroke, heart failure, Re-infarction, or CV death. Covariates included demographics, comorbidities, presentation at admission, and use of secondary CV prevention therapies. In total, 17 868 patients with diabetes were discharged alive after a first event of MI. Their median age was 71 years, 36% were women and their median estimated glomerular filtration rate was 75 mL/min/1.73m2. Of those, 365 (2%) were using GLP-1 RAs. During median 3 years of follow-up, 7005 patients experienced the primary composite outcome. Compared with standard of diabetes care, use of GLP-1 RAs was associated with a lower event risk [adjusted hazard ratio (HR) 0.72; 95% confidence interval (CI): 0.56–0.92], mainly attributed to a lower rate of re-infarction and stroke. Results were similar after propensity score matching or when compared with users of sulfonylurea. There was no suggestion of heterogeneity across subgroups of age, sex, chronic kidney disease, and STEMI.

Conclusion

GLP-1 RAs use, compared with standard of diabetes care, was associated with lower risk for major CV events in healthcare-managed survivors of an MI.
Technical aspects of uniportal video-assisted thoracoscopic double sleeve bronchovascular resections
Authors: Gonzalez-Rivas Diego , Garcia Alejandro , Chen Chang , Yang Yang , Jiang Lei , Sekhniaidze Dmitrii , Jiang Gening , Zhu Yuming
Published: January 2020

Double sleeve, bronchial and vascular reconstructions are challenging procedures indicated for centrally located tumours to avoid pneumonectomy. Traditionally, these resections have been performed by thoracotomy, but thanks to advances in imaging systems, better surgical instruments and the gained experience in video-assisted thoracic surgery (VATS), the scenario now is different. During the last decade, we have seen a rapid evolution of the uniportal VATS technique from simple lobectomies to advanced double sleeve bronchovascular procedures and carinal resections. The advantages of VATS over open surgery for major lung resections in terms of postoperative pain and morbidity, length of hospital stay and quality of life have prompted experienced surgeons to adopt uniportal VATS for cases requiring a sleeve resection. However, when a double bronchial and vascular sleeve resection is required, the adoption rate of minimally invasive surgery is still very low even for very experienced VATS surgeons. The difficulty of tumour mobilization, complexity of the suturing technique and the concern about possible uncontrolled massive bleeding during VATS are the main reasons for this low rate of adoption. In this article, we describe the technical aspects and tricks of this procedure when it is done by the uniportal VATS approach.
The Accumulation of Wealth in Marriage: Over-Time Change and Within-Couple Inequalities
Authors: Kapelle Nicole , Lersch Philipp M
Published: January 2020

This study examines the accumulation of personal wealth of husbands and wives and investigates the development of within-couple wealth inequalities over time in marriage. Going beyond previous research that mostly studied the marriage wealth premium using household-level wealth data and that conceptualized marriage as an instantaneous transition with uniform consequences over time, we argue that entry into marriage is a gendered life-course event that dynamically shapes husbands’ and wives’ wealth accumulation. Using high-quality data from the German Socio-Economic Panel Study (2002, 2007, 2012, and 2017), we apply fixed-effects regression models to describe wealth accumulation within marriage. We find evidence that wealth premiums are lower during early years of marriage, but increase steadily thereafter. The premium is mostly concentrated in housing wealth. Results from supplementary analyses with limited data, however, suggest that the premium may not be causal for men. Regarding within-couple wealth inequalities, we find a pronounced within-couple wealth gap prior to marriage during pre-marital cohabitation. This gap remains stable over time in marriage. In contrast to findings regarding income, our study indicates that the institution of marriage may not amplify within-couple wealth inequalities further.
Roles and recommendations from primary care physicians towards managing low-risk breast cancer survivors in a shared-care model with specialists in Singapore—a qualitative study
Authors: Fok Rose Wai-Yee , Low Lian Leng , Quah Hui Min Joanne , Vasanwala Farhad , Low Sher Guan , Soh Ling Ling , Mohamad Farid , Loh Kiley Wei-Jen , Soong Yoke Lim , Ke Yu , Chan Alexandre , Tan Ngiap-Chuan
Published: January 2020

Background

Breast cancer is prevalent and has high cure rates. The resultant increase in numbers of breast cancer survivors (BCS) may overwhelm the current oncology workforce in years to come. We postulate that primary care physicians (PCPs) could play an expanded role in comanaging survivors, provided they are given the appropriate tools and training to do so.

Objective

To explore the perspectives of PCPs towards managing BCS in a community-based shared-care programme with oncologists.

Methods

Eleven focus groups and six in-depth interviews were conducted with seventy PCPs recruited by purposive sampling. All sessions were audio-recorded, transcribed verbatim and coded by three independent investigators. Thematic data analysis was performed and the coding process facilitated by NVivo 12.

Results

Majority of PCPs reported currently limited roles in managing acute and non-cancer issues, optimizing comorbidities and preventive care. PCPs aspired to expand their role to include cancer surveillance, risk assessment and addressing unmet psychosocial needs. PCPs preferred to harmonize cancer survivorship management of their primary care patients who are also BCS, with defined role distinct from oncologists. Training to understand the care protocol, enhancement of communication skills, confidence and trust were deemed necessary. PCPs proposed selection criteria of BCS and adequacy of their medical information; increased consultation time; contact details and timely access to oncologists (if needed) in the shared-care programme.

Conclusions

PCPs were willing to share the care of BCS with oncologists but recommended role definition, training, clinical protocol, resources and access to oncologist’s consultation to optimize the programme implementation.
Telescope: an interactive tool for managing large-scale analysis from mobile devices
Authors: Brito Jaqueline J , Mosqueiro Thiago , Rotman Jeremy , Xue Victor , Chapski Douglas J , la Hoz Juan De , Matias Paulo , Martin Lana S , Zelikovsky Alex , Pellegrini Matteo , Mangul Serghei
Published: January 2020

Background

In today's world of big data, computational analysis has become a key driver of biomedical research. High-performance computational facilities are capable of processing considerable volumes of data, yet often lack an easy-to-use interface to guide the user in supervising and adjusting bioinformatics analysis via a tablet or smartphone.

Results

To address this gap we proposed Telescope, a novel tool that interfaces with high-performance computational clusters to deliver an intuitive user interface for controlling and monitoring bioinformatics analyses in real-time. By leveraging last generation technology now ubiquitous to most researchers (such as smartphones), Telescope delivers a friendly user experience and manages conectivity and encryption under the hood.

Conclusions

Telescope helps to mitigate the digital divide between wet and computational laboratories in contemporary biology. By delivering convenience and ease of use through a user experience not relying on expertise with computational clusters, Telescope can help researchers close the feedback loop between bioinformatics and experimental work with minimal impact on the performance of computational tools. Telescope is freely available at https://github.com/Mangul-Lab-USC/telescope.
A new method to estimate ocean-bottom-seismometer orientation using teleseismic receiver functions
Authors: Zheng Hong , Fan Jianke , Zhao Dapeng , Li Cuilin , Dong Dongdong , Zhang Guangxu , Wang Xiujuan
Published: January 2020

The orientation of an ocean-bottom-seismometer (OBS) is a critical parameter for analysing three-component seismograms, but it is difficult to estimate because of the uncontrollable OBS posture after its deployment. In this study, we develop a new and effective method to estimate the OBS orientation by fitting the amplitude of direct P wave of teleseismic receiver functions. The reliability of this method is verified using synthetic data and observed waveforms recorded at land seismic stations in Shandong Province, China. Our extensive synthetic tests show that our new method is little affected by a thin sedimentary layer that has a low S-wave velocity. The orientations of OBS stations that we deployed in the Yap subduction zone in the Western Pacific Ocean are estimated and corrected using our new method. After the correction, the direct P waves of teleseismic receiver functions show very good consistency. The effects of white and coloured noise in different levels, epicentral distance and backazimuth are also investigated, and the results show that these factors have small effects on the new method. We also examine the effect of sensor tilting on estimation of the OBS orientation, and find that a tilting correction should be made before the misorientation correction. We compare the OBS orientations determined with the new method and other methods and find that they are generally consistent with each other. We also discuss advantages and shortcomings of various methods, and think that our new method is more robust than the existing methods.
‘Real Toryism’ or Christian democracy? The political thought of Douglas Jerrold and Charles Petrie at the New English Review, 1945–50*
Authors: Love Gary
Published: January 2020

This article uses the periodical press, memoirs and archival sources to examine the Conservative thought of Douglas Jerrold and Charles Petrie at the New English Review between 1945 and 1950. It seeks to argue that while there was more continuity than rupture in their political thought, they were able to make a transition from interwar authoritarianism and ‘fascism’ to postwar anti-communism and democracy. There were similarities between their political Catholicism and Christian democracy, but ultimately their ‘real Toryism’ and continued sympathy for Franco prevented them from fully aligning themselves with British and west European Christian democrats.
Th17 Cells in Inflammatory Bowel Disease: An Update for the Clinician
Authors: Hou Guoqing , Bishu Shrinivas
Published: January 2020

Studies in humans strongly implicate Th17 cells in the pathogenesis of inflammatory bowel disease. Thus, Th17 cells are major targets of approved and emerging biologics. Herein, we review the role of Th17 in IBD with a clinical focus.
17 AZD4205, A SELECTIVE, GI TRACT-ENRICHED SELECTIVE JAK1 INHIBITOR FOR CROHN’S DISEASE: PRECLINICAL EVIDENCE AND PHASE I DATA
Authors: Wang Mei , Zhang Lin , Han Sufang , Xu Yanping , Li Jingrun , Chen Kan , Li Li , Yang Zhenfan , Liang Jie , Wu Kaichun , Rubin David
Published: January 2020

Background

AZD4205 is an oral, ATP-competitive, JAK1 selective inhibitor. Nonclinical data showed its higher drug concentration within the gastrointestinal (GI) tract relative to plasma in the rodents, suggesting its potential as an effective and safe treatment option for patients with Crohn’s disease (CD). AZD4205 also has been evaluated in a phase I study in healthy volunteers (HVs). Here we report AZD4205 preclinical data in cells and animal models, and phase I data from HVs.

Method

Enzymatic activity of AZD4205 was assessed in in vitro biochemical assay with human isolated JAK kinase domains. The cellular activity of AZD4205 was evaluated in human peripheral blood mononuclear cells (PBMCs) by assessing cytokine induced phosphorylated-STATs (pSTATs). The in vivo efficacy of AZD4205 was assessed in a murine CD model induced by subcutaneous injection of 10 mg/kg indomethacin, and AZD4205 was orally administered at different doses for 5 consecutive days. The ileum and bone marrow tissues were collected post AZD4205 treatment to measure pSTAT3 expression using immunohistochemistry. The drug concentration of AZD4205 in ileum and plasma were measured by HPLC.

Phase I study of HVs (Clinicaltrial.gov Identifier: NCT03728023) comprised of single ascending dose (SAD) of 5 to 150 mg, food effect (FE) with 50 mg single dosing with or without food, and multiple ascending dose (MAD) of 25 to 100 mg once daily for 14 days. Safety monitoring was conducted, and blood samples were collected to assess PK of AZD4205.

Result

In enzymatic assay, AZD4205 exhibits greater than 200-400-fold selectivity over other JAK family kinases. AZD4205 inhibits pSTAT1, pSTAT3, and pSTAT5 in human PBMCs with IC50 of 50, 308, and 90 nM, respectively.

The drug concentration of AZD4205 in murine ileum was around 100-fold higher than that in plasma. In murine CD model, the pSTAT3 was inhibited by 93% in ileum but only 43% in bone marrow at 4 hr post AZD4205 treatment. AZD4205 exhibited a dose-dependent effect on improving body weight loss and decreasing colon density in this model.

In the NCT03728023 study, 66HVs were dosed with AZD4205 or matched placebo, SAD (n=20 in AZD4205, n=10 in placebo), FE (n=12 in AZD4205) and MAD (n=18 in AZD4205, n=6 in placebo). AZD4205 was well-tolerated, and no ≥ G3 drug related AE was observed. A dose-proportional increase of AZD4205 plasma concentrations was detected in HVs. The T1/2 was close to 48 hr and hence accumulation of about 3-fold in Cmax and AUC was observed after multiple once daily dosing. No relevant food effect was observed to significantly impact AZD4205 PK.

Conclusion

Based on preclinical murine data, AZD4205 has unique GI-tract enriched PK properties, as a promising agent for CD. Clinical data in HVs demonstrated its favorable safety and PK profiles. A phase II study in moderate to severe CD is planned.
16 CCL11 EXACERBATES COLITIS VIA MODULATION OF EPITHELIAL WOUND REPAIR
Authors: Polosukhina Dina , Singh Kshipra , Barry Daniel , Allaman Margaret , Piazuelo Maria , Washington Mary , Wilson Keith , Coburn Lori
Published: January 2020

Background

We have shown that CCL11 (eotaxin-1), an eosinophil chemoattractant, is significantly increased in the serum of ulcerative colitis and Crohn’s disease patients vs controls and is also increased in dextran sulfate sodium (DSS)-induced murine colitis. In response to azoxymethane (AOM)-DSS, Ccl11–/– mice have significantly decreased colonic tumor number and burden, histologic injury, and colonic eosinophil infiltration vs wild-type (WT) mice. Ccl11 is expressed by isolated colonic epithelial and lamina propria immune cells. Studies in bone marrow chimera mice revealed that both hematopoietic- and epithelial-cell derived CCL11 were important for protection in the AOM-DSS model. Our aim was to assess the role of CCL11 in an injury and recovery colitis model and in an epithelial wound restitution model.

Methods

C57BL/6 WT and Ccl11–/– mice were exposed to 4% DSS in the drinking water for 5 days followed by 5 days of water for an injury and recovery model. Body weights were assessed daily. Histologic injury was assessed by a validated scoring system including depth and percent involvement of inflammation and crypt damage. Colonic eosinophil and neutrophil infiltration was determined by immunohistochemistry for major basic protein (MBP) and myeloperoxidase (MPO), respectively. The number of MBP- and MPO-positive cells per high-powered field (HPF) were quantified in a blinded manner. Young Adult Mouse Colon (YAMC) epithelial cells were stained with antibodies to the CCL11 receptors CCR2, CCR3, and CCR5 for assessment by flow cytometry. YAMC cell monolayers were wounded and followed for 24 h in normal media or with the addition of 100ng/mL recombinant CCL11 (rCCL11). The wound area was measured at 0 and 24 h to determine percent restitution. RNA was isolated and assessed for Ccl11 mRNA expression.

Results

In the injury and recovery model, Ccl11–/– mice exhibited decreased body weight loss (p<0.05), histologic injury (20.6 ± 1.5 vs 12.4 ± 1.6, p<0.001), and fewer MBP-positive cells/HPF (23.5 ± 1.4 vs 2.4 ± 0.2, p<0.001) vs WT mice. While colonic MPO-positive cells were increased with DSS exposure, there was no difference in Ccl11–/– mice. Naïve/unstimulated YAMC cells expressed both CCR3 and CCR5. Wounding the YAMC monolayer alone did not lead to increased Ccl11 mRNA expression. However, in the presence of rCCL11, there was a decrease in wound restitution at 24 h (48.0% ± 1.7 vs 31.3% ± 2.1, p<0.001).

Conclusions

Loss of CCL11 leads to clinical and histologic improvement in an injury and recovery colitis model. Colonic epithelial cells express both CCR3 and CCR5, and epithelial wound restitution is decreased in the presence of rCCL11. These data suggest that CCL11 has a deleterious effect on epithelial restitution leading to an exacerbation of colitis. Therefore, treatment with anti-CCL11 antibodies may be a therapeutic strategy in IBD patients.
P058 ISOLATED TERMINAL ILEITIS AT COLONOSCOPY – WHAT FACTORS PREDICT A NEW DIAGNOSIS OF CROHN’S DISEASE?
Authors: Patel Rajan , Ang Kar Mun , Moledina Saadiq , Musa Saif , Alisa Akeel , Tang Kwok
Published: January 2020

Background

Faecal calprotectin (FC) is a biomarker elevated in active inflammatory bowel disease (IBD). FC is more sensitive in colonic than small bowel IBD. Ileo-colonoscopy is usually performed to confirm a diagnosis of IBD. Isolated non-specific terminal ileitis is often inconclusive despite biopsy. We aimed to assess the factors that predict terminal ileal Crohn’s disease after ileitis is seen at colonoscopy.

Methods

A single centre retrospective study of all endoscopic cases of isolated terminal ileitis diagnosed at colonoscopy over a 4 year period (January 2015 – December 2018) was performed. Data was obtained from the Unisoft Endoscopy reporting software. Statistical analyses included chi-square, student t-test and binary logistic regression. Faecal calprotectin, CRP and histology were noted. >150μg/mg was used as a cut off for elevated FC.

Results

139 cases were identified and exclusion criteria were applied (known Crohn’s disease, colonic disease). 74 cases were included for analysis. The mean age was 43.9. 44 (59.5%) of the cases were women. 38 (51.4%) had FC performed of which 27 (71.1%) had a FC >150μg/mg.

60 (81.1%) cases had macroscopic terminal ileum ulcers, 9 (15%) of these had histological evidence of ulceration. Subsequent diagnoses of Crohn’s disease were made in 15 (20.3%) patients. Odds ratio of 1.28 (p = 0.016, Cl 0.45-0.047) in the TI ulcers + FC >150μg/mg vs. no TI ulcers + FC <150μg/mg.

Conclusion

1 in 5 patients with isolated terminal ileitis were subsequently diagnosed with Crohn’s disease. Almost 90% of these new cases had a faecal calprotectin >150μg/mg. There is poor correlation between endoscopic and histological terminal ileum ulceration. We conclude that terminal ileal ulceration in combination with faecal calprotectin >150μg/mg increases the likelihood of a new diagnosis of Crohn’s disease.
9 METABOLIC UTILIZATION OF PROPANEDIOL BY ADHERENT-INVASIVE E. COLI REGULATES INTESTINAL TISSUE IMMUNITY
Authors: Viladomiu Monica , Metz Maeva , Lima Svetlana , Guo Chun-Jun , Simpson Kenneth , Longman Randy
Published: January 2020

While the molecular mechanisms by which the microbiome modulates mucosal immunity in Crohn’s disease (CD) are still largely unknown, recent data highlight the involvement of specific diet- and bacterial-derived metabolites in the regulation of intestinal immune cell activation and differentiation. We have recently shown that Adherent-Invasive E.coli (AIEC), which are enriched in CD patients, are sufficient to induce intestinal Th17 cells. Although AIEC lack pathogenic factors including type III secretion systems, many CD-derived isolates express virulence-associated metabolic enzymes including propanediol dehydratase (PduC), which enables AIEC to use fucose-derived propanediol as an alternate carbon source in the gut. We found that pduC is enriched in the microbiome and among E. coli genomes in CD patients compared to healthy controls. With fucosylated oligosaccharides on the surface of intestinal epithelial cells, we hypothesized that this propanediol utilization pathway provides AIEC a competitive advantage for epithelial cell adherence and intestinal immune cell activation. To evaluate the physiologic contribution of pduC to mucosal Th17 induction, we generated a pduC-deficient (ΔpduC) mutant of a CD-derived, AIEC isolate. Deletion of pduC resulted in reduced inflammatory Th17 cells and attenuated weight loss following T cell transfer colitis. Using genetic mouse models, we found that CX3CR1+ mononuclear phagocytes are required for this AIEC-mediated Th17 induction and IL-10 is required to restrain pduC-dependent dextran sodium sulfate (DSS)-induced colitis. Using a catalytically-inactive mutant, we determined that PduC metabolic activity was required for this immune phenotype. Cell-free supernatants from WT AIEC (but not the isogenic, pduC-deficient clone) promoted ex vivo Th17 cell polarization and metabolomics analysis (LC-MS) of these supernatants defined PduC-dependent metabolites capable of promoting Th17 polarization. These studies reveal a link between AIEC microbial metabolism and inflammatory Th17 cells with the potential to serve as a therapeutic target in the treatment of Crohn’s disease.
P162 CRITICAL ROLES OF DIETARY SIMPLE SUGARS IN COLITIS PATHOGENESIS
Authors: Godfrey Victoria , Zaki Hasan
Published: January 2020

The incidence of inflammatory bowel disease (IBD) is strikingly high in Western countries, implicating the role of Western diet in its etiology and pathogenesis. Western diet is characterized by high fat, low fiber, and high sugar. Despite clinical evidence of an association between high sugar diet and IBD susceptibility, the precise role of dietary simple sugars such as glucose, fructose, and sucrose in colitis pathogenesis is unknown. Using dextran sodium sulfate (DSS) and IL10-deficient mouse models of colitis, we studied the effect of simple sugars in colitis susceptibility. Mice were given high glucose, fructose or sucrose in their drinking water or left untreated before and during colitis induced by DSS. Sugar-fed mice exhibited increased colitis susceptibility evidenced by higher body weight loss, diarrhea, rectal bleeding, and severe histopathological changes in the colon as compared to those of sugar-untreated colitic mice. Pre-colitis dietary habit of sugar consumption was critical since sugar pretreated mice were susceptible to DSS-induced colitis even without high sugar diet intake during DSS administration. Consistent with these findings, there were higher incidence of spontaneous colitis development in Il10-/- mice following consumption of high sugar. To understand the underlying mechanism, we evaluated the effect of high sugar diet on intestinal epithelial cell death, inflammation, epithelial barrier permeability, and gut microbiota composition in healthy mice. We did not observe any major pathological changes and apoptosis in the colon of sugar-fed mice. Inflammatory responses, activation of inflammatory signaling pathways, and the expression of tight junction proteins were comparable between control and sugar-fed mice. Interestingly, gut microbiota composition of sugar-fed mice was altered as measured by 16S rRNA gene sequencing of DNA isolated from feces. Microbial species richness was reduced and relative abundance of several bacterial species was either increased or decreased in sugar-fed mice. We further confirmed that sugar-induced alteration of gut microbiota is responsible for exacerbated colitis by using antibiotics or germ-free mice. Mice receiving antibiotics during high-sugar intake did not show increased DSS-colitis susceptibility. Similarly, high-sugar diet did not induce overt colitis pathogenesis in germ-free mice. These findings demonstrate a critical role of dietary caloric sugars in the predisposition and promotion of colitis and could be implicated in the treatment and management of IBD.
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LACTOBACILLUS REUTERI SUPPRESSES PRO-INFLAMMATORY DRIVEN REACTIVE OXYGEN SPECIES IN VITRO IN HUMAN INTESTINAL EPITHELIAL CELLS AND IN VIVO IN A TNBS COLITIS MOUSE MODEL
Authors: Ruan Wenly , Engevik Melinda , Chang-Graham Alexandra , Hyser Joseph , Versalovic James
Published: January 2020

Background

Reactive oxygen species (ROS) play a role in maintaining intestinal epithelial homeostasis and are normally kept at low levels via antioxidant compounds. Dysregulation of ROS can lead to intestinal inflammation and contribute to inflammatory bowel disease (IBD). Select gut microbes possess the enzymatic machinery to produce antioxidants whereas others can dysregulate levels of ROS. Our model microbe, Lactobacillus reuteri (ATCC PTA 6475), has been demonstrated to reduce intestinal inflammation in mice models. It contains the genes encoding two distinct GshA-like glutamylcysteine ligases. We hypothesize that L. reuteri can secrete γ-glutamylcysteine to suppress ROS, minimize NFκB activation and regulate secretion of e pithelial cytokines.

Methods & Results

Conditioned media from L. reuteri was analyzed via mass spectrometry to confirm the presence of γ-glutamylcysteine. All cysteine containing products including γ-glutamylcysteine were fluorescently tagged in the conditioned media and then incubated with HT29 cell monolayers as well as human jejunal enteroid (HJE) monolayers. γ-glutamylcysteine was demonstrated to enter intestinal epithelial cells based on microscopy. Next, a Thioltracker assay was used to show increased intracellular glutathione levels by L. reuteri secreted γ-glutamylcysteine. HT29 cells and HJEs were then treated with IL-1β or hydrogen peroxide, and L. reuteri metabolites as well as γ-glutamylcysteine significantly suppressed pro-inflammatory cytokine driven ROS and IL-8 production. L. reuteri secreted products also reduced activity of NFκB as determined by a luciferase reporter assay. γ-glutamylcysteine deficient mutants were generated by targeted mutagenesis of GshA genes, and these mutant L. reuteri strains had a diminished ability to suppress IL-8 production and ROS. To further test the role of L. reuteri secreted γ-glutamylcysteine in vivo, a 2,4,6-Trinitrobenzenesulfonic acid (TNBS)- induced mouse colitis model was used. Adolescent mice were orogavaged with PBS, L. reuteri, L. reuteri GshA2 mutant, or γ-glutamylcysteine for a week after which TNBS was rectally administered to induce colitis. We demonstrate that L. reuteri and γ-glutamylcysteine can suppress histologic inflammation compared to PBS control and L. reuteri GshA2 mutant groups.

Conclusions

Together these data indicate that L. reuteri secretes γ-glutamylcysteine which can enter the intestinal epithelial cells and modulate epithelial cytokine production. It acts via suppression of ROS and NFκB which then decreases IL-8 production. We are able to demonstrate this in vitro in both HT 29 cells and HJEs. We now also demonstrate this in vivo in a mouse colitis model. These experiments highlight a prominent role for ROS intermediates in microbiome-mammalian cell signaling processes involved in immune responses and intestinal inflammation.
P117 CONTENT VALIDITY OF THE SUBCUTANEOUS ADMINISTRATION ASSESSMENT QUESTIONNAIRE (SQAAQ) IN ADULT AND ADOLESCENT PATIENTS WITH MODERATE TO SEVERE CROHN’S DISEASE
Authors: Hunter Theresa , Naegeli April , Delbecque Laure , Al-Zubeidi Tamara , Pegram Hannah , Kitchen Helen
Published: January 2020

Background

Subcutaneous (SC) delivery of biologic therapies is typically via a prefilled syringe or an auto-injector designed to offer a more convenient and more consistent administration of drug product. There is currently no gold standard instrument that is used to assess the usability and patient confidence and preference for an injection device in Crohn’s disease (CD). The Subcutaneous Administration Assessment Questionnaire (SQAAQ) was developed to evaluate ease of use and confidence in SC administrations of therapy. The SQAAQ (Figure 1) features 12 items scored on a 7-point Likert scale (1= Strongly Disagree to 7= Strongly Agree). Higher scores indicate better usage experience. The recall period is immediately after patients auto-inject. This study aimed to establish evidence of the content validity of the SQAAQ among patients with moderate to severe CD.

Methods

A semi-structured interview guide was used to conduct face-to-face qualitative interviews with adult (18+ years) and adolescent (12–17 years) patients from the US with a clinician confirmed diagnosis of CD. Cognitive debriefing interviews were structured into sections that explored patients’ opinions, interpretation, and acceptance of the SQAAQ. Data were coded using Atlas.ti using a framework approach where pre-defined codes were applied to quotes that demonstrated patients’ understanding/interpretation, acceptance, and usage of the instructions, items, response scale/options, and recall period.

Results

Twenty-four patients (adults, n=16; adolescents, n=8) participated in the interviews. Thirteen adults and 3 adolescents experienced moderate disease severity (reported by clinicians). Adult and adolescent groups contained an equal proportion (50%) of male and female participants. Adults had a mean age 50.3 years (range 27.0–75.0), adolescents with mean age 15.6 years (range 14.0–17.0). Overall, two-thirds (n=16, 67%) of patients were white, and the mean duration for CD diagnosis was 8.0 years (range 1.0–23.7). Ten (42%) patients had been on SC treatment for a mean 4.5 years (range 0.2–16.9), all self-administered. Seven (29%) patients had been receiving intravenous treatment for a mean 2.1 years (range 0.2–5.4). The SQAAQ was found to be conceptually relevant, understandable and usable by adults and adolescents with CD. Notably the measurement concepts were confirmed as relevant by patients who currently used SC treatment devices. The patients with no SC experience (n=7, 29%), understood the item wording, response scales, and recall period and had few difficulties completing the measure.

Conclusion

Content validity of the SQAAQ was established in this sample of adolescent and adult patients with CD. The SQAAQ is appropriate for inclusion in CD clinical trials for SC treatment devices with adult and adolescent patients.
P123 THIRTY DAY READMISSIONS AMONG PATIENTS WITH COMPLICATED INFLAMMATORY BOWEL DISEASE AND EFFECTS ON OUTCOMES: RESULTS OF ANALYSIS OF 2016 NATIONWIDE READMISSION DATABASE
Authors: Sharma Sachit , Khan Zubair , Aziz Muhammad , Vohra Ishaan , Acharya Ashu
Published: January 2020

Objective

To determine the rate of hospital readmissions within 30 days of discharge in patients with complicated Inflammatory Bowel Disease (IBD) and its impact on mortality, morbidity, and health care cost in the United States.

Methods

We performed a retrospective study using the Nationwide Readmission Database(NRD) for the year 2016 (Data on 35.6 million discharges). We collected data on hospital readmissions of 29,356 adults who were hospitalized for complicated Inflammatory Bowel Disease(Crohn’s disease and Ulcerative Colitis) and discharged. Complications were defined using ICD 10 codes. Patients with age less than 18, elective admission and admission during December month were excluded. The primary outcome was rate of all-cause readmission within 30 days of discharge. Secondary outcomes were reasons for readmission, readmission mortality rate, morbidity and resource use. Multivariate regression analysis was used to determine the independent predictors of 30-day readmission.

Results

Among patients admitted to US hospitals with complicated IBD, the total time at risk was 407,983 days, with the first readmission occurring at day 1 and the last readmission at day 30. The 30-day rate of readmission was 17.6%, with the most common cause of readmission being Crohn’s disease of small intestine with intestinal obstruction (9%). The mortality rate among patients readmitted to the hospital (0.6%), was higher than that for index admissions (0.3%) (P < .01). Mean length of stay was 5.5 days for index admission and 5.9 days for readmission (p=0.01). Mean total charge for index admission was $44,768 compared to $48,766 for readmission (p<0.01). Mean total cost for index admission was $11,491 compared to $12,704 for readmission (p<0.01). A total of 30,943 hospital days were associated with readmission, and the total health care in-hospital economic burden was $65 million (in costs) and $251 million (in charges). Independent predictors of readmission were age, insurance status, higher Charlson comorbidity score, lower income, teaching status of hospital and longer stays in the hospital. Older age, private insurance, median income more than $48,000 were associated with lower odds for readmission, whereas higher comorbidities and admission to teaching hospital were associated with increased odds for readmission.

Conclusions

In a retrospective study of patients hospitalized for complicated IBD in 2016, 17.6% were readmitted to the hospital within 30 days of discharge. Readmission was associated with higher mortality, morbidity, and resource use. Age, insurance status, higher comorbidity score, lower income and admission to teaching hospital were independent predictors of readmission.