scholarly journals A35 MICROBIAL PROTEOLYTIC SIGNATURE IN ULCERATIVE COLITIS INDUCES AN INFLAMMATORY SIGNATURE IN MICROBIOTA-HUMANIZED MICE

2020 ◽  
Vol 3 (Supplement_1) ◽  
pp. 42-43
Author(s):  
H J Galipeau ◽  
W Turpin ◽  
A Caminero Fernandez ◽  
A Santiago ◽  
J Libertucci ◽  
...  
Keyword(s):  
Ulcerative Colitis ◽  
At Risk ◽  
Proteolytic Activity ◽  
Intestinal Microbiota ◽  
Inflammatory Responses ◽  
Critical Role ◽  
Low Grade ◽  
Polymorphonuclear Cells ◽  
Fecal Samples ◽  
Patients At Risk

Abstract Background Altered gut microbiota composition has been associated with inflammatory bowel diseases (IBD) including ulcerative colitis (UC), but causality and bacterially-driven mechanisms, are unclear. Proteases within the gastrointestinal tract play a critical role in maintaining homeostasis and are tightly regulated by anti-proteases. Host-derived proteolytic imbalances have been described in IBD, including UC, however, the role of intestinal microbiota as a source of proteases and anti-proteases has largely been ignored. Aims To study microbial proteolytic activity and intestinal microbiota profiles in a cohort of individuals at-risk for IBD, and in those individuals that develop UC at follow-up. Methods Fecal samples were collected from healthy individuals at-risk for IBD and who went on to develop UC (pre-UC; n=14) and again after UC diagnosis (post-UC, n=10). Fecal samples from matched at-risk individuals that did not develop UC were used as healthy controls (n=52). Overall fecal proteolytic and elastolytic activity was measured. We performed metagenomics sequencing in 4 UC subjects (pre and post) and 4 matched HC using Illumina Hi-Seq from stool DNA. To investigate bacterial origin and functional significance, pregnant germ-free (GF) mice were colonized with a fecal sample from a selected UC subject (pre and post) and a matched HC. Naturally colonized litters were followed for 12 weeks, after which proteolytic activities and signs of inflammation were measured. Results Fecal proteolytic and elastase activity was increased in pre- and post-UC samples compared to HCs. Metagenomics revealed over 20k genes were significantly different between HC and pre-UC samples, and of these, 440 related to proteases and peptidases. Increased fecal proteolytic activity, higher lipocalin levels, and increased colonic polymorphonuclear cells in colonic H&E sections was observed in pre- and post-UC colonized mice compared to HC colonized mice. Mice colonized with pre-UC microbiota showed increased mRNA expression of genes linked to immunological disease, antimicrobial and inflammatory responses (ie. Tlr2, Tlr5, Nod2, and Il1b) as compared to HC colonized mice. Conclusions These results suggest increased fecal proteolytic activity is observed prior to the onset and clinical diagnosis of UC in patients at-risk for IBD, and upon transfer to mice born from colonized GF dams, low-grade inflammation develops. These pathways could be developed as novel non-invasive biomarkers to monitor at-risk populations. Submitted on behalf of the CCC-GEM Project consortium. Supported by CCC GIA to EF Verdu Funding Agencies CCC

scholarly journals A210 ADLERCREUTZIA IS DEPLETED IN UC PATIENTS, EVEN BEFORE CLINICAL DIAGNOSIS, AND INVERSELY CORRELATES WITH FECAL ELASTOLYTIC ACTIVITY

2020 ◽  
Vol 3 (Supplement_1) ◽  
pp. 82-83
Author(s):  
A Santiago Badenas ◽  
J Libertucci ◽  
W Turpin ◽  
H J Galipeau ◽  
K Croitoru ◽  
...  
Keyword(s):  
Ulcerative Colitis ◽  
At Risk ◽  
Microbial Community ◽  
Proteolytic Activity ◽  
Clinical Diagnosis ◽  
Community Analysis ◽  
Microbial Community Analysis ◽  
Healthy Controls ◽  
Fecal Samples ◽  
Elastolytic Activity

Abstract Background A combination of genetics, environmental, and immune factors contribute to the development of ulcerative colitis (UC). Host proteolytic imbalance has been reported in active UC. Preliminary results from our lab suggest microbial proteolytic activity is increased before as well as after onset of UC, and transfer of this activity to mice contributes to inflammation. Aims Our aim was to correlate the elastolytic activity of fecal samples from individuals at risk for IBD, before and after onset of ulcerative colitis, with their fecal microbiota profiles Methods We first investigated proteolytic activity in fecal samples from individuals at risk to develop UC (pre-UC, n=12) prior to disease onset and after UC diagnosis (post-UC, n=7) and matched healthy controls (n=66). Microbial community analysis was performed by sequencing the V4 region of the 16S rRNA gene region using Illumina MiSeq platform. Sequences were analyzed with QIIMEv1.9.0. We measured bacterial proteolytic activity, using a FITC-elastin assay. Results Microbial community analysis revealed that the overall diversity (both richness and evenness) in UC patients was decreased compared to healthy controls as well as pre-UC patients. The relative abundance of the genus Adlercreutzia was decreased by 3.1 fold in pre-UC patients compared to healthy controls and was further decreased in post-UC (3.8 fold). The presence of Adlercreutzia was also found to be negatively correlated (r=-0.47, p<0.0001) with elastolytic activity in stool supernatant, suggesting a possible protective role in the disease. Conclusions We found novel potentially protective bacteria, Adlercreutzia, which was depleted in UC patients, even before clinical diagnosis and correlated negatively with proinflammatory elastolytic activity described previously in IBD. The protective mechanisms are under investigation. On behalf of the CCC-GEM Project consortiumand Supported by a CCC GIA to EFV Funding Agencies CCC

scholarly journals A29 NOVEL FECAL BIOMARKERS THAT PRECEDE CLINICAL DIAGNOSIS OF ULCERATIVE COLITIS

2021 ◽  
Vol 4 (Supplement_1) ◽  
pp. 268-269
Author(s):  
H J Galipeau ◽  
A CAMINERO FERNANDEZ ◽  
W Turpin ◽  
M Bermudez-Brito ◽  
A Santiago ◽  
...  
Keyword(s):  
Ulcerative Colitis ◽  
At Risk ◽  
Gut Microbiota ◽  
Proteolytic Activity ◽  
Clinical Diagnosis ◽  
Elastase Activity ◽  
Microbial Composition ◽  
Fecal Samples ◽  
And Function

Abstract Background Altered gut microbiota composition and function has been associated with inflammatory bowel diseases (IBD) including ulcerative colitis (UC), but causality and mechanisms remain unknown. Most studies have examined patients with active or treated disease and little is known about microbial compositional or functional changes that occur before disease onset. Aims We studied a longitudinal cohort of subjects at risk for IBD to define the fecal microbial composition and function in subjects prior to UC onset (pre-UC) and at diagnosis (post-UC), and in matched at-risk subjects that remained healthy. Methods Fecal samples were collected from healthy individuals at-risk for IBD (pre-UC; n=13) and subjects were followed longitudinally until UC diagnosis (post-UC, n=9), at which point another fecal sample was collected. Fecal samples from a cohort of matched at-risk individuals that did not develop UC were used as healthy controls (n=48). We applied 16S rRNA gene sequencing, next generation shotgun sequencing, in vitro proteolytic assays and gnotobiotic colonizations to define the microbial composition and proteolytic function in fecal samples. Results The microbiota of post-UC subjects clustered separately from pre-UC and HC subjects, based on bray-curtis and unweighted UniFrac, had reduced alpha-diversity, and had reduced abundance of Aldercreutzia compared to pre-UC and HC. In vitro functional analysis revealed increased fecal proteolytic and elastase activity in pre-UC and post-UC samples compared to HC. Metagenomics identified pathways and gene families related to protein metabolism and proteases/peptides that were significantly different between HC and pre-UC samples, suggesting a bacterial component to the pre-UC proteolytic signature. Elastase activity inversely correlated with the relative abundance of Adlercreutzia, and other potentially beneficial taxa, and directly correlated with Bacteroides vulgatus, a known proteolytic taxon. High elastase activity was confirmed in Bacteroides isolates from fecal samples. Bacterial contribution and functional significance of the proteolytic signature was investigated in germ-free adults and litters born from dams colonized with HC, pre-UC or post-UC microbiota. Mice colonized with pre-UC microbiota at adulthood or neonatally developed higher fecal proteolytic activity and an inflammatory immune tone compared with HC colonized mice. Conclusions We have identified increased fecal proteolytic activity that precedes clinical diagnosis of UC and associates with gut microbiota changes. This may constitute a non-invasive biomarker of inflammation to monitor at-risk populations that can be targeted therapeutically with anti-proteases. Funding Agencies CAG, CCC, CIHR

scholarly journals Non-invasive detection of bladder cancer through the analysis of driver gene mutations and aneuploidy

2017 ◽  
Author(s):  
Simeon Springer ◽  
Maria Del Carmen Rodriguez Pena ◽  
Lu Li ◽  
Christopher Douville ◽  
Yuxuan Wang ◽  
...  
Keyword(s):  
At Risk ◽  
Bladder Cancer ◽  
Gene Mutations ◽  
Urine Samples ◽  
Driver Gene ◽  
Low Grade ◽  
Invasive Approach ◽  
Non Invasive ◽  
Patients At Risk ◽  
Copy Number Changes

AbstractCurrent non-invasive approaches for bladder cancer (BC) detection are suboptimal. We report the development of non-invasive molecular test for BC using DNA recovered from cells shed into urine. This “UroSEEK” test incorporates assays for mutations in 11 genes and copy number changes on 39 chromosome arms. We first evaluated 570 urine samples from patients at risk for BC (microscopic hematuria or dysuria). UroSEEK was positive in 83% of patients that developed BC, but in only 7% of patients who did not develop BC. Combined with cytology, 95% of patients that developed BC were positive. We then evaluated 322 urine samples from patients soon after their BCs had been surgically resected. UroSEEK detected abnormalities in 66% of the urine samples from these patients, sometimes up to 4 years prior to clinical evidence of residual neoplasia, while cytology was positive in only 25% of such urine samples. The advantages of UroSEEK over cytology were particularly evident in low-grade tumors, wherein cytology detected none while UroSEEK detected 67% of 49 cases. These results establish the foundation for a new, non-invasive approach to the detection of BC in patients at risk for initial or recurrent disease.

Abstract 087: Mitochondrial Fission Inducer, Dynamin-Related Protein 1 (DRP1), is Required for Endothelial Inflammatory Responses

Hypertension ◽  
2016 ◽  
Vol 68 (suppl_1) ◽  
Author(s):  
Steven J Forrester ◽  
Tatsuo Kawai ◽  
Katherine J Elliott ◽  
Kunie Eguchi ◽  
Victor Rizzo ◽  
...  
Keyword(s):  
Er Stress ◽  
Inflammatory Responses ◽  
Vascular Dysfunction ◽  
Mitochondrial Fission ◽  
Critical Role ◽  
Dominant Negative ◽  
Low Grade ◽  
Aortic Endothelial Cells ◽  
Endothelial Inflammation ◽  
Inflammatory Stimuli

Among various cardiovascular diseases, hypertension (HTN) is considered to be a disease plagued by chronic low-grade inflammation associated with endothelial dysfunction. Interestingly, recent studies have identified mitochondrial adaptation and/or dysfunction as components to hypertensive vascular dysfunction. While mitochondria are indispensable to maintain cellular metabolism, they also participate in adaptive and maladaptive cell/tissue responses via several retro grade signaling pathways. DRP1 plays a major role in mitochondrial quality control. However, whether DRP1 is involved in mitochondrial dysfunction and endothelial inflammation during development of HTN remains unknown. In the present study, we tested the hypothesis that inflammatory stimuli, through DRP1-dependent mitochondrial alteration, enhance endothelial inflammation. In cultured rat aortic endothelial cells (RAECs), TNFα (10 μg/mL) transiently induced mitochondrial fission maximally at 3h which was inhibited using a mitochondrial fission inhibitor, Mdivi1 (10 μM) (0.16±0.04 vs 0.10±0.02 mitochondria fragmentation count with MitoTracker, p<.01 ). TNFα and FCCP (a fission agonist, 10 μM) increased THP-1 monocyte adhesion to RAECs, which was also inhibited with Mdivi1 (256±17 vs 139±16 for TNFα, 238±30 vs 156±14 for FCCP, attached cells per field scanned, p<.01 ). Likewise, mdivi1 and adenoviruses encoding siRNA for DRP1 or dominant-negative K38A DRP1 (50 moi) attenuated TNFα-induced VCAM-1 induction in RAECs. TNFα increased aerobic respiration, which was prevented by mdivi1 or ER stress inhibitor PBA (10 mM). Inhibition of ER stress, glycolysis or mitochondrial respiration using PBA, 2-DG (1 mg/mL) or oligomycin (1 μM) prevented VCAM-1 induction. However, suppression of TNFα-induced mitochondrial ROS production by mito-Tempo (25 nM) was unable to prevent VCAM-1 induction. In C57BL6 mice receiving AngII (1000 ng/kg/min, 2 weeks) infusion, treatment with Mdivi-1 (25 mg/kg ip every other day) or PBA (1g/kg/day) prevented vascular VCAM-1 induction. In conclusion, our data suggests a critical role for ER stress and subsequent functional and structural remodeling of mitochondria induced by DRP1 in mediating endothelial inflammatory activation in HTN.

scholarly journals Predicting Persistent Opioid Use, Abuse, and Toxicity Among Cancer Survivors

2019 ◽  
Vol 112 (7) ◽  
pp. 720-727 ◽  
Author(s):  
Lucas K Vitzthum ◽  
Paul Riviere ◽  
Paige Sheridan ◽  
Vinit Nalawade ◽  
Rishi Deka ◽  
...  
Keyword(s):  
Risk Factors ◽  
At Risk ◽  
Cancer Survivors ◽  
Critical Role ◽  
Multivariable Analysis ◽  
Area Under The Curve ◽  
Opioid Abuse ◽  
Opioid Use ◽  
Multivariable Logistic Regression Model ◽  
Patients At Risk

Abstract Background Although opioids play a critical role in the management of cancer pain, the ongoing opioid epidemic has raised concerns regarding their persistent use and abuse. We lack data-driven tools in oncology to understand the risk of adverse opioid-related outcomes. This project seeks to identify clinical risk factors and create a risk score to help identify patients at risk of persistent opioid use and abuse. Methods Within a cohort of 106 732 military veteran cancer survivors diagnosed between 2000 and 2015, we determined rates of persistent posttreatment opioid use, diagnoses of opioid abuse or dependence, and admissions for opioid toxicity. A multivariable logistic regression model was used to identify patient, cancer, and treatment risk factors associated with adverse opioid-related outcomes. Predictive risk models were developed and validated using a least absolute shrinkage and selection operator regression technique. Results The rate of persistent opioid use in cancer survivors was 8.3% (95% CI = 8.1% to 8.4%); the rate of opioid abuse or dependence was 2.9% (95% CI = 2.8% to 3.0%); and the rate of opioid-related admissions was 2.1% (95% CI = 2.0% to 2.2%). On multivariable analysis, several patient, demographic, and cancer and treatment factors were associated with risk of persistent opioid use. Predictive models showed a high level of discrimination when identifying individuals at risk of adverse opioid-related outcomes including persistent opioid use (area under the curve [AUC] = 0.85), future diagnoses of opioid abuse or dependence (AUC = 0.87), and admission for opioid abuse or toxicity (AUC = 0.78). Conclusion This study demonstrates the potential to predict adverse opioid-related outcomes among cancer survivors. With further validation, personalized risk-stratification approaches could guide management when prescribing opioids in cancer patients.

scholarly journals Periodontal Disease, Inflammatory Cytokines, and PGE2 in Pregnant Patients at Risk of Preterm Delivery: A Pilot Study

2018 ◽  
Vol 2018 ◽  
pp. 1-7 ◽  
Author(s):  
Catalina Latorre Uriza ◽  
Juliana Velosa-Porras ◽  
Nelly S. Roa ◽  
Stephani Margarita Quiñones Lara ◽  
Jaime Silva ◽  
...  
Keyword(s):  
At Risk ◽  
Pilot Study ◽  
High Risk ◽  
Periodontal Disease ◽  
Preterm Delivery ◽  
Inflammatory Cytokines ◽  
Inflammatory Responses ◽  
Low Risk ◽  
Prostaglandin E ◽  
Patients At Risk

Periodontal disease is an infection that, in pregnant women, can act as a risk factor for preterm delivery by increasing local and systemic inflammatory responses. Objective. To analyze the presence of periodontal disease, proinflammatory cytokines, and prostaglandin E2 (PGE2) in pregnant patients at high risk for preterm delivery. Materials and Methods. Pilot study for a case-control study. We included 46 pregnant patients (23 patients at risk of preterm delivery as cases and 23 patients without risk of preterm delivery as controls). We excluded patients who received periodontal treatment, antibiotics, or antimicrobials over the last 3 months as well as those with infections or diseases such as diabetes or hypercholesterolemia. The patients underwent a periodontal assessment, and their levels of cytokines (interleukin- [IL-] 2, IL-6, IL-10, and tumor necrosis factor- [TNF-] α) and prostaglandin E2 (PGE2) were quantified. Results. Patients with periodontal disease showed higher levels of cytokines (IL-2, IL-6, IL-10, and TNF-α) and PGE2. Patients at high risk for preterm birth showed higher IL levels compared with patients at low risk for preterm delivery. PGE2 increased with the severity of periodontal disease. PGE2 was higher in patients at low risk for preterm delivery, although this difference was not significant. Conclusion. Periodontal disease can increase the systemic inflammatory response as well as the levels of PGE2 and inflammatory cytokines in pregnant patients.

scholarly journals Systemic processes in influenza in fection, as a triggers of the development of somatic pathology in patients at risk

2015 ◽  
Vol 20 (4) ◽  
pp. 47-53
Author(s):  
E. G Deeva ◽  
T. G Zubkova ◽  
N. V Dunaeva ◽  
S. Zh Koltsebaeva ◽  
G. Yu Chelaeva ◽  
...  
Keyword(s):  
At Risk ◽  
Neurological Diseases ◽  
Influenza Infection ◽  
Critical Role ◽  
Risk Groups ◽  
Patients At Risk ◽  
Triggering Factor ◽  
Complicated Course ◽  
Severe Infections ◽  
The One

Pathogenetic mechanisms of influenza infection on the one hand are the triggering factor of certain diseases (asthma, neurological diseases and others.), on the other hand they worsen the course of concomitant somatic pathology, leading to severe, complicated course of infection and lethal outcomes in high-risk groups. The two components of the flu - a syndrome of systemic inflammation, which is manifested in the overproduction of cytokines and generalized vascular thrombosis syndrome (VTS) are the most important components of the pathogenesis of influenza and play a critical role in the development of severe infections, especially in patients with a history ofpremorbid background. Analysis of the pathogenic mechanisms of diseases, at risk, is necessary for the development of a comprehensive targeted tactics prevention, treatment and medical examination that will prevent mortality in these groups.

scholarly journals Intestinal microbiota predicts lung cancer patients at risk of immune-related diarrhea

Immunotherapy ◽  
2019 ◽  
Vol 11 (5) ◽  
pp. 385-396 ◽  
Author(s):  
Tian Liu ◽  
Qi Xiong ◽  
Lingling Li ◽  
Yi Hu
Keyword(s):  
Lung Cancer ◽  
At Risk ◽  
Cancer Patients ◽  
Intestinal Microbiota ◽  
Lung Cancer Patients ◽  
Patients At Risk

scholarly journals Mechanisms of maintenance of intestinal homeostasis by autochthonic microbiota and probiotics

2016 ◽  
Vol 72 (10) ◽  
pp. 611-615 ◽  
Author(s):  
Marian Binek ◽  
Magdalena Kizerwetter-Świda ◽  
Agata Anna Cisek ◽  
Magdalena Rzewuska ◽  
Dorota Chrobak-Chmiel ◽  
...  
Keyword(s):  
Intestinal Microbiota ◽  
Competitive Exclusion ◽  
Inflammatory Responses ◽  
Critical Role ◽  
Probiotic Bacteria ◽  
Secretory Iga ◽  
Epithelial Barrier Function ◽  
Intestinal Microbes ◽  
Innate Defense ◽  
Pathogen Colonization

Intestinal microbes are taxonomically diverse and constitute an ecologically dynamic microbiom interactively performing various physiological and physiopathological processes. It has been proposed that normal intestinal microbiotas play a critical role in the host’s metabolic homeostasis and immune tolerance. The modulation of intestinal microbiota populations by prebiotics, probiotics, and synbiotics may be beneficial for the host’s health. Under certain conditions, the intestinal microbiota and the host’s homeostasis can be restored by introducing bacteria that co-mediate anti-inflammatory responses. Commensal microbes and probiotics exert their beneficial effect by at least three mechanisms. These include - the maintenance of the epithelial barrier function and the attenuation of changes in intestinal permeability through effects on tight junction, decreasing paracellular permeability, providing innate defense against pathogens, and enhancing the physical impediment of the mucous layer, - competitive exclusion by the application of probiotic bacteria stabilizing the indigenous microflora, - immunomodulatory capacity, affecting a variety of signaling pathways with modulation of proper immune, inflammatory and allergic responses. The epithelial gut barrier faces important challenges, since its function is to prevent pathogens and harmful elements of the gut lumen from penetrating into the internal environment. Competitive exclusion treatment can increase resistance to pathogen colonization and control intestinal disturbance. The dominance of symbiotic and probiotic bacteria among the gut microbiota favors a tolerogenic immune response. The release of secretory IgA stabilizes tight junctions between cells of the epithelial layer as well as hampers pathogens and symbionts invading deeper layers. The understanding of these vital processes may help to protect the host against infection, prevent chronic inflammation, and maintain mucosal integrity.

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