scholarly journals P013 The JAK-3 and TYK-2 / STAT pathways are activated in moderate to severe ulcerative colitis

2020 ◽  
Vol 14 (Supplement_1) ◽  
pp. S136-S137
Author(s):  
M Loza ◽  
J M Brea ◽  
C Calviño-Suarez ◽  
I Baston-Rey ◽  
R Ferreiro-Iglesias ◽  
...  
Keyword(s):  
Ulcerative Colitis ◽  
Animal Models ◽  
Inflammatory Response ◽  
Western Blot ◽  
Signalling Pathways ◽  
Target Engagement ◽  
Inclusion Criteria ◽  
Phosphorylated Proteins ◽  
Single Centre Study ◽  
Stat Pathway

Abstract Background Ulcerative colitis (UC) is a chronic, progressive and disabling disease with a complex pathology of unknown aetiology influenced by genetic, environmental and microbiota factors that lead to an immunological and inflammatory response in the colon. Janus Activated Kinase (JAK) family plays a key role in modulating the adaptive and innate inflammatory response. The JAK/STAT pathway involvement in UC has been demonstrated in both animal models and human studies. Thus, overexpressed JAK-3 has been detected in the intestine of patients with UC, suggesting a key role in their pathophysiology and the inhibition of TYK-2 in animal models resulted in an improvement of the disease, which would explain its implication in the inflammatory process. We hypothesise here that there could be an activation of JAK-3 and TYK-2 signalling pathways in UC patients. Thus, we aimed to detect the activation of both signalling pathways by means of western-blot studies in UC patient samples Methods A prospective, observational single-centre study was designed. Inclusion criteria were adult patients with endoscopic active UC (more than Mayo-0) confirmed in a programmed colonoscopy. All patients signed informed consent. Samples were obtained from overstock of routine biopsies in the more severe segment affected of the large bowel. Tissues were homogenised and processed in order to obtain cell lysates by employing RIPA buffer and ultrasounds. The degree of activation of the JAK-3 and TYK-2 pathways was measured by detecting the phosphorylation of both targets as well as of STAT1, STAT3, STAT4, STAT5 and STAT6 through western blot by employing specific antibodies for total and phosphorylated proteins. Results 19 UC patients were consecutively included. Mean age was 46 years old. 53% were female, 47% were extensive colitis (E3) and 53% left-side colitis (E2). Regarding endoscopic activity, 26% had Mayo-1, 53% Mayo-2, and 21% Mayo-3. Immunoreactive bands for both phosphorylated JAK-3 and TYK-2 were detected in the biopsies from UC patients, evidencing that colonic inflammation leads to an activation of both targets. The study of STATs phosphorylation showed immunoreactive bands for phosphorylated forms of STAT1, STAT3, STAT4, STAT5 and STAT6 confirming the activation of both signalling-pathways in these patients (Figure 1). Conclusion The developed translational workflows involving basic/clinical research confirm the activation of both JAK-3 and TYK-2-dependent signalling pathways in UC patients, validating both kinases as targets for treating UC. The developed methodology allows studying the target engagement for future JAK-3/ TYK-2 inhibitors employed in clinical trials.

scholarly journals Methods Used and Application of the Mouse Grimace Scale in Biomedical Research 10 Years On: A Scoping Review

Animals ◽  
2021 ◽  
Vol 11 (3) ◽  
pp. 673
Author(s):  
Alexandra L. Whittaker ◽  
Yifan Liu ◽  
Timothy H. Barker
Keyword(s):  
Animal Models ◽  
Mouse Models ◽  
Biomedical Research ◽  
Scoping Review ◽  
Facial Features ◽  
Inclusion Criteria ◽  
Academic Literature ◽  
Research Fields ◽  
Neuroscience Research ◽  
Wide Range

The Mouse Grimace Scale (MGS) was developed 10 years ago as a method for assessing pain through the characterisation of changes in five facial features or action units. The strength of the technique is that it is proposed to be a measure of spontaneous or non-evoked pain. The time is opportune to map all of the research into the MGS, with a particular focus on the methods used and the technique’s utility across a range of mouse models. A comprehensive scoping review of the academic literature was performed. A total of 48 articles met our inclusion criteria and were included in this review. The MGS has been employed mainly in the evaluation of acute pain, particularly in the pain and neuroscience research fields. There has, however, been use of the technique in a wide range of fields, and based on limited study it does appear to have utility for pain assessment across a spectrum of animal models. Use of the method allows the detection of pain of a longer duration, up to a month post initial insult. There has been less use of the technique using real-time methods and this is an area in need of further research.

scholarly journals Nigella A ameliorates inflammation and intestinal flora imbalance in DSS induced colitis mice

AMB Express ◽  
2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Xingjiang Hu ◽  
Nana Xu ◽  
Xi Yang ◽  
Xi Hu ◽  
Yunliang Zheng ◽  
...  
Keyword(s):  
Ulcerative Colitis ◽  
Chinese Medicine ◽  
Traditional Chinese Medicine ◽  
Inflammatory Response ◽  
Therapeutic Effect ◽  
Antioxidant Activities ◽  
Intestinal Flora ◽  
Signs And Symptoms ◽  
Microbial Composition ◽  
Bacterial Genus

Abstract Nigella A, also named Sieboldianoside A, has been extracted from many kinds of Traditional Chinese Medicine (TCM), such as Nigella glandulifera, Stauntonia chinensis DC., and the leaves of Acanthopanax sieboldianus. Nigella A exhibited potential analgesic, anti-inflammatory, anti-tumor, and antioxidant activities. However, whether Nigella A could treat ulcerative colitis (UC) is still unknown. As saponins always be regarded as the kinds of ingredients that could regulate immunity and intestinal flora. This research aimed to investigate the therapeutic effect of Nigella A on UC and explore its effect on intestinal flora. We noted that Nigella A and Sulfasalazine (SASP) could significantly improve the signs and symptoms, alleviate colonic pathological injury in DSS-induced mice. The changing of many specific bacterial genus such as Lactobacillus, Porphyromonadaceae, Bacteroides and Escherichia might closely related to the recovery of intestinal inflammatory response. This study initially confirmed the therapeutic effect of Nigella A and SASP on DSS-induced colitis by improving the diversity of intestinal microbial composition. Nigella A has the potential to be developed for the treatment of UC and other disorders related to the imbalance of intestinal flora.

Abstract W MP50: Pair Housing Reverses the Detrimental Effect of Social Isolation and Restores BDNF and MBP in Aged Mice after Stroke

Stroke ◽  
2015 ◽  
Vol 46 (suppl_1) ◽  
Author(s):  
Rajkumar Verma ◽  
Nia Harris ◽  
Brett Friedler ◽  
Joshua Crapser ◽  
Anita Patel ◽  
...  
Keyword(s):  
Inflammatory Response ◽  
Western Blot ◽  
Social Isolation ◽  
Enhanced Recovery ◽  
Infarct Volume ◽  
Recognition Task ◽  
Aged Mice ◽  
Post Stroke ◽  
Beneficial Effects ◽  
Novel Object Recognition Task

Background: Age is an important non-modifiable risk factor for stroke. Stroke rates double every decade after age 55. Social isolation (SI) exacerbates behavioural deficits, slows functional recovery and worsens histological injury after stroke in young animals, primarily by increasing the inflammatory response. However, the inflammatory response differs in aging, and whether the detrimental effects of SI are seen in aged animals is unknown. We hypothesize that acute and chronic post stroke SI will worsen stroke pathology and recovery in aged mice and pair housing (PH) will reverse these effects. Methods: Eighteen-month-old male C57BL/6 mice were pair housed (PH) for two weeks prior to stroke and randomly assigned to various housing conditions immediately after stroke. Behavioral analysis was done weekly starting at day 7. Mice were sacrificed either at 72 hours or 4 weeks after 60-minute right MCAO or sham surgery (n=9-10/group). Results: Mice isolated after stroke (ST-ISO) mice had significantly greater hemispheric infarct volume and neurological deficit scores (p<.05. n=13/group) than pair-housed (PH) stroke mice at 72 hours. SI mice that were isolated immediately after stroke showed significantly higher plasma IL-6 levels compared to PH sham (P<.001, n=13/group ) or PH stroke mice (P<.05) after 72 hour, but levels were similar by 4 weeks post-stroke (n=9-14/group). No change in tissue atrophy was seen after 4 weeks, however a significant interaction [F (1, 28) = 259.6, P<0.001] between housing and stroke was found in the Novel Object Recognition Task (NORT) at day 14. PH led to increased expression of Brain-derived neurotrophic factor (BDNF) and myelin basic protein (MBP) by IHC and western blot (n=5/group for IHC and n= 4/ western blot). Conclusions: Social isolation immediately after stroke led to enhanced injury acutely. Despite similar infarcts at 4 weeks, SI mice had delayed behavioral recovery. Pair housing led to increased expression of BDNF and myelin protein expression. Therefore, the beneficial effects of pair housing may be related to BDNF and MBP expression and enhanced recovery after injury in aged animals.

EP.WE.1048Weight Regain After Sleeve Gastrectomy More Common in Male and Less Common in Super-Obese Patients - Results of an 18-Month Follow-up Study

2021 ◽  
Vol 108 (Supplement_7) ◽  
Author(s):  
Ahmed Salman Bodla ◽  
Jenny Abraham ◽  
Neha Shah ◽  
Vinod Menon
Keyword(s):  
Sleeve Gastrectomy ◽  
Revisional Surgery ◽  
Patient Specific ◽  
Obese Patients ◽  
Inclusion Criteria ◽  
Super Obese ◽  
Burn Out ◽  
Single Centre Study ◽  
Male Patients

Abstract Aims Long-term success of Sleeve Gastrectomy (SG) is undermined by weight regain (WR). Several procedure- and patient-specific factors have been proposed in previous studies. Here we look at 18-month follow-up post-SG to investigate WR and patient-specific variables influencing this. Methods A single-centre study involving retrospective analysis of a prospectively-maintained database. Inclusion criteria: Primary non-revisional SG patients with adequate follow-up data to assess WL/WR trends. Patients were divided into two subgroups based on their %EWL between 6-to-12 and 12-to-18 months: weight regainers (WR) and weight losers/maintainers (WL/M). Results Out of 338 SG cases between 2012-2017, 180 met inclusion criteria of which 18.3% were men and 45% were super-obese. All patients lost weight during first 6 months (mean %EWL 52.3%, P &lt; 0.0001). Between 6-to-12 months, 87.6% patients continued WL with a further mean %EWL of 10.35% (P &lt; 0.0001). Between 12-to-18 months, a drastic deceleration/reversal of WL progress was observed with an average of only 0.76% EWL (P = 0.84), with 42% of patients regaining weight in this period (mean EWG 6.8%). Male patients encountered significantly higher WR rate (OR 3.27, P = 0.003), whereas it was much less frequent in pre-operatively super-obese patients (OR 0.48, P = 0.036). Moreover, there was no difference in the 6-month %EWL between WR and WL/M subgroups (P = 0.62), thus negating the possibility of WL burn-out phenomenon. Conclusions Different rates of WR in men and super-obese patients may indicate underlying behavioural and biological differences. More research is needed to investigate them in detail, having implications for revisional surgery and follow-up support.

scholarly journals IL-8 and LYPD8 expression levels are associated with the inflammatory response in the colon of patients with ulcerative colitis

2020 ◽  
Author(s):  
Tomoyuki Okada ◽  
Tsutomu Kanda ◽  
Naoki Ueda ◽  
Yuichiro Ikebuchi ◽  
Keiichi Hashiguchi ◽  
...  
Keyword(s):  
Ulcerative Colitis ◽  
Inflammatory Response ◽  
Expression Levels

scholarly journals Investigation of Potential Genetic Biomarkers and Molecular Mechanism of Ulcerative Colitis Utilizing Bioinformatics Analysis

2019 ◽  
Author(s):  
Jiaqi Zhang ◽  
Xue Wang ◽  
Lin Xu ◽  
Zedan Zhang ◽  
Fengyun Wang ◽  
...  
Keyword(s):  
Ulcerative Colitis ◽  
Gene Therapy ◽  
Inflammatory Response ◽  
Signaling Pathway ◽  
Functional Enrichment ◽  
Hub Genes ◽  
Amyloid A ◽  
Chemokine Signaling ◽  
Chemokine Signaling Pathway ◽  
Potential Biomarkers

Abstract Objectives: To reveal the molecular mechanisms of ulcerative colitis (UC) and provide potential biomarkers for UC gene therapy. Methods: We downloaded the GSE87473 microarray dataset from the Gene Expression Omnibus (GEO) and identified the differentially expressed genes (DEGs) between UC samples and normal samples. Then ,a module partition analysis was performed based on a weighted gene co-expression network analysis (WGCNA),followed by pathway and functional enrichment analyses. Furthermore, we investigated the hub genes . At last, data validation was performed to ensure the reliability of the hub genes. Results: Between UC group and normal group, 988 DEGs were investigated . The DEGs were clustered into 5 modules using WGCNA. These DEGs were mainly enriched in functions such as the immune response, the inflammatory response and chemotaxis, and they were mainly enriched in KEGG pathways such as the cytokine-cytokine receptor interaction , chemokine signaling pathway, and complement and coagulation cascades. The hub genes, including dual oxidase maturation factor 2(DUOXA2), serum amyloid A (SAA) 1 and SAA2, TNFAIP3-interacting protein 3(TNIP3), C-X-C motif chemokine (CXCL1), solute carrier family 6 member 14(SLC6A14) and complement decay-accelerating factor (CD antigen CD55),were revealed as potential tissue biomarkers for UC diagnosis or treatment. Conclusions: This study provides supportive evidence that DUOXA2, A-SAA, TNIP3, CXCL1, SLC6A14 and CD55 might be used as potential biomarkers for tissue biopsy of UC, especially SLC6A14 and CD55, which may be new targets for UC gene therapy. Moreover, the DUOX2/DUOXA2, NF-κB /TNIP3 and CXCL1/CXCR2 pathways might play an important role in the progression of UC through the chemokine signaling pathway and inflammatory response.

scholarly journals FOXD3/FOXD4 is required for the development of hindgut in the rat model of anorectal malformation

2018 ◽  
Vol 243 (4) ◽  
pp. 327-333 ◽  
Author(s):  
Luo-Jia Wang ◽  
Wei-Lin Wang ◽  
Hong Gao ◽  
Yu-Zuo Bai ◽  
Shu-Cheng Zhang
Keyword(s):  
Animal Models ◽  
Western Blot ◽  
Real Time ◽  
Digestive Tract ◽  
Anorectal Malformation ◽  
Aberrant Expression ◽  
Strong Basis ◽  
Rat Embryos ◽  
Ethylene Thiourea

Congenital anorectal malformation is the most common digestive tract malformation in newborns. It has been reported that FOXD3/FOXD4, a forkhead transcription factor, regulates the generation, migration, and differentiation of neural crest cells. However, whether FOXD3/FOXD4 takes part in anorectal malformation remains unclear. In the present study, we used ethylene thiourea to induce the animal models of anorectal malformation in rat embryos and to interrogate the role of FOXD3/FOXD4 in anorectal malformation pathogenesis. Hindgut samples of the animal models were collected at E15, E17, E19, and E21 days of age. The expression of FOXD3/FOXD4 was detected by immunohistochemistry, western blot, and quantitative real-time fluorescence PCR. By immunohistochemical staining, FOXD3/FOXD4 was observed in epithelial cells of the rectum and the anus both in normal and rat embryos with anorectal malformation. Expression level analysis by western blot indicated that FOXD3/FOXD4 expression increased in ethylene thiourea-induced anorectal malformation groups. mRNA expression as determined by quantitative real-time fluorescence PCR analysis was consistent with the western blot results. Tentative conclusions were drawn that FOXD3/FOXD4 is expressed in the hindgut in rat embryos and is upregulated in anorectal malformation. FOXD3/FOXD4 is required for the development of the hindgut, and its aberrant expression may be an important factor leading to the incidence of anorectal malformation. Impact statement Congenital anorectal malformation (ARM) is the most common digestive tract malformation in newborns. The pathophysiological ground remains unclear. In this study, we used animal models of ARM for the first time to interrogate the role of FOXD3/FOXD4 in ARM pathogenesis. The animal models of ARM were successfully induced by ethylene thiourea (ETU) in rat embryos providing a strong basis for pathogenesis study of this disease. Expression analysis of FOXD3/FOXD4 was carried out in these models, and the results shape a deeper understanding of FOXD3/FOXD4 being required for the normal development of the hindgut. The aberrant expression of FOXD3/FOXD4 may be an important factor leading to ARM incidence.

Sign in / Sign up

Export Citation Format

Share Document