Introduction

Constipation is associated with intestinal microbiota and metabolites. Nevertheless, the causal relationship between intestinal microbial and constipation remains to be demonstrated.

Aims & Methods

The current study aimed to research associations between microbiota composition and constipation to understand better how intestinal microbes contribute to gastrointestinal disturbances observed in patients. We assessed the taxonomic using the 16S rRNA sequencing and the metabolic signatures through liquid chromatography-mass spectrometry (LC/MS) of stool microbiota in constipation and healthy donor groups.

Results

The results showed that the α-diversity of the gut microbial population was increased, and at the species level, the abundance of ruminococcus_lactaris was elevated; meanwhile, the significantly differential metabolite indole-3-carboxaldehyde (3-IAId) was markedly elevated in the constipation group.By analysing microbiota and their association with metabolites, we found that the abundance of ruminococcus_lactaris was positively correlated with the abundance of 3-IAId in the constipation group. Then we confirmed ruminococcus_lactaris could produce 3-IAId in vitro. Further studies show that colonisation of SPF mice with fecal samples from patients with constipation significantly prolonged the gastrointestinal transit time of mice. Moreover，ruminococcus_lactaris monocolonization can induce constipation in SPF mice. Besides, we observed that it was sufficient to inhibit the peristaltic contraction of the colon in mice both in vivo and in vitro by treatment with 3-IAId.

Conclusion

In This study, we clarify that ruminococcus_lactaris and its metabolite 3-IAId could induce constipation in mice, and the 3-IAId could be used as a novel biomarker for constipation.

Introduction

Diagnostics of enteropathy with membrane digestion disturbance (EMDD) is based on the evaluation of carbohydrase activity in duodenal mucosa (DM). There is no information about diagnostic value of ileum mucosa (IM) carbohydrases.
Study objective. Assess the diagnostic value of carbohydrases activity in mucous membrane of duodenum and distal ileum in patients with EMDD and irritable bowel syndrome (IBS).

Aims & Methods

The study includes 12 patients with EMDD, 16 patients with IBS and 12 relatively healthy persons (control group). Study design included analysis of food tolerance and evaluation of activity of sucrase (S), glucoamylase (GA), maltase (М) and lactase (L) in mucous membrane of duodenum and ileum based on the method of Dahlquist with N.I.Belostotsky modification. Microsoft Office Excel and STATISTICA 10.0 were used for statistical evaluation of material. Mann-Whitney criterion was used to evaluate the differences between quantitative parameters. The differences were considered to be significant at p <0.05. Control group was used for comparison.

Results

Ratio of carbohydrase activity in duodenum and ileum in control group and patients with IBS and EMDD.

In control group the carbohydrases activity in duodenum mucosa was not significantly statistically different from the one in ileum mucosa.
In patients with IBS glucoamylase and maltase activity parameters in IM were significantly different from activity in DM, which can be interpreted as functional inequality of enzyme activity of various departments of small intestine.
In patients with EMDD GA level in ileum mucosa is 5.2 times higher, maltase level is 3.8 times higher and sucrose level is 2 times higher than duodenum enzyme levels, which demonstrates the possibility of formation of compensatory reaction of adaptive shift type towards distal segments of small intestine, initially described by A.M. Ugolev et al. in 1985

Conclusion

Enzyme activity distribution along small intestine is an adaptor function of its segments. Our study confirmed the need and promising outlook of further investigations in terms of evaluation of proximo-distal gradients of enzyme activity to study compensatory reactions which can be fulfilled by distal segment of small intestine in case of duodenum damage.
The evaluation of carbohydrase activity in ileum mucosa provides additional opportunities for diagnostics of EMDD and its differentiation from IBS, which can be used to improve therapeutic approaches.

Introduction

Clinical trials in coeliac disease require reliable histological metrics to quantify mucosal damage and demonstrate improvement with treatment. Regulatory agencies are recognizing this and expect some form of histological assessment in coeliac disease trials. Qualitative scoring systems are highly variable among readers. It has been suggested that quantitative scoring provides better consistency among different readers.

Aims & Methods

This study aimed to assess the variability that occurs when quantifying villus height (Vh), crypt depth (Cd) and the Vh:Cd ratio, and identified approaches to improve correlation among observers at different institutions. A set of 236 slides stained with haematoxylin and eosin from 28 patients with coeliac disease on a gluten-free diet (NCT03738475) were used. Quantitative scoring of Vh, Cd, Vh:Cd ratio and the sum of Vh and Cd were initially compared between different institutions that prioritized ocular microscopy or digital pathology. Additionally, the change from baseline with gluten ingestion for each of these measurements was compared between institutions. Improvements to inter-reader correlations were assessed by implementing a common digital pathology software and using agreed upon criteria for making quantitative measurements. Data sets were subjected to statistical analysis to understand correlation and significance. Regression slopes were calculated and the coefficient of determination, R², was derived as a measure of linearity. The concordance correlation coefficient was calculated as an index of agreement beyond linearity. Pairwise Pearson correlation coefficients between readers from different institutions were calculated to demonstrate the degree of association between readers.

Results

Initial inter-reader analysis found that Vh:Cd ratios had a similar, linear trend between readers, regardless of ocular or digital pathology usage, but that there was a consistent systematic difference partially due to differences in reader assessment of the location of the crypt–villus border. Use of a common digital pathology platform and clear scoring criteria greatly improved inter-reader correlations, even when scoring was done by a non-pathologist. Importantly, the assessment of change from baseline in the quantitative measurements after gluten exposure was also highly concordant (Table). The software also enabled visual comparison of readers’ individual measurements of the crypt and villous lengths allowing potential reasons for differences in these measurements to be identified.

Table 1. Changes from baseline in Vh and Cd metrics obtained via ocular microscopy and digital pathology after gluten challenge.

Metric	Substudy 1		Substudy 2	Difference from institution 2
	Digital pathology (institution 2)	Ocular microscopy (institution 1)	Digital pathology + training (institution 1)	Substudy 1 (institution 1)	Substudy 2 (institution 1)
Vh, μm	−95.25 (−130.84, −59.66)**	−38.01 (−72.90, −3.12)*	−87.3 (−137.5, −37.0)†	−57.24**	−7.95†
Cd, μm	50.61 (35.87, 65.34)**	7.73 (−0.26, 15.72)†	40.7 (16.6, 64.8)†	42.88**	9.91†
Vh:Cd ratio	−0.79 (−1.03, −0.54)**	−0.38 (−0.61, −0.15)*	−0.7 (−1.0, −0.4)†	−0.41**	−0.09†
Vh+Cd, μm	−44.64 (−74.62, −14.67)*	−30.28 (−67.44, 6.88)†	−46.5 (−83.9, −9.1)†	−14.36†	1.90†
*p < 0.05 for difference between digital and ocular reads. **p < 0.001 for difference from baseline (0). †p values were not significant (> 0.05). Data are mean (95% CI). Vh, Cd, Vh:Cd ratio and Vh+Cd measurements were obtained by ocular microscopy by one reader from institution 1 and by digital pathology by three readers from institution 2. Shown here are the changes in Vh and Cd metrics between baseline and Day 14 after gluten challenge and the changes (improvement) in the performance of the reader (same reader in each substudy) between substudies 1 and 2. Cd, crypt depth; CI, confidence interval; NR, not reported; Vh, villus height; Vh+Cd, sum of the villus height and crypt depth; Vh:Cd, villus height:crypt depth.

Conclusion

The results suggest that quantitative histology in coeliac disease is reliable and can be improved through digital pathology, software measurement aids and attention to selecting optimally oriented sections. Given the ease with which pathologists without training in the use of quantitative histology software can produce results similar to those from experienced central laboratories, quantitative histology may be feasible outside the clinical trial setting.

Disclosure

AV has nothing to disclose. DAL, DR, DC, GS, JM and QL are employees of Takeda Development Center Americas, Inc. and receive stock or stock options. JI is the founder and owner of Jilab, Inc.; is a Full Professor at Tampere University, Medical Faculty; and has a consulting agreement with Takeda. JAM has grants from Nexpep/ImmusanT, National Institutes of Health, Immunogenix, Takeda Pharmaceutical, Allakos, Oberkotter, Cour; and has received consultancy fees from Bionix, Lilly Research Laboratory, Johnson & Johnson, Dr. Schar USA, UCB Biopharma, Celimmune, Intrexon Corporation, Dren Bio, Reistone Pharma, Chugai Pharma, Kanyos, Boehringer Ingelheim, Equillium and Torax Medical. MR is a consultant for Takeda and Bristol Myers Squibb. MZC has nothing to disclose. TTW has nothing to disclose.
Funding statement: This study was sponsored by Takeda Development Center Americas, Inc. Medical writing support was provided by Oxford PharmaGenesis, Oxford, UK and was funded by Takeda Pharmaceuticals.

Introduction

There have been advances in endoscopic hemostasis in acute gastrointestinal bleeding, from the conventional injection of submucosal solutions of dilute epinephrine to the application of thermal energy and, more recently, mechanical hemostasis (1,2). There has been the development of non-contact hemostasis in the form of an endoscopically applied hemostatic powder spray. When applied, these inert compounds absorb water, which concentrates clotting factors on the surface of the particles, thus initiating the clotting cascade. These compounds enhance clot formation, and the coagulum sloughs off after 24 to 72 hours. The primary objective of the study is to evaluate the safety of the Resolv® Endoscopic Hemostat System. The secondary objective is to evaluate the efficacy of the use of the Resolv© Endoscopic Hemostat System in the control of non-variceal GI bleed.

Aims & Methods

This is a single-center, prospective, open-label, single-arm study (CTRI/2021/12/038478) to evaluate the safety and efficacy of a novel plant-based polysaccharide Hemostasis’ Resolv© Endoscopic Hemostat System, from July 2022 to February 2023, in achieving acute hemostasis in adult subjects diagnosed with non-variceal upper gastrointestinal bleeding. Clinically known or confirmed GI bleeding—nonvariceal UGIB with bleeding severity of Forrest 1b, such as post-EMR, ESD, or tumour bleed—underwent standard endoscopy. Patients were evaluated for the proportion of subjects with AE or SAE within 72 hours after hemostat application, the proportion of subjects with device-related SAE within 30 days of the procedure, the proportion of subject mortality within 30 days of the procedure (all-cause) [Primary endpoints], the proportion of subjects with acute procedural hemostasis (index endoscopy), the proportion of subjects with recurrent bleeding within 72 hours of the index procedure/ application, and the need for surgery or an alternative hemostat for failure to achieve hemostasis within 72 hours [Secondary endpoints]. Blood sugars were closely monitored pre- and post-procedure, due to concerns regarding its effect on RBS since the powder is a plant-based polysaccharide.

Results

Sixty patients (M: F- 42/18, Age: 54.7±13.9) underwent the procedure and were considered for analysis. The device was used in the following indications: post-polypectomy (n = 34), peptic ulcer bleed (n = 12), malignant tumour bleed (n = 6), biopsy bleed (n=5), congestive gastropathy bleed (n=2), post-EMR bleed (n = 1). The technical success rate was 100%. Within 72 hours of use, there were no adverse events or side effects related to the device, and there was no mortality (all-cause) secondary to device use (Primary endpoints). The index endoscopy hemostasis was 100%. Within 72 hours of hemostasis, n=1 patients had a rebleeding (malignant ulcer bleed), which was controlled using Argon plasma coagulation (Secondary endpoints). The mean RBS was 150.3 mg/dl in 18 of 60 (30%) diabetic patients. The mean RBS 2 hours and 4 hours post-procedure was 130 and 131 mg/dl.

Conclusion

Resolv® Endoscopic Hemostat System is a safe and highly efficacious device for achieving acute hemostasis in adult subjects diagnosed with non-variceal upper gastrointestinal bleeding.

References

1. Lau JYW, Yu Y, Tang RSY, et al. Timing of Endoscopy for Acute Upper Gastrointestinal Bleeding. N Engl J Med 2020;382:1299-308
2. Laine L, Jensen DM. Management of patients with ulcer bleeding. Am J Gastroenterol 2012;107:345-60; quiz 61.