Biomarkers for Intestinal Permeability in Patients With Constipation

• Sponsor: Mayo Clinic (Other)
• Overall Status: Completed
• CT.gov ID: NCT02246647
• Collaborator: Takeda Pharmaceuticals International, Inc. (Industry)
• Enrollment: 39
• Location: 1
• Duration (Months): 27.2
• Patients Per Site Per Month: 1.4

Study Details

Study Description

Brief Summary

Our overall objective with this study is firstly to provide a comprehensive assessment of intestinal permeability, mucosal barrier function using existing biomarkers and secondly to explore novel biomarkers for measuring intestinal permeability in patients with constipation predominant Irritable Bowel Syndrome (IBS-C).

Condition or Disease	Intervention/Treatment	Phase
Intestinal Diseases; Irritable Bowel Syndrome; Constipation	Diagnostic Test: Permeability measurement; Procedure: Esophagogastroduodenoscopy; Procedure: Flexible sigmoidoscopy

Detailed Description

In order to determine the differences in permeability in IBS-C in comparison with healthy volunteers, the following will be determined: differences in in vivo small intestinal and colonic permeability, differences in small intestinal and colonic mucosal barrier function, differences in effects of fecal supernatants on barrier function of T84 monolayers, and differences in novel biomarkers for intestinal permeability

Study Design

• Study Type: Observational
• Actual Enrollment: 39 participants
• Observational Model: Case-Control
• Time Perspective: Cross-Sectional
• Official Title: Biomarkers for Intestinal Permeability in Patients With Functional Lower Gastrointestinal Disorders Associated With Constipation.
• Study Start Date: Sep 1, 2014
• Actual Primary Completion Date: Dec 8, 2016
• Actual Study Completion Date: Dec 8, 2016

Arms and Interventions

Arm	Intervention/Treatment
Healthy volunteers Permeability measurement: Ingestion of saccharides {mannitol (regular, 12C) 100 mg, lactulose 1 g and labelled (13C mannitol) 100 mg} in 250ml of water Esophagogastroduodenoscopy Flexible sigmoidoscopy	Diagnostic Test: Permeability measurement; Saccharide excretion was compared between IBS-C and healthy volunteers; Other Names: 12C Mannitol 13C Mannitol Lactulose ; Procedure: Esophagogastroduodenoscopy; Duodenal biopsies were collected from IBS-C and healthy volunteers; Procedure: Flexible sigmoidoscopy; Colonic biopsies were collected from IBS-C and healthy volunteers
IBS-C Permeability measurement: Ingestion of saccharides (mannitol (regular, 12C) 100 mg, lactulose 1 g and labelled (13C mannitol) 100 mg} in 250ml of water Esophagogastroduodenoscopy Flexible sigmoidoscopy	Diagnostic Test: Permeability measurement; Saccharide excretion was compared between IBS-C and healthy volunteers; Other Names: 12C Mannitol 13C Mannitol Lactulose ; Procedure: Esophagogastroduodenoscopy; Duodenal biopsies were collected from IBS-C and healthy volunteers; Procedure: Flexible sigmoidoscopy; Colonic biopsies were collected from IBS-C and healthy volunteers

Outcome Measures

Primary Outcome Measures

1. Lactulose:C13 Mannitol Excretion Ratio 8-24hrs. [8-24 hr post test-dose administration]

   In vivo measurement of intestinal permeability using 13C mannitol & lactulose was used. High performance liquid chromatography-tandem mass spectrometry was used to measure concentrations calculated using the overall urine volume excreted in each interval. Concentrations of 13C adjusted for the % of 13C in 12C mannitol (4.98% of 12C mannitol excreted was subtracted from 13C mannitol values; determined by analyzing replicate samples of control urine). All lactulose or 13C mannitol concentrations 8-24hr post-ingestion were used to determine colonic permeability. Lactulose to 13C mannitol excretion ratios, as a measure of dose of saccharide administered, were calculated.

Secondary Outcome Measures

1. Lactose:C13 Mannitol Excretion Ratio 0-2hours [0-2 hr post-test dose administration]

2. Baseline Transmucosal Resistance (TMR) of Duodenal Mucosa [Baseline]

3. Cumulative FITC-Dextran (4kDa) Concentration Across Duodenal Mucosa [3 hours post FITC-Dextran (4kDa) administration]

   This is not a pharmacokinetic or pharmacodynamic measure. Hence only one time assessment is made 3 hours after FITC-Dextran (4kDa) administration.

4. Rate of FITC-Dextran (4kDa) Flux Across Duodenal Mucosa [Over 3 hours post FITC-Dextran (4kDa) administration]

   This is not a pharmacokinetic or pharmacodynamic measure. Hence only one time assessment is made 3 hours after FITC-Dextran (4kDa) administration.

5. Baseline Transmucosal Resistance (TMR) of Colonic Mucosa [Baseline]

6. Cumulative FITC-Dextran (4kDa) Concentration Across Colonic Mucosa [3 hours post FITC-Dextran (4kDa) administration]

7. Rate of FITC-Dextran (4kDa) Flux Across Colonic Mucosa [Over 3 hours post FITC-Dextran (4kDa) administration]

8. Cumulative E.Coli Bio- Particle K12 Concentration Across Duodenal Mucosa [3 hours post E.coli Bio- Particle administration]

9. Rate of E.Coli Bio- Particle K12 Flux Across Duodenal Mucosa [Over 3 hours post E.coli Bio- Particle administration]

10. Cumulative E.Coli Bio- Particle K12 Concentration Across Colonic Mucosa [3 hours post E.coli Bio- Particle administration]

11. Rate of E.Coli Bio- Particle K12 Flux Across Colonic Mucosa [Over 3 hours post E.coli Bio- Particle administration]

12. Duodenal Impedance [Baseline]

13. Mean Serum Endotoxin (Bacterial LPS) Levels [Fasting, one time measurement after 8 hours]

Eligibility Criteria

Criteria

• Ages Eligible for Study: 18 Years to 65 Years
• Sexes Eligible for Study: Female
• Accepts Healthy Volunteers: Yes

Inclusion criteria:

1. 18 - 65 years old

2. IBS-C by Rome III criteria (for IBS-C participants)

3. No abdominal surgery (except appendectomy and cholecystectomy)

Exclusion criteria:

1. History of Inflammatory Bowel Disease (IBD) , microscopic colitis or celiac disease

2. Use of tobacco products within the past 6 months

3. Use of NSAIDs or aspirin within the past week

4. Use of oral corticosteroids within the previous 6 weeks

5. Ingestion of artificial sweeteners such as Splenda (sucralose), Nutrasweet (aspartame), lactulose or mannitol 2 days before the study begins, e.g., foods to be avoided are sugarless gums or mints and diet soda

6. Ingestion of any prescription, over the counter, or herbal medications which can affect gastrointestinal transit 7 days before study begins

7. Any treatment specifically taken for IBS, including loperamide, cholestyramine, alosetron

8. Drugs with a known pharmacological activity at 5-HT4, 5-HT2b or 5-HT3 receptors (e.g, tegaserod, ondansetron, tropisetron, granisetron, dolasetron, mirtazapine);

9. All narcotics (e.g, codeine, morphine, and propoxyphene, either alone or in combination)

10. Anti-cholinergic agents (e.g, dicyclomine, hyoscyamine, propantheline).

11. Ultram

12. GI preparations

• Anti-nausea agents (e.g, trimethobenzamide, promethazine, prochlorperazine, dimenhydrinate, hydroxyzine)

• Osmotic laxative agents (e.g, lactulose, sorbitol or PEG solutions as Miralax and Glycolax)

• Prokinetic agents (e.g, cisapride, metoclopramide, itopride, domperidone);

1. Antimuscarinics;

2. Peppermint oil;

3. Systemic antibiotics, rifaximin, metronidazole.

4. Bleeding disorders or medications that increase risk of bleeding from mucosal biopsies.

5. Score > 8 for anxiety or depression on Hospital anxiety and depression scale.

6. Pregnancy

Contacts and Locations

Locations

	Site	City	State	Country	Postal Code
1	Mayo Clinic in Rochester	Rochester	Minnesota	United States	55905

Sponsors and Collaborators

• Mayo Clinic
• Takeda Pharmaceuticals International, Inc.

Investigators

• Principal Investigator: Madhusudan Grover, MBBS, Mayo Clinic

Study Documents (Full-Text)

None provided.

More Information

Publications

None provided.

• Responsible Party: Madhusudan (Madhu) Grover, MBBS, Principal Investigator, Mayo Clinic
• ClinicalTrials.gov Identifier: NCT02246647
• Other Study ID Numbers: 14-002382
• First Posted: Sep 23, 2014
• Last Update Posted: Aug 21, 2019
• Last Verified: Aug 1, 2019

• Studies a U.S. FDA-regulated Drug Product: Yes
• Studies a U.S. FDA-regulated Device Product: No
• Additional relevant MeSH terms: Irritable Bowel Syndrome; Intestinal Diseases; Constipation; Mannitol; Lactulose

Study Results

Participant Flow

Arm/Group Title	Healthy Volunteers	IBS-C
Arm/Group Description	Healthy subjects ages 18 - 65 Must not have had any abdominal surgeries, IBS, IBD, microscopic colitis or celiac disease. Must not have used tobacco products in the last 6 months. Must not have used corticosteroids in the last 6 weeks. Must not have taken any treatment specifically taken for IBS, including loperamide, cholestyramine, alosetron . Must not be taking any drugs with a known pharmacological activity at 5-HT4, 5-HT2b or 5-HT3 receptors. Must not have taken anti-cholinergic agents (e.g. dicyclomine, hyoscyamine, propantheline), Ultram. No GI preparations Anti-nausea agents (e.g., trimethobenzamide, promethazine, prochlorperazine, dimenhydrinate, hydroxyzine) Osmotic laxative agents (e.g, lactulose, sorbitol or PEG solutions as MiraLAX and GlycoLax) - Prokinetic agents (e.g, cisapride, metoclopramide, itopride, domperidone); Antimuscarinics;Peppermint oil;Systemic antibiotics, rifaximin, metronidazole. Must not have a bleeding disorder or medications that incr	Age (yr) 18 to 65 Must have IBS-C by Rome III criteria No abdominal surgery (except appendectomy and cholecystectomy) Exclusion criteria: Must have no History of IBD (Crohn's disease or ulcerative colitis), microscopic colitis or celiac disease. Use of tobacco products within the past 6 months (since nicotine may affect intestinal permeability) Use of NSAIDs or aspirin within the past week (since NSAIDs affect intestinal permeability) Use of oral corticosteroids within the previous 6 weeks No Ingestion of artificial sweeteners such as SplendaTM (sucralose), NutraSweet TM(aspartame), lactulose or mannitol 2 days before the study begins, e.g., foods to be avoided are sugarless gums or mints and diet soda. Ingestion of any prescription, over the counter, or herbal medications which can affect gastrointestinal transit 7 days before study begins ny treatment specifically taken for IBS, including loperamide, cholestyramine, alosetron No drugs with a known pharmacological activity at 5
Period Title: Overall Study
STARTED	19	20
COMPLETED	18	19
NOT COMPLETED	1	1

Baseline Characteristics

Arm/Group Title	Healthy Volunteers	IBS-C	Total
Arm/Group Description	Healthy volunteers between ages 18 - 65, who have not history of IBS.	Volunteers who have been diagnosed with IBS - Constipation (IBS-C)	Total of all reporting groups
Overall Participants	18	19	37
Age (Count of Participants)
<=18 years	0 0%	0 0%	0 0%
Between 18 and 65 years	18 100%	19 100%	37 100%
>=65 years	0 0%	0 0%	0 0%
Age (years) [Mean (Standard Deviation) ]
Mean (Standard Deviation) [years]	45.37 (2.82)	43.06 (2.78)	44.24 (1.96)
Sex: Female, Male (Count of Participants)
Female	18 100%	19 100%	37 100%
Male	0 0%	0 0%	0 0%
Ethnicity (NIH/OMB) (Count of Participants)
Hispanic or Latino	0 0%	0 0%	0 0%
Not Hispanic or Latino	18 100%	19 100%	37 100%
Unknown or Not Reported	0 0%	0 0%	0 0%
Race (NIH/OMB) (Count of Participants)
American Indian or Alaska Native	0 0%	0 0%	0 0%
Asian	0 0%	0 0%	0 0%
Native Hawaiian or Other Pacific Islander	0 0%	0 0%	0 0%
Black or African American	0 0%	0 0%	0 0%
White	18 100%	19 100%	37 100%
More than one race	0 0%	0 0%	0 0%
Unknown or Not Reported	0 0%	0 0%	0 0%
Region of Enrollment (Count of Participants)
United States	18 100%	19 100%	37 100%

Outcome Measures

1. Primary Outcome

Title	Lactulose:C13 Mannitol Excretion Ratio 8-24hrs.
Description	In vivo measurement of intestinal permeability using 13C mannitol & lactulose was used. High performance liquid chromatography-tandem mass spectrometry was used to measure concentrations calculated using the overall urine volume excreted in each interval. Concentrations of 13C adjusted for the % of 13C in 12C mannitol (4.98% of 12C mannitol excreted was subtracted from 13C mannitol values; determined by analyzing replicate samples of control urine). All lactulose or 13C mannitol concentrations 8-24hr post-ingestion were used to determine colonic permeability. Lactulose to 13C mannitol excretion ratios, as a measure of dose of saccharide administered, were calculated.
Time Frame	8-24 hr post test-dose administration

Outcome Measure Data

Analysis Population Description

[Not Specified]

Arm/Group Title	Healthy Volunteers	IBS - C
Arm/Group Description	Ingestion of saccharides (mannitol (regular, 12C) 100 mg, lactulose 1 g and (non-radioactive) 13C labeled mannitol, 100 mg and sucralose, 50 mg) in 250ml of water Esophagogastroduodenoscopy Flexible sigmoidoscopy Esophagogastroduodenoscopy: Duodenal biopsies will be collected from Healthy Volunteers. Flexible sigmoidoscopy: Colonic biopsies will be collected from Healthy Volunteers.	Ingestion of saccharides (mannitol (regular, 12C) 100 mg, lactulose 1 g and (non-radioactive) 13C labeled mannitol, 100 mg and sucralose, 50 mg) in 250ml of water Esophagogastroduodenoscopy Flexible sigmoidoscopy Esophagogastroduodenoscopy: Duodenal biopsies will be collected from IBS - C cases. Flexible sigmoidoscopy: Colonic biopsies will be collected from IBS - C cases.
Measure Participants	18	19
Mean (Standard Error) [Ratio]	0.01 (0.004)	0.02 (0.005)

Statistical Analysis 1

Statistical Analysis Overview	Comparison Group Selection	Healthy Volunteers, IBS - C
	Comments
	Type of Statistical Test	Other
	Comments
Statistical Test of Hypothesis	p-Value	0.87
	Comments
	Method	Wilcoxon (Mann-Whitney)
	Comments
Method of Estimation	Estimation Parameter	Mean Difference (Final Values)
	Estimated Value	0.01
	Confidence Interval	(2-Sided) %; to
	Parameter Dispersion	Type: Value:
	Estimation Comments

2. Secondary Outcome

Title	Lactose:C13 Mannitol Excretion Ratio 0-2hours
Time Frame	0-2 hr post-test dose administration

Outcome Measure Data

Analysis Population Description

[Not Specified]

Arm/Group Title	Healthy Volunteers	IBS - C
Arm/Group Description	Ingestion of saccharides (mannitol (regular, 12C) 100 mg, lactulose 1 g and (non-radioactive) 13C labeled mannitol, 100 mg and sucralose, 50 mg) in 250ml of water Esophagogastroduodenoscopy Flexible sigmoidoscopy Esophagogastroduodenoscopy: Duodenal biopsies will be collected from Healthy Volunteers. Flexible sigmoidoscopy: Colonic biopsies will be collected from Healthy Volunteers.	Ingestion of saccharides (mannitol (regular, 12C) 100 mg, lactulose 1 g and (non-radioactive) 13C labeled mannitol, 100 mg and sucralose, 50 mg) in 250ml of water Esophagogastroduodenoscopy Flexible sigmoidoscopy Esophagogastroduodenoscopy: Duodenal biopsies will be collected from IBS - C cases. Flexible sigmoidoscopy: Colonic biopsies will be collected from IBS - C cases.
Measure Participants	18	19
Mean (Standard Error) [Ratio]	0.007 (0.0004)	0.01 (0.001)

3. Secondary Outcome

Title	Baseline Transmucosal Resistance (TMR) of Duodenal Mucosa
Time Frame	Baseline

Outcome Measure Data

Analysis Population Description

Two participants ( 1 in Healthy volunteers and 1 in IBS-C ) were not analysed for the Baseline transmucosal resistance (TMR) of duodenal mucosa due to various reasons such inadequate size of the biopsy specimen, participant did not show up for the procedure etc.

Arm/Group Title	Healthy Volunteers	IBS - C
Arm/Group Description	Four biopsies from duodenum from the Healthy Volunteers were mounted in 4 mL Ussing chambers (Physiologic Instruments, San Diego, CA, USA) exposing 0.031 sq.cm area, within 45 min of collection. Chambers were filled with Krebs with 10 mM mannitol (mucosal side) and Krebs with 10 mM glucose (submucosal side).Baseline transmucosal resistance (TMR) and short circuit current (Isc) of each tissue was measured.Average TMR and Isc across 3-4 biopsies/subject were calculated.Paracellular flux across biopsies was measured using 4 kDa FITC-Dextran administered on the mucosal side (1 mg/mL chamber concentration). Cumulative flux at the end of three hours and rate of flux was calculated for FITC-Dextran.	Four biopsies from duodenum from the IBS - C cases were mounted in 4 mL Ussing chambers (Physiologic Instruments, San Diego, CA, USA) exposing 0.031 sq.cm area, within 45 min of collection. Chambers were filled with Krebs with 10 mM mannitol (mucosal side) and Krebs with 10 mM glucose (submucosal side).Baseline transmucosal resistance (TMR) and short circuit current (Isc) of each tissue was measured.Average TMR and Isc across 3-4 biopsies/subject were calculated.Paracellular flux across biopsies was measured using 4 kDa FITC-Dextran administered on the mucosal side (1 mg/mL chamber concentration). Cumulative flux at the end of three hours and rate of flux was calculated for FITC-Dextran
Measure Participants	17	18
Mean (Standard Error) [Ω*sq.cm]	29.8 (1.94)	28.16 (1.96)

4. Secondary Outcome

Title	Cumulative FITC-Dextran (4kDa) Concentration Across Duodenal Mucosa
Description	This is not a pharmacokinetic or pharmacodynamic measure. Hence only one time assessment is made 3 hours after FITC-Dextran (4kDa) administration.
Time Frame	3 hours post FITC-Dextran (4kDa) administration

Outcome Measure Data

Analysis Population Description

Four participants (2 in Healthy volunteers and 2 in IBS-C) were not analysed for the Cumulative FITC-Dextran (4kDa) concentration across duodenal mucosa due to various reasons such inadequate size of the biopsy specimen, participant did not show up for the procedure, etc.

Arm/Group Title	Healthy Volunteers	IBS - C
Arm/Group Description	Four biopsies from each site from the Healthy Volunteers were mounted in 4 mL Ussing chambers (Physiologic Instruments, San Diego, CA, USA) exposing 0.031 sq.cm area, within 45 min of collection. Chambers were filled with Krebs with 10 mM mannitol (mucosal side) and Krebs with 10 mM glucose (submucosal side).Baseline transmucosal resistance (TMR) and short circuit current (Isc) of each tissue was measured.Average TMR and Isc across 3-4 biopsies/subject were calculated.Paracellular flux across biopsies was measured using 4 kDa FITC-Dextran administered on the mucosal side (1 mg/mL chamber concentration). Cumulative flux at the end of three hours and rate of flux was calculated for FITC-Dextran.	Four biopsies from each site from the IBS - C cases were mounted in 4 mL Ussing chambers (Physiologic Instruments, San Diego, CA, USA) exposing 0.031 sq.cm area, within 45 min of collection. Chambers were filled with Krebs with 10 mM mannitol (mucosal side) and Krebs with 10 mM glucose (submucosal side).Baseline transmucosal resistance (TMR) and short circuit current (Isc) of each tissue was measured.Average TMR and Isc across 3-4 biopsies/subject were calculated.Paracellular flux across biopsies was measured using 4 kDa FITC-Dextran administered on the mucosal side (1 mg/mL chamber concentration). Cumulative flux at the end of three hours and rate of flux was calculated for FITC-Dextran
Measure Participants	16	17
Mean (Standard Error) [ng/mL]	123.3 (24.29)	156.8 (33.73)

5. Secondary Outcome

Title	Rate of FITC-Dextran (4kDa) Flux Across Duodenal Mucosa
Description	This is not a pharmacokinetic or pharmacodynamic measure. Hence only one time assessment is made 3 hours after FITC-Dextran (4kDa) administration.
Time Frame	Over 3 hours post FITC-Dextran (4kDa) administration

Outcome Measure Data

Analysis Population Description

Four participants ( 2 in Healthy volunteers and 2 in IBS-C ) were not analysed for the Rate of FITC-Dextran (4kDa) flux across duodenal mucosa due to various reasons such inadequate size of the biopsy specimen, participant did not show up for the procedure etc.

Arm/Group Title	Healthy Volunteers	IBS - C
Arm/Group Description	Four biopsies from each site from the Healthy Volunteers were mounted in 4 mL Ussing chambers (Physiologic Instruments, San Diego, CA, USA) exposing 0.031 sq.cm area, within 45 min of collection. Chambers were filled with Krebs with 10 mM mannitol (mucosal side) and Krebs with 10 mM glucose (submucosal side).Baseline transmucosal resistance (TMR) and short circuit current (Isc) of each tissue was measured.Average TMR and Isc across 3-4 biopsies/subject were calculated.Paracellular flux across biopsies was measured using 4 kDa FITC-Dextran administered on the mucosal side (1 mg/mL chamber concentration). Cumulative flux at the end of three hours and rate of flux was calculated for FITC-Dextran.	Four biopsies from each site from the IBS - C cases were mounted in 4 mL Ussing chambers (Physiologic Instruments, San Diego, CA, USA) exposing 0.031 sq.cm area, within 45 min of collection. Chambers were filled with Krebs with 10 mM mannitol (mucosal side) and Krebs with 10 mM glucose (submucosal side).Baseline transmucosal resistance (TMR) and short circuit current (Isc) of each tissue was measured.Average TMR and Isc across 3-4 biopsies/subject were calculated.Paracellular flux across biopsies was measured using 4 kDa FITC-Dextran administered on the mucosal side (1 mg/mL chamber concentration). Cumulative flux at the end of three hours and rate of flux was calculated for FITC-Dextran
Measure Participants	16	17
Mean (Standard Error) [ng/hr/sq.cm]	4980 (1044)	6528 (1584)

6. Secondary Outcome

Title	Baseline Transmucosal Resistance (TMR) of Colonic Mucosa
Time Frame	Baseline

Outcome Measure Data

Analysis Population Description

One participant in IBS-C was not analysed for the Baseline Transmucosal Resistance (TMR) of Colonic Mucosa due to various reasons such inadequate size of the biopsy specimen.

Arm/Group Title	Healthy Volunteers	IBS - C
Arm/Group Description	Four biopsies from colon from the Healthy Volunteers were mounted in 4 mL Ussing chambers (Physiologic Instruments, San Diego, CA, USA) exposing 0.031 sq.cm area, within 45 min of collection. Chambers were filled with Krebs with 10 mM mannitol (mucosal side) and Krebs with 10 mM glucose (submucosal side).Baseline transmucosal resistance (TMR) and short circuit current (Isc) of each tissue was measured.Average TMR and Isc across 3-4 biopsies/subject were calculated.Paracellular flux across biopsies was measured using 4 kDa FITC-Dextran administered on the mucosal side (1 mg/mL chamber concentration). Cumulative flux at the end of three hours and rate of flux was calculated for FITC-Dextran.	Four biopsies from colon from the IBS - C cases were mounted in 4 mL Ussing chambers (Physiologic Instruments, San Diego, CA, USA) exposing 0.031 sq.cm area, within 45 min of collection. Chambers were filled with Krebs with 10 mM mannitol (mucosal side) and Krebs with 10 mM glucose (submucosal side).Baseline transmucosal resistance (TMR) and short circuit current (Isc) of each tissue was measured.Average TMR and Isc across 3-4 biopsies/subject were calculated.Paracellular flux across biopsies was measured using 4 kDa FITC-Dextran administered on the mucosal side (1 mg/mL chamber concentration). Cumulative flux at the end of three hours and rate of flux was calculated for FITC-Dextran
Measure Participants	18	18
Mean (Standard Error) [Ω*sq.cm]	17.60 (1.70)	19.06 (1.10)

7. Secondary Outcome

Title	Cumulative FITC-Dextran (4kDa) Concentration Across Colonic Mucosa
Time Frame	3 hours post FITC-Dextran (4kDa) administration

Outcome Measure Data

Analysis Population Description

7 participants (2 in Healthy volunteers and 5 in IBS-C) were not analysed for the Cumulative FITC-Dextran (4kDa) Concentration Across Colonic Mucosa due to various reasons such inadequate size of the biopsy specimen, participant did not show up for the procedure, etc.

Arm/Group Title	Healthy Volunteers	IBS - C
Arm/Group Description	Four biopsies from colon from the Healthy Volunteers were mounted in 4 mL Ussing chambers (Physiologic Instruments, San Diego, CA, USA) exposing 0.031 sq.cm area, within 45 min of collection. Chambers were filled with Krebs with 10 mM mannitol (mucosal side) and Krebs with 10 mM glucose (submucosal side).Baseline transmucosal resistance (TMR) and short circuit current (Isc) of each tissue was measured.Average TMR and Isc across 3-4 biopsies/subject were calculated.Paracellular flux across biopsies was measured using 4 kDa FITC-Dextran administered on the mucosal side (1 mg/mL chamber concentration). Cumulative flux at the end of three hours and rate of flux was calculated for FITC-Dextran.	Four biopsies from colon from the IBS - C cases were mounted in 4 mL Ussing chambers (Physiologic Instruments, San Diego, CA, USA) exposing 0.031 sq.cm area, within 45 min of collection. Chambers were filled with Krebs with 10 mM mannitol (mucosal side) and Krebs with 10 mM glucose (submucosal side).Baseline transmucosal resistance (TMR) and short circuit current (Isc) of each tissue was measured.Average TMR and Isc across 3-4 biopsies/subject were calculated.Paracellular flux across biopsies was measured using 4 kDa FITC-Dextran administered on the mucosal side (1 mg/mL chamber concentration). Cumulative flux at the end of three hours and rate of flux was calculated for FITC-Dextran
Measure Participants	16	14
Mean (Standard Error) [ng/mL]	132.2 (24.23)	122.5 (27.44)

8. Secondary Outcome

Title	Rate of FITC-Dextran (4kDa) Flux Across Colonic Mucosa
Time Frame	Over 3 hours post FITC-Dextran (4kDa) administration

Outcome Measure Data

Analysis Population Description

7 participants (2 in Healthy volunteers and 5 in IBS-C) were not analysed for the Rate of FITC-Dextran (4kDa) Flux Across Colonic Mucosa due to various reasons such inadequate size of the biopsy specimen, participant did not show up for the procedure, etc.

Arm/Group Title	Healthy Volunteers	IBS - C
Arm/Group Description	Four biopsies from colon from the Healthy Volunteers were mounted in 4 mL Ussing chambers (Physiologic Instruments, San Diego, CA, USA) exposing 0.031 sq.cm area, within 45 min of collection. Chambers were filled with Krebs with 10 mM mannitol (mucosal side) and Krebs with 10 mM glucose (submucosal side).Baseline transmucosal resistance (TMR) and short circuit current (Isc) of each tissue was measured.Average TMR and Isc across 3-4 biopsies/subject were calculated.Paracellular flux across biopsies was measured using 4 kDa FITC-Dextran administered on the mucosal side (1 mg/mL chamber concentration). Cumulative flux at the end of three hours and rate of flux was calculated for FITC-Dextran.	Four biopsies from colon from the IBS - C cases were mounted in 4 mL Ussing chambers (Physiologic Instruments, San Diego, CA, USA) exposing 0.031 sq.cm area, within 45 min of collection. Chambers were filled with Krebs with 10 mM mannitol (mucosal side) and Krebs with 10 mM glucose (submucosal side).Baseline transmucosal resistance (TMR) and short circuit current (Isc) of each tissue was measured.Average TMR and Isc across 3-4 biopsies/subject were calculated.Paracellular flux across biopsies was measured using 4 kDa FITC-Dextran administered on the mucosal side (1 mg/mL chamber concentration). Cumulative flux at the end of three hours and rate of flux was calculated for FITC-Dextran
Measure Participants	16	14
Mean (Standard Error) [ng/hr/sq.cm]	4931 (1156)	6069 (1030)

9. Secondary Outcome

Title	Cumulative E.Coli Bio- Particle K12 Concentration Across Duodenal Mucosa
Time Frame	3 hours post E.coli Bio- Particle administration

Outcome Measure Data

Analysis Population Description

11 participants (4 in Healthy volunteers and 7 in IBS-C) were not analysed for Cumulative E.Coli Bio- Particle K12 Concentration Across Duodenal Mucosa due to various reasons such inadequate size of the biopsy specimen, participant did not show up for the procedure, etc.

Arm/Group Title	Healthy Volunteers	IBS - C
Arm/Group Description	Four biopsies from duodenum from the Healthy Volunteers were mounted in 4 mL Ussing chambers (Physiologic Instruments, San Diego, CA, USA) exposing 0.031 sq.cm area, within 45 min of collection. Chambers were filled with Krebs with 10 mM mannitol (mucosal side) and Krebs with 10 mM glucose (submucosal side).Baseline transmucosal resistance (TMR) and short circuit current (Isc) of each tissue was measured. Average TMR and Isc across 3-4 biopsies/subject were calculated. Transcellular transport was studied by using translocation of the fluorescently labeled E. coli K-12 Bio-Particles was measured using a 107 CFU/mL chamber concentration on the mucosal side. Cumulative flux at the end of three hours and rate of flux was calculated for E. coli K-12 Bio-Particles.	Four biopsies from duodenum from the IBS - C cases were mounted in 4 mL Ussing chambers (Physiologic Instruments, San Diego, CA, USA) exposing 0.031 sq.cm area, within 45 min of collection. Chambers were filled with Krebs with 10 mM mannitol (mucosal side) and Krebs with 10 mM glucose (submucosal side).Baseline transmucosal resistance (TMR) and short circuit current (Isc) of each tissue was measured. Average TMR and Isc across 3-4 biopsies/subject were calculated. Transcellular transport was studied by using translocation of the fluorescently labeled E. coli K-12 Bio-Particles was measured using a 107 CFU/mL chamber concentration on the mucosal side. Cumulative flux at the end of three hours and rate of flux was calculated for E. coli K-12 Bio-Particles.
Measure Participants	14	12
Mean (Standard Error) [CFU/ml]	1.48*10^4 (2.6*10^3)	1.82*10^4 (6.0*10^3)

10. Secondary Outcome

Title	Rate of E.Coli Bio- Particle K12 Flux Across Duodenal Mucosa
Time Frame	Over 3 hours post E.coli Bio- Particle administration

Outcome Measure Data

Analysis Population Description

11 participants (4 in Healthy volunteers and 7 in IBS-C) were not analysed for Rate of E.Coli Bio- Particle K12 Flux Across Duodenal Mucosa due to various reasons such inadequate size of the biopsy specimen, participant did not show up for the procedure, etc.

Arm/Group Title	Healthy Volunteers	IBS - C
Arm/Group Description	Four biopsies from duodenum from the Healthy Volunteers were mounted in 4 mL Ussing chambers (Physiologic Instruments, San Diego, CA, USA) exposing 0.031 sq.cm area, within 45 min of collection. Chambers were filled with Krebs with 10 mM mannitol (mucosal side) and Krebs with 10 mM glucose (submucosal side).Baseline transmucosal resistance (TMR) and short circuit current (Isc) of each tissue was measured. Average TMR and Isc across 3-4 biopsies/subject were calculated. Transcellular transport was studied by using translocation of the fluorescently labeled E. coli K-12 Bio-Particles was measured using a 107 CFU/mL chamber concentration on the mucosal side. Cumulative flux at the end of three hours and rate of flux was calculated for E. coli K-12 Bio-Particles.	Four biopsies from duodenum from the IBS - C cases were mounted in 4 mL Ussing chambers (Physiologic Instruments, San Diego, CA, USA) exposing 0.031 sq.cm area, within 45 min of collection. Chambers were filled with Krebs with 10 mM mannitol (mucosal side) and Krebs with 10 mM glucose (submucosal side).Baseline transmucosal resistance (TMR) and short circuit current (Isc) of each tissue was measured. Average TMR and Isc across 3-4 biopsies/subject were calculated. Transcellular transport was studied by using translocation of the fluorescently labeled E. coli K-12 Bio-Particles was measured using a 107 CFU/mL chamber concentration on the mucosal side. Cumulative flux at the end of three hours and rate of flux was calculated for E. coli K-12 Bio-Particles.
Measure Participants	14	12
Mean (Standard Error) [CFU/h/sq.cm]	5.42*10^5 (1.2*10^5)	5.70*10^5 (1.90*10^5)

11. Secondary Outcome

Title	Cumulative E.Coli Bio- Particle K12 Concentration Across Colonic Mucosa
Time Frame	3 hours post E.coli Bio- Particle administration

Outcome Measure Data

Analysis Population Description

7 participants (2 in Healthy volunteers and 5 in IBS-C) were not analysed for Cumulative E.coli Bio- Particle K12 Concentration Across Colonic Mucosa due to various reasons such inadequate size of the biopsy specimen, participant did not show up for the procedure, etc.

Arm/Group Title	Healthy Volunteers	IBS - C
Arm/Group Description	Four biopsies from colon from the Healthy Volunteers were mounted in 4 mL Ussing chambers (Physiologic Instruments, San Diego, CA, USA) exposing 0.031 sq.cm area, within 45 min of collection. Chambers were filled with Krebs with 10 mM mannitol (mucosal side) and Krebs with 10 mM glucose (submucosal side).Baseline transmucosal resistance (TMR) and short circuit current (Isc) of each tissue was measured. Average TMR and Isc across 3-4 biopsies/subject were calculated. Transcellular transport was studied by using translocation of the fluorescently labeled E. coli K-12 Bio-Particles was measured using a 107 CFU/mL chamber concentration on the mucosal side. Cumulative flux at the end of three hours and rate of flux was calculated for E. coli K-12 Bio-Particles.	Four biopsies from colon from the IBS - C cases were mounted in 4 mL Ussing chambers (Physiologic Instruments, San Diego, CA, USA) exposing 0.031 sq.cm area, within 45 min of collection. Chambers were filled with Krebs with 10 mM mannitol (mucosal side) and Krebs with 10 mM glucose (submucosal side).Baseline transmucosal resistance (TMR) and short circuit current (Isc) of each tissue was measured. Average TMR and Isc across 3-4 biopsies/subject were calculated. Transcellular transport was studied by using translocation of the fluorescently labeled E. coli K-12 Bio-Particles was measured using a 107 CFU/mL chamber concentration on the mucosal side. Cumulative flux at the end of three hours and rate of flux was calculated for E. coli K-12 Bio-Particles.
Measure Participants	16	14
Mean (Standard Error) [CFU/ml]	1.07*10^4 (1.8*10^3)	2.09*10^4 (6.8*10^3)

12. Secondary Outcome

Title	Rate of E.Coli Bio- Particle K12 Flux Across Colonic Mucosa
Time Frame	Over 3 hours post E.coli Bio- Particle administration

Outcome Measure Data

Analysis Population Description

7 participants (2 in Healthy volunteers and 5 in IBS-C) were not analysed for Rate of E.Coli Bio- Particle K12 Flux Across Colonic Mucosa due to various reasons such inadequate size of the biopsy specimen, participant did not show up for the procedure, etc.

Arm/Group Title	Healthy Volunteers	IBS - C
Arm/Group Description	Four biopsies from colon from the Healthy Volunteers were mounted in 4 mL Ussing chambers (Physiologic Instruments, San Diego, CA, USA) exposing 0.031 sq.cm area, within 45 min of collection. Chambers were filled with Krebs with 10 mM mannitol (mucosal side) and Krebs with 10 mM glucose (submucosal side).Baseline transmucosal resistance (TMR) and short circuit current (Isc) of each tissue was measured. Average TMR and Isc across 3-4 biopsies/subject were calculated. Transcellular transport was studied by using translocation of the fluorescently labeled E. coli K-12 Bio-Particles was measured using a 107 CFU/mL chamber concentration on the mucosal side. Cumulative flux at the end of three hours and rate of flux was calculated for E. coli K-12 Bio-Particles.	Four biopsies from colon from the IBS - C cases were mounted in 4 mL Ussing chambers (Physiologic Instruments, San Diego, CA, USA) exposing 0.031 sq.cm area, within 45 min of collection. Chambers were filled with Krebs with 10 mM mannitol (mucosal side) and Krebs with 10 mM glucose (submucosal side).Baseline transmucosal resistance (TMR) and short circuit current (Isc) of each tissue was measured. Average TMR and Isc across 3-4 biopsies/subject were calculated. Transcellular transport was studied by using translocation of the fluorescently labeled E. coli K-12 Bio-Particles was measured using a 107 CFU/mL chamber concentration on the mucosal side. Cumulative flux at the end of three hours and rate of flux was calculated for E. coli K-12 Bio-Particles.
Measure Participants	16	14
Mean (Standard Error) [CFU/h/sq.cm]	4.26*10^5 (5.3*10^4)	7.67*10^5 (4.19*10^5)

13. Secondary Outcome

Title	Duodenal Impedance
Time Frame	Baseline

Outcome Measure Data

Analysis Population Description

12 participants (2 in Healthy volunteers and 10 in IBS-C) were not analysed for Duodenal impedance due to various reasons such inadequate size of the biopsy specimen, participant did not show up for the procedure, etc.

Arm/Group Title	Healthy Volunteers	IBS - C
Arm/Group Description	Volunteers fasted for eight hours and underwent sedated esophagogastroduodenoscopy. Duodenal impedance measurements were performed using a endoscopically passed catheter that measures electrical impedance of the duodenal epithelium by mucosal contact under direct visualization. A 2 mm diameter catheter was used with two 360° circumferential sensors placed at a separation of 2mm with the end of the distal ring being 1mm away from the catheter tip. The electrodes were connected to an impedance voltage transducer via thin wires, which ran the length of the catheter traversing through the working channel of the upper endoscope. The voltage generated by the transducer was limited to produce 10µA current at a frequency of 2 kHz. Impedance was measured every 90° in duodenal circumference with a decompressed lumen after suctioning of all fluid from the lumen. The catheter was embedded in the mucosa parallel to the two sensors and measurements were taken from a steady baseline for >10s.	IBS - C cases fasted for eight hours and underwent sedated esophagogastroduodenoscopy. Duodenal impedance measurements were performed using a endoscopically passed catheter that measures electrical impedance of the duodenal epithelium by mucosal contact under direct visualization. A 2 mm diameter catheter was used with two 360° circumferential sensors placed at a separation of 2mm with the end of the distal ring being 1mm away from the catheter tip. The electrodes were connected to an impedance voltage transducer via thin wires, which ran the length of the catheter traversing through the working channel of the upper endoscope. The voltage generated by the transducer was limited to produce 10µA current at a frequency of 2 kHz. Impedance was measured every 90° in duodenal circumference with a decompressed lumen after suctioning of all fluid from the lumen. The catheter was embedded in the mucosa parallel to the two sensors and measurements were taken from a steady baseline for >10s.
Measure Participants	16	9
Mean (Standard Error) [Ω]	705.9 (42.73)	729.5 (64.85)

14. Secondary Outcome

Title	Mean Serum Endotoxin (Bacterial LPS) Levels
Time Frame	Fasting, one time measurement after 8 hours

Outcome Measure Data

Analysis Population Description

2 participants (1 in healthy volunteers and 1 in IBS-C) were not analysed for Mean Serum Endotoxin (Bacterial LPS) Levels due to various reasons such as non-availability of the blood sample, technical errors, etc.

Arm/Group Title	Healthy Volunteers	IBS - C
Arm/Group Description	Fasting, EDTA anti-coagulated, whole blood was collected from Healthy Volunteers. Samples were analyzed using an EAA kit (Spectral Diagnostic Inc, Toronto, Ontario, Canada) that utilizes a monoclonal IgM antibody specific for gram-negative bacterial lipopolysaccharide (LPS). Samples were tested in duplicate within one hour of collection using a Berthold SmartLine luminometer (photon-counting). The endotoxin activity level was calculated as follows: chemiluminescence (test sample-negative control)/chemiluminescence (positive-negative control). Average values for each participant were expressed as absolute units.	Fasting, EDTA anti-coagulated, whole blood was collected from IBS - C cases. Samples were analyzed using an EAA kit (Spectral Diagnostic Inc, Toronto, Ontario, Canada) that utilizes a monoclonal IgM antibody specific for gram-negative bacterial lipopolysaccharide (LPS). Samples were tested in duplicate within one hour of collection using a Berthold SmartLine luminometer (photon-counting). The endotoxin activity level was calculated as follows: chemiluminescence (test sample-negative control)/chemiluminescence (positive-negative control). Average values for each participant were expressed as absolute units.
Measure Participants	17	18
Mean (Standard Error) [EU/mL]	0.35 (0.02)	0.36 (0.03)

Adverse Events

Time Frame	15 months-from the first participant screening to the last participants completion date
Arm/Group Title	Healthy Volunteer		IBS-C
Arm/Group Description	Healthy volunteers between ages 18 - 65, who have no history of IBS.		ROME III IBS-C patients
All Cause Mortality
	Healthy Volunteer		IBS-C
	Affected / at Risk (%)	# Events	Affected / at Risk (%)	# Events
Total	0/18 (0%)		0/19 (0%)
Serious Adverse Events
	Healthy Volunteer		IBS-C
	Affected / at Risk (%)	# Events	Affected / at Risk (%)	# Events
Total	0/18 (0%)		0/19 (0%)
Other (Not Including Serious) Adverse Events
	Healthy Volunteer		IBS-C
	Affected / at Risk (%)	# Events	Affected / at Risk (%)	# Events
Total	0/18 (0%)		1/19 (5.3%)
Gastrointestinal disorders
Indigestion/Heartburn	0/18 (0%)	0	1/19 (5.3%)	1

Limitations/Caveats

[Not Specified]

More Information

Certain Agreements

Principal Investigators are NOT employed by the organization sponsoring the study.

The only disclosure restriction on the PI is that the sponsor can review results communications prior to public release and can embargo communications regarding trial results for a period that is less than or equal to 60 days. The sponsor cannot require changes to the communication and cannot extend the embargo.