Identification of Physiological Biomarkers of Gastro-intestinal Discomforts Induced by Milk Consumption

NCT04205045 | Completed

• 1 other identifier: MIDI
• Study Type: observational
• Target: 40
• Locations: 1 country
• Sites: 1

Brief Summary

This study aims to explore digestibility of lactose and milk proteins, gut permeability, microbiota composition and psychological wellbeing status in healthy subjects who are non-habitual milk consumers due to milk-related gastro-intestinal discomfort in comparison with healthy and habitual milk consumers.

Conditions

Healthy Subjects

Keywords

milk digestibility; gastro-intestinal discomfort; milk proteins; lactose; gut permeability; gut microbiota

Outcome Measures

Primary Outcomes (10)

• Changes from baseline in plasmatic concentration of milk-derived peptides following milk ingestion.

  Measure of casein digestion-derived peptides (beta-casomorphins and all known peptides derived from milk proteins) in plasma samples by mean of HPLC-MS/MS.

  baseline and at 0.5, 1, 2, 4 and 6 hours after milk ingestion.

• Changes from baseline of insulin plasmatic levels following milk ingestion.

  Measure of insulin by mean of Luminex kits in plasma samples pre-treated with aprotinin.

  baseline and at 0.5, 1, 2, 4 and 6 hours after milk ingestion.

• Changes from baseline of glucose-dependent insulinotropic peptide (GIP) plasmatic levels following milk ingestion.

  Measure of GIP by mean of Luminex kits in plasma samples pre-treated with aprotinin.

  baseline and at 0.5, 1, 2, 4 and 6 hours after milk ingestion.

• Changes from baseline of glucagon-like peptide 1 (GLP-1) plasmatic levels following milk ingestion.

  Measure of GLP-1 by mean of Luminex kits in plasma samples pre-treated with aprotinin.

  baseline and at 0.5, 1, 2, 4 and 6 hours after milk ingestion.

• Changes from baseline of glucagon plasmatic levels following milk ingestion.

  Measure of glucagon by mean of Luminex kits in plasma samples pre-treated with aprotinin

  baseline and at 0.5, 1, 2, 4 and 6 hours after milk ingestion.

• Changes from baseline of c-peptide plasmatic levels following milk ingestion.

  Measure of c-peptide by mean of Luminex kits in plasma samples pre-treated with aprotinin.

  baseline and at 0.5, 1, 2, 4 and 6 hours after milk ingestion.

• Changes from baseline of ghrelin plasmatic levels following milk ingestion.

  Measure of ghrelin by mean of Luminex kits in plasma samples pre-treated with aprotinin.

  baseline and at 0.5, 1, 2, 4 and 6 hours after milk ingestion.

• Changes from baseline of leptin plasmatic levels following milk ingestion.

  Measure of leptin by mean of Luminex kits in plasma samples pre-treated with aprotinin.

  baseline and at 0.5, 1, 2, 4 and 6 hours after milk ingestion.

• Changes from baseline in serum levels of endocannabinoids and N-acyl-ethanolamines response following milk ingestion.

  Measure of endocannabinoids N-acyl-ethanolamines in serum by mean of HPLC-MS/MS

  baseline and at 0.5, 1, 2, 4 and 6 hours after milk ingestion.

• Increases from baseline in urinary concentration of lactose following milk ingestion.

  Measure of lactose and galactose + glucose excreted in urine samples by mean of high pressure liquid chromatography coupled with tandem mass spectrometry (HPLC-MS/MS).

  baseline and time intervals (0-2, 2-4, 4-6, 6-8, 8-12 and 12-24 hours) following milk ingestion.

Secondary Outcomes (11)

• Changes from baseline in plasmatic concentration of milk-derived aminoacids following milk ingestion.

  baseline and at 0.5, 1, 2, 4 and 6 hours after milk ingestion.

• Changes from baseline of glycaemia following milk ingestion.

  baseline and at 0.5, 1, 2, 4 and 6 hours after milk ingestion.

• Changes from baseline in gastro-intestinal symptoms developed after milk ingestion.

  Baseline and at 0.5, 1, 2, 4, 6, 8, 12 and 24 hours after milk ingestion.

• Concentration of baseline serum DPPIV

  baseline

• Activity of serum DPPIV

  baseline and at 0.5, 1, 2, 4 and 6 hours after milk ingestion.

Study Arms (2)

Non-Habitual Milk Consumers (NHMC)

Healthy subjects who are non-habitual milk consumers because of gastrointestinal discomforts upon milk consumption. Lactose breath test; Gut permeability test; Milk test.

Diagnostic Test: Lactose Breath Test; Other: Gut permeability test; Other: Milk Test

Habitual Milk Consumers (HMC)

Healthy subjects with regular milk consumption. Intervention to be performed: Lactose breath test; Gut permeability test; Milk test.

Diagnostic Test: Lactose Breath Test; Other: Gut permeability test; Other: Milk Test

Interventions

Lactose Breath Test DIAGNOSTIC_TEST

Lactose Breath Test is performed during the recruitment step. Fasting subjects are asked to ingest 20 g of lactose in a 10% water solution. Breath samples are collected at baseline and every 30 min after lactose ingestion, for an overall test duration of 4 h. A cut-off value of 20 ppm over the baseline will be used to identify lactose malabsorbers.

Habitual Milk Consumers (HMC); Non-Habitual Milk Consumers (NHMC)

Gut permeability test OTHER

A 3-sugar probes test is conducted by administering 5 g lactulose, 2 g mannitol and 2 g sucralose in water solution to fasting subjects. Urine are collected at baseline (before sugar ingestion) and for time periods 0-5 h and 5-24 h.

Habitual Milk Consumers (HMC); Non-Habitual Milk Consumers (NHMC)

Milk Test OTHER

Milk test will be performed by administering to fasting subjects a portion of 250 ml of semi-skimmed milk treated with ultra-high temperature (UHT). Glycaemia measurements, blood withdrawals and questionnaires completion will be performed at baseline and repeated after 30 minutes, 1, 2, 4 and 6 hours after milk ingestion. Urine collection will be performed at baseline and for 6 time intervals after milk ingestion (0-2, 2-4, 4-6, 6-8, 8-12, 12-24 hours).

Habitual Milk Consumers (HMC); Non-Habitual Milk Consumers (NHMC)

Eligibility Criteria

• Age: 18 Years - 60 Years
• Sex: all
• Healthy Volunteers: Yes
• Age Groups: Adult (18-64)
• Sampling Method: Non-Probability Sample

Study Population

Community sample. Recruitment will be done through public announcements in the department places, local ambulatories and shops, and invitation through social networks.

You may qualify if:

• Drinking milk (maximum 150 ml/week for non-habitual milk consumers and minimum of 700 ml/week for habitual milk consumers)
• BMI between 18.5 - 30 kg/m2
• Available for three visits (with one week in between).
• Willing to drink 250 ml of milk in fasting condition within 10 min
• Written informed consent
• Negative lactose breath test result (increase in H2 concentrations \< 20 ppm vs basal value) and without symptoms or with any symptoms except "vomiting" and "Loose, mushy or watery stools"
• Lactose malabsorbers (increase in H2 concentrations \> 20 ppm vs basal value but without symptoms)

You may not qualify if:

• Presence of any relevant organic, systemic or metabolic disease or abnormal laboratory values.
• Ascertained intestinal organic diseases, including celiac disease or inflammatory bowel diseases.
• Previous major abdominal surgeries.
• Active malignancy of any type, or history of a malignancy (patients with a history of other malignancies that have been surgically removed and who have no evidence of recurrence for at least five years before study enrolment are also acceptable).
• Untreated food intolerance.
• Lactose intolerant
• Assumption of probiotics or topic and/or systemic antibiotic therapy during the last month.
• Systematical/frequent assumption of contact laxatives.
• Pregnant and lactating women.
• Inability to conform with protocol.
• Treatment with any investigational drug within the previous 30 days.
• Recent history or suspicion of alcohol abuse or drug addiction.
• Subjects having symptoms of "vomiting" and "Loose, mushy or watery stools" at any level of severity following milk consumption and the lactose breath test

Sponsors & Collaborators

• Paola Vitaglione: lead

Study Sites (1)

Department of Agricultural Sciences

Portici, 80055, Italy

Location

Related Publications (1)

• Tagliamonte S, Puhlmann ML, De Filippis F, Guerville M, Ercolini D, Vitaglione P. Relationships between diet and gut microbiome in an Italian and Dutch cohort: does the dietary protein to fiber ratio play a role? Eur J Nutr. 2024 Apr;63(3):741-750. doi: 10.1007/s00394-023-03308-4. Epub 2023 Dec 27.

  PMID: 38151533 DERIVED

Biospecimen

Retention: SAMPLES WITHOUT DNA

serum, plasma, plasma pre-treated with aprotinin, urine, feces

Study Officials

• Paola Vitaglione, PhD

  Federico II University

  STUDY DIRECTOR

Study Design

• Study Type: observational
• Observational Model: CASE CONTROL
• Time Perspective: PROSPECTIVE
• Sponsor Type: OTHER
• Responsible Party: SPONSOR INVESTIGATOR
• PI Title: Associate Professor

Study Record Dates

• First Submitted: December 10, 2019
• First Posted: December 19, 2019
• Study Start: October 1, 2018
• Primary Completion: January 24, 2020
• Study Completion: January 24, 2020
• Last Updated: August 1, 2022

Record last verified: 2022-07

Locations

IT (1)