The Effect of Gluten on Gut Microbiome and Metabolic Health.

Feces samples are collected according to standard operation procedures for subsequent standardized microbial DNA extraction. Microbial DNA will be subjected to sequencing, microbial gene analyses, taxonomy analyses including enterotypes known species and unknown meta-species and functional annotation.

Participants are instructed in swallowing capsules containing different small non-invasive and non-absorbable plastic pellets for 6 consecutive days. On the seventh day they are having an X-ray of the abdomen taken.

Lactulose/mannitol ratio in urine after four hours collection following oral intake of lactulose and mannitol.

Breath hydrogen after intake of standardized breakfast (30,60,90,120,180 minutes)

Systolic and diastolic blood pressure and heart beat rate are measured after 10 min of rest according to current standard operational procedure with an automatic blood pressure meter. Participants are instructed not to talk during the measurements.

Saliva is collected after the participants have taken a small piece of paraffin in their mouth and have chewed until the paraffin has turned into one coherent mass (approximately one minute). Participants are swallowing the produced saliva for that minute. After this, and for the next 3 minutes, saliva is collected in a cup and handed over to the study personnel. Saliva is stored at minus 80 degrees for later studies of saliva microbial flora and saliva biochemistry.

Nasal fluid is collected by a non-invasive methodology for in vivo measurement of an array of immunological signalling molecules in the nasal airway lining. The method is based on a standardized collection of mucosal airway fluid from both nostrils onto small sheets of filter papers with efficient absorption properties. The technique is highly reproducible, and used for measuring immunological mediators representing the immediate response ability of the mucosal immune system.

Glucagon like peptide 1 and 2, Gastric inhibitory polypeptide, peptide YY and Ghrelin.

Levels of gliadin and Immunoglobulin A and G transglutaminase.

Low density Lipoproteins, High density lipoproteins, Total Cholesterol, Very low density lipoproteins and Free fatty acids.

Body composition (lean body mass and fat mass) is measured by bio-electrical impedance using multi frequency Quadscan.

Breath hydrogen measurements are done before the intake of the standardized breakfast (at 30, 60, 90, 120, 180 minutes), as an indicator of colonic fermentation.

Fasting insulin and post prandial at 30, 60, 90, 120 and 180 minutes after standardized meal.

Fasting glucose and post prandial at 30, 60, 90, 120 and 180 minutes after standardized meal.

Fasting.

High sensitive C-reactive protein, Interleukin 1, 6 and 10, Lipopolysaccharide- binding protein, Tumor necrosis factor - alfa.

Weight, height, sagittal height and waist circumference.

Validated method for dietary registration, based on 2 week and 2 weekend days.

Changes from baseline to after intervention in individual gastrointestinal symptoms by validated VAS-questionary.