Whey vs Casein to Combat Post-inflammatory Protein and Muscle Waste in Acute Disease

Study Overview
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Brief Summary
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This study compares three different protein supplements (casein, whey and leucine-enriched whey) and their effect on post-inflammatory muscle waste in a model of acute disease. Each test person will undergo all three interventions. It is believed that leucine is the primary driver of muscle protein synthesis and therefore we hypothesize that leucine-enriched whey and whey are superior to casein in combating post-inflammatory muscle waste, because of its higher leucine content (16%, 11% and 9% leucine, respectively).

Detailed Description
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Background: Acute illness is accompanied by infection/inflammation, anorexia and immobilization all contributing to muscle loss, making nutritional supplement optimization an obvious target for investigation and eventually clinical intervention. In the clinical setting large heterogenicity among patients complicates investigations of muscle metabolism during acute illness. Therefore we introduce a disease model by combining Inflammation + 36 hour fast and bedrest. Inflammation/febrile illness will be initiated by using the well-established human endotoxemia model with a bolus injection of Escherichia coli lipopolysaccharide (LPS), known to cause inflammation comparable with the initial phase of sepsis. The amino acid leucine has shown to be particularly anabolic in performance sports, but little is known about its potential beneficial effects during acute illness. Leucine is a powerful activator of muscle protein synthesis and it seems that protein supplements with the highest leucine content elicit a greater increase in protein synthesis than those with a smaller fraction of leucine. The protein supplements used most in hospitals contain casein derived protein, which has a much lower leucine content than the whey protein compounds typically used in performance sports. This study compares three different protein supplements.The study is an open, randomized crossover trial. Laboratory technicians, test subjects and investigators will be blinded. Interventions: I. LPS (1 ng/kg as bolus) + 36 h fasting + 36 h bedrest + Casein (9% leucine) II. LPS (1 ng/kg as bolus) + 36 h fasting + 36 h bedrest + Whey (11% leucine) III. LPS (1 ng/kg as bolus) + 36 h fasting + 36 h bedrest + Leucine-enriched whey (16% leucine) The test objects will be given 0,6 g protein/kg, 1/3 as a bolus and 2/3 as sipping over a period of 3,5 hour. Muscle metabolism will be investigated by phenylalanine tracer using the forearm model and total protein metabolism using a carbamide tracer. Through muscle biopsies intracellular signalling pathways will be investigated.

Official Title
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Whey vs Casein to Combat Post-inflammatory Protein and Muscle Waste - Combining Endotoxemia, Immobilisation and Fasting in Healthy Young Males in a New Model of Acute Febrile Disease

Conditions
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Muscle Protein Synthesis, Endotoxemia, Nutrition, Milk Protein, Metabolism, Whey, Casein

Intervention / Treatment
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* Dietary Supplement: Casein
* Dietary Supplement: Whey
* Dietary Supplement: Leucine-enriched whey


Participation Criteria
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Eligibility Criteria
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Inclusion Criteria: Healthy Male Age between 20-40 BMI between 20-30 Normal health examination and blood samples Written informed consent Exclusion Criteria: Immobilisation of an extremity, unless a doctor has declared it fully rehabilitated. Allergy against lidocain or latex. The use of anabolic steroids Disease like: Diabetes, epilepsia, infection, cardiovascular disease.

Ages Eligible for Study
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Minimum Age: 20 Years
Maximum Age: 40 Years

Sexes Eligible for Study
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Male

Accepts Healthy Volunteers
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Accepts Healthy Volunteers

Study Plan
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How is the study designed?
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Design Details

Primary Purpose: Basic Science
Allocation: Randomized
Intervention Model: Crossover Assignment
Interventional Model Description: Interventions*: I. LPS (1 ng/kg as bolus) + 36 h fasting + 36 h bedrest + Casein II. LPS (1 ng/kg as bolus) + 36 h fasting + 36 h bedrest + Whey III. LPS (1 ng/kg as bolus) + 36 h fasting + 36 h bedrest + Leucine-enriched whey * LPS will be administered on study day 1 and measurements of metabolism will be performed on study day 2 where we see the secondary effects of acute inflammation. The patient will stay at the hospital over night to ensure continues fast and bedrest. The beverages will be isocaloric and with the same total protein content. The basal period will be 2,5 hour with infusion of tracer. Hereafter a total amount of 0,6 g protein/kg bodyweight will be orally administered, 1/3 as a bolus and 2/3 as sipping over 3,5 hours. Muscle biopsies and blodsampels will be collected during both the basal and the sipping period.
Masking: Double


Arms and Interventions

| Participant Group/Arm | Intervention/Treatment |
| --- | --- |
| Experimental: Casein<br>LPS + 36 hour fast and bedrest + Casein (9% leucine) - 0.6 g protein/kg bodyweight, 1/3 as bolus and 2/3 as sipping. | Dietary Supplement: Casein<br>* see experimental description<br>|
| Experimental: Whey<br>LPS + 36 hour fast and bedrest + Whey (11% leucine) - 0.6 g protein/kg bodyweight, 1/3 as bolus and 2/3 as sipping | Dietary Supplement: Whey<br>* see experimental description<br>|
| Experimental: Leucine-enriched whey<br>LPS + 36 hour fast and bedrest + Leucine-enriched whey (16% leucine) - 0.6 g protein/kg bodyweight, 1/3 as bolus and 2/3 as sipping | Dietary Supplement: Leucine-enriched whey<br>* see experimental description<br>|

What is the study measuring?
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Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Change in muscle phenylalanine netbalance over the forearm muscle | Changes of muscle phenylalanine net balance (= arterio(phe conc)-venous(phe conc) x flow) from baseline to 3.5 hours after intervention using the forearm model | Change from baseline to 3.5 hours after intervention | 

Secondary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Change in whole body protein metabolism measured by a combination of phenylalanine- and tyrosine tracer | Changes in whole body protein synthesis rates (umol/kg/h), breakdown rates (umol/kg/h), phenylalanine to tyrosine conversion rates (umol/kg/h) and net balance (umol/kg/h) | Change from baseline to 3.5 hours after intervention | 
| Blood enrichment of essential amino acids | measures of essential amino acids in the blood | At baseline and every 30 minutes during the intervention period (3.5 hours) | 
| Changes in insulin concentrations | Measures of insulin concentration in blood | At baseline and every 30 minutes during the intervention period (3.5 hours) | 
| Change in Intracellular signalling in muscle measured by western blotting. | Investigating intracellular activity of muscle metabolism pathways by western blotting. | Change from baseline and after 2 hours of intervention | 
| Energy expenditure | Using indirect calorimetry for 15 min | At baseline and after 2.5 hours of intervention | 
| Changes in Glucose, fat and protein oxidation rates | Using indirect calorimetry for 15 min for measuring glucose- (mg/kg/min), fat- (mg/kg/min) and protein oxidation (mg/kg/min) | At baseline and after 2.5 hours of intervention | 
| Change in muscle breakdown and synthesis rates measured by phenylalanine tracer | changes from baseline to 3.5 hours after intervention in Ra(phe)=breakdown (umol/kg/h) and Rd(phe)=synthesis rate (umol/kg/h) | Change from baseline to 3.5 hours after intervention | 
| Changes in Glucagon concentrations | Glucagon concentrations in blood | Change from baseline and to 1 hour and 3.5 hour after the intervention | 
| Changes in GIP concentrations | GIP concentrations in blood | Change from baseline and to 1 hour and 3.5 hour after the intervention | 
| Changes in GLP-1 concentrations | GLP-1 concentrations in blood | Change from baseline and to 1 hour and 3.5 hour after the intervention | 
| Changes in Glucose concentrations | Glucose concentrations in blood | At baseline and every 30 minutes during the intervention period (3.5 hours) | 
| Changes in heart rate profile upon repeated LPS exposure | heart rate (beats/min) | Measured at baseline and 1,2,3,4,5,6 and 24 hours after LPS (6-8 weeks between visit 1,2 and 3) | 
| Changes in temperature profile upon repeated LPS exposure | Axillary temperature (celcius) | Measured at baseline and 1,2,3,4,5,6 and 24 hours after LPS (6-8 weeks between visit 1,2 and 3) | 
| Changes in blood pressure profile upon repeated LPS exposure | blood pressure (mmHg) | Measured at baseline and 1,2,3,4,5,6 and 24 hours after LPS (6-8 weeks between visit 1,2 and 3) | 
| Changes in symptom score profile upon repeated LPS exposure | symptom score (from 0-5) for nausea, back pain, muscle pain, headache and chills. 0=no symptoms, 5=severe symptoms. | Measured at baseline and 1,2,3,4,5,6 and 24 hours after LPS (6-8 weeks between visit 1,2 and 3) | 
| Changes in TNfalfa profile upon repeated LPS exposure | TNfalfa blood concentrations | Measured at baseline and 1, 2, 4, 6 and 24 hours after LPS (6-8 weeks between visit 1,2 and 3) | 
| Changes in IL-1 profile upon repeated LPS exposure | IL-1 blood concentrations | Measured at baseline and 1, 2, 4, 6 and 24 hours after LPS (6-8 weeks between visit 1,2 and 3) | 
| Changes in IL-6 profile upon repeated LPS exposure | IL-6 blood concentrations | Measured at baseline and 1, 2, 4, 6 and 24 hours after LPS (6-8 weeks between visit 1,2 and 3) | 
| Changes in IL-10 profile upon repeated LPS exposure | IL-10 blood concentrations | Measured at baseline and 1, 2, 4, 6 and 24 hours after LPS (6-8 weeks between visit 1,2 and 3) | 

 Terms related to the study
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Keywords Provided by Centre Hospitalier Valida
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milk protein, endotoxemia, muscle protein synthesis, whey, casein