The Effect of a Meatless,Keto Restrictive Diet on Body Composition,Strength Capacity,Oxidative Stress,Immune Response

Effect of a Meatless,Ketogenic Restrictive Diet on Body Composition, Concentration of Trimethylamine N-Oxide (TMAO),Ketone Bodies and Glucose in Blood and Urine, Somatic Disorders, Strength Capacity, Oxidative Stress, Immune Response

The subject of doctoral dissertation: Assessment of the effects of a meatless, ketogenic restrictive diet on body composition, strength capacity, oxidative stress and immune response

During planning of research and topic of the doctoral dissertation, it was considered how to modify a standard ketogenic diet rich in saturated fatty acids so that the use of this model of nutrition has the most anti-inflammatory effect. Therefore, it was decided to conduct a research to check whether a diet rich in omega-3 polyunsaturated fatty acids will show such an effect when following a high-fat diet.

Hypotheses:

1. The ketogenic diet reduces systemic inflammation. 2.The ketogenic diet reduces oxidative stress. 3. The ketogenic diet reduces body fat. 4. A ketogenic diet does not worsen strength performance.

Study Overview

• Status: Completed
• Conditions: Glucose Metabolism Disorders; Body Weight; Insulin Resistance; Lipid Metabolism Disorders; Oxidative Stress; Insulin Sensitivity; Ketosis; Ketoses, Metabolic; Body Fat Disorder; Carbohydrate Metabolism Disorder; Somatic Disorders; Immunologic Factors; Trimethylamine N-oxide; Insulin Tolerance
• Intervention / Treatment: Other: Meatless, restrictive ketogenic diet

Detailed Description

a) main research assumption

Reports in the international literature regarding the importance of the ketogenic diet are ambiguous. Some researchers believe that its use reduces body fat and improves insulin sensitivity or lipid profile, while others question this view and question it. Many authors emphasized the need for further research in this area, and this outline of the research project responds to this postulate.

In the presented experiment, the duration of a single experiment was 14 days, because the individual adaptation period of each participant should be taken into account - the so-called "keto-adaptation".

In order to assess the effectiveness of the nutritional intervention in this experiment, the following were performed: blood sampling for the determination of basic parameters (lipid profile, fasting glucose, fasting insulin, diabetic panel, hormones), determination of glucose and ketone bodies (β ketones) in the blood using a strip test - Optium Xido Neo glucometer, determination of the concentration of ketone bodies (acetoacetic acid) and glucose in the urine using Keto-Diastix - a strip test, examination by means of tests - force (maximum isokinetic force test with the use of Biodex apparatus), determination of inflammatory markers (TNF alpha, pro-inflammatory, anti-inflammatory and pro-or anti-inflammatory interleukins depending on the conditions), determination of oxidative stress markers - related to free radical damage to proteins (carbonyl groups, sulfhydryl groups (SH groups)), body composition measurement using the DEXA method (Dual Energy X-ray Absorptiometry). Choosing this method instead of the very common bioimpedance, due to the fact that using densitometry, it obtained very precise results of adipose tissue.

The above-described procedures were used to check the molecular basis of the phenomenon under study, to determine the parameters in which significant changes are visible and to determine the extent to which they translate into the function of the muscle and the subjective feelings of the subject.

Research methodology:

• body composition (DEXA)- Isometric muscle strength (Biodex)
• blood tests (lipid profile, glucose, insulin, ketone bodies)
• inflammatory markers - Luminex method with the use of BioRad BioPlex 200 reader Pro and anti-inflammatory cytokines (Bio-Plex Pro ™ Human Cytokine 8-plex Assay M50000007A, BIO-RAD, USA) (including IFN-y, IL-6, IL-8, IL-10, TNF-a, IL-2, IL-4)
• metabolic panel - Luminex method with the use of BioRad BioPlex 200 readerm (Bio-Plex Pro ™ Human Diabetes 10-Plex Assay # M171A7001M, BIO-RAD, USA) (including ghrelin, glucagon, insulin, leptin, resistin)
• markers of oxidative stress - Colorimetric method; Plate-based colorimetric measurement (360-385 nm) (concentration of protein carbonyl groups: Protein Carbonyl Colorimetric Assay Kit No. 100005020 (Cayman Chemical, Ann Arbor, MI, USA)
• markers of oxidative stress (concentration of protein thiol groups)
• TMAO (trimethylamine N-oxide)

• Study Type: Interventional
• Enrollment (Actual): 14
• Phase: Not Applicable

Contacts and Locations

Study Locations

• Poland
  • Wielkopolskie
    • Poznań, Wielkopolskie, Poland, 61-871
      • Poznan University of Physical Education

Participation Criteria

Eligibility Criteria

• Ages Eligible for Study: 32 years to 59 years (ADULT)
• Accepts Healthy Volunteers: Yes
• Genders Eligible for Study: All

Description

Inclusion Criteria:

• male or female
• healthy
• age 32 - 59
• non-smoker
• not abusing alcohol
• not subjected to physical exercise for at least 48 hours before the examination

Exclusion Criteria:

• cardiovascular,
• thyroid disease,
• gastrointestinal,
• respiratory
• or any other metabolic diseases adherence to special diets, use of nutritional supplements and use of medication to control blood lipids or glucose

Study Plan

How is the study designed?

Design Details

• Primary Purpose: BASIC_SCIENCE
• Allocation: NA
• Interventional Model: SINGLE_GROUP
• Masking: NONE

Number of Arms

1

Arms and Interventions

Participant Group / Arm

Intervention / Treatment

EXPERIMENTAL: Meatless restrictive ketogenic diet; Diet was designed to be isoproteic (1.8 g x Kg- 1 x body weight- 1 x day-1) with three meals a day., restrictive (EER minus 500 kcal / day). The distribution of macronutrients during the very low carbohydrate ketogenic diet (KD) was: protein 1.8 g x Kg-1 x body weight- 1 x day-1 (~ 25-30%), fats (~ 65-70%, with a strong emphasis on the content of omega 3 fatty acids) and carbohydrate (< 30 g x day- 1; < 10%).

Other: Meatless, restrictive ketogenic diet; Each participant received a nutritional plan that was the same qualitatively but differed quantitatively - a 500 kcal reduction based on Estimated Energy Requirement (EER) was assumed. Before the start of the study, each participant obtained information about the basic principles of the ketogenic diet. Diet was designed to be isoproteic (1.8gxKg-1xbody weight-1xday-1) with three meals a day. The distribution of macronutrients during the very low carbohydrate ketogenic diet (KD) was: protein 1.8 gxKg-1xbody weight-1xday-1(~25-30%), fats (~65-70%,with a strong emphasis on the content of omega 3 fatty acids) and carbohydrate (<30gxday-1;<10%). The food lists encouraged the consumption of fish,raw and cooked vegetables, eggs,fruits with the lowest glycemic index (blueberry, raspberry), plant oils and fats from avocado,olives.Drinks permitted were tea, coffee without sugar and the foods and drinks to be avoided were alcohol,meat (any kind of meat),bread,pasta,rice,milk,dairy and potatoes.

What is the study measuring?

Primary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Body composition	Whole body and regional body composition were measured in the morning after a 12 hours overnight fast by dual energy X-ray absorptiometry (DEXA) Body Composition: Body Fat Mass, Fat Free Mass, and Visceral Body Fat Body Fat Mass [Kg] max 40.143 Kg min 11.129 Kg (higher scores mean a worse outcome) Fat Free Mass [Kg] max 68.527 Kg min 37.72 Kg (higher scores mean a better outcome) Visceral Body Fat [g] max 2139 g min 90 g (higher scores mean a worse outcome)	2 weeks
Body weight	Body weight was measured with an accuracy of 0.1 Kg using an electronic scale (Tanita BC-601, Japan) Body weight (higher scores mean a worse outcome) [Kg] max 95.7 Kg min 60.9 Kg	2 weeks
TMAO	Trimethylamine N-Oxide (measured by Institute of Biochemistry and Biophysics Polish Academy of Sciences, Warsaw Poland) TMAO [ng/ml] (higher scores mean a worse outcome)	2 weeks
Lipids level (TC, HDL, LDL, TG)	Level of lipids - (TC- Total Cholesterol, LDL- low-density lipoprotein, HDL- high-density lipoprotein, TG - triacylglycerol) TC [mg/dl] (higher scores mean a worse outcome) LDL [mg/dl] (higher scores mean a worse outcome) TG [mg/dl] (higher scores mean a worse outcome) HDL [mg/dl] (higher scores mean a better outcome)	2 weeks
glucose level (blood, urine)	glucose level (blood, urine) - measured by Ketodiastix and Optimum Xido Neo higher scores mean a worse outcome	2 weeks
ketone bodies (blood, urine)	ketone bodies (blood, urine) - measured by Ketodiastix and Optimum Xido Neo higher scores mean a better outcome	2 weeks
inflammatory markers	inflammatory markers (including IFN-y, IL-6, IL-8, IL-10, TNF-a, IL-2, IL-4) - Luminex method with the use of BioRad BioPlex 200 reader Pro and anti-inflammatory cytokines (Bio-Plex Pro ™ Human Cytokine 8-plex Assay M50000007A, BIO-RAD, USA) depends - higher scores mean a better or worse outcome	2 weeks
markers of oxidative stress 1	markers of oxidative stress - concentration of protein carbonyl groups - Colorimetric method; Plate-based colorimetric measurement (360-385 nm) (concentration of protein carbonyl groups: Protein Carbonyl Colorimetric Assay Kit No. 100005020 (Cayman Chemical, Ann Arbor, MI, USA) higher scores mean a worse outcome	2 weeks
markers of oxidative stress 2	markers of oxidative stress - concentration of protein thiol group higher scores mean a worse outcome	2 weeks
metabolic panel - diabetes	metabolic panel - diabetes (including ghrelin, glucagon, insulin, leptin, resistin) - Luminex method with the use of BioRad BioPlex 200 readerm (Bio-Plex Pro ™ Human Diabetes 10-Plex Assay # M171A7001M, BIO-RAD, USA) depends - higher scores mean a better or worse outcome	2 weeks
somatic disorders	Somatic disorders measured by questionnaire based on Likert's scale Likert's scale: - Strongly disagree; - Disagree; - Neither agree nor disagree; - Agree; - Strongly agree It depends on question - higher scores mean a better or worse outcome	2 weeks

Collaborators and Investigators

• Sponsor: Katarzyna Siedzik
• Investigators: Principal Investigator: Katarzyna Siedzik, MSc, Poznan University of Physical Education, Poznan, Poland

Publications and helpful links

General Publications

• Phinney SD. Ketogenic diets and physical performance. Nutr Metab (Lond). 2004 Aug 17;1(1):2. doi: 10.1186/1743-7075-1-2.
• Paoli A, Grimaldi K, Toniolo L, Canato M, Bianco A, Fratter A. Nutrition and acne: therapeutic potential of ketogenic diets. Skin Pharmacol Physiol. 2012;25(3):111-7. doi: 10.1159/000336404. Epub 2012 Feb 11.
• Paoli A, Rubini A, Volek JS, Grimaldi KA. Beyond weight loss: a review of the therapeutic uses of very-low-carbohydrate (ketogenic) diets. Eur J Clin Nutr. 2013 Aug;67(8):789-96. doi: 10.1038/ejcn.2013.116. Epub 2013 Jun 26. Erratum In: Eur J Clin Nutr. 2014 May;68(5):641.
• Paoli A, Bosco G, Camporesi EM, Mangar D. Ketosis, ketogenic diet and food intake control: a complex relationship. Front Psychol. 2015 Feb 2;6:27. doi: 10.3389/fpsyg.2015.00027. eCollection 2015.
• Dashti HM, Mathew TC, Hussein T, Asfar SK, Behbahani A, Khoursheed MA, Al-Sayer HM, Bo-Abbas YY, Al-Zaid NS. Long-term effects of a ketogenic diet in obese patients. Exp Clin Cardiol. 2004 Fall;9(3):200-5.
• Veech RL. The therapeutic implications of ketone bodies: the effects of ketone bodies in pathological conditions: ketosis, ketogenic diet, redox states, insulin resistance, and mitochondrial metabolism. Prostaglandins Leukot Essent Fatty Acids. 2004 Mar;70(3):309-19. doi: 10.1016/j.plefa.2003.09.007.
• Balasse EO, Fery F. Ketone body production and disposal: effects of fasting, diabetes, and exercise. Diabetes Metab Rev. 1989 May;5(3):247-70. doi: 10.1002/dmr.5610050304.

Study record dates

Study Major Dates

• Study Start (ACTUAL): October 1, 2019
• Primary Completion (ACTUAL): June 30, 2021
• Study Completion (ACTUAL): August 31, 2022

Study Registration Dates

• First Submitted: September 23, 2022
• First Submitted That Met QC Criteria: September 23, 2022
• First Posted (ACTUAL): September 28, 2022

Study Record Updates

• Last Update Posted (ACTUAL): September 30, 2022
• Last Update Submitted That Met QC Criteria: September 28, 2022
• Last Verified: September 1, 2022

More Information

Terms related to this study

• Keywords: insulin resistance; body composition; ketogenic diet; oxidative stress; ketosis; immune response; strength capacity; omega 3 acids; meatless diet
• Additional Relevant MeSH Terms: Pathologic Processes; Hyperinsulinism; Acid-Base Imbalance; Acidosis; Disease; Insulin Resistance; Body Weight; Metabolic Diseases; Lipid Metabolism Disorders; Ketosis; Glucose Metabolism Disorders
• Other Study ID Numbers: KETOPROJEKT2019*2023

Plan for Individual participant data (IPD)

• Plan to Share Individual Participant Data (IPD)?: NO

Drug and device information, study documents

• Studies a U.S. FDA-regulated drug product: No
• Studies a U.S. FDA-regulated device product: No