REF: Meat Pro SL. Project Principal Investigator Dr. Fernando Naclerio. Centre for Sports Sciences & Human Performance University of Greenwich

Transcription

1 REF: Meat Pro SL Experimental Proposal Effects of meat protein administration on body composition, strength, muscle structure and general markers of health in athletes Project Principal Investigator Dr. Fernando Naclerio Centre for Sports Sciences & Human Performance University of Greenwich

2 Background The protein requirements for athletic populations have been the subject of extensive scientific debate. During the last decade the notion that athletes as well generally active population require greater protein intake than the current RDA recommendation of 0.8 g/kg of body weight per day in healthy adults has become well accepted (ACSMps, 2009; Campbell & Kreider, 2008). In addition, high-protein diets have also become quite popular in general population as part of many weight reduction programs (Layman, Shiue, Sather, Erickson, & Baum, 2003; Volek & Forsythe, 2008). Differences in protein requirements for different types of athletes (i.e., endurance, strength/power) and non-athletes are well documented (Bilsborough & Mann, 2006; BØrsheim, Tipton, Wolfe, & Wolfe, 2002; Paddon-jones et al., 2004; Tipton & Wolf, 2001). Many studies have investigated the effects of protein obtained from different sources such as milk (Elliot, Cree, Sanford, Wolfe, & Tipton, 2006) eggs (Moore et al., 2009), soy (Zhan & Ho, 2005), rice (Joy et al., 2013) or bovine colostrum (Shing, Hunter, & Stevenson, 2009) as well as the technique used to manufacture the protein supplements (Etzel, 2004) or the methodology that is recommended for their ingestion (Campbell et al., 2007). In fact, these factors can be very relevant when the protein supplements are ingested with the scope to optimize training adaptations (Kreider et al., 2010). Although some studies have shown greater strength, muscular hypertrophy and/or lean body mass (LBM) gains when resistance training is combined with milk based protein (Tang et al., 2007) ingested alone or combined with others nutrients such as carbohydrates (Kreider et al., 2007) or creatine (Cribb, Williams, & Hayes, 2007); there are still some inconsistencies regarding the most effective protein sources or doses protocol to maximise training adaptations and outcomes (strength gain, or muscle mass accretion) in athletes. Most of protein extracts are obtained from milk, eggs, bovine colostrum, soy and to less extend from bean, wheat or rice. The quality of these extracts may vary depending on the type and quality of the manufacturing process. A range of processed milk proteins (concentrates, isolates, hydrolyzate or peptides) has been used for different purposes including changes in metabolic health in young or elderly people (McGregor & Poppitt, 2013) or training optimization outcomes in untrained (Hulmi et al., 2009) recreationally active (Reitelseder et al., 2011; West et al., 2011) or welltrained athletes (Cribb & Hayes, 2006; Willoughby, Stout, & Wilborn, 2007). More recently the consumption of high quality protein meat extracts have been proposed as effective nutrition strategy to optimise training adaptation in young athletic and active population (Negro et al., 2014), however, even when meat is a nutrient-rich high-quality protein containing all the essential amino acids in proportions similar to those found in human skeletal muscle, meat protein ponders are a relatively new marketed form of protein that still need to be supported by scientific research. Aim The aim of this project is to investigate the potential benefits of combining a new protein meat hydrolysates extract with a regular resistance training programme on (a) body composition (b) performance (c) muscle structure (d) general markers health and immunity in athletes. As a second objective we will analyse potential differences obtained from the ingestion of the new hydrolysates meat protein extract compared to the ingestion of others commercially available protein sources such as whey or casein.

3 Hypothesis We hypothesise that combining regular resistance training with the ingestion of hydrolysates meat protein extract or others based milk protein sources would maximise training outcomes, help to attenuate post exercise immune suppression and positively affect markers of general health in athletes. Project design This investigation involves a 10 weeks randomised, balanced, double blind parallel group between-subjects design aimed to analyse the effects of combining exercise and a post exercise nutrition strategy (hydrolysates meat protein, whey protein or a isocaloric only carbohydrate placebo) on training outcomes, markers of immunity and health after a period of regular training and feeding intervention. Participants will be divided in three-treatment groups 1) Hydrolysed meat-protein (MEAT); 2) Whey (W) and 3) non-protein iso-energetic placebo (CHO). Once considered eligible for the study, and after an initial familiarisation period and baseline tests, participants will be randomly assigned to one of the intervention groups: MEAT; W and CHO. Each group will follow a 10-week periodised resistance training intervention combined with one of the three specific supplementation treatments (MEAT, W or CHO). Measurements of general markers of health, immunity, body composition, muscle structure and performance will be determined before and after the 10 week training intervention. Once informed consent and health history have been obtained, after the baseline assessment (t1), participants will divided into three similar profile groups, matched by body mass, age, sex and performance. 1) The meat protein group (MEAT n=10) that will take 500 ml of water mixed with 20g of hydrolysates meat protein powder (Meatpro) 2) The whey group (n=10) that will take 500 ml of whey intact isolate powder mixed with 500ml of water and 3) the contrast group (P n=10) that will take an isoenergetic non-protein, only carbohydrate placebo. Participants Thirty participants, recruited from Medway University Campus, will take part in the project. Inclusion criteria are: regularly trained male or female athletes, aged 18 to 45 years old, with at least 3 month of experience in resistance training exercise, volunteers, with no musculoskeletal injuries, metabolic conditions, or diseases; and use of medications, smoking, and nutritional supplements known to affect physical performance, muscle damage or recovery process (e.g., creatine, whey protein, and amino acids) within 6 weeks prior to the start of the study. After being informed of all risk, discomforts, and benefits involved, participants will sign a written informed consent regarding their participation. Procedures will be in accordance with the University Research Ethics Committee. Protocols will be in compliance with the Helsinki Declaration, and be approved by the Ethical Review Board. Intervention Familiarization period: Participants will undertake 3 sessions of familiarization (1 weeks). A qualified strength and conditioning coach will control and assists participants in order to guarantee a correct execution of the selected exercise. All participants will be instructed

4 about the appropriate use of the Rate of Perceived Exertion OMNI-RES (0-10) scale to control the load and training intensity. Resistance Training The resistance-training program will be designed to increase strength and mass of all major muscle groups. The program will be undertaken on three non-consecutive days per week, under the supervision of strength and conditioning coaches, preferable a MSc Strength and Conditioning students at the centre for sport science and human performance (University of Greenwich). Training workout will be performed during the afternoon (between 3pm and 6pm) with at least 48 hours between sessions. Each session will begin with individualized warm-up (5-minute warm-up stretches, followed by one set of eight repetitions of six resistance training exercises without any additional weight). Workout will involve 8 resistance exercises per 3 sets of 6 to 10 repetitions at 70% to 85% of an individual 1RM. Exercises will be performed using free-weights, elastic bands, sticks and machines selected to stress the major muscle groups. Resting periods of around 2 min will be allowed sets. Participants will be instructed to maintain the proper execution along the entire training session. Periodization strategy The intervention will consist of 10-week periodised resistance training programme that will be divided in 3 blocks: First block (1 st to 4 th week) using a moderate load intensity of around 70%1RM; Second block (5 th to 7 th week) where the intensity will increase to >75% to 80% 1MR and the third block (8 th to 10 th week) where the highest loads (>80 to 85% 1RM) will be used. The Rate of Perceive Exertion OMNI-RES (0-10) scale (RPE) will be used to appropriately select the load along the training intervention (Naclerio et al., 2011). Resistance training intervention will be appropriately integrated with the regular sport activity performed by the University athletes along the academic period. Supplementation protocol Training days: Immediately (<10 min) after completing each training session (resistance training or sport specific workout), participants will consume a beverage containing 20g of supplement (meat or whey) or placebo (carbohydrate). Non-training days: before breakfast. Supplements and placebo will be provided in powder form and should be mixed with plenty water at the moment of consumption. The supplements and placebo will looks and tastes identical. Thus a total of one 20g doses will be administered on daily bases. Participant should ingest a total of 70 doses for the total of 10 weeks of study intervention. Supplement will be provided in 14 days based by a blind researcher after participants return back the empty bag of the supplement/placebo consumed during the previous 14 days period.

5 Table 1 details the estimated nutrient composition and caloric content of the supplement and placebo. Energy (kcal) Total protein (g) Total CHO Total fats Meat Supplement CHO Maltodextrine Whey isolate intact 1 dose 20g 1 dose 20 g 1 dose 20g ~ ~ ~ Administration: one 20g bag per day per participant per a period of 10 weeks (70 days in total) Measurements Body composition: - Plestimogrphy Bod Pod (Life Measurements, Concord, CA). Muscular structure - Ultrasound (non-invasive methodology). Performance - Upper and lower body maximal strength - Explosive endurance 60 sec jump test. Oxidative stress, muscle damage and immunosuppression - Salivary Markers* - Antimicrobial peptides (Allgrove et al., 2008, Usui et al., 2011). - Blood markers - Creatine Kinase (CK); creatinine; glycaemia, total cholesterol; HDL and LDL cholesterol, aspartate amino transferasa (AST); alanine amino transferasa (ALT); triglycerides, urea and uric acid ** -Urine Markers Essential Amino acid concentration*** Time of measurements 1. T1: After the familiarisation period and before to start the ten weeks training period 2. T2: At the end of the intervention *Salivary markers will be determined at three times points: before (baseline) immediately after (post) and 60 min after the completing the first and last resistance training session included within the ten weeks intervention period. ** Blood markers will be determined in fasted state before (pre) and at the end of the intervention period. ***Urine samples will be collected prior, and at the end of the intervention period.

6 Workouts Sessions Blood Saliva Body composi(on Performance test Muscle structure Urinesample Familiariza(on Test1 Periodizedresistancetraininginterven(on 1to4weeks (70%1RM) 5to7weeks (>75to80%1RM 8to10weeks (>80to85%1RM Test2 General overview of study Statistical Power of the study and Potential Sample Size From previous publications (Naclerio, Larumbe-Zabala, Cooper, Jimenez, & Goss- Sampson, 2014; Witard et al., 2014); we consider that estimations of sample size should be calculated on a medium effect size expectation. Thus, we assume that partial eta squared will be around 0.5, which is equivalent to f=0.25 in our design. We also assume a desirable ratio of 1:4 between alpha and beta (Cohen, 1988); since alpha will be α=0.05, then β=20 and subsequently the expected statistical power will be 1-β=0.80. After our experience (Cooper et al 2013; Naclerio et al 2014), the three repeated measures will share at least a 50% of variability, which means that correlation among repeated measures will be approximately r=0.7. We will analyse the evolution of physical performance, body composition, muscle structure, general markers of health, muscle damage and immune function in two different times for assessing three conditions (MM; M and P). Using G*Power we calculated that the main between effect needs 22 subjects to achieve the desired statistical power, whereas the interaction effect needs 24 subjects. However, we estimate that a sample of 30 participants will be more conservative in order to allow us to detect even smaller effect sizes and consider the possibility of dropouts.

7 Time line Months Recruitment X familiarisation X Test 1 X Intervention X X X X Test 2 X Data analysis X Results publications X X X Expected outcomes Previous studies demonstrated the beneficial effects of high quality protein supplementation on reducing chemical damage to tissues after exercise and optimize the processes of muscle tissue repair and remodelling in middle-aged athletes or active and healthy men and women (Cockburn, Bell, & Stevenson, 2013; Cockburn, Stevenson, Hayes, Robson-Ansley, & Howatson, 2010). However, results from the current project will demonstrate the potential benefits of combining high quality meat protein extract to maximise training benefits, attenuate post exercise immunosuppression and positively influence general markers of health in athletes. In addition we will be able to analyse potential different outcomes obtained from the ingestion of the new meat protein product or a typical commercially available whey protein extract.. Benefits to the company Our outcomes may show a pattern of behaviour in order to improve training adaptation, and general markers of health in athletes. These results will increase interest in this product among the athletic population. Nowadays, athletes and active people are almost exclusively consuming high quality protein derived from milk sources such as whey and casein protein extract. These types of protein supplements need to be administered in several daily dosses of 10g to 20g). Meat protein extract would require only 1 serving per day. This special characteristic will represent a unique selling point (USP) for the company because it will encourage regular consumption of this product. It would therefore be a constituent of regular diets and not only a supplement required to be consumed under specific protocols.. investigators details Principal Investigator Dr. Fernando Naclerio Mailing address; address; Centre for Sport Science and Human Performance University of Greenwich at Medway Grenville building Central Avenue Chatham Maritime Kent ME4 4TB f.j.naclerio@gre.ac.uk

8 Principals co-investigators Dr. Marco Seijo, University of Greenwich (UK) Dr. Eneko Larumbe Zabala Clinical Research Institute Texas Tech University (USA) Sponsor support In order to conduct this study we require Meat Pro S.L to provide us the meat protein, whey protein intact and maltodextrine packed in 20g doses-bags maintaining the same appearance and flavour between the three products. The company will also encode the bags in order to maintain the double blind characteristics of the study. The company will reveal the composition of the encoded product as requested by the Principal investigation after completing the data collection and analysis. Public disclosure plan Once written up, the finished articles will be submitted for publications to high impact journals. A complete and fully report including all the results will be submitted to the sponsor. Researchers are willing to cooperate with the company in order to establish some marketing and diffusion strategies to communicate the results to nutrition related professionals and general population.

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10 Layman, D. K., Shiue, H., Sather, C., Erickson, D.J., & Baum, J. (2003). Increased dietary protein modifies glucose and insulin homeostasis in adult women during weight loss. J. Nutr., 133, McGregor, R. A., & Poppitt, S. D. (2013). Milk protein for improved metabolic health: a review of the evidence. Nutr Metab (Lond), 10(1), 46. Moore, R. D., Robinson, M. J., Fry, J.L., Tang, J.E., Glover, E. I., Wilkinson, S.B.,... Phillips, S.M. (2009). Ingested protein dose response of muscle and albumin protein synthesis after resistance exercise in young men. Am J Clin Nutr., 89, Naclerio, F., Larumbe-Zabala, E., Cooper, R., Jimenez, A., & Goss-Sampson, M. (2014). Effect of a carbohydrate-protein multi-ingredient supplement on intermittent sprint performance and muscle damage in recreational athletes. Appl Physiol Nutr Metab, 1-8. Naclerio, F., Rodríguez-Romo, G., Barriopedro-Moro, M.I., Jimenez, A., Alavar, B., & N, Travis, Triplett. (2011). Control of resistance training intensiy by the omni perceived exertion scale. J. Strength Cond. Res, 25(7), Negro, M., Vandoni, M., Ottobrini, S., Codrons, E., Correale, L., Buonocore, D., & Marzatico, F. (2014). Protein supplementation with low fat meat after resistance training: effects on body composition and strength. Nutrients, 6(8), Paddon-jones, D., Sheffield-Moore, M., Zhang, X.J., Volpi, E., Wolf, S.E., Aarslan, A.,... Wolfe, R. R. (2004). Amino acid ingestión improves muscles protein síntesis. Am J Physiol Endocrinol Metab, 286(E321-E328). Reitelseder, S., Agergaard, J., Doessing, S., Helmark, I. C., Lund, P., Kristensen, N. B.,... Holm, L. (2011). Whey and casein labeled with L-[1-13C]leucine and muscle protein synthesis: effect of resistance exercise and protein ingestion. Am J Physiol Endocrinol Metab, 300(1), E Shing, C. M., Hunter, D. C., & Stevenson, L. M. (2009). Bovine colostrum supplementation and exercise performance: potential mechanisms. Sports Med, 39(12), Tang, J. E., Manolakos, J. J., Kujbida, G. W., Lysecki, P. J., Moore, D. R., & Phillips, S. M. (2007). Minimal whey protein with carbohydrate stimulates muscle protein synthesis following resistance exercise in trained young men. Appl Physiol Nutr Metab, 32(6), Tipton, K.D, & Wolf, R. (2001). Exercise, protein metabolism and muscle growth. Int J. Sports Nutr. Exc. Metab., 11(1), Volek, J.S., & Forsythe, C. (2008). Very-Low Carbohydrate Diets, Chapter 25. In J. Antonio, D. Kalman, J. R. Stout, M. Greenwood, D. G. Willoughby & G. G. Haff (Eds.), Essential of Sports Nutrition and supplements (pp ): Humana Press. West, D. W., Burd, N. A., Coffey, V. G., Baker, S. K., Burke, L. M., Hawley, J. A.,... Phillips, S. M. (2011). Rapid aminoacidemia enhances myofibrillar protein synthesis and anabolic intramuscular signaling responses after resistance exercise. Am J Clin Nutr, 94(3), Willoughby, D.S., Stout, J.R., & Wilborn, C.D. (2007). Effects of resistance training and protein plus amino acid supplementation on muscle anabolic, mass, and strength. Amino Acids. 32, Witard, O. C., Jackman, S. R., Breen, L., Smith, K., Selby, A., & Tipton, K. D. (2014). Myofibrillar muscle protein synthesis rates subsequent to a meal in response to increasing doses of whey protein at rest and after resistance exercise. Am J Clin Nutr, 99(1),

11 Zhan, S., & Ho, S. C. (2005). Meta-analysis of the effects of soy protein containing isoflavones on the lipid profile. Am J Clin Nutr, 81(2), Dr. Fernando Naclerio Principal Lecturer Strength Training and Sport Nutrition Centre for Sport Science and Human Performance University of Greenwich (UK)