Wednesday, January 2, 2013

The Effect of the Paleolithic Diet on Type 2 Diabetes & Cardiovascular Risk Factors


Section I: Introduction
Over 1.5 million to 10,000 years ago, before cultivated plants and domesticated animals, Homo sapiens evolved and lived nomadic lifestyles in order to survive (Frassetto, Schloetter, Mietus-Snyder, Morris, & Sebastian, 2009; Lindeberg, Cordain, & Eaton, 2003). During the Paleolithic time frame, hunting and gathering of food sources involved traveling on foot an average of 5-10 miles a day (O'Keefe & Cordain, 2004).  Over time, through technological advances and urbanization, humans have evolved to lead drastically different lifestyles due to inactivity and overconsumption of highly caloric nutrient stripped food sources.  Today, the fast food revolution has led to a plethora of chronic diseases, including type 2 diabetes and cardiovascular disease, the most lethal causes of death in the United States (Frassetto et al., 2009).  Decreasing waist circumference, improving energy intake, and increasing energy expenditure has been associated with reducing the rates of type 2 diabetes and improving cardiovascular risk factors (Lindeberg, et al., 2007; Lindeberg et al., 2003).
In 1985, anthropologists Eaton and Konner introduced the Paleontology diet that consists of these outdated food selections: lean meat, fish, nuts, vegetables and fruit (Frassetto et al., 2009; Lindberg et al., 2003).  This diet completely excludes two main food groups, dairy and carbohydrates, which may be detrimental to health over time.  Even, though humans have evolved from our ancestors the genetics have only slightly changed and certain concepts from the Paleolithic period should be adopted, such as increasing energy expenditure and selecting fresh and natural nutrient dense foods (O'Keefe & Cordain, 2004). 
Literature Review
In a crossover 3-month study, the Paleolithic diet and the current dietary guidelines of the Diabetes diet were compared to assess cardiovascular risk factors among participants (n= 13) in Sweden (Jonsson, et al., 2009).  The Paleolithic diet included the following food staples: lean meat, fish, fruit, vegetables, eggs and nuts; while prohibiting grains, dairy, alcohol, and processed or salted foods.  Each participant was randomly assigned to their group and given information on their dietary regimen.  At week six, four-day food records were administered to estimate energy intake.  According to the findings, the Paleolithic diet showed improvements in HDL concentrations by 0.08 mg/dL, diastolic blood pressure by 4 mmHg, and reductions in weight loss of 3 kg, and waist circumference by 4 cm.  There was no significant difference in blood values observed during the oral glucose tolerance test (OGTT). 
In comparison, the Paleolithic diet had lower calorie intake and it consisted of 32+ 7% carbohydrates (CHO) and 24% protein (PRO).  Both calcium and CHO intake did not meet the recommended amounts in the Paleolithic diet.  In general, the daily recommendation for adults between the ages of 19 to 50 is 1,000 mg of calcium and the inadequate consumption of the Paleolithic diet of 356+ 102 mg did not even come close.  Calcium is necessary for bone health and intake has been recently linked to adiposity and weight loss (Steding, Brevard, & Yesilcacy, 2005).  As shown, it is very difficult to obtain the daily calcium recommendations in the Paleontology diet by prohibiting dairy consumption, even if consumption of nutrient rich leafy vegetables is high (Lindeberg et al., 2003).  Limitations in this study would be the short-duration of the intervention, small sample size, and undisclosed physical activity amounts.   
In another controlled study, the effects of the Mediterranean and Paleolithic diet were investigated for 12-weeks on patients (n=29) with ischemic heart disease (IHD) and type 2 diabetes (Lindeberg, et al., 2007).  The male participants were recruited from a Coronary Care Unit in Sweden and randomly assigned to a group.  Then, separately educated on the health benefits of their diet regimen and regular physical activity.  The Mediterranean diet consisted of whole-grains, low-fat dairy, fruit, legumes, fish, and refined monounsaturated fats; while the Paleolithic diet was consistent with the previous parameters.  Similar to Jonsson et al. (2009), a four-day food records were administered after 15+5 days in order to determine the dietary compositions of each group.  In the Paleolithic group, total caloric intake was significantly lower by 25% (p= 0.004) and PRO was 27.9+ 6.8% of the total energy.  This percentage almost constitutes as a high protein diet at 30% and these elevated concentrations have been associated with impaired renal function (Wheeler, et al., 2012; Lindeberg et al., 2003). 
In the findings, glucose response improved by 36% during an OGTT in the Paleolithic group and there was only a 7% improvement the control group.  At week six, both groups decreased AUC insulin by 22%.  After the intervention, the average weight loss was about 4.4 kg and the waist circumference decreased significantly (p= 0.03) in the Paleontology diet by 5.6 cm compared to the control group of 2.9 cm.  However, the glucose tolerance improvement was not associated with weight loss or waist reductions in the Paleolithic group.   Limitations would be the short duration, small sample size, and high drop out rate with the Paleolithic diet (n=3).  Also, physical activity was advised but not recorded and complete nutrient analyses, including calcium, were undocumented. 
In another short-term pilot study, healthy volunteers from Sweden, between the ages of 20-40 years, assessed the effects of an ad libitum Paleolithic diet (Osterdahl, Kocturk, Koochek, & Wandell, 2008).  The diet protocol was presented at a seminar for all male (n=5) and female (n=9) participants, and they recorded their daily intake in a food diary.  However, due to a computer error, complete food registration for only six subjects was documented.  All participants were assessed at phase 1 (baseline), phase two (after normal diet at day 7), and phase 3 (after the Paleolithic intervention at day 21).  According to the food registrations, total energy intake decreased by 36%; CHO significantly decreased by 53% and calcium deceased by 53% (395 mg/d).  Overtime, decreased calcium intake has been associated with increased hip factures (Matković, Kostial, Simonović, Buzina, Brodarec, & Nordin, 1979). 
  In comparison from phase two to phase three, no marked improvements in glucose metabolism were found but there was a significant weight loss of 2.3 kg (p<0.001), waist circumference decreased by 0.05 cm, and systolic blood pressure dropped by 3 mmHg (p= 0.03).  The rapid weight loss between these phases was not shown to be linked to exclusively fat loss.  Therefore, it is possible that reductions in lean body mass may have contributed to the significant weight loss. Limitations to the study would be no control group, small sample size and lack of data on the complete food registrations.    
According to another study by Jönsson et al., the long-term effects of the Paleolithic diet and cereal-based diet was measured in domestic piglets (n= 24) for 15 months (2006).  At two months of age, the pigs were randomly assigned to either diet: cereal (17% PRO, 65% CHO) or Paleolithic (27% PRO, 57% CHO).  Every two weeks body weight was measured and at the end of the study intravenous glucose tolerance test (IVGTT) was performed following a 6-hour fast.  In comparison, the Paleolithic fed pigs observed a reduced weight gain of 22%, lower subcutaneous fat of 43%, shorter length of 6%, and reduced energy intake of 20%.  There was no significant difference in fasting insulin and glucose observed among pigs, but AUC insulin improved by 47%, C-reactive Protein (CRP) was 82% lower (p= 0.0007), and diastolic blood pressure was 13% lower (p= 0.0007) in the Paleolithic diet.  In this particular study, the diets did not match the macronutrients compositions of the other human studies.
In a controlled study by Frassetto et al., nine sedentary healthy volunteers were recruited from the San Francisco area to compare the short-term changes of metabolic variables with the Paleontology diet or usual diet (2009).  All participants were at or below the standardized VO2max for their age and gender and instructed to continue their normal physical activity.  The participants consumed their normal diet for 3-days, followed by 7-day ramp-up diet, and then 10-day intervention of a verified Paleolithic diet.  Contrary to other findings by Lindeberg et al. (2007), no significant change in weight was observed between groups, however the following lipid panels improved: triglycerides (35%), total cholesterol (16%), and LDL concentrations (22%).  Diastolic blood pressure decreased significantly by 3.4+2.7 mmHg and the ratio of AUC plasma insulin to plasma glucose decreased by 40% (p= 0.005) during the glucose tolerance test.  Limitations for this study were the short duration, small sample size, and the absence of a control group. 
In Frassetto et al. (2009) study, the normal diet (18% PRO, 44% CHO, 38% FAT) and prepared Paleolithic diet (30% PRO, 38% CHO, and 32% FAT) had similar calorie composition in a 68-week study by Brinkworth et al. (2004).  The purpose of this longterm study was to observe the impact of cardiovascular risk factors in obese hyperinsulemic participants assigned to one of the following diets: high-protein diet (30% PRO, 40% CHO) or standard protein (15% PRO, 55% CHO).  After the intervention, no changes between groups existed in weight loss, HDL, insulin sensitivity, and CRP levels indicating poor long-term adherence to the intervention.  According to Greenberg et al. (2009) study, the greatest weight loss was observed within 6-months of dieting but the leading factor for long term success was physical activity.  Physical activity was attributed to other healthy behaviors, such as: reduced energy intake and improved metabolism and a decrease in chronic disease prognosis.  In another study by Ross et al., the combination of exercise and diet showed elicited a higher fat reduction of 1.3 kg (p=0.03) than the diet only group (2000).   In addition, the weight loss was due to fat reductions exclusively and it improved metabolic rate, strength, insulin sensitivity, and lipid panels. 
Statement of the Problem
These short-term Paleolithic diet studies may seem promising for cardiovascular risk factors in type 2 diabetics but it neglects a major components of energy balance, physical activity.  The long-term effects of the Paleontology diet on cardiovascular risk factors and bone mineral density (BMD) within a randomly assigned, large sample size, and compared to a control group has not been assessed.  The purpose of the proposed study is to determine the impact of the (P) Paleolithic diet, Paleolithic diet and exercise (P+E), Diabetes diet (D), Diabetes diet and exercise (D+E) on type 2 diabetes and cardiovascular risk factors over 12-months.  The specific null hypothesis is:
1.     There is no difference in fasting blood values (plasma glucose and plasma insulin), body weight, body fat, blood pressure, and cholesterol (total, LDL, and HDL) after the 12-month intervention.     
2.     There is no difference between groups on changes in insulin sensitivity, B-cell function, and bone mineral density after the 12-month study.
3.      There is no difference in dietary intake and nutrient compositions between groups.           
Section II: Method
Participants
The participants were selected using a convenience sample of male and female volunteers recruited from Medical Center of Plano.  All participants were enrolled in the Center of Lifestyle Enhancement Program for Diabetes Education.  There were 200 participants recruited and the inclusion criteria was that the participants were: between 45-60 years old, type 2 diabetic, and not taking any prescribed medication or supplements that would deter from the findings.  The definition of diabetic was consistent with World Health Organization standards, fasting plasma glucose ≥ 7.0mmol/l or 2–h plasma glucose ≥ 11.1mmol/l (Sartorelli et al., 2010). Participants were excluded if they had a history of an acute coronary event or any chronic disease diagnosis, including Osteoporosis that would interfere with the study (n= 50).  Other participants were excluded if they did not adhere to the dietary regimen (n = 25) or had unexpected health complications (n= 25). 
The adult subjects were selected from patients at Medical Center of Plano, enrolled in the Diabetes Education program at the Center of Lifestyle Enhancement, and lived in the Dallas and Fort Worth area.  There were 100 participants that completed the 12-month study. Participants were randomly assigned to the following groups: (P) Paleolithic diet, Paleolithic diet and exercise (P+E), Diabetes diet (D), Diabetes diet and exercise (D+E).  The Institutional Review Board (IRB) approved the study and all participants were informed of the objectives of the study and a written informed consent was obtained prior to the study. 
Instruments and Measurements
A validated self-administered Medical History Questionnaire including family history, alcohol intake, and the use of any medications were recorded for each participant.  It provided demographic, personal history, contact and habitual information about the participants.  In order to control for food restrictions, the groups received written information about their assigned diet regimen.  The Diabetes diet was in accordance with the general recommendations from the American Diabetes Association and the Paleolithic diet was in accordance with the previous parameters from Jonsson, et al. study (2009).  At baseline, each participant met with a Registered Dietician (RD) at Medical Center of Plano to discuss and answer any questions.  Then exercise groups, P+E and D+E, met with an Exercise Physiologist to complete a PAR-Q and receive information on their exercise program.  The exercises consisted of resistance training, flexibility, and aerobic exercises 3-5 d/week at Medical Center’s Cardiac Rehab Center and all were in accordance with ACSM’s Guidelines for Diabetic participants.  The three-day dietary intake was completed by all groups every 2-weeks through a validated computer software program, Fitness Pal, and analyzed by Food Processor SQL (Version 10.0) and Nutrition Software (ESHA Research, OR, USA) (Frassetto et al., 2009).     
After an overnight fast (12-14 hours), blood samples was collected in tubes from the antecubital vein for determination of total cholesterol, HDL, LDL, plasma glucose and insulin.  Plasma insulin was measured by the chemiluminescent assay (Esoterix, Callabasas Hills, CA).  Plasma glucose was determined by the hexokinase method with Abbott Architect analyzer.  Hitachi 912 Chemistry Analyzer by Roche Diagnostics determined total cholesterol, HDL, and LDL content (Sartorelli et al., 2010).  Insulin sensitivity and B-cell function was assessed by the homeostatic model assessment (HOMA) technique by Mattews et al. (1985), which has shown a good correlation to the gold standard euglycemic hyperinsulemic clamp (Bonera et al., 2000; Osterdahl et al., 2008).  Insulin resistance was calculated by the following HOMA-IR formula: fasting serum insulin (μU/ml) x fasting plasma glucose (mmol/l)/ 22.5.  B-cell function was calculated by HOMA-B formula: 20 × fasting insulin (μIU/ml)/fasting glucose (mmol/ml) 3.5 (Bonera et al., 2000).  In accordance with ACSM’s Guidelines for Exercise Testing and Prescription, resting blood pressure, height, and weight were measured.  Resting blood pressure was measured in supine position three separate times and the first Korotoff sound was systolic and the fifth Korotoff sound was diastolic blood pressure.  It was measured using a standard of mercury and 3M Littmann Master Cardiology Stethoscope and Omron Sphygmomanometer.  Salter 200 Academy Mechan Scale, measured weight without shoes or heavy apparel, and height was recorded without shoes and to the nearest 0.5 cm. 
Body fat percentage and bone mineral density (BMD) was measured by the gold standard Dual-energy X-ray absorptiometry (DXA) scan.  Whole body scans were performed on bone densitometer Norland XR-36 and body tissue was analyzed with Software 3.7.4 Version 2.1.0 (Norland Corporation, Ft. Atkinson, WI).  All scans were performed according to the standard protocol with participants in supine alignment.  Following each scan, the tester performed a standard calibration and the precious error for total BMD is 0.75%.  T-scores was calculated and diagnostic criteria was in accordance of the standard deviations (SD) to the norms: <1 is average BMD, 1-2.5 osteopenia, and >2.5 osteoporosis (Kohrt, 2011).   
Procedures
The study was 12-months long and prior to the study, the participants were informed of the study, randomly assigned to a diet group, and consulted a RD.  The exercise groups consulted with the Exercise Physiologist and followed the exercise regimen 3-5 d/week at the Cardiac Rehab Center.  Following an overnight fast (12-14 hours), blood pressure, weight, height, plasma values, insulin sensitivity, B-cell function, and the whole body DXA scan were measured at baseline.  The participants were reminded weekly by email about the study and all received detailed instructions.  Each month, the participants met with the RD to review their diet and three-day food diaries.  The exercise groups met with the Exercise Physiologist each month to address their exercise regimen.  Then, after 12-month interval and following an overnight fast (12-14 hours), blood pressure, weight, height, plasma values, insulin sensitivity, B-cell function, and whole body DXA scans were measured again for follow-up. 
Design and analysis
The results are expressed as means + SDs or medians and interquartile ranges are specified.  The level of significance was set at 0.05 to make decisions about the hypothesis. Statistical analyses were conducted by SPSS version 13 (for Windows).  Associations between changes were assessed with the Spearman correlation coefficients.  The analysis of variance (ANOVA) compared the four groups and changes in insulin sensitivity, B-cell function, and bone mineral density from baseline to after the study.  ANOVA was used to compare the total cholesterol, HDL, LDL, and glucose metabolism in the groups: (P) Paleolithic diet, Paleolithic diet and exercise (P+E), Diabetes diet (D), Diabetes diet and exercise (D+E).  Linear trends were used to test any relationship between calcium intake, changes in BMD and weight.         

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