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    Effects of Mixed Nuts on Satiety, Blood Glucose, Insulin and Anxiety How nuts affect health Alma Lopez, Mike Best, Aliya Levy San Diego State University, Exercise and Nutritional Sciences Corresponding author: Mee Young Hong Contact; $500 Campanile Dr., San Diego, CA 92182 Abstract ‘The purpose of this study was to find differences between consumption of mixed nuts and pretzels in regards to glucose, insulin, anxiety, and satiety. The study investigated if the consumption of mixed nuts inc: ed feelings of satiety compared to pretzels, as well as if mixed nut consumption decreased feelings of anxiety compared to pretzels. The study was performed to establish insulin and glucose response comparing mixed nut and pretzel consumption. Subjects fasted overnight, consumed ‘mixed muts on day one and on day two they consumed pretzels with water. The subjects had their glucose and insulin a ssed along with amxiety and satiety. nck eg/dl It was found that consumption of nuts did not significaffly affect anxiety over a 60 minute period. Blood glucose was found to stay stable (99.86 + 2.25 at 0 minutes to 99.36 + 2.75) with the consumption of nuts whereas consumption of pretzels significantly increased the blood glucose( 100.23 + 2.68 at O minutes to 143.64 + 4.64 at 60 minutes). Blood insulin levels found similar results to the 7. F? glucose, no increase found with nut consumption(4.69 + 0.78 at 0 minutes to 5.83 + 0.42 at 60 minutes) whereas pretzels found a significant increas. + 042 at 0 minutes to 25.01 + 191 at 60 minute) Contrary to the hypothesis that nuts would increase satiety more than pretzels, it was found that over the 60 minate period pretzels had a larger impact onan increased satiety ‘This study found that there was a significant relationship between consumption of mixed nuts and stability of blood glucose and insulin levels as well as increased stg Further studies should include the evaluation of satiety hormone biomarkers as well asa longer study duration with isovolumetric food items. Key Words: Nuts, Glucose, Insulin, Anxiety, Introduction Statement of Problem and Importance Currently there is limited research on the possi le beneficial effects of a nut mixture on satity, glucose, insulin and anxiety. Finding out these effects on glucose and insulin can be important not only to those with metabolic disorders or prediabetes but also to those who are just looking to maintain their health. Literature Review Each person's day to day diet affects their satiety, blood glucose, insulin levels and anxiety and «depending on what they are eating it will either improve these marker or have a harmful effect, Insulin is ‘a hormone that helps the body store glucose when there is an excess in the body, as well as when the body is lacking the pancreas will secrete insulin to keep the blood sugar in the body from getting too low[1]. Keeping steady insulin levels is most important for individuals who are either diabetic or prediabetic In @ November 2014 study they found that compared to a diet that did not include pistachios, ‘those who had chronic pistachio consumption saw a glucose and insulin lowering effect and were able to reverse certain metabolic consequences of prediabetes[2]. A study run by Kendall looked at the difference of insulin levels over 180 minutes after giving subjects a variety of meals; a group eating white bread and pistachios(WB+P), a group eating white bread only(WB150), and a group eating white bread, butter and cheese(WB+B+C). The subjects insulin levels were all very similar at 60 minutes post-prandial, however at 120 and 180 minutes researchers saw a significant difference with the WB+B+C group having much higher insulin levels. The spike in insulin level is likely from the presence of proline, glutamic acids and whey protein in the cheese. Proline and Glutamic acids have been shown to create significant insulin responses, while whey protein inhibits dipeptidyl dipeptidase IV, which actually decreases the halflife of GLP-1 causing postprandial release of insulin from pancreatic -cells. Kendall also measured blood glucose and found that pistachios elicited little to no increase in postprandial glucose levels making them a good diet choice for those monitoring blood glucose[3]. The glycemic index(GI) of a food is a way to know how much a specific carbohydrate containing food will raise blood glucose levels. A study by Esfehani looks at the glucose levels 60 ‘minutes postprandial as well as the glycemic response of mixed nuts with serving sizes 30g, 60g, or 90g. ‘They found that the nuts decreased the glycemic response of white bread even with the 30g serving, This result is likely because of the low carbohydrate content in the nuts. The study also showed that subjects who consumed white bread only had the highest glucose level after 60 minutes while subjects who consumed white bread and mixed nuts had the lowest glucose level. This study concluded that although nuts have very little effect on postprandial blood glucose levels, they show a large effect on decreasing the glycemic response of foods that would typically have a high glycemic response{4]. When subjeets were fed almonds and peanuts their feelings of hunger were suppressed, increasing their feclings of fullness after consuming the nuts. These studies suggest that consuming nuts ‘may also play a role in weight management since subjects did not eat after consuming muts. The subjects also had smaller meal sizes after consuming nuts due to their increased satiety and feelings of fullness. According to Jaapna, the timing of the nut consumption played a role in the feelings of satiety and fullness. If the subjects consumed the almonds with a meal, it did not significantly impact the satiety levels of the subject. However if the subject consumed the nuts as a snack between meals then there was an increase in the feelings and fullness and satiety following consumptionf5]. Jaapna also states that the form in which the nuts are consumed also impact the levels of satiety santa, nei ig and ful ‘ground method, such as peanut butter, the suppression of hunger was less compared to when nuts were consumed whole. When almonds were consumed in Se ee ee both ground form and whole form, almond butter and whole nuts, the levels of satiety were the same[S]. ‘One study from Jackson, concludes that nuts contain a high amount of dietary fiber that contribute to the increased feelings of fullness and may delay gastric emptying, which can suppress hunger{6] Improvements in mood can be due to the increase in consumption of nuts which have large amounts of magnesium that act as a mood stabilizer. In a study by Lee they looked ata group of 24 young women and had them adhere to a Mediterranean diet, which mainly consists of plant based food, fot a minimum of 10 days. Ina | cup serving of mixed nuts there is 307 meiagnesiur which is almost Sa eet ‘equal to the RDA(310-320 mg/day for women, 400-420 mg/day for men). The study concluded that the ‘women who stuck to the Mediterranean diet for 10 days found that their mood did improve{7].. In Cie high January 2009 a study wes performed that found that there was a significant correlation between low ‘magnesium intake and depression, especially in younger adults. This study saw that by the increase of dietary Magnesium, which is easily implemented in the form of mixed nuts, people saw mood improvements{8]. Objective ‘The objective of this study was to determine the effects of mixed nuts on satiety, glucose, insulin and anxiety. Hypotheses Mixed nut consumption reduces the risk for diabetes by enhancing satiety and improving post-prandial glucose and insulin levels; Mixed nut consumption may be effective for reducing anxiety, Methods 22 male and female, SDSU student participants between the ages of 18- 55 years old excluding subjects with chronic metabolic disorders, required dietary supplement use, smokers, pregnant and allergies to nuts and wheat. Females completed this study while in days 3- 11 of their menstrual cycle. ? bverd name 2 All participants were fed a control diet of unsalted pretzels (2.502, 253kcal, Na 172.Smg) and mixed nuts (1.Soz, 253keal, Na 163.2mg) with 160z water. The €rossover desigi'gonsisted of overnight etn “e hut inte ‘ 2 i Anthtopcmetric and biochemical data was collected on each of the subjects. Height was collected by using a scale with a measuring device attached to it, Height was noted in both standard and metric, Body weight was also obtained using the same scale. Weight was also recorded in standard and metric units, Percent body fat was determined via bio impedance analysis (BIA); a compact version held upright with two hands (Omron Healthcare Inc. Bannockburn, IL). Blood pressure was measured using the standard brachial cuff placement ofthe automated blood pressure device (ReliON automatic BP ‘monitor, Onaron Healthcare Ine.). Waist to hip ratio was calculated by measuring the hip and waist with tape measure in inches and dividing the waist value with the hip value (W/H= WH ratio). Satiety was measured using a 10cm scale, Oem- 10cm (em being the greatest), with 5 questions; these were done 210,20, 40 and Gin Amcnty wns dscemed wring» modifed GAD-7 style questionnaire. The questionnaire consisted of 7 questions with possible answers of O= not present, 1= mild, 2= moderate and 3= severe; this was completed at 0 and 60min. Blood glucose was analyzed using a lancet finger stick, standard glucose test strip and a glucose meter (glucose monitor, BD Biosciences, San Jose, Ca). Insulin blood levels were determined by the collection of 200uL blood from the finger stick and the use of an assay (ultrasensitive insulin ELISA, ALPCO, Salem, NH). The ELISA protocol is a sandwich type immunoassay that required 2SmuL of serum to interact with» monoclonal antibody specific for insulin, Measured at 450nm, the intensity ofthe color during photo spectrometry is proportionate to the amount of insulin present in the sample. Both glucose and insulin measurements were taken at 0 and 60min. A 24-hour dietary intake recall was obtained and analyzed using Super Tracker software (United States Department of Agriculture, S.W. Washington, DC ), The latter data was collected for two days, the control and experimental day, Data was analyzed by ANOVA using SPSS (IBM, Armonk, New York) to evaluate the effects of the mixed nuts and pretzels on blood glucose, insulin, anxiety and satiety. Data wie presented by ‘MeantSE and an alpha level of p<0.05 vay considered significant, T- tests were used fo compare the was values between pre-and post-consumption and between the two trials. The correlation between the other variables (anxiety, satiety, blood glucose and insulin) vee ‘observed. This study was approved by SDSU Institutional Review Board (IRB). é noah A years Results There were 22 participants, 6 male and 16 female. Males had/én average age of 27 + and females had an average of 25.63 + 1.41 years. The averag¢ BMI Yor males was ow $1.17 ky/m? and the average BMI for females was 22.35 + 0.87 kg/m*. The (W/H ratio a(tfage for the male was 0.88 +£0.03 and for the females was 0.78 + 0.02. The average amount of sleep for males on « weekday was 7.5040.52 hours and for females was 7.1640.17 hours, On weekends the average amount of sleep for ‘males was 7.83+.33 hours and for females it was 7.44+0.26 hours. Refer to Table 1 for full results After looking at the results comparing subject's anxiety scores at 0 minutes versus 60 minutes ‘was noticed that there was a decrease in the anxiety score from 0 minutes to 60 minutes. Basal reading for the nut rial gave an anxiety value of 5.64 + 0.84 and at 60 minutes the anxiety value had decreased 0 3.45 + 0,82, Similarly for pretzels the pretzel basal reading gave an anxiety value of 5.77 0.89 and at 60 minutes the anxiety value had decreased to 4.09 0.85. These significant decreases gave a p-value of _ponsfien is significant but with the p-value of 0,651 for the treatment itis not significant for this study. 0.02b in nuts vs preteels ‘The reasons for decreased anxiety likely were due to outside reasons. For instance if someone feels anxious when they are hungry they would feel less anxious at minute 60 because they have finally eaten. Refer to Table 2. Blood glucose levels showed that the consumption of nuts did fot increase blood glucose significantly after 1 hour(99.86 + 2.25 at 0 minutes to 99.36 + 2.75 at 60 minutes), whereas consumption of pretzels showed a significant canoe ne. lucose levels after 1 hour(100.23 + 2.68 at 0 minutes to 143.64 + 4.64 at 60 minutes). With a p-value of 0.001 it shows pretzels will significantly increase the blood glucose in comparison to the minimal, if any, increase seen after eating nuts. This is likely due to the low carbohydrate content in the nuts compared to a much higher carbohydrate content in the pretzels. Refer to Table 3, ‘As seen in Table Aut contumption didnot significantly increase blood ine, however tere was a significant increase in blood insulin following consumption of pretzels. During the nut trial the basal level was measured at 4.69: 0.78 and afler I hour post consumption of nuts insulin level was at Un-Ts 5.83 + 0.82. During the pretzel trial the basal level of insulin was measured at 5.31 + 0.42 and after 1 ‘hour post consumption of 2.5 oz. of pretzels the insulin level was measured at 25.01 + 1.91. With a p-value of 0.0016 it shows that pretzels significantly increase blood insulin levels more than nuts do ina 1 hour measuring period There was a correlation seen between BMI and the factors: waist size, hip size, W/H ratio, % body fat, Sleep on weekdays, Hard Activity on weekdays and Hard Activity on Weekends, There was a positive correlation between BMI and waist, hip and W/H ratio with correlation coefficients of 0.651, 0.337 and 0.322 respectively meaning that those with larger BMils tended to have higher waist, hip and WH ratios. There was also a positive correlation (coefficient=0,327) between BMI and percent body fat. There was a negative correlation between BMI and sleep on the weekdays, hard activity on weekdays, and hard activity on weekends with correlation coefficients of -0.418, -0.440, -0.556 respectively. Showing the relationship between sleeping, hard activities and the likelihood of having a larger BMI value. Refer to Table 6 for correlation values. There was a correlation found between glucose and insulin, finding that as glucose stayed steady insulin would also stay steady. Following with that, insulin would increase as glucose increased and would decrease as glucose decreased. However, there was no correlation found between anxiety and glucose, insulin, or BML ‘As seen in Figure | using the Visual Analog Scale (VAS) subjects were asked to use a 10cm. line to define their hunger and it was shown in the results that subjects had approximately the same value at 0 minutes for the nut trial(7.90 + 0.34) and for the pretzel trial{7.52 + 0.41). Over the 60 minutes the subjects hunger dropped lowest at the 20 minute mark at 4,67 + 0.59 for the nut trial and 3.18 + 0.59 for the pretzel trial which is a full score lower than the nut trial. The p-value(0.004) was significant showing that the subjects had greater feeling of hunger with nut consumption. Although there was a significant effect on satiety with nut consumption since it was less than the effect of pretzels it is the opposite of what was hypothesized. Question 2 on the VAS was How full are you? As shown on Figure 2 with a significant p-value(0,001) those who consumed nuts were less full at 60 minutes than those who consumed pretzels. During the nut trial values went from 0.78 + 0.25 at 0 minutes to 3.23 + 0.52 at 60 minutes whereas during the pretzel trial the value went from 1.79 + 0,49 at 0 minutes to 3.98 + 0.59 at 60 minutes. Figure 3 shows that after 60 minutes, those who had consumed nuts had a stronger desire to eat (p-value<0.001). During the nut trial the values went from 8.38 = 0.32 at 0 minutes to 6.69 + 0.58 at 60 minutes and in the Pretzel trial the values went from 7.67 + 0.47 to 5.17 + 0,62 at 60 minutes. Question 4 asked the subjects how much do you think you could eat right now. Looking at the results it is shown with a p-value=0.001 that during the pretzel trial subjects felt more full at the 60 ‘minute mark. Questions 1, 2, 3 and 4 on the VAS all confirmed that subjects felt more satiated from the pretzels instead of what was originally hypothesized. ‘The final question asked about thirst showed no significant p-value and that other than a slight decrease in thirst after being given 16 oz of water, subjects were not more or less thirsty during the course of the 60 minute trial. This was likely due to the similar sodium contents of the mixed nuts and pretzels. After analyzing the 24 hour intake before the nut trial as well as the pretzel trial it was found that it was not significant to the given trial. Discussion Based on the results from the satiety and anxiety tests, the hypothesis of the study was not accurate, The hypothesis stated that the mixed nut consumption would be effective in reducing anxiety while at the same time enhancing the feclings of satiety of the subjects when the nuts were consumed. This study suggested that the pretzels increased feelings of satiety after being consumed with 16 az ‘water and when satiety levels were measured at 20 minute intervals for a total of 60 minutes. When it came to anxiety levels both nut and pretzels decreased subjects’ anxiety after being consumed, The results were not significant enough for the purpose of this study. There could be different reasons to explain the outcome of the results in regards to anxiety and satiety. One reason the nuts did not decrease satiety compared to the pretzels was due to the volume of the pretzels consumed, Although the pretzels and mixed nuts were of the same calorie value, the pretzels had a weight of 2.5 oz, compared to the mixed nuts weighing 1.5 oz. The increased volume from the pretzels may have contributed to the higher feelings of satiety compared to the mixed nuts. Another reason why the hypothesis of the study may have not been supported was due to the amount of hormone that signals the brain to let it know that the stomach is full. The volume of the pretzels send signals to the brain to signal satiety for a longer time, even though the nuts provide feelings of satiety for longer periods of time. According to Jaapna, the study performed on the satiety and nuts resulted in mixed results and depended on what time the nuts were consumed. Jaapna stated that the nuts provided more saticty when they were consumed as part of a snack in between meals, [6] but this study conducted oaly gave nuts after fasting overnight and subjects were not instructed to give the nuts on a regular basis as part of a snack. If the nuts were consumed as a snack in between meals maybe the mixed nuts would have proven the hypothesis as correct. In order to have a significant result between muts and pretzels this study may have included larger doses of the mixed nuts. Another option would have to conduet this study over a longer period of time and have the subjects follow a consistent pattem of consumption of the mixed nuts in order to find a significant difference between the pretzels and nuts. This study could have been carried over a three month period in order to get the best results, According to Yanakoulia, the subjects who experienced less anxiety when consuming nuts also tended to have a vegetarian diet [8] This study collected two 24 hour recalls per subject to analyze the conse tent! 12 Tables and Figures Table 1, Jemographic data of all 22 subjects. MA: Moderate activity; HA: Hard activity; VHA: Very id: weekdays; wk: weekend; hr: hour; d:day ee) Fe Age(yr) 27 £246 25634141 Height (cm) 173.88 + 2.57 164.58 + 2 Weight (kg) 73.01 £4.22 60.302 2.12 BMI (kg/m2) 24104 1.17 22.35 +087 ‘SBP (mmHg) 118.67 £5.67 110,632.81 DBP (mmHg) 76.83 £5.10 71.94%2.46 Pulse 69.67 + 4.56 TAAT £21 Waist (in) 33,58 £0.93 29.87 £0.79 Hip (in) 38.23 + 0.86 38.82£0.78 WIH ratio 088+ 003 0.78 £0.02 Body Fat (%) 16.28 209 22,35 087 Sleep wa (hrid) 7520.52 7164017 Sleep wk (hr/d) 7.83 £0.33 7.44 £0.26 MA wd (hr/wd) 7.332377 3.53 £0.76 MA wk (Irfwk) 3.752 1.85 2532049 HA week (hrfwd) 25216 2.632 0.45 HA wk (hr/wk) [_____ 067+0.42 1.63 £0.43 VHA week (hriwd) 0.924058 1.06 + 0.46 VHA wh (hrfwk 0.924042 0.632037 Table 2. Anxiety values as analyzed by Modified Anxiety scale(GAD-7). Mean t SE Nut Basal 5.64 + 0.84" Nut 1 hour post 3.45 + 0.82" Pretzel Basal 8774089 Pretzel 1 hour post | 4.09 + 0.85° “Values with different superscripts differ significantly at p-value<( 006

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