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In agreement with the present study, Payahoo et al. [16] showed that daily administration of 30 mg zinc gluconate for 1 month reduced body weight, BMI and waist circumferences in the healthy obese adults.

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Study design and participants

This double-blind randomized clinical trial was conducted from December 2015 to April 2016. In order to detect a difference of 4.5 kg/m2 in the BMI and with respect to a pooled standard deviation of 26.21 kg/m2, obtaining from the study by Payahoo et al. [16], the sample size was calculated 20 subjects for each group. In this two-arm parallel study with two-tailed testing, a power (1–β) of 80% and α = 0.05 was used. Fifty healthy adults (men and women) with obesity and BMI more than 30 kg/m2 in the age range of 18–45 years were selected using convenience sampling from the Specialized Clinic of Nutrition & Diet Therapy located at the Faculty of Nutrition Sciences and Food Technology of Shahid Beheshti University of Medical Sciences in Tehran, Iran. In our study, exclusion criteria were the presence of pregnancy or lactation, chronic kidney or hepatic disease, autoimmune and infectious disease, chronic inflammatory diseases, recent surgery, smoking, having weight loss diets in the last 2 months, the use of Zn, calcium, or iron supplements in the last 2 months, and taking anticoagulant drugs, lipid-lowering or beta-blocker drugs. The primary outcomes were anthropometric measurements, and secondary outcome were appetite score, serum levels of inflammatory markers, apelin, NPY, glucose, Zn and insulin, and IR. The study protocol was approved by the Ethics Committee of the National Nutrition and Food Technology Research Institute of Iran (IR.SBMU.nntri.Rec.1394.407). The study was in adherence with the Declaration of Helsinki. Written informed consent was obtained from all subjects before initiating the study. This clinical trial was registered at clinicaltrials.gov at the U.S. National Library of Medicine (NCT02516475).

Randomization

The subjects were randomly allocated to either a Zn or placebo group by block randomization. A trained dietitian completed the block randomization with a block size of 4 and possible balanced combinations with 2 P (placebo) and 2 Z (Zn supplement) subjects, calculated as 6 blocks (ZZPP, PZPZ, PZZP, ZPZP, PPZZ, ZPPZ). Then, blocks were randomly chosen, using a simple random sampling method to determine the assignment of all the participants into the groups.

Intervention

During this study, subjects in the Zn group received 30 mg zinc sulfate as 1 capsule (between meals) while those in the placebo group received corresponding placebo capsules containing starch (also between meals). All capsules were produced by Dineh Iran Company, Tehran, Iran. According to the literature, zinc supplement is safe at a dose of 30 mg/day [31, 32]. Blinding was performed by a trained dietician, and the patients and researchers were kept blinded to the allocation. In addition, subjects in both Zn and placebo groups received a restricted calorie diet (RCD) with ~ 300 kcal lower than the estimated energy requirement based on the Mifflin-St Jeor equation in order to reduce their weight about 1 kg per month, and this RCD contained ~ 55% carbohydrate, ~ 15% protein and ~ 30% fat [33]. Adherence to the diet was monthly assessed by a registered dietitian. Participants were followed twice a month via telephone calls in order to ensure their compliance and were asked to maintain their usual physical activity level. They were also asked to return the remaining capsules, and based on the number of returned capsules by each subject and adherence to the diet, their degree of compliance was determined and the data of individuals with the degree of compliance more than 90% were analyzed at the end of the study.

Dietary intakes and appetite assessments

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Dietary intakes of participations were assessed using a 3-day dietary recall (2 weekdays and 1 weekend day) at baseline and at the end of week 15. Individuals’ diets were analyzed by Nutritionist IV software (N Squared Computing, San Bruno, CA, USA). Basal metabolic rate (BMR) was calculated based on Mifflin and St Jeor et al. [34]. Underreporting was defined as a ratio reported energy intake by 3-day dietary recall/BMR < 1.1 [35]. Simplified nutritional appetite questionnaire (SNAQ), a valid 4-item questionnaire recommended for clinical purposes [36], were used to assess the appetite at baseline and week 15. The SNAQ items were as follows: #1, Appetite; #2, Feeling full; #3, Food tastes; #4, Feeling hunger, and the sum of the 4 items scores constitutes the total SNAQ score which ranges from 4 to 20. The total score of 4 to 14 and 15 to 20 indicates low and normal appetite, respectively [36].

Anthropometric assessments

Weight was measured with minimum clothes and without shoes using a calibrated scale (Seca, CA, USA) and precision of 100 g. Height was measured using a wall-mounted stadiometer with the precision of 0.5 cm. Hip and waist circumference were also measured using an inflexible tapeline with the precision of 0.5 cm, in the narrowest circumference below the rib cage and above the umbilicus and the largest circumference between the waist and knees, respectively [37]. BMI was calculated as the ratio of weight (kg)/height2 (m2). Anthropometric parameters were measured at baseline and at the end of weeks 7 and 15.

Physical activity assessment

Physical activity level was estimated using a valid and reliable physical activity questionnaire [38] and calculating metabolic equivalent (MET) at baseline and the end of the study.

Blood samples and biochemical assessments

A sample of 5 ml blood was collected from all participants after a 12 to 14 h fast, at baseline and at the end of week 15. These samples were centrifuged at 4000 rpm for 15 min. The samples of serum were separated into small aliquots and were frozen at − 80 °C. For Zn analysis, all tubes were washed by acid and rinsed with distilled water, then atomic absorption spectrometry (variant Chemthech Analytical 2000) was used to determine serum Zn concentration [39, 40]. Serum concentration of high-sensitivity C-reactive protein (hs-CRP) was determined by enzyme-linked immunosorbent assay (ELISA) kits (Diagnostics Biochem Canada, Ontario, Canada) with an intra-assay coefficient of variation (CV) of 7.2%. Serum TNF-α was measured by ELISA kits (Diaclone, Besancon, France). Intra-assay CV for serum TNF-αwas 6.5%. Serum apelin concentration was assessed by ELISA kits (ZellBio GmbH, Ulm, Germany), with an intra-assay CV of 7.2%. Serum insulin was determined by ELISA kits (Monobind, USA), with an intra-assay CV of 7.4%. Serum glucosewas measured by commercial kits (Pars Azemoon, Tehran, Iran) with the aid of a Selectra 2 Autoanalyzer (Vital Scientific, Spankeren, The Netherlands). Homeostatic Model Assessment of Insulin Resistance (HOMA-IR) was determined using the following equation: $${ ext{HOMA-IR}}, = ,left[ {{ ext{Fasting serum glucose }}left( {{ ext{mg}}/{ ext{dL}}} ight), imes ,{ ext{Insulin }}left( {upmu{ ext{U}}/{ ext{L}}} ight)} ight]/405$$

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Statistical analysis

Intention-to-treat principle was applied for anthropometric and dietary intake variables. Per-protocol analysis (PPA) was performed for analyzing the biochemical data. Data analysis was performed using SPSS version 20. The results are presented as mean (± SD) and frequency (percent) for quantitative and qualitative variables, respectively. The Kolmogorov–Smirnov test was used to assess normal distribution of data. None normal data distribution has been presented as 25/75 IQR. Natural log transformations on plasma Zn, insulin, TNF-α, NPY, apelin and HOMA-IR were transformed through Box-Cox transformation. To compare qualitative variables between the two groups, the Chi square test was used. We used a t test and paired t-test to compare quantitative parameters between and within groups, respectively. In addition, because anthropometric parameters were measured 3 times during the study, analysis of variance for repeated measurements was used to compare data between various times. Analysis of covariance was performed in order to remove the effect of confounding factors. In this study, P values of less than 0.05 were considered statistically significant.

dmsjournal.biomedcentral.com - Zinc supplementation improves body weight management, inflammatory ...

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