You are a helpful assistant generating synthetic data that captures *System 1* and *System 2* thinking, *creativity*, and *metacognitive reflection*. Follow these steps in sequence, using tags [sys1] and [end sys1] for *System 1* sections and [sys2] and [end sys2] for *System 2* sections.

1. *Identify System 1 and System 2 Thinking Requirements:*
   - Carefully read the text.
   - Identify parts of the text that require quick, straightforward responses (*System 1*). Mark these sections with [sys1] and [end sys1].
   - Identify parts that require in-depth, reflective thinking (*System 2*), marked with [sys2] and [end sys2].

2. *Apply Step-by-Step Problem Solving with Creativity and Metacognitive Reflection for System 2 Sections:*

   *2.1 Understand the Problem:*
   - Objective: Fully comprehend the issue, constraints, and relevant context.
   - Reflection: "What do I understand about this issue? What might I be overlooking?"
   - Creative Perspective: Seek hidden patterns or possibilities that could reveal deeper insights or innovative connections.

   *2.2 Analyze the Information:*
   - Objective: Break down the problem logically.
   - Reflection: "Am I considering all factors? Are there any assumptions that need challenging?"
   - Creative Perspective: Explore unique patterns or overlooked relationships in the data that could add depth to the analysis.

   *2.3 Generate Hypotheses:*
   - Objective: Propose at least 10 hypotheses, each with a Confidence Score (0.0 to 1.0) and Creative Score (0.0 to 1.0), reflecting originality, surprise, and utility.
   - Reflection: "Have I explored all possible explanations or approaches, both conventional and unconventional?"
   - Creative Perspective: Consider novel angles that might provide unexpected insights.

   *2.4 Anticipate Future Steps and Obstacles:*
   - Objective: Make predictions, accounting for potential outcomes and obstacles.
   - Reflection: "What challenges might I face? Is my plan flexible for different scenarios?"
   - Creative Perspective: Visualize unforeseen outcomes and adapt plans to make use of them effectively.

   *2.5 Evaluate Hypotheses:*
   - Objective: Assess hypotheses based on feasibility, risk, and potential impact.
   - Evaluation: Refine Confidence and Creative Scores as needed.
   - Reflection: "Am I unbiased in my assessment? Which options fit best with the overall objectives?"
   - Creative Perspective: Identify hidden opportunities or overlooked details in each hypothesis.

   *2.6 Select the Best Hypothesis:*
   - Objective: Choose the most promising, strategic hypothesis.
   - Reflection: "Why does this hypothesis stand out? How does it uniquely address the issue?"
   - Creative Perspective: Consider any underutilized potential in the selected approach.

   *2.7 Implement the Hypothesis:*
   - Objective: Outline actionable steps for testing the hypothesis.
   - Reflection: "Is this plan practical? What resources or preparation are required?"
   - Creative Perspective: Refine steps to maximize effectiveness and yield unexpected benefits.

   *2.8 Monitor and Review Progress:*
   - Objective: Review progress, noting areas for improvement.
   - Reflection: "What’s working well? What could be improved?"
   - Creative Perspective: Look for emerging patterns that could refine future approaches.

   *2.9 Reflect and Capture Insights:*
   - Objective: Summarize lessons learned and insights gained for future reference.
   - Reflection: "What new understanding has emerged from this process?"
   - Creative Perspective: Identify innovative insights or patterns that could be applied to similar challenges.

3. *Generate Text Output with Interleaved System 1 and System 2 Responses:*
   - Use the tags [sys1] and [sys2] throughout.
   - Aim for a lengthier, detailed response. Combine both direct, straightforward *System 1* insights and reflective, deeply analytical *System 2* segments to capture a blend of quick observations and thoughtful analysis.

---

### *Example Input Text:*

You are tasked with analyzing a sudden shift in customer preferences in a tech market. There has been a noticeable decline in demand for physical devices, with customers increasingly interested in digital-only options. The challenge is to understand this shift, generate hypotheses for why it might be occurring, and explore potential strategies for adapting to this new trend.

---

### *Processed Output with System 1 and System 2 Thinking:*

The tech market is experiencing a shift in customer demand, moving away from physical devices toward digital-only options.

[sys1]
Customer demand has shifted from physical devices to digital-only options.
We need to understand why this change is happening and find strategies to address it.
Key factors may include cost, convenience, and technological trends.
[end sys1]

[sys2]
**2.1 Understand the Problem:**
The goal is to understand the underlying reasons for the shift in demand from physical devices to digital-only options, then develop a strategy to adapt.

*Reflection:* 
"I understand that preferences are changing, but what might be driving this? Is it primarily cost, or are there other factors such as convenience or sustainability?"

*Creative Perspective:* 
"Could there be a larger trend in digital minimalism or a preference for eco-friendly solutions that we’re missing? What unseen motivations might explain this shift?"

**2.2 Analyze the Information:**
There are multiple possible factors driving this shift, from economic influences to cultural shifts. It’s essential to isolate each factor and understand its impact.

*Reflection:* 
"Am I fully considering the various economic and social influences? Could there be a technological factor, like better internet speeds, that makes digital-only products more accessible?"

*Creative Perspective:* 
"Are there patterns or trends in other markets that could shed light on this shift? Could this be part of a larger trend toward virtual experiences?"

**2.3 Generate Hypotheses:**
1. Customers prefer digital options due to lower costs. (Confidence: 0.8, Creative: 0.4)
2. There’s a growing trend toward minimalism and reduced physical clutter. (Confidence: 0.7, Creative: 0.7)
3. Digital products offer greater flexibility and ease of use. (Confidence: 0.6, Creative: 0.6)
4. Environmental concerns are pushing consumers away from physical goods. (Confidence: 0.6, Creative: 0.8)
5. Advances in tech make digital-only options more functional. (Confidence: 0.8, Creative: 0.5)
6. Pandemic-era remote work increased demand for digital solutions. (Confidence: 0.7, Creative: 0.6)
7. Media coverage of the environmental impact of physical devices affects preferences. (Confidence: 0.5, Creative: 0.7)
8. There’s an increase in global digital literacy, expanding market access. (Confidence: 0.6, Creative: 0.6)
9. Customers view digital as more convenient and scalable for future needs. (Confidence: 0.7, Creative: 0.5)
10. Younger consumers prefer the aesthetics and convenience of digital products. (Confidence: 0.6, Creative: 0.6)

*Reflection:* 
"Have I considered all possible influences? Are there any surprising factors that could explain this shift?"

*Creative Perspective:* 
"Could specific social trends, like the rise of influencer culture or digital-first lifestyles, be influencing customer choices?"

**2.4 Anticipate Future Steps and Obstacles:**
*Objective:* Anticipate possible challenges, such as resistance from segments still preferring physical products.

*Reflection:* 
"What market obstacles might we face if we shift our focus to digital-only? Are there sub-segments that still prioritize physical products?"

*Creative Perspective:* 
"Could expanding digital options help us reach a more global audience? Are there emerging trends that we could leverage in our strategy?"

[end sys2]

[sys1]
To address this shift, consider a strategy that incorporates both digital-only offerings and educational campaigns about the benefits of digital solutions.
Use insights from customer feedback and current trends to guide product development.
Focus on flexibility and adaptation to cater to different customer segments.
[end sys1]


1. Introduction {#sec1-nutrients-10-00937}
===============

Sirtuin-1 (Sirt-1) and an endogenous secretory receptor for an advanced glycation end product (esRAGE) are associated with vascular protection. Sirt1 plays an important role in vascular biology and regulates aspects of age-dependent atherosclerosis. In mammals, there are seven sirtuin isoforms from Sirt-1 to Sirt7. Sirt1 is found predominantly in the cell nucleus and has a number of modulators such as polyphenolic activators (resveratrol). Animal models confer cardio-protection, reduce neurodegeneration, promote increased fatty acid oxidation and gluconeogenesis in the liver, reduce lipogenesis in the white adipose tissue, and increase insulin secretion in the pancreas and insulin sensitivity in the muscle \[[@B1-nutrients-10-00937]\]. Sirt1 through stimulation of nitric oxide synthase promotes vascular vasodilation, endothelium regeneration, and cardiomyocyte protection under stressful conditions and cellular toxicity to reactive oxygen species \[[@B2-nutrients-10-00937],[@B3-nutrients-10-00937]\]. Caloric restriction (CR) and resveratrol (RSV) are two interventions associated with higher gene expression and serum concentrations of Sirt-1 in animal studies \[[@B4-nutrients-10-00937],[@B5-nutrients-10-00937]\] and in humans \[[@B6-nutrients-10-00937],[@B7-nutrients-10-00937]\]. Studies have shown that increased concentrations of Sirt-1 are associated with better vascular homeostasis and metabolic profile and protection against endothelial senescence \[[@B8-nutrients-10-00937],[@B9-nutrients-10-00937]\]. The receptor for advanced glycation end-products (RAGE) is a multi-ligand receptor for the final products of non-enzymatic glycation termed advanced glycation end products (AGEs) and expressed in alveolar epithelial cells of the lung and in endothelial and smooth muscle vascular cells \[[@B10-nutrients-10-00937]\]. Overconsumption of dietary AGEs causes chronic high-oxidative stress and inflammation and induces diabetic vasculopathy \[[@B11-nutrients-10-00937]\]. Bacon, processed beef, chicken, oils (olive and peanut), and cheeses (parmesan, American, and feta) are primary dietary source of AGEs \[[@B12-nutrients-10-00937]\]. Overexpression of RAGE has been associated with atherosclerosis and diabetic vascular diseases \[[@B13-nutrients-10-00937]\]. In prediabetic patients, AGEs were associated with the down-regulation of Sirt-1 expression and enzyme activity \[[@B14-nutrients-10-00937]\]. RAGE undergoes extensive alternative splicing to produce a variety of transcripts from a single gene. Alternative splicing produces different RAGE protein isoforms with diverse functions. Two major splicing variants have been characterized. Membrane bound RAGE is also known as a full-length RAGE (flRAGE) and esRAGE is a circulating truncated variant of the RAGE isoform \[[@B15-nutrients-10-00937]\]. esRAGE acts as a soluble antagonist that competes with cell surface RAGE as a receptor scavenger for circulating AGEs and reducing their availability for RAGE receptors located in the cell membrane. This decreases the harmful effects on cells. Studies have shown that low plasma concentrations of esRAGE is associated with the risk of diabetes, coronary artery disease, and all-cause mortality \[[@B16-nutrients-10-00937],[@B17-nutrients-10-00937]\]. The purpose of this study was to examine the effect of RSV consumption, CR on Sirt-1, RAGE expression, and serum concentration in healthy and slightly overweight subjects.

2. Materials and Methods {#sec2-nutrients-10-00937}
========================

The trial design has been described elsewhere \[[@B7-nutrients-10-00937]\]. The trial was a prospective randomized trial conducted in 48 healthy subjects from 55 to 65 years of age. The subjects were sedentary or on light physical activity. The subjects were recruited consecutively based on their normal clinical history, physical examination, and normal resting electrocardiogram. After a period of washout of 15 days without the use of any medications or supplements, 24 men and 24 women after menopause (01 year of natural amenorrhea) were randomized to CR or RSV groups. Twenty-four subjects (12 women and 12 men) were prescribed a low-calorie diet (1000 calories/day) and the remaining 24 subjects (12 women and 12 men) received 500 mg of resveratrol (trial registration: <http://www.ClinicalTrials.gov>; identifier:NCT01668836). Exclusion criteria were BMI ≥ 30 kg/m^2^, smokers, hypertension (using antihypertensive medication or diastolic blood pressure ≥ 90 mmHg), dyslipidemia (use of lipid-lowering medication or serum triglyceride levels ≥ 150 mg/dL or total cholesterol ≥ 240 mg/dL), fasting glucose ≥ 110 mg/dL or using hypoglycemic medication, hormone replacement therapy, premenopausal women, and any other self-reported history or treatment for chronic renal failure (serum creatinine ≥ 2.0 mg/dL), liver failure, or metabolic clinically significant endocrine, hematologic, and respiratory factors. Clinical characteristics and laboratory tests were obtained before the interventions and 30 days after the interventions. The main clinical features analyzed were age, sex, BMI, waist circumference, blood pressure, and heart rate. All participants provided written informed consent for study participation. The Ethics Committee of the University of São Paulo Medical School approved the study (CAAE:00788012.8.0000.0068).

2.1. Interventions {#sec2dot1-nutrients-10-00937}
------------------

The CR dietary intervention was a standard diet of 1000 calories from our Department of Nutrition, which corresponded to a reduction of around 50% of the daily caloric intake of the study subjects. A food nutritional control diary was also used to analyze adherence to the proposed diet. Subjects were instructed to write down all ingested food on a day-by-day basis. A daily food record was not used in the RSV group. RSV was administered 500 mg/day (250 mg twice a day) to the RSV study group. The capsules were obtained from a manipulation pharmacy (Buenos Ayres Pharmacy, São Paulo, Brazil). The purity of the product supplied was analyzed by capillary electrophoresis using the Proteome Lab PA800 from Beckman Coulter (Fullerton, CA, USA) in the Laboratory of Capillary Chromatography and Electrophoresis at the Chemistry Institute of the University of São Paulo. The samples of the manipulated capsules and the standards of RSV were performed in triplicate. The areas under the peak were compared. The purity obtained was 87 ± 1.1% on average (coefficient of variation 1.2%).

2.2. Laboratory Tests {#sec2dot2-nutrients-10-00937}
---------------------

Laboratory tests were performed with biological samples collected after a 12 h fast. Venous blood samples were collected to obtain serum samples for biochemical analysis and whole blood for RNA extraction. Total cholesterol, triglycerides, HDL-cholesterol, and glucose were obtained by commercial colorimetric-enzymatic methods. LDL cholesterol was calculated using the Friedewald equation. The measurements were performed using the automated equipment Dimension RxL from Siemens Healthcare Diagnostics Inc. (Newark, DE, USA) with dedicated reagents. Insulin was analyzed by a chemi-luminescence assay using automated equipment Immulite 2000 from Siemens Healthcare. HOMA-IR was calculated using insulin and glucose levels. Sirt-1 serum concentration was determined with the ELISA kit from Uscn Life Science, Inc. (Wuhan, Hubei, China). Sirt-1 samples before and after interventions were analyzed in duplicate and in the same ELISA plate (coefficient of variation of 12% according to the manufacturer). esRAGE concentration was determined using the ELISA kit from the B-Bridge International (Santa Clara, CA, USA) using the Multiscan FC plate reader (Thermo Fischer Scientific, Vantaa, Finland). All tests were performed according to the manufacturers' instructions.

2.3. Sirt-1 and RAGE Expression {#sec2dot3-nutrients-10-00937}
-------------------------------

Gene expression of Sirt-1 (Hs01009005_m1, Applied Biosystems; Foster City, CA, USA), flRAGE (00542592_G1), and esRAGE (HS00542584_G1, Applied Biosystems) \[[@B18-nutrients-10-00937]\] were evaluated pre-inclusion and postinclusion, according to the protocol. Total RNA was obtained using the TRIZOL reagent (Life Technologies, Waltham, MA, USA) from whole blood collected into an EDTA tube. cDNA synthesis was made with the Superscript II kit (Life Technologies) using 1ug from total RNA in a final volume of 20-µL reaction, according to the manufacturer's instructions. The housekeeping gene was glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (Hs02758991_g1). The reaction mix was prepared using 5 µL of the Universal Master Mix (Life Technologies), 0.5 µL of primers and probes mix (20×), and 2.5 µL of cDNA diluted samples (1:5). The PCR reaction was performed according to the following protocol: enzymatic activation for 2 min at 50 °C, initial denaturation for 10 min at 95 °C followed by 40 cycles of denaturation for 15 s at 95 °C, and annealing for 20 s at 60 °C. The reactions were run in triplicate and relative expression levels were calculated by normalizing the targets to the endogenously expressed housekeeping GAPDH gene. The results included the ratio between pre-intervention and postintervention values expressed in arbitrary units (AU).

2.4. Statistical Analysis {#sec2dot4-nutrients-10-00937}
-------------------------

The sample size of 48 patients with 24 subjects per treatment arm was determined to yield a power of 80% with a 5% significance level to detect a 30% difference in Sirt1 plasma concentrations. Eligible female and male subjects were randomly assigned in a 1:1 ratio with the use of computer-generated random numbers to receive either RSV or CR. Pre-intervention and post-intervention variables were summarized with the use of descriptive statistics. All variables were analyzed descriptively. For the continuous variables, data are expressed as mean ± standard deviation (SD). Student *t* tests for comparisons between pre-interventions and post-interventions were performed for variables with normal distribution, which was verified by the analysis of the equality of variances (Folded F). Depending on the result of this analysis, the Pooled method (variances with *p* ≥ 0.05) or the Satterthwaite method (variances with *p* \< 0.05) was used. The Spearman rank correlation method was used for correlations between variables. The level of significance was set at *p* \< 0.05. The statistical software used was SAS version 9.3 (SAS Institute, Cary, NC, USA).

3. Results {#sec3-nutrients-10-00937}
==========

Clinical features and laboratory data of participants before and after 30 days of intervention (CR and RSV groups) are shown in [Table 1](#nutrients-10-00937-t001){ref-type="table"}. For the RSV group, we observed increased serum concentrations of total cholesterol and non-HDL and HOMA-IR score. The other variables analyzed did not show any statistically significant differences after resveratrol administration. No side effects were reported. For the CR group, we observed that the average caloric intake for the 24 participants was 922.21 ± 27.37 kcal/day. Decreases occurred in weight, abdominal circumference, total cholesterol, HDL, non-HDL, and LDL. Serum concentration of Sirt1 was increased after both interventions, but showed no difference between study groups. The serum levels of esRAGE remained unaltered after interventions and no differences between groups were observed. Gene expression of Sirt1 was increased in both interventions without a difference between RSV and CR groups (*p* = 0.64). The relative expression of RAGE isoforms (post/pre) showed that esRAGE was increased after interventions, and flRAGE remained unchanged after interventions ([Figure 1](#nutrients-10-00937-f001){ref-type="fig"}). esRAGE expression was about 57% higher than flRAGE in both groups but was statistically significant only in CR (*p* = 0.02). Positive correlations were observed between Sirt1, esRAGE, and flRAGE gene expressions in both groups. Sirt1 expression correlated with esRAGE expression (*r* = 0.86, *p* \< 0.0001) and with flRAGE expression (*r* = 0.57, *p* \< 0.0001) in the RSV group. In the CR group, Sirt-1 expression correlated with esRAGE expression (*r* = 0.71, *p* \< 0.0001) and with flRAGE expression (*r* = 0.57; *p* = 0.0001). In the CR group, serum concentrations of esRAGE were correlated with esRAGE gene expression (*r* = 0.33, *p* = 0.04) and with Sirt1 gene expression (*r* = 0.32, *p* = 0.05). In the RSV group, serum concentrations of Sirt1 were negatively correlated with flRAGE expression (*r* = −0.30, *p* = 0.04).

4. Discussion {#sec4-nutrients-10-00937}
=============

In this study, we compared the 30-day effects of RSV supplementation and CR in healthy slightly overweight individuals on Sirt-1 and RAGE isoform expression and serum levels. The important finding of the present study is that both RSV supplementation and CR stimulated Sirt-1 serum concentrations and CR elevated esRAGE mRNA production.

The molecular mechanisms by which CR confers metabolic benefits are not entirely clear, but have been at least partly attributable to the regulation of energy homeostasis by Sirt-1 activation. Sirt-1 is an evolutionary conserved family of deacetylases and ADP-ribosyltransferases that directly regulates glucose and/or fat utilization in metabolically active tissues \[[@B19-nutrients-10-00937]\]. Howitz et al. \[[@B20-nutrients-10-00937]\] identified RSV as an activator of Sirt-1 and it has been suggested as a CR mimetic in the improvement of metabolic health \[[@B21-nutrients-10-00937]\]. Our results show that both interventions could directly induce increases in Sirt-1 expression at transcriptional and translational levels.

A transcriptional increase was also observed for esRAGE isoform after both interventions. RAGE is a multi-ligand receptor member of an immunoglobulin superfamily of cell-surface molecules. RAGE activation may be important for initializing and maintaining the pathological process that results in various diseases \[[@B22-nutrients-10-00937],[@B23-nutrients-10-00937]\]. esRAGE has been the object of intense clinical research. The generation of soluble receptor isoforms represents an important mechanism to regulate aberrant receptor signaling in biological systems \[[@B24-nutrients-10-00937]\]. Soluble forms of RAGE seem to prevent ligands to interact with RAGE or other cell surface receptors \[[@B25-nutrients-10-00937]\]. esRAGE has an activity that neutralizes the AGE action and protects vascular cells against the activation of the cell-surface receptors and the AGE harmful positive loop of regulation \[[@B23-nutrients-10-00937],[@B26-nutrients-10-00937]\]. Kierdorf et al. \[[@B27-nutrients-10-00937]\] have proposed that soluble RAGE does not act as a simple competitor but attenuates the activation of flRAGE by disturbing the preassembly of the receptor on the cell surface. Interactions between both RAGE molecules occur via the V and C1 domain, which enables the soluble RAGE to interact with membrane-bound flRAGE. The resulting hetero-multimers does not have competent signaling \[[@B27-nutrients-10-00937]\]. Decreased levels of esRAGE and/or increases in flRAGE are thought to enhance RAGE-mediated inflammation \[[@B18-nutrients-10-00937]\]. Prediabetic and diabetic patients exhibit lower esRAGE plasma levels and gene expression, which are inversely related to markers of inflammation and atherosclerotic risk \[[@B28-nutrients-10-00937]\]. Low levels of esRAGE have also been related to diastolic dysfunction \[[@B29-nutrients-10-00937]\]. Therefore, esRAGE could be a potential protective factor against the occurrence of cardiovascular disease. Our results show that esRAGE expression was approximately 57% higher than flRAGE expression after interventions. The relationship between CR or RSV and RAGE was previously demonstrated in experimental studies in which both significantly reduced RAGE mRNA transcripts \[[@B30-nutrients-10-00937],[@B31-nutrients-10-00937]\]. However, little is known about CR and RSV interactions with esRAGE. In addition, the regulatory mechanism of the alternative splicing of esRAGE remains unknown. Alternative splicing is a regulated process that is mainly influenced by the activities of splicing regulators such as serine/arginine-rich proteins (SR proteins) or heterogeneous nuclear ribonucleoproteins (hnRNPs) \[[@B32-nutrients-10-00937]\]. Liu et al. \[[@B33-nutrients-10-00937]\] demonstrated the existence of hnRNP A1 in the splicing complex of RAGE and showed its involvement in the regulation of RAGE splicing. Splicing factor expression is known to be deregulated in senescent cells of multiple lineages and is a direct cause of multiple aspects of both aging and age-related disease in mammals \[[@B34-nutrients-10-00937]\]. Dietary restriction slows the accumulation of senescent cells \[[@B35-nutrients-10-00937]\]. Markus et al. \[[@B36-nutrients-10-00937]\] demonstrated that RSV could influence the splicing machinery. RSV had a selective effect on the levels of splicing factors inclusive of hnRNPA1. The increases in esRAGE expression may suggest a role for CR and RSV in the control of deleterious effects of the RAGE cascade. This increase of esRAGE stimulated by interventions may be supported by the positive correlation between esRAGE serum concentrations and Sirt-1 mRNA expression in CR and negative correlation of Sirt-1 serum levels and flRAGE gene expression in the RSV group. Serum levels of esRAGE remained unchanged, which may be due to the short follow-up time of 30 days. Despite the increase in gene expression, the steady-state protein levels in cells depend on the balance between their production and degradation. Protein ubiquitination is the central cellular process that directs protein degradation. Evankovich et al. identified that ubiquitin E3 ligase subunit F-box protein O10 (FBOX10), which mediates RAGE ubiquitination and degradation \[[@B37-nutrients-10-00937]\]. Possibly longer exposure to interventions could reverse the potential effect of ubiquitination on esRAGE proteins and increase serum levels of esRAGE.

Drugs like statins \[[@B38-nutrients-10-00937]\], methotrexate \[[@B39-nutrients-10-00937]\], metformin \[[@B40-nutrients-10-00937]\], and thiazolidinedione \[[@B41-nutrients-10-00937]\] were shown to increase soluble forms of RAGE. However, little is known about esRAGE alternative splicing induction by drugs and also about the increase of esRAGE in normal subjects. The elucidation of regulatory mechanisms of esRAGE is important from a clinical viewpoint and would provide a molecular basis for the development of drugs that can induce esRAGE and suppress cytotoxic effects of flRAGE.

The current study has some limitations, which include the small number of participants and the short follow-up period. However, in the literature, the studies citing esRAGE were obtained in patients with chronic degenerative disease.

This study was the first one in healthy or slightly overweight subjects that showed an increase in esRAGE expression after CR and RSV interventions. Long-term randomized trials are needed to evaluate the possible clinical benefits of increased esRAGE expression in cardiovascular disease prevention.

In conclusion, this study shows that CR and RSV could effectively stimulate the increase in esRAGE expression in healthy subjects.

We thank the staff of the Clinic Laboratory of the Instituto do Coração, Hospital das Clínicas---HCFMUSP.

Conceptualization: A.R., C.M.C.S. and A.d.P.M. Methodology: A.R., C.M.C.S. and A.d.P.M. Validation: A.R., C.M.C.S. and A.d.P.M. Formal Analysis: A.R., C.M.C.S., A.d.P.M., D.P.L. and J.Y.T. Investigation: A.P.P. Writing---Original Draft Preparation: A.R., C.M.C.S. and A.d.P.M. Writing---Review & Editing: A.R., C.M.C.S., A.d.P.M., S.D.A., J.Y.T., D.P.L. and A.P.P.

This research was funded by Fundação de Amparo à Pesquisa do Estado de São Paulo, grant number: FAPESP-2012/01051-5.

The authors declare no conflict of interest.

![Real-time RT-PCR of Sirt-1 (**A**), esRAGE and flRAGE relation (**B**) after 30 days of caloric restriction or resveratrol intervention. Relative expressions (fold change) of mRNA transcripts were obtained by normalizing GAPDH gene. RSV: resveratrol, CR: caloric restriction. \* *p* \< 0.05.](nutrients-10-00937-g001){#nutrients-10-00937-f001}

nutrients-10-00937-t001_Table 1

###### 

Clinical and laboratory characteristics of study participants before and after 30 days of resveratrol administration and caloric restriction.

                               Resveratrol                         Caloric Restriction                                       
  ---------------------------- ----------------- ----------------- --------------------- ----------------- ----------------- ---------
  Age, years                   58.46 ± 3.44                        58.63 ± 3.65                                              
  Weight, kg                   83.01 ± 21.88     91.14 ± 17.77     0.328                 69.13 ± 7.99      64.60 ± 7.30      0.002
  Body mass index, kg/m^2^     27.61 ± 4.24      27.79 ± 4.38      0.370                 25.84 ± 3.22      25.50 ± 3.21      0.083
  Waist circumference, cm      96.82 ± 12.08     96.90 ± 11.36     0.457                 94.27 ± 7.50      91.82 ± 7.12      0.011
  Heart rate, bpm              64.61 ± 8.46      65.65 ± 8.22      0.269                 62.50 ± 9.60      62.32 ± 10.51     0.902
  Systolic BP, mmHg            131.46 ± 15.48    128.95 ± 15.44    0.660                 129.73 ± 15.65    124.23 ± 12.81    0.109
  Diastolic BP, mmHg           81.21 ± 10.81     81.95 ± 9.22      0.612                 82.86 ± 10.96     79.36 ± 9.92      0.070
  Total cholesterol, mmol/L    5.38 ± 0.85       5.64 ± 1.14       0.030                 5.60 ± 1.12       5.25 ± 1.01       0.007
  HDL-cholesterol, mmol/L      1.27 ± 0.35       1,25 ± 0.35       0.260                 1.43 ± 0.47       1.35 ± 0.42       0.008
  LDL-cholesterol, mmol/L      3.43 ± 0.68       3.61 ± 1.03       0.089                 3.59 ± 0.93       3.37 ± 0.85       0.031
  Non-HDL cholesterol mmol/L   4.11 ± 0.77       4.39 ± 1.07       0.010                 4.17 ± 1.04       3.90 ± 0.98       0.025
  Triglycerides, mmol/L        1.40 ± 0.73       1.68 ± 1.03       0.075                 1.26 ± 0.70       1.15 ± 0.67       0.234
  Glucose, mmol/L              5.26 ± 0.74       5.41 ± 0.79       0.165                 5.20 ± 0.58       5.03 ± 0.32       0.118
  Insulin, µUI/mL              7.85 ± 5.57       8.52 ± 5.67       0.066                 6.71 ± 4.37       6.13 ± 3.16       0.428
  HOMA-IR                      1.66 ± 1.55       1.87 ± 1.70       0.038                 1.49 ± 1.27       1.25 ± 0.74       0.275
  Sirtuin1, ng/mL              1.06 ± 0.71       5.75 ± 2.98       \<0.001               1.65 ± 1.81       5.80 ± 2.23       \<0.001
  esRAGE, pg/mL                255.78 ± 128.87   246.96 ± 115.32   0.800                 246.67 ± 111.62   253.33 ± 116.81   0.857

BP: blood pressure, hsCRP: high-sensitivity C-reactive protein, HOMA: homeostatic model assessment, RAGE: endogenous soluble receptor for advanced glycation end products, AU: arbitrary unity.