Reduces Free Fatty Acid by Increasing Plasma Leptin and Adiponectin Levels

Lunasin-Enriched Soy Extract (LunaRich XTM), in Combination with the Dietary Supplement Reliv Now, Reduces Free Fatty Acid by Increasing Plasma Leptin and Adiponectin Levels in LDL-Receptor Mutant Pigs

Alfredo F. Galvez1*, Hosea Matel1, Jan Ivey2 and Doug Bowles2

1Missouri Plant Science Center and 2University of Missouri, Columbia

ABSTRACT

Elevated levels of free fatty acids (FFA) are associated with obesity and insulin resistance and can lead to diabetes, metabolic syndrome and cardiovascular disease. Daily supplementation using 500 mg of lunasin-enriched soy extract (LES) and one serving (18.3 gm) of the dietary supplement Reliv Now® has been found to significantly reduce FFA levels by 74% and 65% after 4 and 6 weeks, respectively, on obese LDL-receptor mutant (Rapacz) pigs. The decrease in FFA levels can be explained by the coordinate increase in plasma levels of leptin by 52% and 64% and adiponectin by 20% and 60% after 4 and 6 weeks, respectively. The decrease in FFA and the increase in leptin and adiponectin correlates with slower weight gain in the obese pigs. Removal of LES and Reliv Now supplements from the basic diet of the pigs increases FFA and decreases leptin and adiponectin to the baseline, unsupplemented levels. These results suggest that supplementation with LES and Reliv Now has the potential to reduce FFA and increase leptin and adiponectin levels, which are associated with preventing obesity and insulin resistance and the related disease modalities arising from these health conditions. However, human clinical trials are needed to validate these preliminary results.

INTRODUCTION

According to the US Centers for Disease Control (CDC), there is a rising trend in obesity levels today with a parallel increase in type 2 diabetes. Elevated levels of free fatty acids (FFA) have been associated with insulin resistance in obese patients [1] and can increase inflammation [2]. Insulin resistance and pro-inflammatory response are clinically important because they can lead to several diseases like type 2 diabetes, hypertension and cardiovascular disease [3]. Although there is a close association of obesity, insulin resistance and type 2 diabetes to elevated FFA levels, there is a lack of effective treatments to lower free fatty acids [1].

Lunasin is a bioactive component in soy with a novel chromatin-binding property and epigenetic effects on gene expression [4, 5]. The soy peptide is heat stable, water soluble and found in significant amounts in select soy protein preparations [6]. It can get inside mammalian epithelial cells through its RGD cell adhesion motif, bind preferentially to deacetylated histones and inhibit histone H3 and H4 acetylation [4]. There is growing evidence that responses to dietary and environmental effects involve epigenetic changes in gene expression, which are modulated by the reversible processes of DNA methylation-demethylation and histone acetylation-deacetylation [7, 8].

Lunasin has been shown to have an anti-mitotic effect when expressed inside mammalian cells [5] and prevents cancer formation when applied exogenously in cell culture and in mice skin cancer model [4]. The lunasin peptide has also been shown to have anti-inflammatory [9] and anti-oxidant effects [10], to improve innate immunity (unpublished results), and to prevent metastasis of colon cancer cells to the liver [11]. The multiple health effects of the lunasin soy peptide have been attributed to its ability to bind to chromatin and affect gene expression associated with cellular health [12]. The elucidation of its mechanism of action makes lunasin an important molecule for research studies to understand the emerging role of diet on epigenetic mechanisms that can impact the development of chronic diseases, such as obesity, diabetes and metabolic syndrome.

We have previously shown the effect of lunasin-enriched soy extract in lowering LDL cholesterol [13] and we now report a novel effect of the lunasin-enriched soy extract in combination with a soy- based dietary supplement, Reliv Now, on a supplement feeding study using a LDL-receptor mutant pig model [14]. Reliv Now contains optimal blend of vitamins, minerals and herbal agents and is primarily used to lower risk factors for cardiovascular disease. Our study shows that supplementation with lunasin in combination with Reliv Now leads to a significant decrease in free fatty acid levels and the coordinated increase of leptin and adiponectin plasma levels in LDL-receptor mutant pigs

MATERIALS AND METHODS

We chose the Rapacz pig model [15] to test the biological effect of the lunasin-enriched soy extract (LES) extract on obesity, cardiovascular disease risk and metabolic syndrome because their weight and liver function are closer to humans than any other animal models. Also, the pigs have mutations to their LDL receptor gene that predispose them to high cholesterol, obesity and increased risk for heart disease [15]. The 5 Rapacz pigs used for this study were approximately 1.5 years old and considered obese, weighing more than 20% from normal. The experiment was done with animal use permit at the Veterinary hospital at the University of Missouri, Columbia, MO.

In order to administer Reliv Now, the 18.3 gms powder was measured from a 29.6 cc scooper, mixed with 10 cc of water and formed into a dough ball that the pigs ate happily. The 500 mg dose of LES was selected based on previous study showing that 250 mg dose of LES was enough to reduce LDL cholesterol after 4 weeks in 1 -year old Rapacz pigs. We increased the dose to 500 mg to account for the increased weight of the pigs at the start of the study when they were 1.5 years of age. The 500 mg LES was put into two capsules of 250 mg each and was fed to the pigs by inserting them into a snack bar that the pigs like to eat.

To determine the bioavailability of lunasin after ingestion of 500 mg LES, we supplemented the diet of one pig with 500 mg LES and then blood draws were taken at 10 min, 30 min, 60 min and 24 hr after ingestion. The blood plasma was separated and the amount of lunasin was detected and quantitated by ELISA using a polyclonal lunasin antibody (Pacific BioLabs) and standard ELISA protocol. 

To test the effects of the lunasin-enriched soy extract (LES) in combination with Reliv Now, we fed five Rapacz pigs daily with at least one kg of standard casein diet (BV233) and supplemented with 18.3 gm of Reliv Now and 500 mg of LES . The pigs were maintained on a soy-free diet (BV233 pig chow) throughout the treatment. Pigs were fed their regular diet (BV233) once per day in the morning at approximately 9:00 AM. Later in the afternoon, at approximately 4:00 PM, the Reliv Now and LES treatments were administered. Weight and blood draws were taken at pre-treatment (0), at 4 weeks, 6 weeks and 8 weeks after treatment had begun. At the end of 8 weeks, there was an additional 4 weeks washout period without treatment and weight and blood draws were also taken at the end of washout period (12 weeks). Blood draws were taken after an overnight fast (approximately 15h). Blood samples were tested for lipid panel including plasma levels of free fatty acids (FFA) by the Analytical Laboratory at the Veterinary School of the University of Missouri. Blood samples were collected into monoject tubes with 15% EDTA, centrifuged for 20 min at 3300 rpm to separate the blood plasma, which were transferred to 1 ml cryogenic vials and stored at -70oC to test for leptin and adiponectin levels. Plasma levels of leptin and adiponectin were determined using porcine leptin and adiponectin ELISA kits obtained from USCN Life Sciences, Inc. (Houston, TX USA). Standard protocols from the manufacturer were followed, including the use of 3 replicate measurements for each data point, to detect and quantify the amounts of leptin and adiponectin in the plasma.

RESULTS

Before conducting the supplementation study, we first determined whether lunasin can be detected in the blood plasma of a Rapacz pig at different time points after ingesting a 500 mg dose of lunasin-enriched soy extract (LES). Figure 1 shows that lunasin can be detected from the blood plasma as early as 10 min after ingestion and increased significantly after 30 and 60 min before peaking at 24h after ingestion. The total amount of lunasin detected in blood plasma after 24h was approximately 0.11 mg (Fig.1), which indicates that lunasin becomes bioavailable after ingestion of 500 mg LES. 

 

Reliv Holds Worldwide Rights to Bioactive Lunasin!

By now you’ve probably heard the news: Reliv now holds the exclusive rights to the intellectual property behind lunasin!  

What are the basics of the agreement?

Reliv International has entered into an exclusive license for the intellectual property of Soy Labs LLC related to the nutritional ingredient lunasin. The license covers an issued patent and several patent applications related to lunasin and soy-related peptides, proprietary information and manufacturing processes. Further, the arrangement allows certain Soy Labs employees to join the Reliv corporate staff.

So is Reliv the only place you can get lunasin?

Yes and no. Lunasin is a naturally occurring peptide found in seed-bearing plants. Since soy is such an excellent source of protein, it is also a superior source of lunasin. Lunasin is part of nature, so holding the exclusive rights to all lunasin would be like holding the rights to oxygen.

But not all lunasin is created equal – not even close. The concentration and bioavailability of lunasin among soy foods (soy milk, tofu, etc.) and other sources varies dramatically. And no other source can match the concentration and bioavailability of the lunasin found in Reliv products.

What Reliv does have exclusive rights to is the lunasin produced through a proprietary extraction process developed by Soy Labs. No one else has been able to effectively extract bioactive lunasin of such quality and in such concentration. Reliv is now way ahead of the curve in harnessing the power of this clinically proven superfood.

What lunasin patent is now transferred to Reliv?

Lunasin’s mechanism of action for reducing LDL cholesterol, first identified by Dr. Alfredo Galvez, has received a U.S. patent. That patent is now the intellectual property of Reliv. This means that no other company can promote soy’s lunasin-based cholesterol-lowering properties like Reliv can.

The agreement also delivered to Reliv multiple patents pending in the U.S. and abroad.

What else does Reliv get out of the deal?

Reliv gets exclusive rights and direct control over the entire LunaRich X production process and technologies. Reliv also now owns a proprietary bioassay that effectively determines the amount of bioactive lunasin available after digestion – the only test of its kind. It is this test that gives Reliv the unique ability to guarantee the bioavailability of lunasin in our LunaRich products.

Looking forward, the agreement opens up opportunities for further research and development with lunasin and other plant-based ingredients.

Which Soy Labs employees join Reliv?

All members of Soy Labs’ lunasin innovation team have broken away from Soy Labs to form a new business entity: SL Technologies. SL Technologies is now a subsidiary under the Reliv corporate umbrella. SL Technologies staff includes:

  • Lead Scientific Advisor Dr. Alfredo Galvez
  • President Ryan Schmidt
  • Chief Operating Officer Andrew Birney
  • Executive Assistant Tina Van Horn
  • Administrative Assistant Johnda Darby
  • Special Projects Coordinator Tamara Shenouda

‘Beyond Excited’

We’ve always endeavored to remain on the cutting edge of nutrition science in formulating our products at Reliv. And with the acquisition of the intellectual property behind lunasin, we are now leading the industry in the budding field of nutritional epigenetics. Lunasin is one of the first nutritional compounds identified to affect gene expression and promote optimal health at the epigenetic level.  

New research shows that through nutrition people can influence which genes are expressed. Reliv is committed to building on this emerging science and developing nutritional solutions that help people take control of their health.

Epigenetics has hit the mainstream, and we believe nutritional epigenetics will soon follow. We’re beyond excited about what Reliv’s acquisition of lunasin-related technologies will mean for our Distributors and consumers around the globe.

To your health,

Carl W. Hastings

Vice Chairman and Chief Scientific Officer

Reliv's Dr. Carl Hastings is rated #9 out of 5000

Out of more than 5000 Chief Scientific Officers in the United States, Reliv's Dr. Carl Hastings is rated #9.  

Reliv’s Dr. Carl Hastings Named Top 10 Chief Scientific Officer
ExecRank lists Hastings ninth among more than 5,000 CSOs in the United States

CHESTERFIELD, MO, April 3, 2013 – Reliv International (NASDAQ: RELV) today announced that ExecRank executive ranking service has named Reliv’s Dr. Carl Hastings number nine on their Top Chief Scientific Officers Ranking list for 2012. Hastings joins a top ten of the nation’s leading CSOs, many of them from Fortune 100 companies. According to ExecRank, there are over 5,000 Chief Scientific Officers and C-Level Scientific Executives in the United States and this list represents “the best of the best in their field.”“This honor is hard-earned and well-deserved,” said Robert L. Montgomery, Reliv Chairman and CEO. “Carl is a man of science who has devoted his life to developing breakthrough nutritional solutions to help meet the world’s biggest health needs. The top ten ranking only confirms what his peers and we at Reliv have known for years: Dr. Carl Hastings is one of the leading nutrition scientists in the world.” 

A Very Good Year   The ExecRank listing comes following a year in which Hastings earned two product patents (his sixth and seventh) and introduced LunaRich® soy powder and LunaRich X™. LunaRich products optimize bioactive lunasin, the soy peptide research shows works at the epigenetic level to promote cellular health.GlucAffect®, Reliv’s blood sugar management formula, earned a patent in February 2012 and ProVantage®, Reliv’s athletic performance enhancement product, followed with its own patent in May. Both products now contain LunaRich, a Reliv-exclusive ingredient that delivers more bioactive lunasin — many times more — than any other soy powder available today.

 

Soy and Your Health

Soy and Women's Health

An important recent study found that soy food consumption did not increase the risk of cancer recurrence or death among survivors of breast cancer. Women in the highest intake category of soy foods had a 9 percent reduced risk of mortality and a 15 percent reduced risk for recurrence compared to those who had the lowest intake level. Researchers used data from a multi-institution collaborative study called the After Breast Cancer Pooling Project. Breast cancer outcomes were assessed, on average, nine years after cancer diagnosis.

For postmenopausal women with hormone-sensitive breast cancer, another recent study showed those who consumed high amounts of soy isoflavones had a lower risk of recurrence. In addition, evidence suggests that eating soy during childhood and/or adolescence reduces breast cancer later in life

Soy and Men's Health

Researchers at Northwestern University have found that a new, nontoxic drug made from soy’s isoflavone genistein could prevent cancer cells in the prostate from spreading to the rest of the body. So far, the cancer therapy drug has worked in preclinical animal studies and now shows benefits in humans with prostate cancer.

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Reliv Announces Breakthough in Metabolic Syndrome

Metabolic syndrome — a cluster of symptoms including high blood sugar, obesity and cardiovascular problems — has reached epidemic proportions. According to the American Heart Association, 47 million Americans now have metabolic syndrome. The syndrome afflicts 40 percent of people in their sixties and seventies. That number is expected to rise along with our aging population. The CDC also predicts that one-third of Americans will develop diabetes if current trends continue.

A recent CDC study published in Pediatrics shows the epidemic isn’t limited to older adults. Obesity in teens has skyrocketed in the past decade — 23 percent of teens are now classified as diabetic or prediabetic.

A person with metabolic syndrome is twice as likely to develop heart disease and five times more likely to develop diabetes. Other silent symptoms include eye, kidney and nervous system damage over time.

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Fox News: 8 foods to keep your heart healthy

Have you heard of lunasin?  It’s in every soy product in America but researchers now think it may be the secret sauce that gives soy its heart health power punch, including cholesterol reduction and general cellular health. Research shows lunasin may be among the first bioactive compounds to affect gene expression and promote optimal health at the epigenetic level.