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. 

 

For Lunasin Research

Lunasin is a peptide found naturally in soy that scientists have identified as the key to many of soy's documented health benefits.  More than 40 peer-reviewed and published studies have documented lunasin's health benefits.  This bioactive soy component is the subject of research by more than 25 research institutions around the world.  This research has been supported by 27 public and private funding sources and has resulted in 9 patents.  Bodies of research include: cancer prevention, cardiovascular, inflammation, skin health and anti-aging.
(Click on here to bring you to over 40 studies on lunasin.com)

 

 

Soy Protein Protects Mice from Skin Cancer Lotion wards off dangerous DNA changes, says study

By Adam Marcus
HealthDay Reporter MONDAY, Oct. 15  (HealthDayNews) -- California researchers say a cream rich in a particular soy protein delays and even prevents skin cancer in mice.

The compound, called lunasin, helps ward off cancerous changes to DNA, the long molecule that holds all the biological instructions for life, says a report in today's issue of the journal Cancer Research.

Study co-author Alfredo Galvez, president and CEO of FilGen BioSciences of Fairfield, Calif., which helped fund the latest work, says his company hopes to create a "cosmeceutical" form of lunasin that can be marketed as a skin cream to fight both skin tumors and breast cancer. Food and Drug Administration (FDA) regulations for topical medications are less stringent than they are for other prescription drugs, meaning FilGen could have a lotion on the market in several years as opposed to a decade or so for a medication taken internally, he says.

Soy has achieved cult status as a cure-all for everything from cardiovascular disease to cancer, with advocates pointing to lower rates of these illnesses in Asia, where soy is a staple.

Two years ago the FDA said it would permit food labels to say that eating 25 milligrams a day of soy protein may help cut the risk of heart disease.

A number of compounds in soybeans, including isoflavones and a molecule called BBI, have shown promise in treating cancer, and mounting research suggests that lunasin also may have potent anti-cancer properties.

In a study two years ago, Galvez and his colleagues showed that when the protein is injected into tumor cells, the cells stop dividing and die in a process known as apoptosis.

Lunasin behaves like a backup tumor suppressor, stepping in when the genes that normally quash runaway cell growth fail, as they do when cancer forms. "It seems like lunasin is selectively killing cells that are being transformed or are in the process of transformation into cancerous tissue," says Ben de Lumen, professor of food science at the University of California at Berkeley and a co-founder of FilGen.

De Lumen says the protein in soybeans appears to help seeds churn out DNA without having to divide. Since mammalian cells can't perform this trick, when the plant protein enters them "it messes up the cell."

In the new study, Galvez, de Lumen and others tested a topical lotion laced with lunasin on the skin of mice, applying it regularly for 19 weeks. They also exposed the animals to chemical carcinogens known to promote skin tumors.

Mice that received the largest dose of the cream -- 125 micrograms (millionths of a gram) twice a week -- were 70 percent less likely to develop skin tumors than untreated rodents, the researchers say. And when they did get cancer, they tended to have fewer tumors, and those appeared about two weeks later than those in the animals that didn't receive the lotion.

Early work with the protein shows it can penetrate into the basal layer of skin, where cell division is most rapid. Galvez says whether lunasin works against tumors triggered by exposure to ultraviolet radiation, which is the leading cause of skin cancer in humans, remains to be seen.

Although the compound seems safe for mice, when the gene for lunasin was injected into tumors, it killed both diseased and healthy tissue, says Galvez. FilGen is preparing to conduct toxicity studies with the protein.

As for other tumors, such as breast cancer, Galvez says the trick will be to find a way to make lunasin digestible. "The only stumbling block is to develop orally delivered lunasin that can get into the blood and deposit it into the tissue where it can be a preventative agent," Galvez says.

Carol MacLeod, a professor of medicine at the University of California, San Diego, Cancer Center, is planning a study to test the effectiveness of lunasin against breast cancer in mice. MacLeod is receiving funding for the work from the company and the state of California, an "arm's length" pairing she says will help enhance the credibility of the findings. MacLeod says she's not expecting any data from the study for many months.

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

Clinical Trial: Cancer Related Fatigue

Radiation Fatigue and the Use of a Nutritional Supplement

View Full Clinical Study

Edward H.Gilbert, M.D., Denise Lindley R.N.,BSN,OCN, Michelle Karlin, RN, AOCNP,FNP-BC

Northpoint Cancer Center, Dallas Texas and Verity Cancer Center, Plano Texas
January 2010

Radiation induced fatigue is a specific syndrome. It is much more defined than “cancer related fatigue.” It is usually not multifactorial, and typically arises within 1-5 weeks after the beginning of radiation treatments. There is usually resolution in the 3-4 weeks post treatment. Radiation fatigue is less intense than chemotherapy caused fatigue.

The nutritional supplement trial with Reliv was based on a number of anecdotal stories of remarkable reversals in various symptoms, and in disease processes such as arthritis, diabetes, fibromyalgias, etc. These responses included reduced pain, increased energy, and feelings of wellbeing. This trial had some degree of simplicity. The fatigue symptom was limited in etiological causes, since the criteria for trial entry was fatigue presenting after the start of radiation treatment. The response was to measure the change in a short period of 1-4 weeks on the product. Patients were at different stages in their radiation treatment. Many would be finishing, and in the 2-4 weeks after completion fatigue normally improves on its own.

It was appreciated that this was essentially a “feasibility study,” and its scientific accuracy would needed to be documented in a controlled trial. Yet, if any responses were seen, then this would validate doing a next phase. The placebo effect itself was recognized as a limitation. Also, a major issue was compliance in taking the powder twice a day, and thereby giving credibility to any non-responders. This was not adequately measured, so that in the 9 patients who had no response, lack of compliance was possible. However, the majority were seemingly honest in their reporting.

Material and Methods:

The product used was Classic Reliv and for half of the study, Innergize was added to the Classic Reliv. Availability of the product determined whether Innergize was added. The dose was 1 scoop twice a day in liquid (it should be noted that a minimum of 2 scoops twice a day is a more standard recommendation). The lower dose was decided upon to help compliance. There was enough product available for a 4 weeks trial for each patient.

The assessment tool was “The Fatigue Severity Scale,” and this can been seen in the figure 1 below. It may not be totally appropriate for radiation caused fatigue, but it was at least a rough validation of response. Absolute scores were less important than relative change of the score over time. Compliance of returning the questionnaire was very varied, but all had at least 2 responses. The attempt was for 1 response a week. Further, patients were interviewed as to how they perceived their responses to be. For this study, credibility was given to the patient subjective reporting.

Patient selection was their declaration of significant fatigue, starting after the onset of radiation treatment. There was no restriction on when the fatigue began and when they started on the Reliv.

It should be noted that there were no other variables introduced to the patients. The fatigue began, and rather than giving any further interventions, the Reliv was started. Further, the acute results are what are being reported. Still, as noted in the individual reports, there were
some patients with longer lasting effects, with these patients willing to continue the supplement on their own.

Excellent response was a combination of significant change in the “Fatigue Severity Scale” and their self reporting of a major improvement. A good response was from a +8 to +15 change and also the self report of good improvement. No response could be their stopping the product (one because of vomiting), compliance problems, and most frequently the report of no response and no score improvement.

Discussion:

Radiation induced fatigue is a well defined symptom linked to radiation therapy. There has not been an adequate treatment approach, with little having a significant effect. Physical exercise is considered very important, but patients are often too tired to have a consistent regimen. Mind-body therapies can be helpful, but most patients do not pursue these (acupuncture, meditation, message therapies, etc). And there are the issues of a more pronounced “cancer related fatigue,” complex in etiology and suggested intervention techniques. Further, many of these patients are on concurrent chemotherapy, are anemic, and report other cancer treatment related side effects.

Nutritional recommendations are controversial and very complicated. A general statement of taking vitamin supplements, or extra specific minerals, or various therapies such as juicing, is often made. Patients are typically left to their own pursuits or discoveries, yet most health care professionals would encourage nutritional approaches to help the fatigue problem. Using a product such as Reliv, with a 22 year history, was a relatively simplistic attempt to study whether a response was possible.

This study was a “feasibility study” in the sense that all we were looking for was some effect. The number of variables to do a proper scientific analysis is large, and would represent the next phase of research. The possibility of the placebo effect requires a randomized trial, and was inappropriate to this feasibility study. However, this is recognized in reporting, as well as compliance difficulties, and subjectivity of the questionnaires. All that said, there were the very definite improvements, and in some situations, dramatic changes.
Reliv contains a variety of vitamins, minerals and herbs in a powder base excellent for absorption. It is an easy product to prescribe, with dosing fairly standard. Further, there has been much experience with some studies and many testimonials for a positive result with Reliv. We personally were encouraged by these reports of significant improvement with a range of symptoms from other diseases. Testing a nutritional supplement for relief from fatigue was the motivation for this study. Further, few nutritional products have been formally recommended by the scientific experts in oncology.

The result was a certainly positive response. Approximately 75% of the patients with radiation fatigue had relatively immediate results, many within 2 weeks of starting the product. Roughly 25% of the cohort reported excellent results in reversal of radiation fatigue, and improvement or relief from other symptoms. There was no harm done to those who reported no response (except one with a few episodes of vomiting). The results warrant a more extensive trial with all the scientific rigors applied.
Reliv is distributed through a multilevel sales channel. Some clinicians and administrators have high resistance and negative preconceptions of products distributed via multilevel marketing. It is crucial to point out that all product in this study was provided free of charge and no attempt was made to enlist patients in further purchase or marketing the product. A few members of the cohort did buy additional product after the initial response but none were asked to participate in distribution.

Cancer related symptoms often are the most significant issue in quality of life. Treatments such as radiation can be highly symptom producing, with fatigue perhaps the most generic problem. Having the possibility to reverse this problem during the course of radiation is a significant benefit helping both the patient, their family, and the healthcare professionals managing their treatments.

Summary:

A feasibility study on 37 patients undergoing or just having completed radiation and having significant radiation fatigue was conducted. The patients were placed on a nutritional supplement program named Reliv, and measured for response over 2-4 week continuous treatment. Results show that 75% of participants considered their result as helping their fatigue, and 25% of the entire group reported an “excellent” response. The feasibility study’s result warrants a second phase clinical study conducted under rigorous scientific methodology, and should be considered for the syndrome of “cancer related fatigue.”

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.

Click here for entire article

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.

Click here for entire article

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.