The Underlying Mechanisms of

Chlorella, AFA, Echinacea, Cordyseps Sinensis,

Beetroot and Liver Anhydrate on the Potentiality

of Enhancement of Athletic Performance.


Preliminary definitions 

  • In vitro: (of a biological process) made to occur in a laboratory vessel of other controlled experimental environment rather than within a living organism or natural setting.
  • Transgenic: Of, relating to, or denoting an organism that contains genetic material into which DNA from an unrelated organism has been artificially introduced.
  • PCR: Polymerase Chain Reaction is a process used in molecular biology to amplify a single copy of a few copies of a piece of DNA across several orders of magnitude, generating thousands to millions of copies of a particular DNA sequence.
  • Placebo: A substance that does not contain active ingredients and is made to be physically indistinguishable (that is, it looks and tastes identical) from the actual drug being studied.
  • Double-blinded clinical trial: A type of study in which two or more parties involved in the clinical trial do not know which participants have been assigned which interventions. Typically, the parties include the investigator and participants.
  • Cross-over design: describes a clinical trial in which groups of participants receive two or more interventions in a particular order. For example, one group receives drug A during the initial phase of the trial, followed by drug B during a later phase.
  • NMR: Nuclear magnetic resonance spectroscopy is a research technique that exploits the magnetic properties of certain atomic nuclei. It determines the physical and chemical properties of atoms or the molecules in which they are contained.
  • In vivo: studies in which the effects of various biological entities are tested on whole, living organisms, usually animals or humans, as opposed to a partial or dead organism.
  • Pilot study: a small scale preliminary study conducted in order to evaluate the feasibility, time, cost, adverse events and effect size in an attempt to predict an appropriate sample size and improve upon the study design prior to performance of a full scale study.
  • Meta-analysis: a statistical technique for combining the findings from independent studies. It is most often used to assess the clinical effectiveness of healthcare interventions; it does this by combining data from two or more randomized control trials.

Aphanizomenon flos-aquae (AFA)
How it is believed to support, protect and build blood properties, enhance mood, and muscle stem cell repain

General Overview of AFA
Aphanizomenon flos-aquae (AFA) is a fresh water unicellular blue-green algae that is rich in phycocyanin, which has antioxidant and anti-inflammatory properties. AFA grows spontaneously in Upper Klamath Lake in Oregon, USA.

How It Can Protect & Detoxify The Blood
In normal cells, when exposed to free radicals, the free radicals attack red blood cell (erythrocyte) membrane components, resulting in oxidative stress or lipid peroxidation; this eventually leads to red blood cells undergoing hemolysis. However, in the presence of AFA, hemolysis was significantly delayed (1) This study further showed that AFA exhibits a dose-dependent protection against erythrocyte oxidative damage.

One study found that human consumption of AFA increased blood levels of monocytes and lymphocytes (2) Monocytes are white blood cells, also known as leukocytes, and are importance cells for the innate immune cells. A lymphocyte is one of the subtypes of white blood cells, and include natural killer (NK) cells, T-cells and B-cells.

Consumption of AFA has been shown to have an effect on Natural Killer (NK) cells. NK are an important cell involved in the immune system and act as a primary defense against viral and bacterial infections. AFA contain cell wall components, such as peptidoglycans, which are known to induce NK cell activation (4). In addition, after consuming AFA, a large amount of NK cells migrated into the tissues from the bloodstream, indicating that they were helping to fight the body against foreign matter (5). NK cells can distinguish between infected cells and uninfected cells by recognizing molecules on the surface of cells.

AFA has also been shown to exert detoxification properties. In a study, the detoxification method of AFA to cadmium was investigated. When AFA was exposed to cadmium, intracellular cadmium increased within the phytoplankton, indicating that AFA was absorbing the heavy metal (3). Heavy metals, if left unchecked, can cause negative effects on growth, cell division, and can result in an increase in reactive oxygen species, leading to oxidative stress.


  1. Benedetti S, et al. Antioxidant properties of a novel phycocyanin extract from the blue-green alga: Aphanizomenon flos-aquae. Life Sciences. 2004; 2353-2362

Number of citations: 69

Red blood cells had reduced oxidative hemolysis and lipid peroxidation when treated with AFA. AFA also exhibited a dose and time dependent effect.

  1. Jensen GS, et al. Consumption of Apanizomenon flos-aquae has rapid effects on the circulation and function of immune cells in humans. JANA. 2000; 2: 50-58

Number of citations: Information not available

  1. Ran X, et al. The response and detoxification strategies of three freshwater phytoplankton species, Aphanizomenon flos-aquae, Pediastrum simplex, and Synedra acus, to cadmium. Environ Sci Pollut Res Int. 2015; 22(24): 19596-606

Number of citations: Information not available

This in vivo study was conducted to determine the ability of AFA to detoxify when exposed to high levels of cadmium. After exposure, adsorbed cadmium and intracellular cadmium increased significantly in AFA

  1. Nakao Y, et al. Surface-expressed TLR6 participates in the recognition of diacylated: li-poprotein and peptidoglycan in human cells. J Immunol. 2005; 1566-1573

Number of citations: 77

This in vitro study showed that diacylated lipoprotein and peptidoglycan are recognized by macrophages through the coexpression of TLR2 and TLR6.

  1. Hart AN, et al. Natural killer cell activiation and modulation of chemokine receptor profile in vitro by an extract from the cyanophyta Aphanizomenon flos-aquae. Journal of Medicinal Food. 2007; 10(3): 435-441.

Number of citations: 4

This in vitro study was conducted to show that AFA modulates the activity of NK cells. The results showed that AFA induced the expression of chemokine receptors and activated NK cells.

How it supports muscle stem cell repair and recovery
AFA has been shown to increase the number of bone marrow derived stem cells by 25% within one hour of oral consumption (1). AFA is able to increase the number of circulating stem cells via the following mechanism: Interference with the adhesion of stem cells to the bone marrow via L-selectin. This allows the stem cells to circulate into the blood stream to regions of tissue damage or areas that need repair. AFA contains a compound that specifically binds to L-selectin. L-selectin is a cell adhesion molecule that is involved in cellular migration during normal immunosurveillance and inflammatory conditions. Specifically, L-selectin is involved in releasing and retaining bone marrow stem cells into the blood stream (2). By AFA binding to L-selectin, it triggers the process of releasing stem cells to allow for repair and recovery of muscle cells.

In one study where muscle cells were injury-induced by cardiotoxins, there was greater regeneration of the anterior tibialis muscle when treated with AFA (3). In addition, with AFA treatment, there was a greater migration of stem cells to the area of injury, thereby enhancing recovery after muscle injury. Furthermore, stem cells migrated to a lesser extent to non-injured regions, supporting the view that AFA-derived stem cells primarily migrate towards areas of injury or tissue damage.

During exercise, free radicals can form which causes oxidative damage to the muscle cells. AFA is well known to have oxygen radical absorbance capacity (ORAC) and has a strong ability to scavenge peroxyl radicals. In an in vivo study, it was shown that an AFA compound called Phycocyanin had significantly higher antioxidant activity compared to other well-known antioxidant molecules, including ascorbic acid, uric acid, alpha-tocopherol, and Trolox (4). Because AFA has a strong antioxidant capacity, it prevents the free radicals that are formed after exercise or over-exertion from damaging muscle cells, thus, allowing them to recovery and repair. It should also be noted that in conditions where there is an excessive formation of free radicals (which result in tissue damage), the body’s natural antioxidants, such as superoxide dismutase, are decreased. In such a case, treatment with AFA was able to exert the anti-oxidant activity by decreasing the production of free radical derivatives as well as enhancing the anti-oxidant activity (5). In addition, it was shown that AFA treatment reduced lipid peroxidation, and enhanced the activity of glutathione levels and superoxide dismutase, which is an enzymatic anti-oxidant. Superoxide dismutase breaks down damaging free radicals (e.g. superoxide radical O2) into non-damaging oxygen molecules (O2) or hydrogen peroxide (H2O2). Enzymes such as superoxide dismutase remove damaging superoxide radicals by adding or removing an electron with the following reactions:

  • Cu2+-SOD + O2→ Cu+-SOD + O2
  • Cu+-SOD + O2+ 2H+ → Cu2+-SOD + H2O2


  1. Jensen GS, et al. Mobilization of human CD34+ CD133+ and CD34+ CD133(-) stem cells in vivo by consumption of an extract from Aphanozimenon flos-aquae – related to modulation of CXCR4 expression by an L-selection ligand? Cardiovasc Revasc Med. 2007; 8: 189-202.

 Number of citations: Information not available

This study evaluated the effects on human stem cells in vitro and in vivo of AFA. The results show that AFA contains a novel ligand for CD62L that modulates CXCR4 expression on CD34+ bone marrow cells in vitro.

  1. Frenette PS, et al. Endothelial selectins and vascular cell adhesion molecule-1 promote hematopoietic progenitor homing to bone marrow. Proc Natl Acad Sci USA. 1998; 95(24): 14423-8

Number of citations: 236

This study investigated the role of endothelial selectins in the process of stem cells from the bone marrow. The results from this study demonstrate the selectins are important in the process of stem cell homing to the bone marrow.

  1. Drapeau C, et al. Mobilization of bone marrow stem cells with StemEnhance improves muscle regeneration in cardiotoxin-induced muscle injury. Cell Cycle. 2010; 9(9): 1819-1823

Number of citations: 6

In this study, a water soluble extract of AFA, known as StemEnhance, induced mobilization of stem cells from the bone marrow to allow for muscle regeneration.

  1. Benedetti S, et al. Oxygen radical absorbance capacity of phycocyanin and phycocyanobilin from the food supplement Aphanizomenon flos-aquae. Journal of Mecicinal Food. 2010; 13(1): 223-227

Number of citations: 10

The purpose of this study was to evaluate the ability of AFA to protect erythrocytes from oxidative damage. With AFA treatment, oxidative hemolysis and lipid peroxidation was significantly lower.

  1. Kuriakose GC. Evaluation of Renoprotective effect of Aphanizomenon flos-aquae on cisplatin-induced renal dysfunction in rats. Renal Failure. 2008; 30:717-725

Number of citations: 8

This study showed that chronic treatment of AFA significantly and dose-dependently restored renal functions, reduced lipid peroxidation, and enhanced reduced glutathione levels, superoxide dismutase and catalase activities.

Mood Enhancer
Phenylethylamine (PEA) is a compound that is found in the brain and has been shown to be a neuromodulator of aminergic synapses and improves energy and elevates mood. In addition, when PEA was administered to depressed patients, mood was improved (1).  AFA has been found to contain the compound PEA (2).

PEA is able to affect mood, cognition, emotion, reward, leaning and attention through modulation of the brain monoaminergic system. Monoamines include neurotransmitters, such as dopamine, noradrenaline, and serotonin, which affect a wide range of physiological and homeostatic functions. For example, dopamine is important in cognitive processes, such as attention, working memory and motivational behavior. Trace amounts of PEA alters the monoamine transporter function in the brain (3). This suggests that PEA plays a modulatory role for this receptor in the presynaptic regulation of monoaminergic activity.


  1. Sabelli HC, et al. Phenylethylamine modulation of affect: therapeutic and diagnostic implications. Journal of Neuropsychiatry and Clinical Neurosciences. 1995; 7(1): 6-14

Number of citations: Information not available

This paper reviews the literature regarding PEA as a mood enhancer. Treatment with PEA improved mood in depressed patients.

  1. Apsley, JW. The Genesis effect: spearheading regeneration with wild blue green algae. Volume 1 (2nd edition). Genesis Communications. 1996.

Number of citations: Information not available

  1. Xie Z, et al. Trace amine-associated receptor 1 as a monoaminergic modulator in brain. Biochemical Pharmacology. 2009; 78(9): 1095-1104.

Number of citations: 26

This paper discusses the recent findings in receptors that appear to modulate brain monoaminergic systems.

Nootropic Effect
As mentioned in the previous section, AFA contains PEA. Several studies have shown the effect that PEA has on regulating catecholaminergic neurotransmitter release in the brain. Phenylethylamine (PEA) was shown to play a role in modifying the release of acetylcholine in dopaminergic neurons (1). In this study, when PEA was administered systemically, there was an increased release of acetylcholine in the striatum. This increase in acetylcholine is believed to involve the nigrostriatal dopaminergic system, specifically the dopamine D2 receptor, although the mechanism still remains unclear.

Studies have shown that acetylcholine and the cholinergic system is an important neurotransmitter in the brain that controls activities that depend on selective attention and conscious awareness. For example, in degenerative diseases of the brain, such as Alzheimer’s Disease, decreases in consciousness is associated with a deficit in the cholinergic system (2).  In addition, a study conducted on patients with Alzheimer’s found that treating with an acetylcholine precursor was able to improve scores in cognitive ability (3), showing the effect that acetylcholine has on mental clarity and mental capacity.


  1. Kota M, et al. B-Phenylethylamine modulates acetylcholine release in the rat striatum: involvement of a dopamine D2 receptor mechanism. European Journal of Pharmacology. 2001; 418:65-71.

Number of citations: Information not available

This in vivo study examined the effects of PEA on striatal acetylcholine release and showed that systemic administration of PEA increased acetylcholine release, whereas locally applied PEA decreased striatal acetylcholine release.

  1. Perry E, et al. Acetylcholine in mind: a neurotransmitter correlate of consciousness? Trends Neurosci. 1999; 22: 273-280.

Number of citations: Information not available

This paper reviews the relationship between the cholinergic system and consciousness.

  1. Moreno M. Cognitive improvement in mild to moderate Alzheimer’s dementia after treatment with acetylcholine precursor choline alfoscerate: a multicentre, double-blind randomized, placebo controlled trial. Clin Ther. 2003; 25: 178-193.

Number of citations: Information not available

This double-blind, randomized, placebo controlled trial evaluated the effect of acetylcholine on Alzheimer’s patients.


Chlorella is quite small (approx. 2 to 10 microns in size) and has unique properties that make it a useful detoxification tool. It has a molecular structure that allows bonding to metals, chemicals and even some pesticides.

When chlorella is taken into your body, its natural action will bind it to heavy metals, chemicals, and pesticides found within the digestive tract. The digestive tract is the body’s pathway to your bloodstream, where harmful chemicals and toxins are delivered and deposited into the body’s cells. Chlorella vulgaris has the ability to remove metal ions in a short time, by biosorption. Chlorella’s chemical structure is also uniquely designed so that it doesn’t bind to other beneficial minerals in the body like calcium, magnesium and zinc.

Chlorella species have been shown to remove heavy metals such as cadmium (Cd), Copper (Cu) and lead (Pb) from the blood and from cells (1,2,3).  Chlorella and other algae also contain high concentrations of phytochelatins, capable of binding strongly to heavy metal ions (6). Phytochelatins are present in the chlorella’s tough outer wall and act as chelators, which is crucial for heavy metal detoxification.  Chelation is multiple coordination bonds between chlorella and the bound heavy metal or toxin. Chlorella then sequesters the heavy metal, transports it through the body, and then excretes it. This metal binding property (i.e. phytochelatins) is related to the presence of chloroplasts in the cellular wall of Chlorella, which is rich in sulphur, potassium, calcium and phosphorus (7). In particular, C. vulgaris can accumulate >70% of chromium in the cell, and thus, has been used in various stress investigations (5).

Heavy metals, if left unchecked, can cause negative effects on growth, cell division, and can result in an increase in reactive oxygen species, leading to oxidative stress. In addition to chelating and sequestering heavy metals, aquatic plants, such as Chlorella, have evolved both enzymatic (superoxide dismutase, catalase, ascorbate peroxidase) and non-enzymatic (ascorbate, glutathione) antioxidant mechanisms to prevent the oxidative stress (4). These antioxidant mechanisms break down damaging free radicals (e.g. superoxide radical O2) into non-damaging oxygen molecules (O2) or hydrogen peroxide (H2O2). Enzymes such as superoxide dismutase remove damaging superoxide radicals by adding or removing an electron with the following reactions:

  • Cu2+-SOD + O2→ Cu+-SOD + O2
  • Cu+-SOD + O2+ 2H+ → Cu2+-SOD + H2O2

Thus, chlorella uses two main mechanisms to cleanse and detoxify the blood:

  1. The cell wall of chlorella is able to specifically bind to toxins and heavy metals to sequester, transport and remove them from the body, preventing heavy metals and toxins to be harmful.
  2. Chlorella contains antioxidant properties, to allow the conversion of free radicals like superoxide dismutase into non-damaging molecules.


  1. Wilde KL, Stauber JL, Markich S, et al. The effect of pH on the uptake and toxicity of copper and zinc in a tropical freshwater alga (Chlorella sp). Arch Environ. Contam. Toxicol. 2006; 51: 174-185.

Number of citations: 127

An in vitro study that demonstrated that Chlorella sp. is more sensitive to Cu than U at a pH of 6.5 and 5.7, and that the toxicity of Cu and U is highly dependent on the pH.

  1. Gin KY, Tangm YZ, Aziz MA. Derivagion and application of a new model for heavy metal biosorption by algae. Water Res. 2002; 36:1313-1323.

Number of citations: 58

An in vitro study that was conducted to describe the relationship between specific parameters and heavy metal sorption by algae. It was found that for Chlorella, the removal efficiency of heavy metal and metal adsorption was a ratio of the algal biomass concentration to the initial heavy metal concentration.

  1. Lourie E, Patil V, Gjengedal. Efficient purification of heavy metal contaminated water by micro-algae activated pine bark. Water Air Soil Poll. 2010; 210: 493-500.

Number of citations: Information not available

An in vitro experiment conducted to determine the biosorption of Cu, Pb, Zn, Cd, Co, and Ni in algae-treated bark. Metal sorption efficiency was greatest in bark that was treated with microalgae.

  1. Lei AP, Hu ZL, Wong YS, et al. Antioxidant responses of microalgal species to pyrene. J Appl. Phycol. 2006; 18:67-78.

Number of citations: 5

An in vitro study to test the antioxidant response of Chlorella. Results indicated that microalgae alter their antioxidant systems in response to pyrene exposure.

  1. Rai UN, Singh NK, Upadhyay AK, et al. Chromate tolerance and accumulation in Chlorella vulgaris L: role of antioxidant enzymes and biochemical changes in detoxification of metals. Bioresource Technology. 2013; 136: 604-609.

Number of citations: 4

An in vitro study to determine the effects of chromium toxicity on Chlorella vulgaris. The results reveal that Chlorella responds better under chromium stress in terms of tolerance, growth and metal accumulation.

  1. Cobbett C, Goldsbrough P. Phytochelatins and metallothioneins: roles in heavy metal detoxification and homeostatis. Annu Rev Plant Biol. 2002; 53: 159-182.

Number of citations: 990

This paper reviews the various literature relating to heavy metal binding in plant cells and scientific advances in heavy metal detoxification.

  1. Lopez CE, Castro JM, Gonzales V, et al. Determination of metal ions in algal solution samples by capillary electrophoresis. J. Chromatogr. Sci. 1998; 36: 352-356.

Number of citations: 7

Capillary electrophoresis as used to separate metal ions in an algal solution sample to determine the metals that bind to Chlorella vulgaris. The metal ions, Cr, Mn, Cd, and Cu had high peaks indicating that they were bound to Chlorella.

How it is believed to support muscle stem cell repair and recovery
Lead has been shown to suppress stem cells in the bone marrow, which leads to negative effects to the progenitor cells and lower resistance to a variety of pathogens. Chlorella vulgaris, however, induces a significant recovery in the reduced number of myeloid progenitor cells in stressed mice (1). This study also demonstrated that the bone marrow protective effects of Chlorella vulgaris was partly due to its chelating effect, as seen in the significant reduction of blood lead levels (mechanism as explained above). Recent findings show that mice that were lead exposed and then treated with Chlorella vulgaris modulated the reduced ability of bone marrow stromal cells to display myeloid progenitor cells and restored the overall numbers of myeloid progenitor cells in the long term (2).

Lab studies have demonstrated that Chlorella vulgaris may have a direct myelostimulating effect by inducing the production of endogenous cytokines (3, 4). Cytokines are a broad category of small proteins that are involved in cell signalling, which are released and affect the behaviour of other cells, such as the inflammatory or immune response. Chlorella vulgaris increases the production of IFN-γ, IL-1α and TNF-α – these are mediators of the inflammatory response (5) and can stimulate macrophages which produce pro-inflammatory cytokines (3). The presence of anti-inflammatory macrophages has been shown to prevent the loss of protein content and muscle atrophy. Macrophages, in particular, prevent the decrease in myotube size and myosin heavy chain content (9). It is thought that macrophages have a positive effect on muscle growth as anabolic growth factors are released by macrophages, and they also stimulate endothelial cell proliferation and migration (10). These are the underlying mechanisms that results in the cell recovery process.

Blue-green algae have been shown to have an effect on the functioning of immune cells, and increases the number of adult stem cells in the body (12).  In addition, blue-green algae contain a component termed the “Immolina fraction” by sciences. This fraction increases the concentration of cytokines in the bloodstream. Cytokines are a set of molecules that are involved in cell signalling, which can lead to a cascade of biological events in the body, including signalling cell repair. Cytokines, in this case, stimulate leukocyte movement within the body and the migration of leukocytes from blood to tissues (13). Leukocytes are white blood cells fight foreign substances and disease and free radicals in tissues, including muscles, and help to protect your body against diseases and injury. After exercise or high muscle performance, these free radicals can form in the muscles, which can damage the muscle tissues. As blue-green algae can increase leukocytes into the muscles, the leukocytes can help to reduce the recovery time by inhibiting free radical damage (14).

In terms of muscle repair & recovery
Muscle damage and atrophy is thought to be a result of oxidative stress, which has been indicated by an accumulation of several oxidative stress markers. Chlorella elicits various immunopharmacological effects and functions as an antioxidant (6), likely because of its high concentration of carotenoids, lutein, B-carotene, alpha-tocopherol and ascorbic acid.

Long-term usage of Chlorella has shown decreased disintegration of muscle and suppressed oxidative stress in the quadriceps muscle (7). This study suggests an underlying mechanism: Chlorella prevents the accumulation of lipid peroxides in the muscle cells and prevents mitochondrial dysfunction by protecting the cytochrome c oxidase activity.  Cytochrome c oxidase is a large enzyme found in the mitochondria that is involved in the synthesis of ATP. By protecting this enzyme, Chlorella is able to maintain synthesis of ATP and energy production in the cells. In addition, chlorella stimulates the action of anti-inflammatory macrophages. Macrophages release anabolic growth factors which lead to endothelial cell proliferation, aiding in recovery. Macrophages also promote the growth of muscle by increasing myotube size and myosin heavy chain content.  Note: Mitochondrial dysfunction has been shown to be related to age-related muscle degeneration such as sarcopenia, and aged rats have had a loss in cytochrome c oxidase activity (11).


  1. Dantas DCM, Queiroz MLS. Effects of Chlorella vulgaris on bone marrow progenitor cells of mice infected with Listeria monocytogenes. Int. J. Immunopharmacol. 1999; 21: 499-508.

Number of citations: 20

An animal study with mice infected with L. monocytogenes, a Chlorella vulgaris extract was shown to reduce the number of CFU (colony forming units) of L. monocytogenes at 48h and 72h after administration. Chlorella was shown to produce a significant increase in the resistance of animals infected with L. monocytogenes.

  1. Queiroz ML, Torello CO, Perhs SM, et al. Chlorella vulgaris up-modulation of myelosuppression induced by lead: the role of stromal cells. Food Chem Toxicol. 2008; 46: 3147-3154.

Number of citations: 8

An in vitro study was done to determine the chelating effects of Chlorella vulgaris on the bone marrow. Chlorella treatment was shown to significantly reduce lead levels in blood and tissues.

  1. Hasegawa T, Kimura Y, Hiromatsu K, et al. Effect of hot water extract of Chlorella vulgaris on cytokine expression patterns in mice with murine acquired immunodeficiency syndrome after infection with Listeria monocytogenes. Immunopharmacology. 1997; 35: 273-282.

Number of citations: 48

This was an animal study where mice were orally administered a hot water extract of Chlorella vulgaris (CVE). CVE lead to mechanisms that enhanced host defense against Listeria infection by augmenting cell-mediated immunity.

  1. Hasegawa, T, Kumamoto S, Ando Y, Yamada A, et al. Oral administration of hot water extracts of Chlorella vulgaris reduces IgE production against milk casen in mice. Int. J. Immunopharmacol. 1999; 21: 311-323.

Number of citations: 25

In this animal study, mice were orally administered a hot water extract of Chlorella vulgaris (CVE). This was shown to suppress the production of immunoglobulin.

  1. Schuerwegh AJ, Dombrecht EJ, Stevens WJ, et al. Influence of pro-inflammatory (IL-1 alpha, IL-6, TNF-alpha, IFN-gamma) and anti-inflammatory (IL-4) cytokines on chondrocyte function. Osteroarthritis Cartilate. 2003; 11: 681-687.

Number of citations: 80

An in vitro study was conducted to investigate the influence of pro-inflammatory cytokines. Pro-inflammatory cytokines were able to induce apoptosis, and anti-inflammatory cytokines inhibit NO production and proliferation of bovine chondrocytes.

  1. Lee HS, Choi CY, Cho C, et al. Attenuating effect of chlorella supplementation on oxidative stress and NFkappaB activation in peritoneal macrophages and liver of C57BL/6 mice fed on an atherogenic diet. Biosci Biotechnol Biochem. 2003; 67: 2083-2090.

Number of citations: 18

This study was conducted on mice to determine if chlorella supplementation helps to reduce oxidative stress. The results from this study show that chlorella supplementation attenuates oxidative stress by reducing reactive oxygen production and increasing anti-oxidative processes.

  1. Nakashima Y, Ohsawa I, Nishimaki K, et al. Preventive effects of Chlorella on skeletal muscle atrophy in muscle-specific mitochondrial aldehyde dehydrogenase 2 activity-deficient mice. BMC Complementary & Alternative Medicine. 2014; 14: 390.

Number of citations: Information not available.

This was an animal study with transgenic mice (gene that results in small body size, muscle atrophy, high muscle impairment marker) that were fed a Chlorella-supplemented diet. Chlorella was shown to improve skeletal muscle atrophy.

  1. Cao Z, Wanagat J, McKiernan SH, et al. Mitochondrial DNA deletion mutations are concomitant with ragged red regions of individual, aged muscle fibers; analysis by laser-capture microdissection. Nucleic Acids Res. 2001; 29:4502-4508.

Number of citations: 111

PCR (polymerase chain reaction) was used to identify the mitochondrial genotype of aged muscle fibers. This study showed that large deletion mutations are associated with abnormalties in aged rat muscle.

  1. Dumina N, Frenette J. Macrophages protect against muscle atrophy and promote muscle recovery in vivo and in vitro. Am J Pathol. 2010; 176(5): 2228-2235.

Number of citations: 16

The role of macrophages in muscle atrophy and regrowth was investigated using in vitro models. Anti-inflammatory macrophages prevented the loss of protein content and atrophy after 2 days.

  1. Schubert SY, Benarroch A, Ostvang J, et al. Regulation of endothelial cell proliferation by primary monocytes. Arterioscler Thromb Vasc Biol. 2008; 28: 97-104.

Number of citations: 17

An in vitro study was done to wound repair after binding of monocytes to endothelial cells. The interaction between endothelial cells and monocytes is critical for the regulation of endothelial cell proliferation.

  1. Hiona A, Leeuwenburgh C. The role of mitochondrial DNA mutations in aging and sarcopenia: Implications for the mitochondrial viscous cycle theory of aging. Experimental Gerontology. 2008; 43(1): 24-33.

Number of citations: 87

This review paper discusses skeletal muscle mass and strength and how it likely involves mitochondrial DNA (mtDNA) mutations or mitochondrial dysfunction.

  1. Jensen G, Ginsberg D, Huerta P, et al. A novel approach to nutritional mobilization of the immune system. JANA. 2000; 2: 50-58.

Number of citations: Information not available

A placebo-controlled, randomized, double-blinded study, this study examined the effects of consuming blue green algae on the immune system. This study revealed that consumption of algae lead to fast changes in immune without activating lymphocytes.

  1. Grzanna R, Polotsky A, Phan PV, et al. Immolina, a high molecular weight polysaccharide fraction of spirulina, enhances chemokins expression in human monocytic thp-1 cells. J Altern Complement Med. 2006; 12: 429-35.

Number of citations: 14

This study was conducted to determine the immunostimulatory effect of Immolina, a fraction derived from spirulina. Immolina activated human monocytic THP-1 cells.

  1. Scheett TP, Avery NG, Sharman MJ, et al. Leukocyte subpopulation responses during recovery from repeated days of resistance exercise: Effects of supplementing with polyunsaturated phosphatidylcholine. Medicine & Science in Sports & Exercise. 2002; 34: S232.

Number of citations: Information not available


Cordyseps Sinesis
How it is believed to increase oxygen carrying capabilities and Adenosine Triphosphate (ATP) production 

General Overview of Cordyseps Sinesis
Studies have shown that mice treated with CS (cordyceps sinensis) consistently demonstrate an increase in the liver β-ATP to inorganic phosphate (Pi) ratio (1). This ratio represents the high energy state in the liver and suggests an increase in blood flow in the liver and other organs. In addition, CS studied in vitro showed that it inhibited tracheal twitch contractions and relaxed persistent contractions of the trachea and the aorta, resulting in more blood flow (2). Increased blood flow allows increased oxygen to reach the liver and other organs.

The mechanism underlying the high hepatic energy state as a result of CS administration remains unknown. However, studies have shown that there is a link between liver blood flow and liver ATP levels (3). It was suggested that ATP levels and induction of ATP synthesis as a result of CS administration is caused by activated adenine translocase activity and/or mitochondrial respiratory-chain function in the liver. The adenine translocase is a transporter protein in the mitrochondrial membrane that enables ATP to be transported into the cell where it becomes the cell’s primary energy source.

A decrease in ATP consumption as a result of depressed Na pump function can produce a high energy state in the liver (4). The Na pump involves the synthesis of ATP via a transmembrane ATPase pump. This pump is responsible for exchanging ATP (into the cell) and for K ions (out of the cell). During this exchange of ions, ATP is hydrolysed (broken down) into ADP. With a depression in the function of the Na pump, ATP is not broken down and the concentrations become higher, making energy more available to cells.

However, further studies are in progress to identify the exact mechanism.

A study provided findings that indicate that CS increases hepatic blood flow in mice that had a condition associated with reduced blood flow (5). In this study, CS relaxes the contractions in the liver blood vessel system and increases hepatic blood flow, causing liver ATP levels to increase. Increased blood flow allows oxygen to enter cells. Oxygen is then used in the process of aerobic respiration for the production of ATP. Increased ATP levels contributes to the acceleration and improvement of liver function.

Note: In addition, increased oxygen to the organs (as a result of increased blood flow) is required for the synthesis of ATP. Aerobic respiration is a well-known mechanism where oxygen is required to generate ATP in the body.


  1. Manabe N, Sugimoto M, Azuma Y, et al. Effect of the mycelial extract of cultured Cordycep sinensis on in vivo hepatic energy metabolism in the mouse. Japanese Journal of Pharmacology. 1996; 70: 85-88.
  2. Tsunoo A, Takemoto N, Kamijo M. Pharmacological effects of the mycelial extract of cultured Cordyceps sinensis on airways and aortae of the rat. In Science and Cultivation of Edible Fungi, pp: 425-431.
  3. Whalen M, Shapiro JI. Controlled ventilation during NMR spectroscopic studies: hemodynamic and biochemical consequences. Magnetic Resonance Imaging. 1991; 9: 229-234.
  4. Takahashi H, Geoffrion Y, Butler KW, et al. In vivo hepatic energy metabolism during the progression of alcoholic liver diseases: a noninvasive P nuclear magnetic resonance study in rate. Hepatology. 1990; 11:65-73. Can’t find this exact source but there are some that are similar
  5. Manabe N, Azuma Y, Sugimoto M, et al. Effects of the mycelial extract of cultured Cordyceps sinensis on in vivo hepatic energy metabolism and blood flow in dietary anaemic mice. British Journal of Nutrition. 2000; 83: 197-204.


How it is believed to support the natural stimulation of Red Blood Cells (RBCs) and support the immune system

General Overview of Echinacea
Intake of Echinacea increases the expression of the leucocyte heat shock protein 70 (hsp70) and boosts white cell counts (1). HSPs are typically induced when cells are exposed to environmental stressors, such as heat, inflammation and oxidative stress. HSPs play a role in the assembly and transport of newly synthesized protein within cells, and also remove denatured proteins, and thus, are important molecules for preventing damage and repair after injury. HSPs are essential for maintaining cellular functions by preventing the misfolding and aggregation of proteins (8,9). They are also important in antigen presentation, and the activation of lymphocytes and macrophages, thus, stimulating the immune system (10,11). Antigen presentation is especially important after a viral infection, as it signals to the body that an antigen is “foreign” and “nonself”. It has been proposed that HSPs are an adaptive immune response, whereby their circulation in the bloodstream signals danger to the host.

Compounds within Echinacea have been suggested to modify the immune response, although the mechanism is still unclear. The result of this study (1) showed that the immunomodulatory effects of Echinacea are via a non-specific activation of the immune system, including an increase in white cell count (which also indicates an enhanced immune response) and increase in HSP70 response. Erythrocyte haemolysis (rupture or destruction of red blood cells) was also decreased.

Echinacea boosts the immune system by stimulating phagocytosis. Phagocytosis is the process by which white blood cells and lymphocytes attack invading organisms, such as parasites and bacteria. Echinacea also stimulates the production of T-cells and macrophages in the bloodstream. It also enhances the concentration of Interferon, Interleukin, Immunoglobulin and other natural immune chemicals in the blood. Some evidence suggests that non-specific activation of the immune system by Echinacea may be mediated by increased monocyte secretion of cytokines (2,3), and these monocytes then synthesize increased amounts of HSP70.

Echinacea intake also provided protection against oxidative damage to erythrocytes (red blood cells). The membranes of the red blood cells contain high concentrations of polyunsaturated fatty acids that are very susceptible and sensitive to free radical induced peroxidation (4). Echinacea is a good source of antioxidants and thus, can provide free radical scavenging activities, protecting red blood cells from oxidative damage (5,6,7). So, instead of increasing the action or production or RBCs, Echinacea is able to protect existing RBCs to maintain them at an optimal level.

Echinacea has been shown to stimulate the macrophage activity, which can result in an increase in PGE2 (prostaglandin E2) secretion from active macrophages (13). PGE2 is a protein that stimulates the production of serum erythropoietin (EPO)(14). EPO is a hormone that is secreted from the kidney and stimulates the stem cells to develop into Red Blood Cells from the bone marrow.  In addition, PGE2 has been shown to stimulate granulocyte-macrophage-colony-stimulating factor (GM-CSF) release from smooth muscle cells, and also acts as a growth factor for RBCs (15).  Echinacea also increases the activity level of T cells in the body that also stimulates GM-CSF, further enhancing the growth factors of RBCs. In all, Echinacea helps to stimulate RBCs by activating PGE2, GM-CSF, T-cells, which all have an impact on EPO and RBC production.


  1. Agnew LL, Guffogg SP, Matthias A, et al. Echinacea intake induces an immune response through altered expression of leucocyte hsp70, increased white cell counts and improved erythrocyte antioxidant defences. Journal of Clinical Pharmacy and Therapeutics. 2005; 30: 363-369.
  2. Rininger J, Kickner S, Chigurupati P, et al. Immunopharmacological activity of Echinacea preparations following simulated digestion on murine macrophages and human peripheral blood mononuclear cells. Journal of Leukocyte Biology. 2000; 68:503-510.
  3. Goel V, Chang C, Slama J. Echinacea stimulates macrophage function in the lung and spleen of normal rats. Journal of Nutritional Biochemistry. 2002; 13:487-492.
  4. Miki M, Tamai H, Mino M, et al. Free radical chain oxidation of rat red blood cells by molecular oxygen and its inhibition by alpha-tocopheral. Archives of Biochemistry and Biophysics. 1987; 258: 373-380.
  5. Pellati F, Benvenuti S, Magro L, et al. Analysis of phenolic compounds and radical scavenging activity of Echinacea spp. Journal of Pharmaceutical and Biomedical Analysis. 2004; 35: 289-301. I could not find this.
  6. Hu C, Kitts DD. Studies on the antioxidant activity of Echinacea root extract. Journal of Agricultural and Food Chemistry. 2000; 48:1466-1472.
  7. Sloley BD, Urichuk LJ, Tywin C, et al. Comparison of chemical components and antioxidants capacity of different Echinacea species. Journal of Pharmacy and Pharmacology. 2001; 53:849-857.
  8. Fink AL. Chaperone-mediated protein folding. Physiol. Rev. 1999; 79: 425-449.
  9. Hartl FU, Hayer-Hartl M. Molecular chaperones in the cytosol: from nascent chain to folded protein. Science. 2002; 295: 1852-1858
  10. Li Z, Menoret A, Srivastava P. Roles of heat shock proteins in antigen presentation and cross presentation. Curr. Opin. Immunol. 2002; 14: 45-51.
  11. Wallin RP, Lundqvist A, More S, et al. Heat shock proteins as activators of the innate immune system. Trends Immunol. 2002; 23:130-135.
  12. Whitehead MT, Martin TD, Scheett TP, et al. The effect of 4 wk of oral Echinacea supplementation on serum erythropoietin and indices of erthropoietic status. International Journal of Sport Nutrition and Exercise Metabolism. 2007; 17: 378-390.
  13. Burger RA, Torres AR, Warren RP, et al. Echinacea-induced cytokine production by human macrophages. Int J Immunopharmacol. 1997; 19:379-379.
  14. Boer AK, Drayer AL, Rui H, et al. Prostaglandin-E2 enhances EPO-mediated STAT5 transcriptional activity by serine phosphorylation or CREB. Blood. 2002; 100: 467-473.
  15. Lazzeri N, Belvisi MG, Patel HJ, et al. Effects of prostaglandin E2 and cAMP elevating drugs on GM-CSF release by cultured human airway smooth muscle cells relevance to asthma therapy. Am J Respir Cell. Mol. Biol. 2001; 24: 44-48.


Beet Root
How it is believed to support vasodilatation (expansion of blood vessel walls for increased blood flow)

General Overview of Echinacea
Beet Root contains dietary nitrate, which has been reported to lower blood pressure through a sequence of actions, starting from the reduction of nitrate to nitrite, then to NO in the circulation

(1). In the circulation, NO is formed from nitrite via several nitrite reductases such as deoxyhemoglobin and xanthan oxidoreductase (2).

The results of a study (1) showed that beetroot had an effect on microvascular vasodilation and arterial stiffness. Results also showed that there were significant increases in plasma and urinary nitrate and nitrite concentrations, indicating that this is involved in the vasodilation. This study also found a significant increase in endothelium independent vasodilation, supporting the hypothesis that beet root exerts it effect via the endothelium-independent pathway. In general, an increase in vascular function occurred in association with peak increases in plasma nitrate and nitrite. In basic terms, the endothelium (inner lining) of blood vessels uses nitric oxide to signal the surrounding smooth muscle to relax, resulting in vasodilation of the blood vessels, allowing more blood flow.

The proposed mechanism by which nitrite from beet root improves microvascular vasodilation and lowers DBP (diastolic blood pressure) is by its bioconversion to nitrate by oral bacteria, and further to NO in vivo, resulting in blood vessel dilation, increased microvascular vasodilation, and BP lowering (3,4).

Other clinical studies have shown that dietary nitrate from beetroot juice lowers blood pressure, inhibits platelet aggregation and prevents endothelial dysfunction (5,6).

Beet root also contains other potential bioactive compounds such as potassium, which can also enhance vascular function.  Many vegetables, especially root vegetables, grow within the soil and collect high concentrations of nitrates. Vegetables high in nitrates include Beet Root.


  1. Hobbs DA, Goulding MG, Nguyen A, et al. Acute ingestion of beetroot bread increases endothelium-independent vasodilation and lowers diastolic blood pressure in healthy men: a randomized controlled trial. Journal of Nutrition. 2013; 143(9): 1399-1405.
  2. Lundberg JO, Weitzberg E, Lundberg JM, Alving K. Intragastric nitric oxide production in humans: measurements in expelled air. Gut. 1994: 35:1543-6.
  3. Lundberg JO, Weitzberg E, Cole JA, et al. Nitrate, bacteria and human health. Nat Rev Microbiol. 2004; 2: 593-602.
  4. Gladwin MT. Does eNOS stand for erythrocytic NO synthase? Blood. 2006; 107:2595-6.
  5. Kapil V, Milsom AB, Okorie M, et al. Inorganic nitrate supplementation lowers blood pressure in humans: role for nitrite derived NO. Hypertension. 2010; 56: 274-81.
  6. Webb AJ, Patel N, Loukogeorgakis S, et al. Acute blood pressure lowering, vasoprotective, and antiplatelet properties of dietary nitrate via bioconversion to nitrite. Hypertension. 2008; 51:784-90.

Argentinian Grass fed/Vegan fed Liver Anhydrate
Nutritional benefits to athletes, in particular the B suite of vitamins (especially B12, 6 & 3 ), Iron, Copper, Zinc, etc., and the macro and micro minerals

General Overview of Vegan/Grass fed, Argentinean,  Liver Anhydrate
Liver anhydrate is a natural source of iron and is rich in vitamin A, Vitamin B-complex, Vitamin C and Vitamin D. It also contains copper, phosphorus, calcium, folic acid, and protein.

Its iron content and B vitamins help with general bone development and muscle development. In one study, it was shown that extracellular iron modulates the differentiation of osteroclast cells (bone stem cells), and increases the proliferation of osteoclast progenitor cells (1). This means that these stem cells are able to proliferate to replace and repair old or damaged cells in the body. In addition, the expression of a macrophage colony-stimulating factor was increased, a process involved in the development and activation of osteoclasts and bone resorption. In this course of action, the osteoclasts break down damaged bone and release the minerals from the bone into the blood; as a result, calcium is transferred from the bone to blood, making calcium available to other parts of the body.

B vitamins are involved in the gene encoding of the folate metabolising enzyme called methylenetetrahydrofolate reductase (MTHFR). MTHFR is an important enzyme in the body as deficiency of this enzyme has been associated with development delay, motor and gait dysfunction, seizure and neurological impairments. In this reaction, Vitamin B6 acts as a cofactor in the trans-sulphuration pathway, leading to MTHFR (2). Low expression of MTHFR is associated with a deficiency in B vitamins, suggesting that this is the pathway that B vitamins are involved in. Further, the final product of the reaction that MTHFR is responsible for catalysing is the biologically active form of folic acid called Levomefolic acid.

Previous studies have also shown that elevated homocysteine (Hcy) concentrations are associated with a decline in physical function and decreased muscle strength, especially as you age. Studies have shown that elevated Hcy concentrations can be reduced with Vitamin B12 and folic acid (3), suggesting that Vitamin B12 and folic acid are involved in the Hcy pathways.

In addition, Vitamin B is believed to be involved in increasing bone mineral density, collagen cross-linking, osteoclast activity and/or methylation capacity associated with increased bone strength (4).

Zinc and copper have also been shown to be involved in several antioxidant systems, including metalloproteins, such as the copper-zinc enzyme superoxide dismutase and the zinc and copper-binding protein metallothionein (5,6). Superoxide dismutase reduces the toxicity of oxygen free radicals, turning them into less harmful oxygen and hydrogen peroxide molecules. Metallothionein is a free-radical scavenging protein that depends on an adequate supply of zinc (7). As exhausting exercise and physical performance consumes oxygen, a higher concentration of oxygen free radicals can result, leading to oxidative damage. The increase in oxygen reactive species stimulates the antioxidant mechanisms (8,9), which include the copper/zinc dependent metalloproteins, such as superoxide dismutase and metallothionein in erythrocytes and in the plasma. Metallothionein has a cell protective role against oxidation, and is related to the reduced cysteinyl residues that scavenge peroxyl and hydroxyl radicals (10). Metallothionein expression is regulated mainly by zinc (7) and its synthesis is associated with an increase in zinc binding within cells (5).

Copper metabolism also plays a role in physical activity as it is involved in energy metabolism and synthesis of haemoglobin, myoglobin, cytochromes and peptide hormones (11). Copper is also a component of superoxide dismutase.

Also, it helps to stimulate appetite but at the same time it adds more lean body mass, and ensures that the body has a good and healthy red blood cell count at all times.


  1. Xie W, Lorenz S, Dolder S, et al. Extracellular iron is a modulator of the differentiation of osteoclast lineage cells. Calcif Tissue Int. 2016; 98(3): 275-83.
  2. Clarke M, Ward M, Strain JJ, et al. B-vitamins and bone in health and disease: the current evidence. Proceedings of the Nutrition Society. 2014; 73: 330-339.
  3. Homocysteine Lowering Trailists’ Collaboration. Dose dependent effects of folic acid on blood concentrations of homocysteine: a meta-analysis of the randomized trials. Am J Clin Nutr. 2005; 82:806-812.
  4. Swart KMA, van Schoor NM, Lips P. Vitamin B12, folic acid, and bone. Curr Osteoporos Rep. 2013; 11: 213-218.
  5. Hamer DH. Metallothionein. Ann Rev Biochem. 1986; 55: 913-951.
  6. McCord JM, Fridovich I. Superoxide dismutase. An enzymatic function for erythrocuprein (hemocuprein). J Biol Chem. 1969; 244: 6049-6055.
  7. Dehnert C, Hutler M, Liu Y, et al. Erythropoiesis and performance after two weeks of living high and training low in well trained triathletes. Int J Sports Med. 2002; 23: 561-566.
  8. Sen CK. Antioxidants in Exercise Nutrition. Sports Med. 2001; 31: 891-908.
  9. Smith JA, Martin DT, Telford RD, et al. Greater erythrocyte deformability in world-class endurance athletes. Am J Physiol. 1999; H2188-H2193.
  10. Miura T, Muraoka S, Ogiso T. Antioxidant activity of metallothionein compared with reduced glutathione. Life Sci. 1997; 60: 301-309.
  11. Speich M, Pineau A, Ballereau F. Minerals, trace elements and related biological variables in athletes during physical activity. Clin. Chim. 2001; 312:1-11.

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