Ingredients References

NEEDS REFERENCES

1. Acetyl-L-Carnitine
2. Betaine Anhydrous
3. Chromium
4. Conjugated Linoleic Acid (CLA)
5. Creatine
6. Isomaltulose
7. Malic Acid
8. Selenium
9. Vanadium
10. Vitamin B6 (Pyridoxine)
11. Methylcobalamin

HAS REFERENCES

1. 5-Hydroxytryptophan (5-HTP)
2. Beta-1,3-glucan
3. Beta-Alanine
4. Caffeine
5. Casein Hydrolysate
6. CDP Choline
7. Citrulline Malate
8. Cranberry
9. Curcumin
10. Cyanidin 3-Glucoside (C3G)
11. DMAE
12. Epigallocatechin-3-gallate
13. Electrolytes & Nutrient Timing
14. Eurycoma longifolia
15. Forskolin
16. Highly Branched Cyclic Dextrin (Cluster Dextrin)
17. L-Leucine
18. L-Theanine
19. L-Tyrosine
20. Lycopene
21. Magnesium
22. Omega-3 Fatty Acids
23. PhGABA
24. Phosphatidic Acid
25. Phosphatidylcholine
26. Piperine
27. Pomegranate
28. Protein
29. Raspberry Ketone
30. Resveratrol
31. Sleep
32. Superfood
33. Tribulus terrestris
34. Vitamin D3
35. Yohimbine
36. Zinc
37. ZMA

5-Hydroxytryptophan (5-HTP)

1. Maes M et al. Effects of serotonin precursors on the negative feedback effects of glucocorticoids on hypothalamic-pituitary-adrenal axis function in depression. Psychoneuroendocrinology. 1995;20(2):149-67. doi: 10.1016/0306-4530(94)00049-g.
2. Birdsall TC. 5-Hydroxytryptophan: a clinically-effective serotonin precursor. Altern Med Rev. 1998 Aug;3(4):271-80.
3. Maffei ME. 5-Hydroxytryptophan (5-HTP): Natural Occurrence, Analysis, Biosynthesis, Biotechnology, Physiology and Toxicology. Int J Mol Sci. 2020 Dec 26;22(1):181. doi: 10.3390/ijms22010181.

Beta-1,3-glucan

1. Moorlag S et al. β-Glucan Induces Protective Trained Immunity against Mycobacterium tuberculosis Infection: A Key Role for IL-1 Cell Reports 31, 107634, May 19, 2020. DOI: doi.org/10.1016/j.celrep.2020.107634. ABSTRACT: β-glucan is a potent inducer of epigenetic and functional reprogramming of innate immune cells, a process called “trained immunity,” resulting in an enhanced host response against secondary infections. We investigate whether β-glucan exposure confers protection against pulmonary Mycobacterium tuberculosis (Mtb) infection. β-glucan induces trained immunity via histone modifications at gene promoters in human monocytes, which is accompanied by the enhanced production of proinflammatory cytokines upon secondary Mtb challenge and inhibition of Mtb growth. Mice treated with β-glucan are significantly protected against pulmonary Mtb infection, which is associated with the expansion of hematopoietic stem and progenitor cells in the bone marrow and increased myelopoiesis. The protective signature of β-glucan is mediated via IL-1 signaling, as β-glucan shows no protection in mice lacking a functional IL-1 receptor (IL1R−/−). The administration of β-glucan may be used as a novel strategy in the treatment of mycobacterial infections and possibly as an adjuvant to improve anti-tuberculosis vaccines.
2. Del Cornò M et al. Shaping the Innate Immune Response by Dietary Glucans: Any Role in the Control of Cancer? Cancers 2020, 12, 15. DOI: 10.3390/cancers12010155. ABSTRACT: β-glucans represent a heterogeneous group of naturally occurring and biologically active polysaccharides found in many kinds of edible mushrooms, baker’s yeast, cereals and seaweeds, whose health-promoting effects have been known since ancient times. These compounds can be taken orally as food supplements or as part of daily diets, and are safe to use, nonimmunogenic and well tolerated. A main feature of β-glucans is their capacity to function as biological response modifiers, exerting regulatory effects on inflammation and shaping the effector functions of different innate and adaptive immunity cell populations. The potential to interfere with processes involved in the development or control of cancer makes β-glucans interesting candidates as adjuvants in antitumor therapies as well as in cancer prevention strategies. Here, the regulatory effects of dietary β-glucans on human innate immunity cells are reviewed and their potential role in cancer control is discussed.
3. Borchani C et al. Structural Characterization, Technological Functionality, and Physiological Aspects of Fungal B-D-glucans: A Review. Crit Rev Food Sci Nutr, 56(10:1746-52, PMIC 25830657, Jul 2016. ABSTRACT: Quote: “Thus, they [(1-3)(1-6)-B-glucans] are effective in inhibiting growth of cancer cells and metastasis and preventing bacterial infection. In humans, B-glucans reduce blood cholesterol, improve glucose absorption by body cells, and so help wound healing."
4. Vetvicka V et al. Glucans and Cancer: Comparison of Commercially Available B-glucans – Part IV. Anticancer Res, 38(3):1327-1333, Mar 2018. ABSTRACT: Quote: "Among the well-studied effects of B-glucans, we can mention stimulation of both humoral and cellular immunity, metabolic control of diabetes, stimulation of wound healing, stress reduction, attenuation of chronic fatigue syndrome, lowering cholesterol levels, and inhibition of cancer. …Chronic respiratory problems. In Japan, glucan has been widely used, for over 30 years, in the treatment of gastrointestinal cancer."
5. Davis JM et al. Effects of oat beta-glucan on innate immunity and infection after exercise stress. Med Sci Sports Exerc. 2004, Aug;36(8):1321-7. ABSTRACT: These data suggest that daily ingestion of beta glucan may offset the increased risk of upper respiratory tract infections (URTI) associated with exercise stress, which may be mediated, at least in part, by an increase in macrophage antiviral resistance.
6. Volman JJ et al. Dietary modulation of immune function by β-glucans. Physiology & Behavior, Volume 94, Issue 2, 23 May 2008, Pages 276-284. ABSTRACT: The immune response can be modulated by nutrients like β-glucans, which are glucose polymers that are major structural components of the cell wall of yeast, fungi, and bacteria, but also of cereals like oat and barley. In this review the current status concerning possibilities to modulate immune function by β-glucans is discussed. In vitro as well as in vivo studies in animals and humans show that especially β-glucans derived from fungi and yeast have immune modulating properties. Most frequently evaluated are effects on leukocyte activity, which has been suggested to contribute to the increased resistance against infections observed after β-glucan interventions.
7. Rondanelli M et al. The biological activity of beta-glucans. Minerva Med. 2009 Jun;100(3):237-45. ABSTRACT: This review summarizes the recent knowledge about the positive effect of beta glucans on human health. Quote: "A growing body of science indicates that beta-glucans promote health in a number of important ways. For instance, several studies have also shown that oat beta-glucans blunt the glycemic and insulin response. Moreover, beta-1,3-glucans improve the body's immune system defense against foreign invaders by enhancing the ability of macrophages, neutrophils and natural killer cells to respond to and fight a wide range of challenges such as bacteria, viruses, fungi, and parasites."
8. Vetvicka V et al. β-Glucan Improves Conditions of Chronic Fatigue in Mice by Stimulation of Immunity. The Open Biochemistry Journal, 2-18-2020. ABSTRACT: This study found that glucan supplementation strongly improved the suppressed phagocytosis and changes in cytokine and levels of oxidative stress markers caused by fatigue. In addition, glucan supplementation also increased the motor functioning of tested animals.
9. Akramiene D et al. Effects of ß-glucans on the immune system. Medicinia, 11 August 2007. ABSTRACT: This paper explains how ß-Glucans are naturally occurring polysaccharides. These glucose polymers are constituents of the cell wall of certain pathogenic bacteria and fungi. These substances increase host immune defense by activating the complement system, enhancing macrophages and natural killer cell function. ß-Glucans also show anti-carcinogenic activity. They can prevent oncogenesis due to the protective effect against potent genotoxic carcinogens. As immunostimulating agents, which act through the activation of macrophages and NK cell cytotoxicity, ß-glucan can inhibit tumor growth in promotion stage, too.
10. Graugaum HJ et al. A double-blind, randomized, placebo-controlled nutritional study using an insoluble yeast beta-glucan to improve the immune defense system. Dood Nutr Sci, 3(6):738-746, June 2012. ABSTRACT: In a placebo-controlled, double-blind, randomized clinical trial, the effect of an insoluble yeast beta-glucan preparation on the incidences of common colds and its effect on common cold symptoms were compared to placebo. …the beta-glucan group had significantly less infections compared to placebo. Beta-glucan significantly reduced the typical cold symptoms ('sore throat and/or difficulty swallowing', 'hoarseness and/or cough' and 'runny nose') as opposed to placebo. The present study demonstrated a prophylactic [preventative] effect of yeast beta-glucan on the occurrence of common colds as opposed to placebo. In addition, when these episodes occurred, they were from the beginning less pronounced and subsided faster.
11. Carlos AF et al. β-Glucan successfully stimulated the immune system in different jawed vertebrate species. Comparative Immunology, Microbiology and Infectious Diseases, Volume 62, February 2019, Pages 1-6. ABSTRACT: Several reports have shown the positive effects of β-glucans on the immune system. For 28 days, scientists, fed four different vertebrate species: mice, dogs, piglets and chicks, with two β-glucan molecules (BG01 and BG02). They measured the serum interleukin 2 as an indicator of innate immune response, the neutrophils and monocytes phagocytosis index as a cellular response, and antibody formation as an adaptive response. The results clearly showed that the different β-glucan molecules exhibited biologically differently behaviors, but both molecules stimulate the immune system in a similar pattern in these four species. This finding suggests that vertebrates shared similar mechanisms/patterns in recognizing the β-glucans and confirms the benefits of β-glucans across different vertebrate species.
12. Garcia-Valtanen p et al. Evaluation of trained immunity by β-1, 3 (D)-glucan on murine monocytes in vitro and duration of response in vivo. Immunology and Cell Biology (2017) 95, 601–610; DOI: 10.1038/icb.2017.13. ABSTRACT: The β-1, 3 (D)-glucan (β-glucan) present in the cell wall of Candida albicans induces epigenetic changes in human monocytes resulting in primed macrophages exhibiting increased cytokine responsiveness to reinfection. This phenomenon is referred to as trained immunity or innate immune memory. However, whether β-glucan can reprogramme murine monocytes in vitro or induce lasting effects in vivo has yet to be elucidated. Thus, purified murine spleen-derived monocytes were primed with β-glucan in vitro and assessed for markers of differentiation and survival. Important macrophage cell markers during monocyte-to- macrophage differentiation were downregulated and survival enhanced due to partial inhibition of apoptosis. Increased survival and not the β-glucan training effect explained the elevated production of tumour necrosis factor-α (TNFα) and interleukin-6 (IL-6) induced by subsequent lipopolysaccharide (LPS) challenge. In vivo, 4 days after systemic administration of β-glucan, mice were more responsive to LPS challenge as shown by the increased serum levels of TNFα, IL-6 and IL-10, an effect shown to be short lived as enhanced cytokine production was lost by day 20. Here, we have characterised murine macrophages derived from β-glucan-primed monocytes based on their surface marker expression and for the first time provide evidence that the training effect of β-glucan in vivo declines within a 3-week period.
13. Paris S et al. β-Glucan-Induced Trained Immunity in Dogs. Front. Immunol. 09 October 2020. 11:566893. DOI: 10.3389/fimmu.2020.566893. ABSTRACT: Several observations in the world of comparative immunology in plants, insects, fish and eventually mammals lead to the discovery of trained immunity in the early 2010’s. The first demonstrations provided evidence that innate immune cells were capable of developing memory after a first encounter with some pathogens. Trained immunity in mammals was initially described in monocytes with the Bacille Calmette-Guerin vaccine (BCG) or prototypical agonists like β-glucans. This phenomenon relies on epigenetic and metabolic modifications leading to an enhanced secretion of inflammatory cytokines when the host encounters homologous or heterologous pathogens. The objective of our research was to investigate the trained immunity, well-described in mouse and human, in other species of veterinary importance. For this purpose, we adapted an in vitro model of trained innate immunity in dogs. Blood enriched monocytes were stimulated with β-glucans and we confirmed that it induced an increased production of pro-inflammatory and anti-microbial compounds in response to bacterial stimuli. These results constitute the first demonstration of trained immunity in dogs and confirm its signatures in other mammalian species, with an implication of cellular mechanisms similar to those described in mice and humans regarding cellular epigenetics and metabolic regulations.
14. Byrne K et al. Differential induction of innate memory in porcine monocytes by b-glucan or bacillus Calmette-Guerin. Innate Immunity. 0(0) 1–13. 2020. DOI: 10.1177/1753425920951607. ABSTRACT: Innate immunomodulation via induction of innate memory is one mechanism to alter the host’s innate immune response to reduce or prevent disease. Microbial products modulate innate responses with immediate and lasting effects. Innate memory is characterized by enhanced (training) or depressed (tolerance) innate immune responses, including pro- inflammatory cytokine production, to secondary exposure following a priming event. To investigate the ability of b-glucans and bacillus Calmette-Guerin to induce innate training or tolerance in pig cells, porcine monocytes were cultured with priming agonist (b-glucans or bacillus Calmette-Guerin) then re-stimulated 5 d later with a heterologous microbial agonist to determine induction of innate memory. Priming with b-glucan from Saccharomyces cerevisiae depressed IL-1b and TNF-a cytokine responses to re-stimulation with LPS, indicative of a tolerized state. However, bacillus Calmette-Guerin priming induced a trained state in porcine monocytes, as LPS re-stimulation enhanced IL-1b and TNF-a gene expression and protein production. We present the first evidence of innate memory in pig monocytes, with bacillus Calmette-Guerin (training) or Saccharomyces cerevisiae b-glucan (tolerance). Induction of a trained or tolerized state in vitro is a first step to identify agonists to alter the innate immune system at the animal level with the intent of enhancing disease resistance.
15. Petit J et al. Long-lived effects of administering b-glucans: Indications for trained immunity in fish. Developmental and Comparative Immunology 64 (2016) 93e102. 2016. DOI: dx.doi.org/10.1016/j.dci.2016.03.003. ABSTRACT: Over the past decades, it has become evident that immune-modulation of fish with b-glucans, using injection, dietary or even immersion routes of administration, has stimulating but presumed short-lived effects on both intestinal and systemic immunity and can increase protection against a subsequent pathogenic challenge. Although the exact effects can be variable depending on, among others, fish species and administration route, the immune-stimulating effects of b-glucans on the immune system of fish appear to be universal. This review provides a condensed update of the most recent literature describing the effects of b-glucans on the teleost fish immune system. We shortly discuss possible mechanisms influencing immune-stimulation by b-glucans, including microbial composition of the gut, receptor recognition and downstream signalling. Of interest, in mammalian monocytes, b-glucans are potent inducers of trained immunity. First, we screened the literature for indications of this phenomenon in fish. Criteria that we applied include indications for at least one out of three features considered characteristic of trained immunity; (i) providing protection against a secondary infection in a T- and B- lymphocyte independent manner, (ii) conferring increased resistance upon re-infection and, (iii) relying on key roles for innate immune cell types such as natural killer cells and macrophages. We conclude that several indications exist that support the notion that the innate immune system of teleost fish can be trained. Second, we screened the literature for indications of long-lived effects on innate immunity of fish after administering b-glucans, a criterion which could help to identify key roles for macrophages on resistance to infection. We discuss whether b-glucans, as well-known immune-stimulants, are able to train the immune system of fish and argue in favour of further studies designed to specifically investigate this phenomenon in fish.
16. Kwanghook K et al. Algae-derived β-glucan enhanced gut health and immune responses of weaned pigs experimentally infected with a pathogenic E. coli. Animal Feed Science and Technology, Volume 248, February 2019. ABSTRACT: Quote: "Feed supplementation of algae-derived β-glucan alleviated diarrhea of F18 E. coli infected pigs by enhancing gut integrity. Feeding β-glucan also boosted host immune response against E. coli infection."
17. Abraham A et al. A novel vaccine platform using glucan particles for induction of protective responses against Francisella tularensis and other pathogens. Clinical and Experimental Immunology, 198: 143–152. 2019. DOI: 10.1111/cei.13356. ABSTRACT: accines are considered the bedrock of preventive medicine. However, for many pathogens, it has been challenging to develop vaccines that stimulate protective, long-lasting immunity. We have developed a novel approach using β-1,3-D-glucans (BGs), natural polysaccharides abundantly present in fungal cell walls, as a biomaterial platform for vaccine delivery. BGs simultaneously provide for receptor-targeted antigen delivery to specialized antigen-presenting cells together with adjuvant properties to stimulate antigen-specific and trained non-specific immune responses. This review focuses on various approaches of using BG particles (GPs) to develop bacterial and fungal vaccine candidates. A special case history for the development of an effective GP tularaemia vaccine candidate is highlighted.

Beta-Alanine

1. Harris RC et al. The absorption of orally supplied beta-alanine and its effect on muscle carnosine synthesis in human vastus lateralis. Amino Acids. 2006 May;30(3):279-89.
2. Stout JR et al. Effects of beta-alanine supplementation on the onset of neuromuscular fatigue and ventilatory threshold in women. Amino Acids. 2007;32(3):381-6.
3. Abe H. Role of histidine-related compounds as intracellular proton buffering constituents in vertebrate muscle. Biochemistry (Mosc). 2000 Jul;65(7):757-65.
4. Suzuki Y et al. High level of skeletal muscle carnosine contributes to the latter half of exercise performance during 30-s maximal cycle ergometer sprinting. Jpn J Physiol. 2002 Apr;52(2):199-205.
5. Derave W et al. Beta-Alanine supplementation augments muscle carnosine content and attenuates fatigue during repeated isokinetic contraction bouts in trained sprinters. J Appl Physiol. 2007 Nov;103(5):1736-43.
6. Hill CA et al. Influence of beta-alanine supplementation on skeletal muscle carnosine concentrations and high intensity cycling capacity. Amino Acids. 2007 Feb;32(2):225-33.

Caffeine

1. Nehlig A eta al. Caffeine and sports activity: a review. Int J Sports Med. 1994 Jul;15(5):215-23. doi: 10.1055/s-2007-1021049.
2. Yokokawa T et al. Caffeine increases myoglobin expression via the cyclic AMP pathway in L6 myotubes. Physiol Rep. 2021 May;9(9):e14869. doi: 10.14814/phy2.14869.
3. Caffeine for the Sustainment of Mental Task Performance: Formulations for Military Operations. Institute of Medicine (US) Committee on Military Nutrition Research. Washington (DC): National Academies Press (US); 2001.
4. Tabrizi R et al. The effects of caffeine intake on weight loss: a systematic review and dos-response meta-analysis of randomized controlled trials. Crit Rev Food Sci Nutr. 2019;59(16):2688-2696.
5. Ruiz-Moreno C et al. Caffeine increases whole-body fat oxidation during 1 h of cycling at Fatmax. Eur J Nutr. 2021 Jun;60(4):2077-2085.
6. Costill DL et al. Effects of caffeine ingestion on metabolism and exercise performance. Med Sci Sports. Fall 1978;10(3):155-8.

Casein Hydrolysate

1. Iwasa M et al. The milk casein hydrolysate-derived peptide enhances glucose uptake through AMP-activated protein kinase signaling pathway in skeletal muscle cells. Exp Physiol. 2020 Dec 28. PMID: 33369793 DOI: 10.1113/EP088770. New findings: What is the central question of this study? Previously, we have shown that supplementation of Lactobacillus helveticus-fermented milk and milk casein hydrolysate (MCH) improves glucose metabolism in exercised mice and humans. Common active ingredients contained in both Lactobacillus helveticus-fermented milk and MCH may enhance glucose metabolism, but the details are yet to be clarified. What is the main finding and its importance? MCH enhanced glucose uptake in skeletal muscle cells by stimulating AMP-activated kinase (AMPK), but not insulin signaling. Moreover, the MCH-derived specific peptide, Ile-Pro-Pro, mimicked this effect, suggesting a mechanism for the MCH-induced metabolic improvement. ABSTRACT: Improvement of glucose metabolism in the skeletal muscle has a key role in exercise performance and prevention of metabolic diseases. In our previous study, we had shown that intake of milk casein hydrolysate improves glucose metabolism in humans, but the mechanism of action was not elucidated. In this study, we aimed to investigate the mechanism of action of milk casein hydrolysate and its derived peptides on glucose uptake and glucose metabolic signaling in cultured skeletal muscle cells. Differentiated C2C12 myotubes were used for the experiments. The differentiated cells were incubated with milk casein hydrolysate, valine-proline-proline, and isoleucine-proline-proline. Subsequently, the rate of 2-deoxy-glucose uptake and the phosphorylation levels of insulin-dependent and -independent signaling factors were examined. We found that the rate of 2-deoxy-glucose uptake in both milk casein hydrolysate and isoleucine-proline-proline-treated cells was higher than that in the control cells. Immunoblotting assays showed that the phosphorylation levels of AMP-activated protein kinase, a rate-limiting factor in insulin-independent signaling, and that of liver kinase B1, an upstream factor of AMP-activated protein kinase, in both milk casein hydrolysate and isoleucine-proline-proline-treated cells were higher than those in the control cells. Such significant effects were not observed after treatment with valine-proline-proline. Moreover, the insulin-dependent signaling was not significantly affected under the different conditions. The findings of our study suggest that milk casein hydrolysate enhances glucose uptake by activating insulin-independent AMP-activated protein kinase signaling in skeletal muscle cells, which might be mediated by a milk casein hydrolysate-derived peptide, namely, isoleucine-proline-proline.
2. Saunders MJ et al. Carbohydrate and protein hydrolysate coingestions improvement of late-exercise time-trial performance. Int J Sport Nutr Exerc Metab. 2009 Apr;19(2):136-49. PMID: 19478339 DOI: 10.1123/ijsnem.19.2.136. ABSTRACT: This study examined whether a carbohydrate + casein hydrolysate (CHO+ProH) beverage improved time-trial performance vs. a CHO beverage delivering approximately 60 g CHO/hr. Markers of muscle disruption and recovery were also assessed. Thirteen male cyclists (VO2peak = 60.8 +/- 1.6 ml . kg-1 . min-1) completed 2 computer-simulated 60-km time trials consisting of 3 laps of a 20-km course concluding with a 5-km climb (approximately 5% grade). Participants consumed 200 ml of CHO (6%) or CHO+ProH beverage (6% + 1.8% protein hydrolysate) every 5 km and 500 ml of beverage immediately postexercise. Beverage treatments were administered using a randomly counterbalanced, double-blind design. Plasma creatine phosphokinase (CK) and muscle-soreness ratings were assessed immediately before and 24 hr after cycling. Mean 60-km times were 134.4 +/- 4.6 and 135.0 +/- 4.0 min for CHO+ProH and CHO beverages, respectively. All time differences between treatments occurred during the final lap, with protein hydrolysate ingestion explaining a significant (p < .05) proportion of between-trials differences over the final 20 km (44.3 +/- 1.6, 45.0 +/- 1.6 min) and final 5 km (16.5 +/- 0.6, 16.9 +/- 0.6 min). Plasma CK levels and muscle-soreness ratings increased significantly after the CHO trial (161 +/- 53, 399 +/- 175 U/L; 15.8 +/- 5.1, 37.6 +/- 5.7 mm) but not the CHO+ProH trial (115 +/- 21, 262 +/- 88 U/L; 20.9 +/- 5.3, 32.2 +/- 7.1 mm). Late-exercise time-trial performance was enhanced with CHO+ProH beverage ingestion compared with a beverage containing CHO provided at maximal exogenous oxidation rates during exercise. CHO+ProH ingestion also prevented increases in plasma CK and muscle soreness after exercise.
3. Demling RH et al. Effect of a hypocaloric diet, increased protein intake and resistance training on lean mass gains and fat mass loss in overweight police officers. Ann Nutr Metab. 2000;44(1):21-9. PMID: 10838463 DOI: 10.1159/000012817. ABSTRACT: We compare the effects of a moderate hypocaloric, high-protein diet and resistance training, using two different protein supplements, versus hypocaloric diet alone on body compositional changes in overweight police officers. A randomized, prospective 12-week study was performed comparing the changes in body composition produced by three different treatment modalities in three study groups. One group (n = 10) was placed on a nonlipogenic, hypocaloric diet alone (80% of predicted needs). A second group (n = 14) was placed on the hypocaloric diet plus resistance exercise plus a high-protein intake (1.5 g/kg/day) using a casein protein hydrolysate. In the third group (n = 14) treatment was identical to the second, except for the use of a whey protein hydrolysate. We found that weight loss was approximately 2.5 kg in all three groups. Mean percent body fat with diet alone decreased from a baseline of 27 +/- 1.8 to 25 +/- 1.3% at 12 weeks. With diet, exercise and casein the decrease was from 26 +/- 1.7 to 18 +/- 1.1% and with diet, exercise and whey protein the decrease was from 27 +/- 1.6 to 23 +/- 1.3%. The mean fat loss was 2. 5 +/- 0.6, 7.0 +/- 2.1 and 4.2 +/- 0.9 kg in the three groups, respectively. Lean mass gains in the three groups did not change for diet alone, versus gains of 4 +/- 1.4 and 2 +/- 0.7 kg in the casein and whey groups, respectively. Mean increase in strength for chest, shoulder and legs was 59 +/- 9% for casein and 29 +/- 9% for whey, a significant group difference. This significant difference in body composition and strength is likely due to improved nitrogen retention and overall anticatabolic effects caused by the peptide components of the casein hydrolysate.
4. Fangmann D et al. Differential effects of protein intake versus intake of a defined oligopeptide on FGF-21 in obese human subjects in vivo. Clin Nutr. 2020 Jun 17;S0261-5614(20)30296-X. PMID: 32600859 DOI: 10.1016/j.clnu.2020.06.006. ABStrACT: Background: FGF-21 is described as a powerful metabolic regulator with beneficial effects including glucose-lowering and improvement of insulin sensitivity without hypoglycaemia. On the other hand, FGF-21 is activated when muscle and other tissues are stressed by external effects or internal cellular pathogens that lead to shortcomings in metabolic balance. Previous results suggested that FGF-21 could be a promising target to develop future metabolic therapeutics. Purpose: The present study was performed to gain deeper insight into the regulation of FGF-21 by protein metabolism in obese human subjects. Methods: FGF-21 serum concentrations were measured in a cohort of n = 246 obese humans ± type 2 diabetes mellitus (T2DM) (median age 53.0 [46.0; 60.0] years and BMI 40.43 [35.11; 47.24] kg/m2) and related to the nutritional protein intake. In addition, the effect of a novel oligopeptide purified from a β-casein hydrolysate on FGF-21 was examined in vitro in liver cells and in vivo in a human intervention study with the main focus on metabolic inflammation including 40 mainly obese subjects (mean age 41.08 ± 9.76 years, mean BMI 38.29 ± 9.4 kg/m2) in a randomized 20 weeks double-blind cross-over design. Main findings: In the cohort analysis, FGF-21 serum concentrations were significant lower with higher protein intake in obese subjects without T2DM but not in obese subjects with T2DM. Furthermore, relative methionine intake was inversely related to FGF-21. While global protein intake in obesity was inversely associated with FGF-21, incubation of HepG2 cells with a β-casein oligopeptide increased FGF-21 expression in vitro. This stimulatory effect was also present in vivo, since in the clinical intervention study treatment of obese subjects with the β-casein oligopeptide for 8 weeks significantly increased FGF-21 serum levels from W0 = 23.86 pg/mL to W8 = 30.54 pg/mL (p < 0.001), while no increase was found for placebo. Conclusion: While the total nutritional protein intake is inversely associated with FGF-21 serum levels, a purified and well characterised oligopeptide is able to induce FGF-21 serum levels in humans. These findings suggest a differential role of various components of protein metabolism on FGF-21, rather than this factor being solely a sensor of total nutritional protein intake.

CDP Choline

1. Cavun S et al. CDP-choline increases plasma ACTH and potentiates the stimulated release of GH, TSH and LH: the cholinergic involvement. Fundam Clin Pharmacol. 2004 Oct;18(5):513-23. ABSTRACT: In the present study, we investigated the effect of intracerebroventricular (i.c.v.) administration of cytidine-5'-diphosphate (CDP) choline on plasma adrenocorticotropin (ACTH), serum growth hormone (GH), thyroid stimulating hormone (TSH), follicle stimulating hormone (FSH) and luteinizing hormone (LH) levels in conscious rats. The involvement of cholinergic mechanisms in these effects was also determined. In basal conditions, CDP-choline (0.5, 1.0 and 2.0 micromol, i.c.v.) increased plasma ACTH levels dose- and time-dependently, but it did not affect the TSH, GH, FSH and LH levels. In stimulated conditions, i.c.v. administration of CDP-choline (1 micromol, i.c.v.) produced an increase in clonidine-stimulated GH, thyrotyropin-releasing hormone (TRH)-stimulated TSH, LH-releasing hormone (LHRH)-stimulated LH, but not FSH levels. Injection of equimolar dose of choline (1 micromol, i.c.v.) produced similar effects on hormone levels, but cytidine (1 micromol, i.c.v.) failed to alter plasma levels of these hormones. Pretreatment with hemicholinium-3, a neuronal high affinity choline uptake inhibitor, (20 microg, i.c.v.) completely blocked the observed hormone responses to CDP-choline. The increase in plasma ACTH levels induced by CDP-choline (1 micromol, i.c.v.) was abolished by pretreatment with mecamylamine, a nicotinic receptor antagonist, (50 microg, i.c.v.) but not atropine, a muscarinic receptor antagonist, (10 microg, i.c.v.). The increase in stimulated levels of serum TSH by CDP-choline (1 micromol, i.c.v.) was blocked by atropine but not by mecamylamine pretreatment. However, CDP-choline induced increases in serum GH and LH levels were greatly attenuated by both atropine and mecamylamine pretreatments. The results show that CDP-choline can increase plasma ACTH and produce additional increases in serum levels of TSH, GH and LH stimulated by TRH, clonidine and LHRH, respectively. The activation of central cholinergic system, mainly through the presynaptic mechanisms, was involved in these effects. Central nicotinic receptors solely mediated the increase in plasma ACTH levels while the activation of central muscarinic receptors was involved in the increase in TSH levels. Both muscarinic and nicotinic receptor activations, separately, mediated the increases in serum GH and LH levels after CDP-choline.
2. Ceda GP et al. Effects of cytidine 5'-diphosphocholine administration on basal and growth hormone-releasing hormone-induced growth hormone secretion in elderly subjects. Acta Endocrinol (Copenh). 1991 May;124(5):516-20. ABSTRACT: The basal and GH-releasing hormone-stimulated secretion of GH declines in the elderly. We tested the ability of cytidine 5'-diphosphocholine, a drug used in the treatment of stroke and Parkinson's disease, to alter GH secretion in 11 healthy elderly volunteers, aged 69-84. Each subject received an iv infusion of 2 g of cytidine 5'-diphosphocholine or normal saline. GHRH and TRH were also administered during cytidine 5'-diphosphocholine infusions. The infusion of cytidine 5'-diphosphocholine induced a 4-fold (p less than 0.05) increase in serum GH levels over basal values. A small increase in GH was seen after GHRH administration. However, the addition of GHRH to the cytidine 5'-diphosphocholine infusion resulted in a GH response which was significantly greater than that seen after GHRH alone; the integrated concentration of GH was more than 2-fold greater in the cytidine 5'-diphosphocholine treated group (706.85 +/- 185.1 vs 248.9 +/- 61.4 micrograms.l-1.(120 min)-1; p = 0.01). The PRL and TSH responses to TRH were not significantly affected by cytidine 5'-diphosphocholine infusion, indicating that dopaminergic mechanisms are not involved. These studies demonstrate that cytidine 5'-diphosphocholine can enhance basal and GHRH-stimulated GH release in the elderly, but the mechanism of action of the drug remains unclear.
3. Secades JJ et al. Citicoline: pharmacological and clinical review, 2006 update. Methods Find Exp Clin Pharmacol. 2006 Sep;28 Suppl B:1-56. ABSTRACT: Cytidine 5'-diphosphocholine, CDP-choline, or citicoline is an essential intermediate in the biosynthetic pathway of structural phospholipids in cell membranes, particularly phosphatidylcholine. Following administration by both the oral and parenteral routes, citicoline releases its two main components, cytidine and choline. Absorption by the oral route is virtually complete, and bioavailability by the oral route is therefore approximately the same as by the intravenous route. Once absorbed, citicoline is widely distributed throughout the body, crosses the blood-brain barrier and reaches the central nervous system (CNS), where it is incorporated into the membrane and microsomal phospholipid fraction. Citicoline activates biosynthesis of structural phospholipids of neuronal membranes, increases brain metabolism, and acts upon the levels of different neurotransmitters. Thus, citicoline has been experimentally shown to increase norepinephrine and dopamine levels in the CNS. Owing to these pharmacological mechanisms, citicoline has a neuroprotective effect in hypoxic and ischemic conditions, decreasing the volume of ischemic lesion, and also improves learning and memory performance in animal models of brain aging. In addition, citicoline has been shown to restore the activity of mitochondrial ATPase and membrane Na+/K+ATPase, to inhibit activation of certain phospholipases, and to accelerate reabsorption of cerebral edema in various experimental models. Citicoline has also been shown to be able to inhibit mechanisms of apoptosis associated to cerebral ischemia and in certain neurodegeneration models, and to potentiate neuroplasticity mechanisms. Citicoline is a safe drug, as shown by the toxicological tests conducted, that has no significant systemic cholinergic effects and is a well tolerated product. These pharmacological characteristics and the action mechanisms of citicoline suggest that this product may be indicated for treatment of cerebral vascular disease, head trauma (HT) of varying severity, and cognitive disorders of different causes. In studies conducted in the treatment of patients with HT, citicoline was able to accelerate recovery from post-traumatic coma and neurological deficits, achieving an improved final functional outcome, and to shorten hospital stay in these patients. Citicoline also improved the mnesic and cognitive disorders seen after HT of minor severity that constitute the so-called post-concussional syndrome. In the treatment of patients with acute ischemic cerebral vascular disease, citicoline accelerates recovery of consciousness and motor deficit, achieves a better final outcome, and facilitates rehabilitation of these patients. The other major indication of citicoline is for treatment of senile cognitive impairment, either secondary to degenerative diseases (e.g. Alzheimer disease) or to chronic cerebral vascular disease. In patients with chronic cerebral ischemia, citicoline improves scores in cognitive rating scales, while in patients with senile dementia of the Alzheimer type it stops the course of disease, and neuroendocrine, neuroimmunomodulatory, and neurophysiological benefits have been reported. Citicoline has also been shown to be effective in Parkinson disease, drug addictions, and alcoholism, as well as in amblyopia and glaucoma. No serious side effects have occurred in any series of patients treated with citicoline, which attests to the safety of treatment with citicoline.
4. Serra F et al. Effect of CDP-choline on senile mental deterioration. Multicenter experience on 237 cases. Minerva Med. 1990 Jun;81(6):465-70. ABSTRACT: The efficacy of CDP-choline (1000 mg/die) administered for two 21-day treatment cycles, with a one-week wash-out period between them, was evaluated in out and in-patients suffering from mild to moderate brain aging. The study was performed on 237 fully evaluable patients with the use of the reduced geriatric scale of Plutchik and al., for clinical evaluation of the symptomatology. The clinical data obtained demonstrate that treatment with CDP-choline is able to determine an improvement of symptomatology since the 1st cycle of therapy (p less than 0.001), and a further improvement in the 2nd cycle (p less than 0.001). Particularly, the therapeutic effect of the 1st cycle is persistent in the intermediate wash-out period (suspension of treatment) with a further decrease, of symptomatology regarding some items of Plutchik's scale (p less than 0.01). Finally, treatment with CDP-choline 1000 mg/die for two 21-day cycles in 237 patients suffering from brain aging determined a statistically significant improvement of the cognitive and behavioural parameters taken into consideration: independence/autonomous life; human relations/social life; interest and attentive capacity; individual behaviour. Therefore citicoline is confirmed as a valid therapeutic remedy for the clinical, functional and social recovery of these patients.
5. Di Trapani G et al. Citicoline in the treatment of cognitive and behavioral disorders in pathologic senile decline. Clin Ter. 1991 Jun 30;137(6):403-13. ABSTRACT: A three months study was performed on 150 aging patients with primary memory deficits in order to verify the effectiveness of CDP-Choline, administered in repeated cycles of four weeks, with an interval of one week between cycles, in improving patients' cognitive and behavioral efficiency and in stabilizing their cognitive decline. Objective measures of memory and attention, and a behavioral rating scale were used to assess treatment effects. CDP-Choline treatment demonstrated both symptomatic efficacy and a long lasting effect on cognition and behavior of these patients. Level of activation and attention responsiveness improved during treatment cycles and no further changes were identified of these variables in the follow-up period. Measures related to specific memory functioning showed, besides improvements during treatment, after-effects still active in the follow-up period, suggesting a long lasting change of the cognitive decline trend characteristic of these patients.
6. Petkov VD et al. Effect of CDP-choline on learning and memory processes in rodents. Methods Find Exp Clin Pharmacol. 1992 Oct;14(8):593-605. ABSTRACT: The effects of cytidine (5') diphosphocholine (CDP-choline) on learning and memory were studied using conditioned reflex methods for passive avoidance and active avoidance with punishment reinforcement (step-through, step-down, shuttle box and maze), for active avoidance with alimentary reinforcement (staircase maze), and the Morris water maze. The majority of experiments involved comparative studies of the nootropic drugs meclofenoxate and/or piracetam. CDP-choline was administered orally, in some of the experiments also intraperitoneally, at doses of 10-500 mg/kg body weight once or twice daily for 5 or 7 days. In separate cases only single doses were administered. Trainings started one hour after the last dose of the drugs. Retention tests were given 3 h, 24 h, 7 days or 10 days after training. The results obtained with the different methods document CDP-choline's ability to improve learning and memory in rats and mice. No essential differences in the effects of CDP-choline were established upon oral and intraperitoneal administration of the drug. The learning- and memory-facilitating effects of CDP-choline were similar to those of meclofenoxate and piracetam. The results of the present study permit us to define CDP-choline as a substance capable of improving cognitive levels.
7. Spiers PA et al. Citicoline improves verbal memory in aging. Arch Neurol. 1996 May;53(5):441-8. ABSTRACT: Objective: To test the verbal memory of older volunteers given citicoline. Design: A randomized, double-blind, placebo-controlled, parallel group design was employed in the initial study. After data analysis, a subgroup was identified whose members had relatively inefficient memories. These subjects were recruited for a second study that used a crossover design. The subjects took either placebo or citicoline, 1000 mg/d, for 3 months in the initial study. In the crossover study, subjects took both placebo and citicoline, 2000 mg/d, each for 2 months. SUBJECTS: The subjects were 47 female and 48 male volunteers 50 to 85 years old. They were screened for dementia, memory disorders, and other neurological problems. Of the subjects with relatively inefficient memories, 32 participated in the crossover study. Main Outcome Measure: Verbal memory was tested at each study visit using a logical memory passage. Plasma choline concentrations were measured at baseline; at days 30, 60, and 90 in the initial study; and at day 60 of each treatment condition in the crossover study. Plasma choline concentrations and memory scores were analyzed using repeated-measures analysis of variance and covariance, followed by planned comparisons when appropriate. RESULTS: In the initial study, citicoline therapy improved delayed recall on logical memory only for the subjects with relatively inefficient memories. In the crossover study, the higher dosage of citicoline was clearly associated with improved immediate and delayed logical memory. CONCLUSIONS: Citicoline therapy improved verbal memory functioning in older individuals with relatively inefficient memories. Citicoline may prove effective in treating age-related cognitive decline that may be the precursor of dementia.
8. Fioravanti M et al. Cytidinediphosphocholine (CDP-choline) for cognitive and behavioural disturbances associated with chronic cerebral disorders in the elderly. Cochrane Database Syst Rev. 2004;(2):CD000269. doi: 10.1002/14651858.CD000269.pub2. BACKGROUND: CDP-choline (cytidine 5'-diphosphocholine) is a precursor essential for the synthesis of phosphatidylcholine, one of the cell membrane components that is degraded during cerebral ischaemia to free fatty acids and free radicals. Animal studies suggest that CDP-choline may protect cell membranes by accelerating resynthesis of phospholipids. CDP-choline may also attenuate the progression of ischaemic cell damage by suppressing the release of free fatty acids. CDP-choline is the endogenous compound normally produced by the organism. When the same substance is introduced as a drug it can be called citicoline.CDP-choline is mainly used in the treatment of disorders of a cerebrovascular nature. The many years of its presence in the clinical field have caused an evolution in dosage, method of administration, and selection criteria of patients to whom the treatments were given. Modalities of the clinical studies, including length of observation, severity of disturbance, and methodology of evaluation of the results were also heterogeneous. In spite of uncertainties about its efficacy due to these complexities, CDP-choline is a frequently prescribed drug for cognitive impairment in several European countries, especially when the clinical picture is predominantly one of cerebrovascular disease, hence the need for this review. Due to its effects on the adrenergic and dopaminergic activity of the CNS, CDP-choline has also been used as an adjuvant in the treatment of Parkinson's disease. OBJECTIVES: To assess the efficacy of CDP-choline (cytidinediphosphocholine) in the treatment of cognitive, emotional, and behavioural deficits associated with chronic cerebral disorders in the elderly. SEARCH STRATEGY: The trials were identified from a last updated search of the Specialized Register of the Cochrane Dementia and Cognitive Improvement Group on 22 April 2004 using the terms CDP-choline, CDP, citicoline, cytidine diphosphate choline or diphosphocholine. The Register contains records from all major health-care databases and many ongoing trials databases and is updated regularly. SELECTION CRITERIA: All relevant unconfounded, double-blind, placebo-controlled, randomized trials of CDP-choline for cognitive impairment due to chronic cerebral disorders were considered for inclusion in the review. DATA COLLECTION AND ANALYSIS: Two reviewers independently reviewed the included studies, extracted the data, and pooled it when appropriate and possible. The pooled odd ratios (95% Confidence Interval (CI)) or the average differences (95% CI) were estimated. No intention-to-treat data were available from the studies included. MAIN RESULTS: Fourteen studies were included in this review. Some of the included studies did not present numerical data suitable for analysis. Description of participants varied over the years and by type of disorders and severity, and ranged from aged individuals with subjective memory disorders to patients with Vascular Cognitive Impairment (mild to moderate), Vascular Dementia or Senile Dementia (mild to moderate). Seven of the included studies observed the subjects for a period between 20 to 30 days, one study was of 6 weeks duration, four studies used periods extending over 2 and 3 months, one study observed continuous administration over 3 months and one study was prolonged, with 12 months of observation. The studies were heterogeneous in dose, modalities of administration, inclusion criteria for subjects, and outcome measures. Results were reported for the domains of attention, memory testing, behavioural rating scales, global clinical impression and tolerability. There was no evidence of a beneficial effect of CDP-choline on attention. There was evidence of benefit of CDP-choline on memory function and behaviour. The drug was well tolerated. AUTHORS' CONCLUSIONS: There was some evidence that CDP-choline has a positive effect on memory and behaviour in at least the short to medium term. The evidence of benefit from global impression was stronger, but is still limited by the duration of the studies. Further research with CDP-choline should focus on longer term studies in subjects who have been diagnosed with currently accepted standardised criteria, especially Vascular Mild Cognitive Impairment (VaMCI) or vascular dementia.
9. D'Orlando KJ et al.Citicoline (CDP-choline): mechanisms of action and effects in ischemic brain injury. Neurol Res. 1995 Aug;17(4):281-4. ABSTRACT: Citicoline is approved in Europe and Japan for use in stroke, head trauma and other neurological disorders. It is presently being evaluated in phase II/III stroke trials in the United States. Exogenous administration of CDP-choline provides both choline and cytidine which access the brain and serve as substrates for the synthesis of phosphatidylcholine, a primary neuronal membrane component; the choline also enhances brain acetylcholine synthesis. Membrane repair and regeneration is necessary for recovery from stroke. Furthermore, citicoline may alleviate free fatty acid-induced toxicity which accompanies ischemic insult. Data from many pre-clinical and clinical trials support the hypothesis that citicoline may be a safe and effective treatment for stroke.

Citrulline Malate

1. Bendahan D et al. Citrulline/malate promotes aerobic energy production in human exercising muscle. Br J Sports Med. 2002 Aug;36(4):282-9. doi: 10.1136/bjsm.36.4.282.
2. Callis A et al. Activity of citrulline malate on acid-base balance and blood ammonia and amino acid levels. Study in the animal and in man. Arzneimittelforschung. 1991 Jun;41(6):660-3.
3. Bendahan D et al. Citrulline/malate promotes aerobic energy production in human exercising muscle. Br J Sports Med. 2002 Aug;36(4):282-9.

Cranberry

1. Vidlar A et al. The effectiveness of dried cranberries ( Vaccinium macrocarpon) in men with lower urinary tract symptoms. Brit J Nutr, 2010 Oct;104(8):118-9. Summary: The objective of this study was to evaluate the efficacy and tolerability of cranberry (Vaccinium macrocarpon) powder in men at risk of prostate disease with lower urinary tract symptoms (LUTS), elevated prostate-specific antigen (PSA), negative prostate biopsy and clinically confirmed chronic non-bacterial prostatitis. The results of the present trial are the first firm evidence that cranberries may ameliorate LUTS, independent of benign prostatic hyperplasia or C-reactive protein level.
2. Wang CH et al. Cranberry-Containing Products for Prevention of Urinary Tract Infections in Susceptible Populations: A Systematic Review and Meta-analysis of Randomized Controlled Trials. Arch Intern Med. 2012;172(13):988-996. Summary: The results of the present meta-analysis support that consumption of cranberry-containing products may protect against UTIs in certain populations.
3. Luis A et al. Can Cranberries Contribute to Reduce the Incidence of Urinary Tract Infections? A Systematic Review with Meta-Analysis and Trial Sequential Analysis of Clinical Trials. The Journal of Urology. Volume 3 198. Issue 3. September 2017. Pages 614-621. Summary: The findings clearly showed the potential use of cranberries for the clinical condition of urinary tract infection. Cranberry products significantly reduced the incidence of urinary tract infections as indicated by the weighted risk ratio (0.6750, 95% CI 0.5516–0.7965, p < 0.0001). The results could be used by physicians to recommend cranberry ingestion to decrease the incidence of urinary tract infections, particularly in individuals with recurrent urinary tract infections. This would also reduce the administration of antibiotics, which could be beneficial since antibiotics can lead to the worldwide emergence of antibiotic resistant microorganisms.

Curcumin (Lipidized)

1. Gota VS et al. Safety and pharmacokinetics of a solid lipid curcumin particle formulation in osteosarcoma patients and healthy volunteers. J Agric Food Chem. 2010 Feb 24;58(4):2095-9. doi: 10.1021/jf9024807. ABSTRACT: Curcumin is the lipid-soluble antioxidant compound obtained from the rhizome of Curcuma longa Linn, also known as turmeric. Curcumin targets multiple chemotherapeutic and inflammatory pathways and has demonstrated safety and tolerability in humans, supporting its potential as a therapeutic agent; however, the clinical literature lacks conclusive evidence supporting its use as a therapeutic agent due to its low bioavailability in humans. The purpose of this study was to quantify plasma levels of free curcumin after dosing of a solid lipid curcumin particle (SLCP) formulation versus unformulated curcumin in healthy volunteers and to determine its tolerability and dose-plasma concentration relationship in late-stage osteosarcoma patients. Doses of 2, 3, and 4 g of SLCP were evaluated in 11 patients with osteosarcoma. Plasma curcumin levels were measured using a validated high-performance liquid chromatography method. The limit of detection of the assay was 1 ng/mL of curcumin. In healthy subjects, the mean peak concentration of curcumin achieved from dosing 650 mg of SLCP was 22.43 ng/mL, whereas plasma curcumin from dosing an equal quantity of unformulated 95% curcuminoids extract was not detected. In both healthy individuals and osteosarcoma patients, high interindividual variability in pharmacokinetics and nonlinear dose dependency was observed, suggesting potentially complex absorption kinetics. Overall, good tolerability was noted in both healthy and osteosarcoma groups.
2. Cox KHM et al. Further evidence of benefits to mood and working memory from lipidated curcumin in healthy older people: A 12-week, double-blind, placebo-controlled, partial replication study. Nutrients. 2020 Jun 04. 12(6): 1678. doi: 10.3390/nu12061678. ABSTRACT: A partial replication study by researchers at Swinburne University reveals [lipidated curcumin] improves aspects of mood, memory, and working memory in a healthy older cohort. The pattern of results is consistent with improvements in hippocampal function and may hold promise for alleviating cognitive decline in some populations. This study examined a similar population with slightly elevated cognitive abilities, while eliciting similar results to the first clinical published in 2014 - see Cox KH et al, 2014.
3. Esfahani K et al. A phase I open prospective cohort trial of curcumin plus tyrosine kinase inhibitors for EGFR-mutant advanced non-small cell lung. J Clin Oncol. 2019. 37(15_suppl): e20611-e20611. doi: 10.1200/JCO.2019.37.15_suppl.e20611. ABSTRACT: This study further provides evidence that short-term use of Longvda® curcumin in patients is feasible and safe. Researchers report high treatment adherence and improved quality of life with curcumin. These findings, as well as efficacy data and the effect of curcumin on other inflammation-associated biomarkers, warrant investigation in a larger phase 2 study.
4. Scholey A et al. Curcumin improves hippocampal function in healthy older adults: A three month randomized controlled trial. Poster Presentation in: 13th European Nutrition Conference - Malnutrition in an Obese World: European Perspectives (FENS). Dublin, Ireland. 2019: P3-01-02. ABSTRACT: Additional results confirm that [lipidated curcumin] improves aspects of mood, memory, and working memory in a healthy older cohort. The pattern of results is consistent with improvements in hippocampal function and may hold promise for alleviating cognitive decline in some populations.
5. Scholey A et al. A highly bioavailable curcumin extract improves neurocognitive function and mood in healthy older people: A 12-week randomized, double-blind, placebo-controlled trial (OR32-05-19). Current Dev Nut. 2019 Jun. Poster Presentation. Volume 3(Issue Supplement 1): nzz052.OR32–05–19. doi: 10.1093/cdn/nzz052.OR32-05-19. ABSTRACT: Previously, researchers at Swinburne University showed significant improvements in measures of memory, attention, fatigue, stress, and mood (Cox KH et al, 2015). This trial was a follow up to the results previously seen in 1 and 3 hrs and in 4-weeks. The results of this second trial further confirm that a single daily dose of 400mg of [lipidated curcumin] improves aspects of mood and working memory in healthy older adults, with measures at 12-weeks.
6. Gupte PA et al. Evaluation of the efficacy and safety of capsule solid lipid curcumin particles in knee: A pilot clinical study. J Inflamm Res. 2019. 12: 145-152. doi: 10.2147/JIR.S205390. ABSTRACT: A comparative examination of (solid lipid curcumin particles) showed that administration was not only faster-acting and safe, but had equal efficacy to the control.
7. Koronyo, Y et al. Retinal amyloid pathology and proof-of-concept imaging trial. JCI Insight. 2017. 2(16). doi: 10.1172/jci.insight.93621. ABSTRACT: A proof-of-concept retinal imaging trial showing increased fluorescent intensity in retinal amyloid deposits and the highest brain concentrations of free curcumin obtained with [lipidated curcumin]. This trial confirmed one more time the ability of [lipidated curcumin] to deliver free curcumin to targeted tissues, more specifically the brain and retina, and to support cognitive and complete neuronal health. *Winner of NutraIngredients-USA Nutrition Research Project of the Year 2019 for ground-breaking initiatives as "most innovative and impactful nutrition research project pushing the boundaries of nutritional science." Read more here.
8. Santos-Parker JR et al. Curcumin supplementation improves vascular endothelial function in healthy middle-aged and older adults by increasing nitric oxide bioavailiability and reducing oxidative stress. Aging. 2017 Jan. 3. Vol 9(No1): 187-208.
9. McFarlin et al. Reduced inflammatory and muscle damage biomarkers following oral supplementation with bioavailable curcumin. University of North Texas. BBA Clinical. 2016 Feb 18. 5: 72-78. doi: 10.1016/j.bbacli.2016.02.003. ABSTRACT: Collectively, the findings demonstrated that consumption of [lipidated curcumin] (400mg/day) reduced key inflammatory biomarkers during recovery after exercise-induced muscle damage (EIMD). The observed improvements in biological inflammation may translate to faster recovery and improved functional capacity during subsequent exercise sessions.
10. Santos-Parker JR et al. Curcumin supplement improves vascular endothelial function in middle-aged and older adults. Geront. 2015 Dec. 55(Suppl 2): 195. doi: 10.1093/geront/gnv554.01. ABSTRACT: [Lipidated curcumin] administered at a dose of 2000mg/day (n=16), or placebo (n=13) for 12 weeks increased brachial artery flow-mediation dilation (FMDba) by 34% and forearm blood flow in response to incremental brachial artery infusions of acetylcholine (FBFach) by 44% in middle-aged and older (MA/O) adults (45-74 yrs). Findings support supplementation with [lipidated curcumin] improves endothelial-dependent dilation (EDD) in MA/O adults mediated, in part, by an increase in nitric oxide bioavailability.
11. Rafii MS et al. The biomarker initiative DSBI pilot: Proof of concept for deep phenotyping of biomarkers. Front Behav Neurosci. 2015. 9: 1-11. ABSTRACT: Retina, being part of the CNS, has previously been difficult to analyze directly; however, retinal amyloid imaging could now be a tool to demonstrate the presence of plaques in the brain in a non-invasive manner. In line with previous findings, this study supports [lipidated curcumin] quickly labeling retinal beta amyloid and inducing fluorescent plaque in the neural layers of the retina of humans.
12. Cox KH et al. Investigation of the effects of solid lipid curcumin on cognition and mood in a healthy older population. Centre for Human Psychopharmacology, Swinburne University. J Psychopharmacol. 2015 May. Vol 29(No 5): 642-651. doi: 10.1177/0269881114552744. ABSTRACT: This landmark study is one of the first to show a curcumin supplement improves cognitive function in healthy subjects. The trial recruited 60 subjects aged 60-80, and found daily [lipidated curcumin] (400mg) supplementation led to significant improvements in cognitive function versus the placebo group. Excellent safety was reported, including no dropouts or reports of gastrointestinal upset. Significant improvements were observed in measures for memory, attention, fatigue, stress, and mood in as little as one hour after the first dose.
13. Hazarey VK et al. Efficacy of curcumin in the treatment for oral health – A randomized clinical trial. Government Dental College and Hospital. Nagpur, Maharashtra, India. J Oral Maxillofac Pathol. 2015. 19: 145-52. doi: 10.4103/0973-029X.164524. ABSTRACT: A randomized, controlled clinical trial in 30 clinically diagnosed patients with OSF concluded that [lipidated curcumin] lozenges could be effective in combination strategies for the management of OSF in comparison to single therapeutic modality. In this study, 15 OSF patients in each group (test & control) were treated with either [lipidated curcumin] lozenges (400 mg lozenges for total daily dose of 2 g) or Tenovate ointment (clobetasol propionate (0.05%)). The treatment was given for 3 months and follow-up was done for 6 months.
14. Machida N et al. Effects of Solid, Lipid Curcumin Particles on alcohol metabolism - An expiatory and a randomized, double-blind, placebo-controlled, parallel-group crossover study. Jpn Pharmacol Ther. 2020 Apr. 48(5): 867-873. ABSTRACT: This study further provides evidence that [lipidated curcumin] curcumin is safe and efficacious. Previously examined in 2014, and recently published, researchers report reduced side effects typically associated with alcohol consumption and suggest that [lipidated curcumin] may offer liver health support through the acceleration of ethanol and acetaldehyde metabolism.
15. Frost S et al. Retinal amyloid fluorescence imaging predicts cerebral amyloid burden. Alz Dement. 2014. 10(4): P234-P235. ABSTRACT: Retinal Aβ plaques are similar to plaques in the brain. [lipidated curcumin's] ability to cross the BBB and its affinity for binding to amyloid beta have led to its use as a novel, more cost-effective alternative and imaging tool for screening through the eyes.
16. DiSilvestro et al. Diverse effects of a low-dose supplement of lipidated curcumin in healthy middle-aged people. The Ohio State University. Nutr. J. 2012 Sep 26. 11(79). doi: 10.1186/1475-2891-11-79. ABSTRACT: This study is believed to be the first curcumin trial in healthy people to show improvement in a number of key biomarkers related to healthy aging. Randomized, placebo-controlled study in 39 subjects showing excellent safety as well as significant improvements in markers supporting cognitive health, cardiovascular health, and anti-aging versus placebo.
17. Khattry N et al. Curcumin decreases cytokine levels involved in mucositis in autologous transplant setting: A pharmacokinetic-pharmacodynamic study. Poster presented at 54th American Society of Hematology (ASH) Annul Meeting. Atlanta, GA. 2012 Dec 08. Blood. 120(21): 3039. ABSTRACT: The absorption and efficacy of [lipidated curcumin] in lozenge form in a common oral inflammatory and fibrotic condition was tested compared to the standard of care (clobetasol steroid ointment). Subjects taking [lipidated curcumin] observed improvements in endpoints significantly better than those receiving steroid treatment; and therapeutic plasma levels were detected through buccal absorption.
18. Shah et al. Acute human pharmacokinetics of a lipid-dissolved turmeric extract. Planta Med. 2012. 48-PH5. ABSTRACT: This study concluded that a dose as low as 200mg of [lipidated curcumin] reaches blood levels of free curcumin required for healthy brain aging. Analyzed blood samples with and without the use of glucuronidase enzyme, finding very little of the glucuronidated form compared to previous studies on curcumin.
19. Pharmacokinetics of [lipidated curcumin]: Dose-concentration correlation. Unpublished, UCLA 2011-2012. ABSTRACT: Pilot studies demonstrating absorption and metabolism of [lipidated curcumin] using various dosage forms.
20. Gota et al. Safety and pharmacokinetics of a solid lipid curcumin particle formulation in patients and healthy volunteers. Tata Memorial Cancer Centre. J Ag Food Chem. 2010. 58(4): 2095-2099. ABSTRACT: Human bioavailability study demonstrating significantly greater plasma levels of free (unconjugated) curcumin after a single dose of [lipidated curcumin] in both healthy and disease states with 65x greater Cmax and >100x greater AUC than 95% curcuminoids.
21. [Lipidated curcumin] binds to amyloid in human CNS after a single dose. Unpublished.
22. A phase 1 open-label prospective cohort trial of curcumin plus tyrosin kinase inhibitors for EGFR-mutant advanced NSCLC. McGill U & Jewish General Hospital, Canada, Ongoing.

Cyanidin 3-Glucoside (C3G)

1. Guo H et al. Cyanidin-3-O-β-glucoside regulates fatty acid metabolism via an AMP-activated protein kinase-dependent signaling pathway in human HepG2 cells. Lipids Health Dis. 2012 Jan 13;11:10. doi: 10.1186/1476-511X-11-10. ABSTRACT: Background: Hepatic metabolic derangements are key components in the development of fatty liver disease. AMP-activated protein kinase (AMPK) plays a central role in controlling hepatic lipid metabolism through modulating the downstream acetyl CoA carboxylase (ACC) and carnitine palmitoyl transferase 1 (CPT-1) pathway. In this study, cyanidin-3-O-β-glucoside (Cy-3-g), a typical anthocyanin pigment was used to examine its effects on AMPK activation and fatty acid metabolism in human HepG2 hepatocytes. Results: Anthocyanin Cy-3-g increased cellular AMPK activity in a calmodulin kinase kinase dependent manner. Furthermore, Cy-3-g substantially induced AMPK downstream target ACC phosphorylation and inactivation, and then decreased malonyl CoA contents, leading to stimulation of CPT-1 expression and significant increase of fatty acid oxidation in HepG2 cells. These effects of Cy-3-g are largely abolished by pharmacological and genetic inhibition of AMPK. Conclusion: This study demonstrates that Cy-3-g regulates hepatic lipid homeostasis via an AMPK-dependent signaling pathway. Targeting AMPK activation by anthocyanin may represent a promising approach for the prevention and treatment of obesity-related nonalcoholic fatty liver disease.
2. Wei X et al. Cyanidin-3-O-β-glucoside improves obesity and triglyceride metabolism in KK-Ay mice by regulating lipoprotein lipase activity. J Sci Food Agric. 2011 Apr;91(6):1006-13. doi: 10.1002/jsfa.4275. ABSTRACT: Background: Cyanidin-3-O-β-glucoside (Cy-3-g)-rich foods have been reported to inhibit the onset of obesity, but whether the pure anthocyanin supplementation affects obesity remains uncertain. Results: Cy-3-g supplementation significantly reduced obesity, accumulation of fat in visceral adipose and liver tissues, and plasma triglyceride levels. Furthermore, adenosine monophosphate (AMP)-activated protein kinase phosphorylation (pAMPK) in the skeletal muscle and visceral adipose were significantly increased by Cy-3-g consumption. This was followed by the activation of lipoprotein lipase (LPL) in plasma and skeletal muscle but the suppression of this enzyme in visceral adipose. LPL activation in skeletal muscle cells and its suppression in adipocytes by Cy-3-g were blocked by inhibition of pAMPK. Conclusion: Our present data thus demonstrate that Cy-3-g improves obesity and triglyceride metabolism in KK-Ay mice. The underlying mechanism is found to be partly related to the activation of LPL in plasma and skeletal muscle, and inhibition of LPL in adipose tissue following the activation of pAMPK.
3. Guo H et al. Cyanidin 3-glucoside attenuates obesity-associated insulin resistance and hepatic steatosis in high-fat diet-fed and db/db mice via the transcription factor FoxO1. J Nutr Biochem. 2012 Apr;23(4):349-60. doi: 10.1016/j.jnutbio.2010.12.013. ABSTRACT: Obesity is a major risk factor for the development of type 2 diabetes, and both conditions are now recognized to possess significant inflammatory components underlying their pathophysiologies. Here, we hypothesized that cyanidin 3-glucoside (C3G), a typical anthocyanin reported to possess potent anti-inflammatory properties, would ameliorate obesity-associated inflammation and metabolic disorders, such as insulin resistance and hepatic steatosis in mouse models of diabesity. Male C57BL/6J obese mice fed a high-fat diet for 12 weeks and genetically diabetic db/db mice at an age of 6 weeks received dietary C3G supplementation (0.2%) for 5 weeks. We found that dietary C3G lowered fasting glucose levels and markedly improved the insulin sensitivity in both high-fat diet fed and db/db mice as compared with unsupplemented controls. White adipose tissue messenger RNA levels and serum concentrations of inflammatory cytokines (tumor necrosis factor-α, interleukin-6, and monocyte chemoattractant protein-1) were reduced by C3G, as did macrophage infiltration in adipose tissue. Concomitantly, hepatic triglyceride content and steatosis were alleviated by C3G. Moreover, C3G treatment decreased c-Jun N-terminal kinase activation and promoted phosphorylation and nuclear exclusion of forkhead box O1 after refeeding. These findings clearly indicate that C3G has significant potency in antidiabetic effects by modulating the c-Jun N-terminal kinase/forkhead box O1 signaling pathway and the related inflammatory adipocytokines.
4. Sasaki R et al. Cyanidin 3-glucoside ameliorates hyperglycemia and insulin sensitivity due to downregulation of retinol binding protein 4 expression in diabetic mice. Biochem Pharmacol. 2007 Dec 3;74(11):1619-27. doi: 10.1016/j.bcp.2007.08.008. ABSTRACT: Adipocyte dysfunction is strongly associated with the development of obesity and insulin resistance. It is accepted that the regulation of adipocytokine expression is one of the most important targets for the prevention of obesity and improvement of insulin sensitivity. In this study, we have demonstrated that anthocyanin (cyanidin 3-glucoside; C3G) which is a pigment widespread in the plant kingdom, ameliorates hyperglycemia and insulin sensitivity due to the reduction of retinol binding protein 4 (RBP4) expression in type 2 diabetic mice. KK-A(y) mice were fed control or control +0.2% of a C3G diet for 5 weeks. Dietary C3G significantly reduced blood glucose concentration and enhanced insulin sensitivity. The adiponectin and its receptors expression were not responsible for this amelioration. C3G significantly upregulated the glucose transporter 4 (Glut4) and downregulated RBP4 in the white adipose tissue, which is accompanied by downregulation of the inflammatory adipocytokines (monocyte chemoattractant protein-1 and tumor necrosis factor-alpha) in the white adipose tissue of the C3G group. These findings indicate that C3G has significant potency in an anti-diabetic effect through the regulation of Glut4-RBP4 system and the related inflammatory adipocytokines.
5. Takanori T et al. Anthocyanin enhances adipocytokine secretion and adipocyte-specific gene expression in isolated rat adipocytes. Biochem Biophys Res Commun. 2004 Mar 26;316(1):149-57. doi: 10.1016/j.bbrc.2004.02.031. ABSTRACT: Adipocyte dysfunction is strongly associated with the development of obesity and insulin resistance. It is accepted that the regulation of adipocytokine secretion or the adipocyte-specific gene expression is one of the most important targets for the prevention of obesity and amelioration of insulin sensitivity. In this study, we demonstrated that anthocyanins (cyanidin or cyanidin 3-glucoside) have the potency of a unique pharmacological function in isolated rat adipocytes. Treated adipocytes with anthocyanins enhanced adipocytokine (adiponectin and leptin) secretion and up-regulated the adipocyte specific gene expression without activation of PPARgamma in isolated rat adipocytes. The gene expression of adiponectin was also up-regulated in white adipose tissue in mice fed an anthocyanin supplemented diet. As one of the possible mechanisms, AMP-activated protein kinase activation would be associated with these changes, nevertheless, the AMP:ATP ratio was significantly decreased by administration of the anthocyanins. These data suggest that anthocyanins have a potency of unique therapeutic advantage and also have important implications for preventing obesity and diabetes.
6. Guo H et al. Cyanidin 3-glucoside protects 3T3-L1 adipocytes against H2O2- or TNF-alpha-induced insulin resistance by inhibiting c-Jun NH2-terminal kinase activation. Biochem Pharmacol. 2008 Mar 15;75(6):1393-401. doi: 10.1016/j.bcp.2007.11.016. ABSTRACT: Anthocyanins are naturally occurring plant pigments and exhibit an array of pharmacological properties. Our previous study showed that black rice pigment extract rich in anthocyanin prevents and ameliorates high-fructose-induced insulin resistance in rats. In present study, cyanidin 3-glucoside (Cy-3-G), a typical anthocyanin most abundant in black rice was used to examine its protective effect on insulin sensitivity in 3T3-L1 adipocytes exposed to H(2)O(2) (generated by adding glucose oxidase to the medium) or tumor necrosis factor alpha (TNF-alpha). Twelve-hour exposure of 3T3-L1 adipocytes to H(2)O(2) or TNF-alpha resulted in the increase of c-Jun NH(2)-terminal kinase (JNK) activation and insulin receptor substrate 1 (IRS1) serine 307 phosphorylation, concomitantly with the decrease in insulin-stimulated IRS1 tyrosine phosphorylation and cellular glucose uptake. Blocking JNK expression using RNA interference efficiently prevented the H(2)O(2)- or TNF-alpha-induced defects in insulin action. Pretreatment of cells with Cy-3-G reduced the intracellular production of reactive oxygen species, the activation of JNK, and attenuated H(2)O(2)- or TNF-alpha-induced insulin resistance in a dose-dependent manner. In parallel, N-acetyl-cysteine, an antioxidant compound, did not exhibit an attenuation of TNF-alpha-induced insulin resistance. Taken together, these results indicated that Cy-3-G exerts a protective role against H(2)O(2)- or TNF-alpha-induced insulin resistance in 3T3-L1 adipocytes by inhibiting the JNK signal pathway.
7. Tsuda T et al. Microarray profiling of gene expression in human adipocytes in response to anthocyanins. Biochem Pharmacol. Biochem Pharmacol. 2006 Apr 14;71(8):1184-97. doi: 10.1016/j.bcp.2005.12.042. ABSTRACT: Adipocyte dysfunction is strongly associated with the development of obesity and insulin resistance. It is accepted that the regulation of adipocytokine secretion or the adipocyte specific gene expression is one of the most important targets for the prevention of obesity and amelioration of insulin sensitivity. Recently, we demonstrated that anthocyanins, which are pigments widespread in the plant kingdom, have the potency of anti-obesity in mice and the enhancement adipocytokine secretion and its gene expression in adipocytes. In this study, we have shown the gene expression profile in human adipocytes treated with anthocyanins (cyanidin 3-glucoside; C3G or cyanidin; Cy). The human adipocytes were treated with 100 microM C3G, Cy or vehicle for 24 h. The total RNA from the adipocytes was isolated and carried out GeneChip microarray analysis. Based on the gene expression profile, we demonstrated the significant changes of adipocytokine expression (up-regulation of adiponectin and down-regulation of plasminogen activator inhibitor-1 and interleukin-6). Some of lipid metabolism related genes (uncoupling protein2, acylCoA oxidase1 and perilipin) also significantly induced in both common the C3G or Cy treatment groups. These studies have provided an overview of the gene expression profiles in human adipocytes treated with anthocyanins and demonstrated that anthocyanins can regulate adipocytokine gene expression to ameliorate adipocyte function related with obesity and diabetes that merit further investigation.
8. Tsuda T et al. Gene expression profile of isolated rat adipocytes treated with anthocyanins. Biochim Biophys Acta. 2005 Apr 15;1733(2-3):137-47. doi: 10.1016/j.bbalip.2004.12.014. ABSTRACT: Adipocyte dysfunction is strongly associated with the development of obesity and insulin resistance. It is accepted that the regulation of adipocytokine secretion or the adipocyte specific gene expression is one of the most important targets for the prevention of obesity and amelioration of insulin sensitivity. Recently, we demonstrated that anthocyanins, which are pigments widespread in the plant kingdom, have the potency of anti-obesity in mice and the enhancement adipocytokine secretion and adipocyte gene expression in adipocytes. In this study, we have shown for the first time the gene expression profile in isolated rat adipocytes treated with anthocyanins (cyanidin 3-glucoside; C3G or cyanidin; Cy). The rat adipocytes were treated with 100 muM C3G, Cy or vehicle for 24 h. The total RNA from the adipocytes was isolated and carried out GeneChip microarray analysis. A total of 633 or 427 genes was up-regulated (> 1.5-fold) by the treatment of adipocytes with C3G or Cy, respectively. The up-regulated genes include lipid metabolism and signal transduction-related genes, however, the altered genes were partly different between the C3G- and Cy-treated groups. Based on the gene expression profile, we demonstrated the up-regulation of hormone sensitive lipase and enhancement of the lipolytic activity by the treatment of adipocytes with C3G or Cy. These data have provided an overview of the gene expression profiles in adipocytes treated with anthocyanins and identified new responsive genes with potentially important functions in adipocytes related with obesity and diabetes that merit further investigation.
9. Grace M et al. Hypoglycemic activity of a novel anthocyanin-rich formulation from lowbush blueberry, Vaccinium angustifolium Aiton. Phytomedicine. 2009 May;16(5):406-15. doi: 10.1016/j.phymed.2009.02.018. ABSTRACT: Blueberry fruits are known as a rich source of anthocyanin components. In this study we demonstrate that anthocyanins from blueberry have the potency to alleviate symptoms of hyperglycemia in diabetic C57b1/6J mice. The anti-diabetic activity of different anthocyanin-related extracts was evaluated using the pharmaceutically acceptable self-microemulsifying drug delivery system: Labrasol. Treatment by gavage (500 mg/kg body wt) with a phenolic-rich extract and an anthocyanin-enriched fraction formulated with Labrasol lowered elevated blood glucose levels by 33 and 51%, respectively. The hypoglycemic activities of these formulae were comparable to that of the known anti-diabetic drug metformin (27% at 300 mg/kg). The extracts were not significantly hypoglycemic when administered without Labrasol, demonstrating its bio-enhancing effect, most likely due to increasing the bioavailability of the administered preparations. The phenolic-rich extract contained 287.0+/-9.7 mg/g anthocyanins, while the anthocyanin-enriched fraction contained 595+/-20.0 mg/g (cyanidin-3-glucoside equivalents), as measured by HPLC and pH differential analysis methods. The greater hypoglycemic activity of the anthocyanin-enriched fraction compared to the initial phenolic-rich extract suggested that the activity was due to the anthocyanin components. Treatment by gavage (300 mg/kg) with the pure anthocyanins, delphinidin-3-O-glucoside and malvidin-3-O-glucoside, formulated with Labrasol, showed that malvidin-3-O-glucoside was significantly hypoglycemic while delphinidin-3-O-glucoside was not.
10. Tsuda T et al. Dietary cyanidin 3-O-beta-D-glucoside-rich purple corn color prevents obesity and ameliorates hyperglycemia in mice. J Nutr. 2003 Jul;133(7):2125-30. doi: 10.1093/jn/133.7.2125. ABSTRACT: Anthocyanins, which are used as a food coloring, are widely distributed in human diets, suggesting that we ingest large amounts of anthocyanins from plant-based foods. Mice were fed control, cyanidin 3-glucoside-rich purple corn color (PCC), high fat (HF) or HF + PCC diet for 12 wk. Dietary PCC significantly suppressed the HF diet-induced increase in body weight gain, and white and brown adipose tissue weights. Feeding the HF diet markedly induced hypertrophy of the adipocytes in the epididymal white adipose tissue compared with the control group. In contrast, the induction did not occur in the HF + PCC group. The HF diet induced hyperglycemia, hyperinsulinemia and hyperleptinemia. These perturbations were completely normalized in rats fed HF + PCC. An increase in the tumor necrosis factor (TNF)-alpha mRNA level occurred in the HF group and was normalized by dietary PCC. These results suggest that dietary PCC may ameliorate HF diet-induced insulin resistance in mice. PCC suppressed the mRNA levels of enzymes involved in fatty acid and triacylglycerol synthesis and lowered the sterol regulatory element binding protein-1 mRNA level in white adipose tissue. These down-regulations may contribute to triacylglycerol accumulation in white adipose tissue. Our findings provide a biochemical and nutritional basis for the use of PCC or anthocyanins as a functional food factor that may have benefits for the prevention of obesity and diabetes.
11. Guo H et al. Cyanidin 3-glucoside attenuates high-fat and high-fructose diet-induced obesity by promoting the thermogenic capacity of brown adipose tissue. J Nutr Biochem. 2012 Apr;23(4):349-60. doi: 10.1016/j.jnutbio.2010.12.013.
12. Shi M et al. The effect of cyanidin-3-O-β-glucoside and peptides extracted from yoghurt on glucose uptake and gene expression in human primary skeletal muscle myotubes from obese and obese diabetic participants. Journal of Functional Foods. 2018 Dec 51:55-64. DOI:10.1016/j.jff.2018.10.012
13. Grace M et al. Hypoglycemic activity of a novel anthocyanin-rich formulation from lowbush blueberry, Vaccinium angustifolium Aiton. Phytomedicine. 2009 May;16(5):406-15. doi: 10.1016/j.phymed.2009.02.018.

DMAE (Dimethylaminoethanol)

1. Dimpfel W et al. Efficacy of dimethylaminoethanol (DMAE) containing vitamin-mineral drug combination on EEG patterns in the presence of different emotional states. Eur J Med Res. 2003 May 30;8(5):183-91. ABSTRACT: The psychophysiological model of provoking different emotional states by watching film excerpts with various emotional contents was used to characterize drug action in 80 subjects (male/female=50%) with threshold emotional disturbance within a randomized, group-parallel, double-blind, placebo-controlled study. Analyzing the brain's electrical reaction during presentation of 5 videoclips of 7 min duration followed by 3 minutes pause revealed a content specific representation of topographical frequency changes. This procedure was repeated after 6 and 12 weeks of daily intake of a vitamin-mineral drug combination containing dimethylaminoethanol (DMAE) (Vitagerin Geistlich N) or placebo. Subjects taking the active drug for 3 months developed significant less theta and alpha1 power in sensomotoric areas of the cortex. The grade of change and statistical significance was dependend on the content of the excerpt, but the pattern of changes in general remained the same. Since decreases in theta and alpha1 electrical power have been associated with increased vigilance and attention, subjects taking the drug combination obviously were more active and felt better. - Analysis of the emotional change in mood profile as induced by the TV session was achieved by completing two different quenstionaires (POMS and Bf-S). Both scores revealed a better mood for the active drug group thus corroborating the results from EEG analysis. Therefore the vitamine-mineral drug combination containing DMAE can be interpreted to induce a psychophysiological state of better feeling of wellbeing on both levels of analysis mood and electrical pattern of brain activity in subjects suffering from borderline emotional disturbance.
2. Danysz A et al. The influence of 2-dimethylaminethanol (DMAE) on the mental and physical efficiency in man. Act Nerv Super (Praha). 1967 Nov;9(4):417.
3. Rugginenti A. Effects of dimethylaminoethanol acetyl glutamate on the attentive capacity of a group of soccer players. Arch Maragliano Patol Clin. 1974 Jul-Dec;30(2):189-98.
4. Sabourin H et al. Dimethylaminoethanol and personality disorders of the young adult. Electro-clinical correlations. Ann Med Psychol (Paris). 1966 Apr;124(4):579-84.
5. Boutillier H et al. Apropos of some cases of personality and behavior disorders in children treated by dimethylaminoethanol. Gaz med fr. 1963 oct 10;70:2929-33.
6. Knobel M et al.2-Dimethylaminoethanol in behavior disorders of childhood. Sem Med. 1961 Sep 18;119:939-44.

Electrolytes & Nutrient Timing

1. Cribb PJ et al. Effects of supplement timing and resistance exercise on skeletal muscle hypertrophy. Med Sci Sports Exerc. 2006 Nov;38(11):1918-25.
2. Yuill KA et al. The administration of an oral carbohydrate-containing fluid prior to major elective upper-gastrointestinal surgery preserves skeletal muscle mass postoperatively - a randomized clinical trial. Clin Nutr. 2005 Feb;24(1):32-7.
3. Welsh RS et al. Carbohydrates and physical/mental performance during intermittent exercise to fatigue. Med Sci Sports Exerc. 2002 Apr;34(4):723-31.
4. Winnick JJ et al. Carbohydrate feedings during team sport exercise preserve physical and CNS function. Med Sci Sports Exerc. 2005 Feb;37(2):306-15.
5. Davis JM et al. Carbohydrate drinks delay fatigue during intermittent, high-intensity cycling in active men and women. Int J Sport Nutr. 1997 Dec;7(4):261-73.
6. Merson SJ et al. Rehydration with drinks differing in sodium concentration and recovery from moderate exercise-induced hypohydration in man. Eur J Appl Physiol. 2008 Jul;103(5):585-94.
7. Mudambo SM et al. Body fluid shifts in soldiers after a jogging/walking exercise in the heat: effects of water and electrolyte solution on rehydration. Cent Afr J Med. 2001 Sep-Oct;47(9-10):220-5.
8. Shirreffs SM et al. Rehydration and recovery of fluid balance after exercise. Exerc Sport Sci Rev. 2000 Jan;28(1):27-32.
9. Galloway SD. Dehydration, rehydration, and exercise in the heat: rehydration strategies for athletic competition. Can J Appl Physiol. 1999 Apr;24(2):188-200.
10. Brouns F et al. The effect of different rehydration drinks on post-exercise electrolyte excretion in trained athletes. Int J Sports Med. 1998 Jan;19(1):56-60.
11. Sugiura K et al. Effect of carbohydrate ingestion on sprint performance following continuous and intermittent exercise. Med Sci Sports Exerc. 1998 Nov;30(11):1624-30.
12. Ali A et al. The influence of carbohydrate-electrolyte ingestion on soccer skill performance. Med Sci Sports Exerc. 2007 Nov;39(11):1969-76.
13. Kamel KS et al. Treatment of hyponatremia: a quantitative analysis. Am J Kidney Dis. 1993 Apr;21(4):439-43.
14. Murray R. The effects of consuming carbohydrate-electrolyte beverages on gastric emptying and fluid absorption during and following exercise. Sports Med. 1987 Sep-Oct;4(5):322-51.

Epigallocatechin-3-gallate (EGCG)

1. Chu C et al. Green Tea Extracts Epigallocatechin-3-gallate for Different Treatments. Biomed Research International. Volume 2017, 13 Aug. ABSTRACT: Epigallocatechin-3-gallate (EGCG), a component extracted from green tea, has been proved to have multiple effects on human pathological and physiological processes, and its mechanisms are discrepant in cancer, vascularity, bone regeneration, and nervous system. This review focuses on effects of EGCG, including anti-cancer, antioxidant, anti-inflammatory, anticollagenase, and antifibrosis effects, to express the potential of EGCG and necessity of further studies in this field.
2. Kishimoto Y et al. Pleiotropic preventive effects of dietary polyphenols in cardiovascular diseases. European Journal of Clinical Nutrition, vol. 67, no. 5, pp. 532–535, 2013. ABSTRACT: The purpose of this study was to review recent findings highlighting daily dietary polyphenol intake and the diverse function of polyphenols and their possible relationships to cardiovascular disease (CVD). their previous findings provide that Japanese people intake polyphenols mainly from beverages, especially coffee and green tea (in descending order of polyphenol content). Many kinds of polyphenols act as an antioxidant against low-density lipoprotein oxidation, which is known to promote atherosclerosis. Recent accumulating evidence suggests that dietary polyphenols could exert their cardioprotective actions through their potential to improve metabolic disorder and vascular inflammation. These findings raise the possibility that polyphenols have a wide variety of roles in the intestine, liver and vascular tissue.
3. Qian Yi Eng et al. Molecular understanding of Epigallocatechin gallate (EGCG) in cardiovascular and metabolic diseases. Journal of Ethnopharmacology, Volume 210, 10 January, 2018, pp. 296-310. ABSTRACT: EGCG was found to exhibit a wide range of therapeutic properties including anti-atherosclerosis, anti-cardiac hypertrophy, anti-myocardial infarction, anti-diabetes, anti-inflammatory and antioxidant. These therapeutic effects are mainly associated with the inhibition of LDL cholesterol (anti-atherosclerosis), inhibition of NF-κB (anti-cardiac hypertrophy), inhibition of MPO activity (anti-myocardial infarction), reduction in plasma glucose and glycated haemoglobin level (anti-diabetes), reduction of inflammatory markers (anti-inflammatory) and the inhibition of ROS generation (antioxidant).
4. Xu Y et al. Inhibition of Tobacco-specific Nitrosamine-induced Lung Tumorigenesis in A/J Mice by Green Tea and Its Major Polyphenol as Antioxidants. Cancer Research, Published July 1992. ABSTRACT: This study examined the effects of green tea and its major components, (-)-epigallocatechin gallate (EGCG) and caffeine, on the tobacco-specific nitrosamine 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK)-induced lung tumorigenesis in A/J mice. They also studied the effects of green tea and EGCG on O6-methylguanine and 8-hydroxydeoxyguanosine (8-OH-dGuo) formation in lung tissues caused by NNK treatment. Mice were given 2% tea, 560 ppm EGCG, or 1120 ppm caffeine in drinking water for 13 weeks. The inhibition of 8-OH-dGuo formation in lung DNA by green tea and EGCG is consistent with their ability to inhibit lung tumorigenesis by NNK.
5. Jung YD et al. EGCG, a major component of green tea, inhibits tumour growth by inhibiting VEGF induction in human colon carcinoma cells. British Journal of Cancer, vol. 84, no. 6, pp. 844–850, 2001. ABSTRACT: Catechins are key components of teas that have anti-proliferative properties. This study investigated the effects of green tea catechins on intracellular signaling and VEGF induction in vitro in serum-deprived HT29 human colon cancer cells and in vivo on the growth of HT29 cells in nude mice. In the in vitro studies, (-)-epigallocatechin gallate (EGCG), the most abundant catechin in green tea extract, inhibited Erk-1 and Erk-2 activation in a dose-dependent manner. Treatment with EGCG inhibited tumour growth (58%), microvessel density (30%), and tumor cell proliferation (27%) and increased tumor cell apoptosis (1.9-fold) and endothelial cell apoptosis (3-fold) relative to the control condition (P < 0.05 for all comparisons). EGCG may exert at least part of its anticancer effect by inhibiting angiogenesis through blocking the induction of VEGF.
6. Guang-Jian Du et al. Epigallocatechin Gallate (EGCG) Is the Most Effective Cancer Chemopreventive Polyphenol in Green Tea. Nutrients 2012, 4(11), 1679-1691. ABSTRACT: Previous studies have shown that some polyphenol compounds from green tea possess anticancer activities. In this study, they determined the cancer chemopreventive potentials of 10 representative polyphenols. Among the 10 polyphenols, EGCG showed the most potent antiproliferative effects, and significantly induced cell cycle arrest in the G1 phase and cell apoptosis.
7. Mandel S et al. Multifunctional Activities of Green Tea Catechins in Neuroprotection. Neurosignals. 2005;14(1-2):46-60. DOI: 10.1159/000085385. ABSTRACT: Many lines of evidence suggest that oxidative stress resulting in reactive oxygen species (ROS) generation and inflammation play a pivotal role in the age-associated cognitive decline and neuronal loss in neurodegenerative diseases including Alzheimer's (AD), Parkinson's (PD) and Huntington's diseases. One cardinal chemical pathology observed in these disorders is the accumulation of iron at sites where the neurons die. The buildup of an iron gradient in conjunction with ROS (superoxide, hydroxyl radical and nitric oxide) are thought to constitute a major trigger in neuronal toxicity and demise in all these diseases. Thus, promising future treatment of neurodegenerative diseases and aging depends on availability of effective brain permeable, iron-chelatable/radical scavenger neuroprotective drugs that would prevent the progression of neurodegeneration. Tea flavonoids (catechins) have been reported to possess potent iron-chelating, radical-scavenging and anti-inflammatory activities and to protect neuronal death in a wide array of cellular and animal models of neurological diseases. Recent studies have indicated that in addition to the known antioxidant activity of catechins, other mechanisms such as modulation of signal transduction pathways, cell survival/death genes and mitochondrial function, contribute significantly to the induction of cell viability. This review will focus on the multifunctional properties of green tea and its major component (-)-epigallocatechin-3-gallate (EGCG) and their ability to induce neuroprotection and neurorescue in vitro and in vivo. In particular, their transitional metal (iron and copper) chelating property and inhibition of oxidative stress.
8. Biasibetti R et al. Green tea (-)epigallocatechin-3-gallate reverses oxidative stress and reduces acetylcholinesterase activity in a streptozotocin-induced model of dementia. Behav Brain Res. 2013 Jan 1. DOI: 10.1016/j.bbr.2012.08.039. ABSTRACT: Alzheimer's disease (AD) is the most prevalent form of dementia. Intracerebroventricular (ICV) infusion of streptozotocin (STZ) provides a relevant animal model of chronic brain dysfunction that is characterized by long-term and progressive deficits in learning, memory, and cognitive behavior, along with a permanent and ongoing cerebral energy deficit. Numerous studies on green tea epigallocatechin gallate (EGCG) demonstrate its beneficial effects on cognition and memory. As such, this study evaluated, for the first time, the effects of sub-chronic EGCG treatment in rats that were submitted to ICV infusion of STZ (3mg/kg). Male Wistar rats were divided into sham, STZ, sham+EGCG and STZ+EGCG groups. EGCG was administered at a dose of 10mg/kg/day for 4 weeks per gavage. Learning and memory was evaluated using Morris' Water Maze. Oxidative stress markers and involvement of the nitric oxide (NO) system, acetylcholinesterase activity (AChE) and glucose uptake were evaluated as well as glial parameters including S100B content and secretion and GFAP content. Our results show that EGCG was not able to modify glucose uptake and glutathione content, although cognitive deficit, S100B content and secretion, AChE activity, glutathione peroxidase activity, NO metabolites, and reactive oxygen species content were completely reversed by EGCG administration, confirming the neuroprotective potential of this compound. These findings contribute to the understanding of diseases accompanied by cognitive deficits and the STZ-model of dementia.
9. Rasoolijazi H et al. The Beneficial Effect of (-)-Epigallocatechin-3-Gallate in an Experimental Model of Alzheimer’s Disease in Rat: a Behavioral Analysis. Iranian Biomedical Journal. October 2007. ABSTRACT: rogressive cognitive decline is one of the hallmark symptoms of Alzheimer’s disease (AD) which can be modeled by β-amyloid injection into specific regions of brain. Since epigallocatechin-3-gallate (EGCG) is a potent antioxidant agent which its role against oxidative stress and inflammation has been shown in prior studies, we tried to determine whether EGCG administration protects against β-amyloid-induced memory and coordination impairment in rats. Methods: Animals (male Wistar rats) were divided into four groups: sham operated, EGCG-pretreated sham operated (sham + EGCG), untreated lesion (lesion), and EGCG-pretreated lesion (lesion + EGCG). Animals in lesion, lesion + EGCG, and sham + EGCG groups received sterile saline or saline plus EGCG (10 mg/kg) intraperitoneally one day pre-surgery and every other day for three weeks. The lesion was induced one day after EGCG pretreatment by injection of 4 μl of sterile saline or water containing 2 nmol/μl β-amyloid (1-40) into the hippocampal fissure. For behavioral analysis, psychomotor coordination (PMC) index and spontaneous alternation behavior were assessed using Rota-rod Treadmill and Y-maze, respectively at the third week post-lesion. Results: We found that β-amyloid (1-40) injection into hippocampus can decrease these behavioral indexes in lesion group in comparison with sham group which is similar to behavioral changes in AD. On the other hand, pretreatment with EGCG can improve the PMC index and spatial Y-maze alternation in the lesion + EGCG group in comparison with lesion group. Conclusion: We concluded that EGCG can be effective in restoring β-amyloid-induced behavioral derangements in rats regarding coordination and memory abilities.

Eurycoma longifolia

1. Ezzat SM et al. Rho-Kinase II Inhibitory Potential of Eurycoma longifolia New Isolate for the Management of Erectile Dysfunction. Evidence Based Complementary and Alternative Medicine, 15 May 2019. Background: Eurycoma longifolia Jack (Fam.: Simaroubaceae), known as Tongkat Ali (TA), has been known as a symbol of virility and sexual power. The aim of the study was to screen E. longifolia aqueous extract (AE) and isolates for ROCK-II inhibition. Results: The AE (1-10 μg/ml) showed a significant inhibition for ROCK-II activity (62.8-81%) at P < 0.001 with an IC50 (651.1 ± 32.9 ng/ml) compared to Y-27632 ([(+)-(R)-trans-4-(1-aminoethyl)-N-(4-pyridyl)cyclohexanecarboxamide dihydrochloride]) (68.15-89.9 %) at same concentrations with an IC50 (192 ± 8.37 ng/ml). Chromatographic purification of the aqueous extract (AE) allowed the isolation of eight compounds; stigmasterol T1, trans-coniferyl aldehyde T2, scopoletin T3, eurycomalactone T4, 6α- hydroxyeurycomalactone T5, eurycomanone T6, eurycomanol T7, and eurycomanol-2-O-β-D-glucopyranoside T8. This is the first report for the isolation of T1 and T3 from E. longifolia and for the isolation of T2 from genus Eurycoma. The isolates (at 10 μg/ml) exhibited maximum inhibition % of ROCK-II 82.1 ± 0.63 (T2), 78.3 ± 0.38 (T6), 77.1 ± 0.11 (T3), 76.2 ± 3.53 (T4), 74.5 ± 1.27 (T5), 74.1 ± 2.97 (T7), 71.4 ± 2.54 (T8), and 60.3 ± 0.14 (T1), where the newly isolated compound trans-coniferyl aldehyde T2 showed the highest inhibitory activity among the tested isolated compounds and even higher than the total extract AE. The standard Y-27632 (10 μg/ml) showed 89.9 ± 0.42 % inhibition for ROCK-II activity when compared to control at P < 0.0001. Conclusion: The traditional use of E. longifolia as aphrodisiac and for male sexual disorders might be in part due to the ROCK-II inhibitory potential.
2. Low BS et al. Standardized quassinoid-rich Eurycoma longifolia extract improved spermatogenesis and fertility in male rats via the hypothalamic-pituitary-gonadal axis. J Ethnopharmacol. 2013 Feb 13;145(3):706-14. Ethnopharmacological Relevance: Eurycoma longifolia Jack, a small Simaroubaceae tree, known locally as 'Tongkat Ali' is popularly used as a sexual tonic in traditional medicine for aphrodisiac activity and improvement of fertility and male libido. Aim of the Study: To investigate the effects of the standardized bioactive fraction of E. longifolia and its chemical constituents on the male fertility and the mechanisms of action involved. Material and Methods: The powdered roots of E. longifolia were extracted separately with methanol and water. The organic extract upon further fractionation on HP 20 resin and elution with the methanol/water mixture afforded four fractions (F1-F4). These fractions, together with the crude aqueous (W) and organic extracts were standardized following their respective major quassinoid content and profile. The effects of the fractions on the rat spermatogenesis were compared with that of the aqueous extract (W) to determine the bioactive fraction. The effects of the bioactive fraction on the sperm count and quality, the histological morphometric changes on the spermatogenesis cycle, fertility and hormonal changes of plasma testosterone, luteinizing hormone (LH), follicle stimulating hormone (FSH) and estrogen in the animals upon oral administration were determined. The effects of the bioactive quassinoids on the testosterone release from the isolated testicular interstitial cells rich in Leydig cells, were also described. Results: The male rats orally administered with 25mg/kg of F2 and 250mg/kg of W, significantly increased the sperm concentration when compared with that of the control animals (P < 0.05). High performance liquid chromatography analysis revealed that 25mg/kg of F2 and 250mg/kg of W were almost similar in concentration of eurycomanone, the major and most potent quassinoid. Microscopic morphometrical analysis of the rat testis following treatment with F2, showed significant increase in the number of spermatocytes and round spermatids at Stage VII of the spermatogenesis cycle when compared to that of the control (P < 0.05). The estimated spermatozoa production rate and the number of Leydig cells were also elevated (P < 0.001). The fertility index, fecundity index and the pup litter size delivered from the females after mating with the males treated with F2 were increased. The plasma testosterone level of the animals given 25mg/kg of F2 orally was significantly different at day-26 (P < 0.05) and day-52 (P < 0.01) from those of control but was not different at day-104. The testicular testosterone also peaked in the animals treated with 25mg/kg F2 and was higher than that in the plasma. The plasma LH and FSH levels of the rats treated with 25mg/kg of F2 were higher than those of the control (P < 0.001). In contrast, the plasma estrogen level was significantly lower than that of the untreated control. Amongst the isolated quassinoids of F2, eurycomanone and 13α(21)-dihydroeurycomaone significantly increased the testosterone level from the Leydig cells of the testicular interstitial cells cultured in vitro (P < 0.05). Conclusion: The standardised extract F2 of E. longifolia and its major quassinoids especially eurycomanone improved the rat spermatogenesis by affecting the hypothalamic-pituitary-gonadal axis and the potential efficacy may be worthy of further investigation.
3. Muhamad AS et al. Eurycoma longifolia Jack: Medicinal properties and its effects on endurance exercise performance. Asian Journal of Exercise & Sport Science, 2009, Vol. 6 (No. 1). ABSTRACT: This review article discusses the medicinal properties of a herbal plant, Eurycoma longifolia Jack, and its possible ergogenic effect on endurance exercise performance. To date, herbs or plant products that have been investigated as ergogenic aids for enhancing endurance performance are caffeine, ginseng, mahuang, ephedrine and related alkaloids. Eurycoma longifolia Jack is one of the herbs found in Malaysia. It is commonly known as ‘Tongkat Ali’ in Malaysia and as ‘Pasak Bumi’ in Indonesia. It is also referred to as ‘Malaysian ginseng’ since it is well-known among various ethnic groups in Malaysia for treating various diseases and enhancing health. Eurycoma longifolia Jack is a tall, slender shrubby tree found on sandy soil. It belongs to the Simaroubaceae family and grows wildly in Southeast Asian countries, i.e. Malaysia, Indonesia, Thailand, and Myanmar. To our knowledge, studies on Eurycoma longifolia Jack as an ergogenic aid for enhancing endurance performance is lacking. Thus, this review article highlights the available studies on its purported medicinal properties as well as studies that have been carried out to investigate its effects on physiological responses and endurance exercise performance.
4. Rehman SU et al. Review on a Traditional Herbal Medicine, Eurycoma longifolia Jack (Tongkat Ali): Its Traditional Uses, Chemistry, Evidence-Based Pharmacology and Toxicology. Molecules, 2016, 21(3), 331. ABSTRACT: Eurycoma longifolia Jack (known as tongkat ali), a popular traditional herbal medicine, is a flowering plant of the family Simaroubaceae, native to Indonesia, Malaysia, Vietnam and also Cambodia, Myanmar, Laos and Thailand. E. longifolia, is one of the well-known folk medicines for aphrodisiac effects as well as intermittent fever (malaria) in Asia. Decoctions of E. longifolia leaves are used for washing itches, while its fruits are used in curing dysentery. Its bark is mostly used as a vermifuge, while the taproots are used to treat high blood pressure, and the root bark is used for the treatment of diarrhea and fever. Mostly, the roots extract of E. longifolia are used as folk medicine for sexual dysfunction, aging, malaria, cancer, diabetes, anxiety, aches, constipation, exercise recovery, fever, increased energy, increased strength, leukemia, osteoporosis, stress, syphilis and glandular swelling. The roots are also used as an aphrodisiac, antibiotic, appetite stimulant and health supplement. The plant is reported to be rich in various classes of bioactive compounds such as quassinoids, canthin-6-one alkaloids, β-carboline alkaloids, triterpene tirucallane type, squalene derivatives and biphenyl neolignan, eurycolactone, laurycolactone, and eurycomalactone, and bioactive steroids. Among these phytoconstituents, quassinoids account for a major portion of the E. longifolia root phytochemicals. An acute toxicity study has found that the oral Lethal Dose 50 (LD50) of the alcoholic extract of E. longifolia in mice is between 1500-2000 mg/kg, while the oral LD50 of the aqueous extract form is more than 3000 mg/kg. Liver and renal function tests showed no adverse changes at normal daily dose and chronic use of E. longifolia. Based on established literature on health benefits of E. longifolia, it is important to focus attention on its more active constituents and the constituents' identification, determination, further development and most importantly, the standardization. Besides the available data, more evidence is required regarding its therapeutic efficacy and safety, so it can be considered a rich herbal source of new drug candidates. It is very important to conserve this valuable medicinal plant for the health benefit of future generations.
5. Solomon MC et al. In vivo effects of Eurycoma longifolia Jack (Tongkat Ali) extract on reproductive functions in the rat. Andrologia. 2014 May;46(4):339-48. ABSTRACT: An aqueous extract of Eurycoma longifolia (Tongkat Ali; TA) roots is traditionally used to enhance male sexuality. Because previous studies are limited to only few sperm parameters or testosterone concentration, this study investigated the in vivo effects of TA on body and organ weight as well as functional sperm parameters in terms of safety and efficacy in the management of male infertility. Forty-two male rats were divided into a control, low-dose (200 mg kg(-1) BW) and high-dose (800 mg kg(-1) BW) group (n = 14). Rats were force-fed for 14 days and then sacrificed. Total body and organ weights of the prostate, testes, epididymides, gastrocnemius muscle and the omentum were recorded. Moreover, testosterone concentration, sperm concentration, motility, velocity, vitality, acrosome reaction and mitochondrial membrane potential (MMP) were assessed. Whilst TA decreased BW by 5.7% (P = 0.0276) and omentum fat by 31.9% (P = 0.0496), no changes in organ weights were found for the prostate, testes and epididymides. Testosterone concentration increased by 30.2% (P = 0.0544). Muscle weight also increased, yet not significantly. Whilst sperm concentration, total and progressive motility and vitality increased significantly, MMP improved markedly (P = 0.0765) by 25.1%. Because no detrimental effect could be observed, TA appears safe for possible treatment of male infertility and ageing male problems.

Forskolin

1. Godard MP et al. Body composition and hormonal adaptations associated with forskolin consumption in overweight and obese men. Obesity Research. August 2005. DOI: 10.1038/oby.2005.162. Objective: This study examined the effect of forskolin on body composition, testosterone, metabolic rate, and blood pressure in overweight and obese (BMI > or = 26 kg/m(2)) men. Research methods and procedure: Thirty subjects (forskolin, n = 15; placebo, n = 15) were studied in a randomized, double-blind, placebo-controlled study for 12 weeks. Results: Forskolin was shown to elicit favorable changes in body composition by significantly decreasing body fat percentage (BF%) and fat mass (FM) as determined by DXA compared with the placebo group (P < or = 0.05). Additionally, forskolin administration resulted in a change in bone mass for the 12-week trial compared with the placebo group (P < or = 0.05). There was a trend toward a significant increase for lean body mass in the forskolin group compared with the placebo group (p = 0.097). Serum free testosterone levels were significantly increased in the forskolin group compared with the placebo group (P < or = 0.05). The actual change in serum total testosterone concentration was not significantly different among groups, but it increased 16.77 +/- 33.77% in the forskolin group compared with a decrease of 1.08 +/- 18.35% in the placebo group. Discussion: Oral ingestion of forskolin (250 mg of 10% forskolin extract twice a day) for a 12-week period was shown to favorably alter body composition while concurrently increasing bone mass and serum free testosterone levels in overweight and obese men. The results indicate that forskolin is a possible therapeutic agent for the management and treatment of obesity.
2. Scarpace PJ, Matheny M. Thermogenesis in brown adipose tissue with age: post-receptor activation by forskolin. Pflugers Arch. 1996 Jan;431(3):388-94. ABSTRACT: beta3-Adrenergic-stimulated thermogenesis in brown adipose tissue (BAT) is diminished with age. beta3-Adrenergic receptors are positively coupled to adenylyl cyclase in BAT. To determine whether thermo- genesis, in response to direct activation of adenylyl cyclase, is also impaired with age, we examined whole body oxygen consumption, mitochondrial guanosine diphosphate (GDP) binding and BAT mitochondrial uncoupling protein (UPC) mRNA levels in 4- and 24-month-old F-344 rats following forskolin administration. We also examined the forskolin-induced change in body temperature in 4-month-old rats. In some instances, the results were compared with administration of the specific beta3-adrenergic agonist, CGP-12177. Forskolin (3.5 mg/kg) increased oxygen consumption but decreased body temperature. In subsequent experiments the BAT was unilaterally denervated. In these rats, the forskolin-(1.8 mg/kg) stimulated increase in oxygen consumption was similar in young and old rats. Forskolin increased GDP binding and UCP mRNA levels in both the denervated and innervated BAT pads. The increases were equal or greater in the BAT from senescent rats. These findings, coupled with our previous report of an impaired CGP-12177-stimulated increase in GDP binding in senescent rats, suggests beta3-adrenergic-stimulated, but not post-receptor-stimulated, thermogenesis is diminished with age.
3. Majeed M et al. Diterpene forskolin: A possible new compound for reduction of body weight by increasing lean body mass. Nutraceuticals, March/April 2002, pp. 6-7. ABSTRACT: Maintaining or increasing lean body mass should be one of the important considerations of any weight loss strategy for the following reasons: 1. increase in lean body mass is proportionate to an increase in the body’s thermogenic response to food and the basic metabolic rate (BMR); 2. food induced thermogenesis controls body weight by an increase in catabolism of body fat (thermogenesis is preferentially fueled by fatty acids derived from body fat and/or from food); and 3. enhanced thermogenesis contributes to a buildup of lean body mass. An extract of Coleus forskohlii root, Benth. (Fam. Labiatae) standardized for diterpene forskolin was tested in an open-field study for weight loss and lean body mass increase. The study’s hypothesis was based on the recognized role of diterpene forskolin as the plant derived compound which stimulates enzyme adenylate cyclase and subsequently cyclic AMP (3’5’adenosine monophosphate) (1,2). Cyclic AMP may release fatty acids from the adipose tissue depots which may result in enhanced thermogenesis (3), loss of body fat, and theoretically increased lean body mass. Six overweight, but otherwise healthy, women were selected for the trial. Each participant was informed about the purpose of the study and was asked to sign an informed consent before entering the study. Each participant was examined by a physician at the inception and after 4 and 8 weeks of the study. The body composition was determined by bioelectrical impedance analysis. The forskolin formula was prepared in the form of two piece hard shell capsules. Each capsule contained 250 mg of the extract standardized for 10% forskolin, and each bottle contained 60 capsules. Participants were instructed to take one capsule in the morning and one in the evening, half an hour before a meal. Each participant was asked to maintain their previous daily physical exercise habits and eating habits. In addition, physical activity was monitored based on a questionnaire before and during the trial. The study was performed in an outpatient bariatric clinic at Hilton Head, S.C. and supervised by a physician specializing in bariatric medicine for over 30 years. During the eight week trial the mean values for body weight, and fat content were significantly decreased, whereas lean body mass was significantly increased as compared to the baseline (Wilcoxon matched pairs test). Weight loss was statistically significant.
4. Kamath. Efficacy and Safety of Forslean in Increasing Lean Body Mass. Department of Ayurvedic Medicine, Kasturba Medical College, Manipal, India, 2005.
5. Tsugiyoshi. Clinical report on root extract of Perilla Plant (coleus forskohlii) in reducing body fat. Asanto Institute, Tokyo, Japan, 2001.

Highly Branched Cyclic Dextrin (Cluster Dextrin)

1. Shiraki T et al. Evaluation of exercise performance with the intake of highly branched cyclic dextrin in athletes. Food Science and Technology Research. 2015 Volume 21 Issue 3 Pages 499-502. DOI: 10.3136/fstr.21.499. ABSTRACT: Highly branched cyclic dextrin (HBCD) is a novel type of maltodextrin with a narrow molecular weight distribution that is produced from starch. In this study, we investigated the effects of HBCD administration on endurance performance. Seven elite swimmers participated in three trials, conducted in random order. In each trial, the subjects received either HBCD, glucose (1.5 g carbohydrate/kg body weight) or water (as a control), and immediately carried out 10 cycles of intermittent swimming consisting of 5 min of swimming at 75% followed by 3 min of rest, and subsequent swimming at 90% to exhaustion. The time to fatigue was about 70% longer in the HBCD trial than that in the glucose and control trials, a significant difference. Plasma glucose in the HBCD group was maintained at higher levels during pre-swimming cycles than that in the glucose or water group. These results suggest that HBCD administration enhances endurance performance.
2. Furuyashiki T et al. Effects of ingesting highly branched cyclic dextrin during endurance exercise on rating of perceived exertion and blood components associated with energy metabolism. Biosci Biotechnol Biochem. 2014;78(12):2117-9. DOI: 10.1080/09168451.2014.943654. ABSTRACT: We compared the effect of relatively low doses (15 g) of highly branched cyclic dextrin (HBCD) with that of maltodextrin during endurance exercise on the rating of perceived exertion (RPE) in a crossover, double-blind study of healthy volunteers. The RPE increased during exercise and its increase was significantly less at 30 and 60 min after ingesting HBCD than maltodextrin.
3. Suzuki K et al. Effect of a sports drink based on highly-branched cyclic dextrin on cytokine responses to exhaustive endurance exercise. J Sports Med Phys Fitness. 2014 Oct;54(5):622-30. PMID: 25270782. ABSTRACT: Background: Aim of the present study was to compare the effects of highly branched cyclic dextrin (HBCD) drink with a glucose-based control drink on immunoendocrine responses to endurance exercise. Methods: Using a randomized, double-blind placebo-controlled cross-over design, seven male triathletes participated in two duathlon races separated by one month, consisting of 5 km of running, 40 km of cycling and 5 km of running. In the first race, four athletes consumed the HBCD-based drink and three athletes consumed the glucose-based drink. In the second race, three athletes consumed the HBCD-based drink and four athletes consumed the glucose-based drink. We collected blood and urine samples before and after the races to analyze leukocyte count and concentrations of hormones and cytokines. Results: Lymphocyte and neutrophil counts increased significantly after exercise in both trials (P < 0.05), but were not significantly different between the trials. Plasma noradrenalin concentration increased significantly (P < 0.05) during exercise in the glucose trial, but not in the HBCD trial. Plasma concentrations of interleukin (IL)-8 and IL-10 increased significantly during exercise in both trials (P < 0.05) but were not significantly different between the trials. Post-race urinary IL-8, IL-10 and IL-12p40 concentrations were significantly lower in the HBCD trial compared with the glucose trial (P < 0.05), although the plasma concentrations of these cytokines were not significantly different between both trials. Conclusion: These results suggest that the HBCD-based drink may attenuate the stress hormone response, and reduce the urinary cytokine levels following exhaustive exercise.
4. Takii H et al. Fluids containing a highly branched cyclic dextrin influence the gastric emptying rate. Int J Sports Med. 2005 May;26(4):314-9. doi: 10.1055/s-2004-820999. ABSTRACT: The rates of gastric emptying for highly branched cyclic dextrin (HBCD) and other carbohydrate (CHO) solutions were examined using ultrasonograph techniques. Ten healthy volunteers ingested water, physiological saline, or solutions containing various CHO, such as HBCD, glucose, maltose, sucrose, and commercially available dextrin. After a subject drank one of the solutions, the relaxed cross-sectional area of the pylorus antrum was measured at rest by real-time ultrasonography. The time required for gastric emptying was correlated with the relaxed cross-sectional area of the pylorus antrum. Among all of the solutions tested, physiological saline was transferred fastest from the stomach to the small intestine. For solutions of the same CHO, 5% solution was transferred faster than 10% solution. For CHO solutions other than HBCD, a low osmotic pressure was associated with rapid transfer from the stomach. The gastric emptying time (GET) of HBCD solution increased with an increase in its concentration. A shorter GET was observed for the CHO solutions at 59 to 160 mOsm regardless of their concentration. A sports drink based on 10% HBCD adjusted to 150 mOsm by the addition of various minerals, vitamins, and organic acids was evacuated significantly (p < 0.05) faster than a 10% HBCD solution or a sports drink based on 10% commercially available dextrin (DE16), which has a higher osmotic pressure (269 mOsm). Our results suggest that a shorter GET could be achieved with CHO solutions with osmotic pressures of 59 - 160 mOsm. Therefore, a sports drink based on 10% HBCD adjusted to 150 mOsm by the addition of minerals, vitamins, and organic acids could supply adequate quantities of CHO, fluid, and minerals simultaneously in a short time, without increasing GET.
5. Takii H et al. Enhancement of swimming endurance in mice by highly branched cyclic dextrin. Biosci Biotechnol Biochem. 1999 Dec;63(12):2045-52. DOI: 10.1271/bbb.63.2045. ABSTRACT: We investigated the ergogenic effect in mice of administering highly branched cyclic dextrin (HBCD), a new type of glucose polymer, on the swimming endurance in an adjustable-current swimming pool. Male Std ddY mice were administered a HBCD, a glucose solution or water via a stomach sonde 10 min before, 10 min after or 30 min after beginning swimming exercise, and were then obliged to swim in the pool. The total swimming period until exhaustion, an index of the swimming endurance, was measured. An ergogenic effect of HBCD was observed at a dose of 500 mg/kg of body weight, whereas it had no effect at a dose of 166 mg/kg of body wt (p < 0.05). The mice administered with the HBCD solution 10 min after starting the exercise were able to swim significantly longer (p < 0.05) than the mice who had ingested water or the glucose solution. The rise in mean blood glucose level in the mice administered with HBCD, which was measured 20 min after starting swimming, was significantly lower (p < 0.05) than that in the mice administered with glucose, although it was significantly higher (p < 0.05) than that in the mice administered with water. The mean blood insulin rise in the mice given HBCD was significantly lower (p < 0.05) than that in the mice given glucose. The mice administered with HBCD 30 min after starting the exercise swam significantly longer (p < 0.05) than the mice who had ingested water, although the enhancement of swimming time was similar to that of the glucose-ingesting mice. The gastric emptying rate of the HBCD solution was significantly faster (p < 0.05) than that of the glucose solution. However, this glucose polymer must have spent more time being absorbed because it has to be hydrolyzed before absorption, reflecting a lower and possibly longer-lasting blood glucose level. We conclude that the prolongation of swimming endurance in mice administered with HBCD depended on its rapid and longer-lasting ability for supplying glucose with a lower postprandial blood insulin response, leading to a delayed onset of fatigue.

L-Leucine

1. Rieu I et al. Leucine supplementation improves muscle protein synthesis in elderly men independently of hyperaminoacidaemia. J Physiol. 2006 Aug 15;575(Pt 1):305-15
2. Drummond MJ et al. Leucine-enriched nutrients and the regulation of mammalian target of rapamycin signalling and human skeletal muscle protein synthesis. Curr Opin Clin Nutr Metab Care. 2008 May;11(3):222-6.
3. Meijer AJ et al. Amino acid signalling and the integration of metabolism. Biochem Biophys Res Commun. 2004 Jan 9;313(2):397-403.
4. Protein sparing produced by proteins and amino acids. Nutr Rev. 1976 Jun;34(6):174-6.
5. Stipanuk MH. Leucine and protein synthesis: mTOR and beyond. Nutr Rev. 2007 Mar;65(3):122-9.
6. Crowe MJ et al. Effects of dietary leucine supplementation on exercise performance. Eur J Appl Physiol. 2006 Aug;97(6):664-72.
7. Mero A. Leucine supplementation and intensive training. Sports Med. 1999 Jun;27(6):347-58.

L-Theanine

1. Park SK et al. A combination of green tea extract and L-theanine improves memory and attention in subjects with mild cognitive impairment: a double-blind placebo-controlled study. J Med Food. 2011 Apr;14(4):334-43. ABSTRACT: A combination of green tea extract and L-theanine (LGNC-07) has been reported to have beneficial effects on cognition in animal studies. In this randomized, double-blind, placebo-controlled study, the effect of LGNC-07 on memory and attention in subjects with mild cognitive impairment (MCI) was investigated. Ninety-one MCI subjects whose Mini Mental State Examination-K (MMSE-K) scores were between 21 and 26 and who were in either stage 2 or 3 on the Global Deterioration Scale were enrolled in this study. The treatment group (13 men, 32 women; 57.58 ± 9.45 years) took 1,680 mg of LGNC-07, and the placebo group (12 men, 34 women; 56.28 ± 9.92 years) received an equivalent amount of maltodextrin and lactose for 16 weeks. Neuropsychological tests (Rey-Kim memory test and Stroop color-word test) and electroencephalography were conducted to evaluate the effect of LGNC-07 on memory and attention. Further analyses were stratified by baseline severity to evaluate treatment response on the degree of impairment (MMSE-K 21-23 and 24-26). LGNC-07 led to improvements in memory by marginally increasing delayed recognition in the Rey-Kim memory test (P .0572). Stratified analyses showed that LGNC-07 improved memory and selective attention by significantly increasing the Rey-Kim memory quotient and word reading in the subjects with MMSE-K scores of 21-23 (LGNC-07, n = 11; placebo, n = 9). Electroencephalograms were recorded in 24 randomly selected subjects hourly for 3 hours in eye-open, eye-closed, and reading states after a single dose of LGNC-07 LGNC-07 (n = 12; placebo, n = 12). Brain theta waves, an indicator of cognitive alertness, were increased significantly in the temporal, frontal, parietal, and occipital areas after 3 hours in the eye-open and reading states. Therefore, this study suggests that LGNC-07 has potential as an intervention for cognitive improvement.
2. Giesbrecht T et al. The combination of L-theanine and caffeine improves cognitive performance and increases subjective alertness. Nutr Neurosci. 2010 Dec;13(6):283-90. ABSTRACT: The non-proteinic amino acid L-theanine and caffeine, a methylxanthine derivative, are naturally occurring ingredients in tea. The present study investigated the effect of a combination of 97 mg L- theanine and 40 mg caffeine as compared to placebo treatment on cognitive performance, alertness, blood pressure, and heart rate in a sample of young adults (n = 44). Cognitive performance, self-reported mood, blood pressure, and heart rate were measured before L-theanine and caffeine administration (i.e. at baseline) and 20 min and 70 min thereafter. The combination of moderate levels of L-theanine and caffeine significantly improved accuracy during task switching and self-reported alertness (both P < 0.01) and reduced self-reported tiredness (P < 0.05). There were no significant effects on other cognitive tasks, such as visual search, choice reaction times, or mental rotation. The present results suggest that 97 mg of L-theanine in combination with 40 mg of caffeine helps to focus attention during a demanding cognitive task.
3. Einšther SJ et al. L-theanine and caffeine improve task switching but not intersensory attention or subjective alertness. Appetite. 2010 Apr;54(2):406-9. ABSTRACT: Tea ingredients L-theanine and caffeine have repeatedly been shown to deliver unique cognitive benefits when consumed in combination. The current randomized, placebo-controlled, double-blind, cross-over study compared a combination of L-theanine (97 mg) and caffeine (40 mg) to a placebo on two attention tasks and a self-report questionnaire before, and 10 and 60 min after consumption. The combination of L-theanine and caffeine significantly improved attention on a switch task as compared to the placebo, while subjective alertness and intersensory attention were not improved significantly. The results support previous evidence that L-theanine and caffeine in combination can improve attention.
4. Kim TI et al. L-Theanine, an amino acid in green tea, attenuates beta-amyloid-induced cognitive dysfunction and neurotoxicity: reduction in oxidative damage and inactivation of ERK/p38 kinase and NF-kappaB pathways. Free Radic Biol Med. 2009 Dec 1;47(11):1601-10. Epub 2009 Sep 16. ABSTRACT: Amyloid beta (Abeta)-induced neurotoxicity is a major pathological mechanism of Alzheimer disease (AD). In this study, we investigated the inhibitory effect of l-theanine, a component of green tea (Camellia sinensis), on Abeta(1-42)-induced neuronal cell death and memory impairment. Oral treatment of l-theanine (2 and 4 mg/kg) for 5 weeks in the drinking water of mice, followed by injection of Abeta(1-42) (2 microg/mouse, icv), significantly attenuated Abeta(1-42)-induced memory impairment. Furthermore, l-theanine reduced Abeta(1-42) levels and the accompanying Abeta(1-42)-induced neuronal cell death in the cortex and hippocampus of the brain. Moreover, l-theanine inhibited Abeta(1-42)-induced extracellular signal-regulated kinase (ERK) and p38 mitogen-activated protein kinase as well as the activity of nuclear factor kappaB (NF-kappaB). l-Theanine also significantly reduced oxidative protein and lipid damage and the elevation of glutathione levels in the brain. These data suggest that the positive effects of l-theanine on memory may be mediated by suppression of ERK/p38 and NF-kappaB as well as the reduction of macromolecular oxidative damage. Thus, l-theanine may be useful in the prevention and treatment of AD.
5. Gomez-Ramirez M et al. The effects of L-theanine on alpha-band oscillatory brain activity during a visuo-spatial attention task. Brain Topogr. 2009 Jun;22(1):44-51. ABSTRACT: Background/Objectives Ingestion of the non-proteinic amino acid L-theanine (gamma-glutamylethylamide) has been shown to influence oscillatory brain activity in the alpha band (8-14 Hz) in humans during resting electroencephalographic (EEG) recordings and also during cognitive task performance. We have previously shown that ingestion of a 250-mg dose of L-theanine significantly reduced tonic (background) alpha power during a demanding intersensory (auditory-visual) attentional cueing task. Further, cue-related phasic changes in alpha power, indexing the shorter-term anticipatory biasing of attention between modalities, were stronger on L-theanine compared to placebo. This form of cue-contingent phasic alpha activity is also known to index attentional biasing within visual space. Specifically, when a relevant location is pre-cued, anticipatory alpha power increases contralateral to the location to be ignored. Here we investigate whether the effects of L-theanine on tonic and phasic alpha activity, found previously during intersensory attentional deployment, occur also during a visuospatial task. Subjects/Methods 168-channel EEG data were recorded from thirteen neurologically normal individuals while engaged in a highly demanding visuo-spatial attention task. Participants underwent testing on two separate days, ingesting either a 250-mg colorless and tasteless solution of L-theanine mixed with water, or a water-based solution placebo on each day in counterbalanced order. We compared the alpha-band activity when subjects ingested L-Theanine vs. Placebo. Results We found a significant reduction in tonic alpha for the L-theanine treatment compared to placebo, which was accompanied by a shift in scalp topography, indicative of treatment-related changes in the neural generators of oscillatory alpha activity. However, L-theanine did not measurably affect cue-related anticipatory alpha effects. Conclusions This pattern of results implies that L- theanine plays a more general role in attentional processing, facilitating longer-lasting processes responsible for sustaining attention across the timeframe of a difficult task, rather than affecting specific moment-to-moment phasic deployment processes.
6. Owen GN et al. The combined effects of L-theanine and caffeine on cognitive performance and mood. Nutr Neurosci. 2008 Aug;11(4):193-8. ABSTRACT: The aim of this study was to compare 50 mg caffeine, with and without 100 mg L-theanine, on cognition and mood in healthy volunteers. The effects of these treatments on word recognition, rapid visual information processing, critical flicker fusion threshold, attention switching and mood were compared to placebo in 27 participants. Performance was measured at baseline and again 60 min and 90 min after each treatment (separated by a 7-day washout). Caffeine improved subjective alertness at 60 min and accuracy on the attention-switching task at 90 min. The L- theanine and caffeine combination improved both speed and accuracy of performance of the attention-switching task at 60 min, and reduced susceptibility to distracting information in the memory task at both 60 min and 90 min. These results replicate previous evidence which suggests that L-theanine and caffeine in combination are beneficial for improving performance on cognitively demanding tasks.
7. Nobre AC et al. L-theanine, a natural constituent in tea, and its effect on mental state. Asia Pac J Clin Nutr. 2008;17 Suppl 1: 167-8. ABSTRACT: Tea is the most widely consumed beverage in the world after water. Tea is known to be a rich source of flavonoid antioxidants. However tea also contains a unique amino acid, L-theanine that may modulate aspects of brain function in humans. Evidence from human electroencephalograph (EEG) studies show that it has a direct effect on the brain (Juneja et al. Trends in Food Science & Tech 1999;10;199-204). L-theanine significantly increases activity in the alpha frequency band which indicates that it relaxes the mind without inducing drowsiness. However, this effect has only been established at higher doses than that typically found in a cup of black tea (approximately 20mg). The aim of the current research was to establish this effect at more realistic dietary levels. EEG was measured in healthy, young participants at baseline and 45, 60, 75, 90 and 105 minutes after ingestion of 50mg L-theanine (n=16) or placebo (n=19). Participants were resting with their eyes closed during EEG recording. There was a greater increase in alpha activity across time in the L-theanine condition (relative to placebo (p+0.05). A second study replicated this effect in participants engaged in passive activity. These data indicate that L- theanine, at realistic dietary levels, has a significant effect on the general state of mental alertness or arousal. Furthermore, alpha activity is known to play an important role in critical aspects of attention, and further research is therefore focussed on understanding the effect of L-theanine on attentional processes.
8. Bryan J et al. Psychological effects of dietary components of tea: caffeine and L-theanine. Nutr Rev. 2008 Feb;66(2):82-90. ABSTRACT: This review summarizes the literature on the association between two dietary components of tea, caffeine and L-theanine, and the psychological outcomes of consumption; it also identifies areas for future research. The studies reviewed suggest that caffeinated tea, when ingested at regular intervals, may maintain alertness, focused attention, and accuracy and may modulate the more acute effects of higher doses of caffeine. These findings concur with the neurochemical effects of L-theanine on the brain. L- theanine may interact with caffeine to enhance performance in terms of attention switching and the ability to ignore distraction; this is likely to be reflective of higher-level cognitive activity and may be sensitive to the detrimental effects of overstimulation. Further research should investigate the interactive effects of caffeine, L-theanine, and task complexity, utilize a range of ecologically valid psychological outcomes, and assess the neuroprotective effects of L-theanine using epidemiological or longer-term intervention studies among individuals at risk of neurodegenerative disease.
9. Haskell CF et al. The effects of L-theanine, caffeine and their combination on cognition and mood. Biol Psychol. 2008 Feb;77(2):113-22. ABSTRACT: L-Theanine is an amino acid found naturally in tea. Despite the common consumption of L-theanine, predominantly in combination with caffeine in the form of tea, only one study to date has examined the cognitive effects of this substance alone, and none have examined its effects when combined with caffeine. The present randomised, placebo-controlled, double-blind, balanced crossover study investigated the acute cognitive and mood effects of L-theanine (250 mg), and caffeine (150 mg), in isolation and in combination. Salivary caffeine levels were co-monitored. L-Theanine increased 'headache' ratings and decreased correct serial seven subtractions. Caffeine led to faster digit vigilance reaction time, improved Rapid Visual Information Processing (RVIP) accuracy and attenuated increases in self-reported 'mental fatigue'. In addition to improving RVIP accuracy and 'mental fatigue' ratings, the combination also led to faster simple reaction time, faster numeric working memory reaction time and improved sentence verification accuracy. 'Headache' and 'tired' ratings were reduced and 'alert' ratings increased. There was also a significant positive caffeine x L- theanine interaction on delayed word recognition reaction time. These results suggest that beverages containing L-theanine and caffeine may have a different pharmacological profile to those containing caffeine alone.
10. Nathan PJ et al. The neuropharmacology of L-theanine(N-ethyl-L-glutamine): a possible neuroprotective and cognitive enhancing agent. J Herb Pharmacother. 2006;6(2):21-30. ABSTRACT: L-theanine (N-ethyl-L-glutamine) or theanine is a major amino acid uniquely found in green tea. L-theanine has been historically reported as a relaxing agent, prompting scientific research on its pharmacology. Animal neurochemistry studies suggest that L- theanine increases brain serotonin, dopamine, GABA levels and has micromolar affinities for AMPA, Kainate and NMDA receptors. In addition has been shown to exert neuroprotective effects in animal models possibly through its antagonistic effects on group 1 metabotrophic glutamate receptors. Behavioural studies in animals suggest improvement in learning and memory. Overall, L-theanine displays a neuropharmacology suggestive of a possible neuroprotective and cognitive enhancing agent and warrants further investigation in animals and humans.
11. Kimura K et al. L-Theanine reduces psychological and physiological stress responses. Biol Psychol. 2007 Jan;74(1):39-45. ABSTRACT: L-theanine is an amino acid contained in green tea leaves which is known to block the binding of L-glutamic acid to glutamate receptors in the brain. Because the characteristics of L-Theanine suggest that it may influence psychological and physiological states under stress, the present study examined these possible effects in a laboratory setting using a mental arithmetic task as an acute stressor. Twelve participants underwent four separate trials: one in which they took L-Theanine at the start of an experimental procedure, one in which they took L-Theanine midway, and two control trials in which they either took a placebo or nothing. The experimental sessions were performed by double-blind, and the order of them was counterbalanced. The results showed that L-Theanine intake resulted in a reduction in the heart rate (HR) and salivary immunoglobulin A (s-IgA) responses to an acute stress task relative to the placebo control condition. Moreover, analyses of heart rate variability indicated that the reductions in HR and s-IgA were likely attributable to an attenuation of sympathetic nervous activation. Thus, it was suggested that the oral intake of L- Theanine could cause anti-stress effects via the inhibition of cortical neuron excitation.

L-Tyrosine

1. Owasoyo JO et al. Tyrosine and its potential use as a countermeasure to performance decrement in military sustained operations. Aviat Space Environ Med. 1992 May;63(5):364-9.
2. Roelands B et al. The effects of acute dopamine reuptake inhibition on performance. Med Sci Sports Exerc. 2008 May;40(5):879-85. doi: 10.1249/MSS.0b013e3181659c4d.
3. Shurtleff D et al. Tyrosine reverses cold-induced working memory deficit in humans. Pharmacol Biochem Behav. 1994 Apr;47(4):935-41. doi: 10.1016/0091-3057(94)90299-2.
4. Struder HK et al. Influence of paroxetine, branched-chain amino acids and tyrosine on neuroendocrine system responses and fatigue in humans. Horm Metab Res. 1998 Apr;30(4):188-94. doi: 10.1055/s-2007-978864.
5. Sutton EE et al. Ingestion of tyrosine: Effects on endurance, muscle strength, and anaerobic performance. Int. J. Sport Nutr. Exerc. Metab. 15:173– 185. doi: 10.1123/ijsnem.15.2.173.
6. Tumilty L et al. Oral tyrosine supplementation improves exercise capacity in the heat. Eur J Appl Physiol. 2011 Dec;111(12):2941-50. doi: 10.1007/s00421-011-1921-4.
7. Attipoe S et al. Tyrosine for Mitigating Stress and Enhancing Performance in Healthy Adult Humans, a Rapid Evidence Assessment of the Literature. Mil Med. 2015 Jul;180(7):754-65. doi: 10.7205/MILMED-D-14-00594.
8. Jongkees BJ et al. Effect of tyrosine supplementation on clinical and healthy populations under stress or cognitive demands--A review. J Psychiatr Res. 2015 Nov;70:50-7. doi: 10.1016/j.jpsychires.2015.08.014.
9. Rasmussen DD et al.Effects of tyrosine and tryptophan ingestion on plasma catecholamine and 3,4-dihydroxyphenylacetic acid concentrations. J Clin Endocrinol Metab. 1983 Oct;57(4):760-3. doi: 10.1210/jcem-57-4-760.

Lycopene

1. Chen P et al. Lycopene and Risk of Prostate Cancer. A Systematic Review and Meta-Analysis. Medicine (Baltimore). 2015 Aug;94(33): 31260. Summary: This study demonstrates that higher lycopene consumption/circulating concentration is associated with a lower risk of PCa.
2. Lane JA et al. ProDiet: A Phase II Randomized Placebo-controlled Trial of Green Tea Catechins and Lycopene in Men at Increased Risk of Prostate Cancer. Cancer Pres Res. 2018 Nov;11(1): 687-696. Summary: This study aimed to establish the feasibility and acceptability of dietary modification in men at increased risk of prostate cancer. Men were invited with a PSA level of 2.0-2.95 ng/mL or 3.0-19.95 ng/mL with negative prostate biopsies. Randomization (3 × 3 factorial design) to daily green tea and lycopene: green tea drink (3 cups, unblinded) or capsules [blinded, 600 mg flavan-3-ol ()-epigallocatechin-3-gallate (EGCG) or placebo] and lycopene-rich foods (unblinded) or capsules (blinded, 15 mg lycopene or placebo) for 6 months. All interventions were acceptable and well tolerated although men preferred the capsules. Dietary prevention is acceptable to men at risk of prostate cancer. This intervention trial demonstrates that a chemoprevention clinical trial is feasible.
3. Wang Y et al. Lycopene, tomato products and prostate cancer-specific mortality among men diagnosed with nonmetastatic prostate cancer in the Cancer Prevention Study II Nutrition Cohort. Int J Cancer. 2016 June 15;138 (12):2846-55. Summary: Among men with high-risk cancers (T3-T4 or Gleason score 8-10, or nodal involvement), consistently reporting lycopene intake ≥ median on both post diagnosis surveys was associated with lower prostate-cancer specific mortality.

Magnesium

1. Cinar V et al. Effects of magnesium supplementation on testosterone levels of athletes and sedentary subjects at rest and after exhaustion. Biol Trace Elem Res. 2011 Apr;140(1):18-23.
2. Maguire D et al. Telomere Homeostasis: Interplay with Magnesium. Int J Mol Sci. 2018 Jan 5;19(1):157. doi: 10.3390/ijms19010157.
3. Ford ES et al. Magnesium Intake in a National Sample of US Adults. J Nutr. 2003 Sep;133(9):2879-82. doi: 10.1093/jn/133.9.2879.
4. Zofková I et al. The relationship between magnesium and calciotropic hormones. Magnes Res. 1995 Mar;8(1):77-84.
5. B T Altura, et al. Magnesium dietary intake modulates blood lipid levels and atherogenesis. Proc Natl Acad Sci U S A. 1990 March; 87(5): 1840–1844.
6. Cohen H et al. Atherogenesis inhibition induced by magnesium-chloride fortification of drinking water. Biol Trace Elem Res. Winter 2002;90(1-3):251-9. doi: 10.1385/BTER:90:1-3:251.
7. Ma B et al. Dairy, Magnesium, and Calcium Intake in Relation to Insulin Sensitivity: Approaches to Modeling a Dose-dependent Association. Am J Epidemiol. 2006 Sep 1;164(5):449-58. doi: 10.1093/aje/kwj246. Epub 2006 Jul 21.
8. Huerta MG et al. Magnesium deficiency is associated with insulin resistance in obese children. Diabetes Care. 2005 May;28(5):1175-81. doi: 10.2337/diacare.28.5.1175.
9. Song Y et al. Dietary magnesium intake in relation to plasma insulin levels and risk of type 2 diabetes in women. Diabetes Care. 2004 Jan;27(1):59-65. doi: 10.2337/diacare.27.1.59.
10. Lopez-Ridaura R et al. Magnesium intake and risk of type 2 diabetes in men and women. Diabetes Care. 2004 Jan;27(1):134-40. doi: 10.2337/diacare.27.1.134.
11. Balon TW et al. Magnesium supplementation reduces development of diabetes in a rat model of spontaneous NIDDM. Am J Physiol. 1995 Oct;269(4 Pt 1):E745-52. doi: 10.1152/ajpendo.1995.269.4.E745.
12. Nadler JL et al. Magnesium deficiency produces insulin resistance and increased thromboxane synthesis. Hypertension. 1993 Jun;21(6 Pt 2):1024-9. doi: 10.1161/01.hyp.21.6.1024.
13. Rodríguez-Morán M et al. Oral magnesium supplementation improves insulin sensitivity and metabolic control in type 2 diabetic subjects: a randomized double-blind controlled trial. Diabetes Care. 2003 Apr;26(4):1147-52. doi: 10.2337/diacare.26.4.1147.
14. Mooren FC et al. Oral magnesium supplementation reduces insulin resistance in non-diabetic subjects - a double-blind, placebo-controlled, randomized trial. Diabetes Obes Metab. 2011 Mar;13(3):281-4.
15. Guerrero-Romero F et al. Oral magnesium supplementation improves insulin sensitivity in non-diabetic subjects with insulin resistance. A double-blind placebo-controlled randomized trial. Diabetes Metab. 2004 Jun;30(3):253-8. doi: 10.1016/s1262-3636(07)70116-7.
16. Chakraborti S et al. Protective role of magnesium in cardiovascular diseases: a review. Mol Cell Biochem. 2002 Sep;238(1-2):163-79. doi: 10.1023/a:1019998702946.
17. Maier JAM. Low magnesium and atherosclerosis: an evidence-based link. Mol Aspects Med. Feb-Jun 2003;24(1-3):137-46.
18. Bohn T et al. Phytic acid added to white-wheat bread inhibits fractional apparent magnesium absorption in humans. Am J Clin Nutr. 2004 Mar;79(3):418-23. doi: 10.1093/ajcn/79.3.418.

Omega-3 Fatty Acids

1. Barbadoro P et al. Fish oil supplementation reduces cortisol basal levels and perceived stress: A randomized, placebo-controlled trial in abstinent alcoholics. Mol Nutr Food Res. 2013 Jun;57(6):1110-4. doi: 10.1002/mnfr.201200676.
2. Tabbaa T et al. Docosahexaenoic acid, inflammation, and bacterial dysbiosis in relation to periodontal disease, inflammatory bowel disease, and the metabolic syndrome. Nutrients. 2013 Aug 19;5(8):3299-310. doi: 10.3390/nu5083299.
3. Sarfarinejad MR. Effect of omega-3 polyunsaturated fatty acid supplementation on semen profile and enzymatic anti-oxidant capacity of seminal plasma in infertile men with idiopathic oligoasthenoteratospermia: a double-blind, placebo-controlled, randomised study. Andrologia. 2011 Feb;43(1):38-47. doi: 10.1111/j.1439-0272.2009.01013.x.
4. Dyall SC. Long-chain omega-3 fatty acids and the brain: a review of the independent and shared effects of EPA, DPA and DHA. Front Aging Neurosci. 2015 Apr 21;7:52. doi: 10.3389/fnagi.2015.00052.
5. Esmaeili V et al. Dietary fatty acids affect semen quality: a review. Andrology. 2015 May;3(3):450-61. doi: 10.1111/andr.12024.
6. So J et al. EPA and DHA differentially modulate monocyte inflammatory response in subjects with chronic inflammation in part via plasma specialized pro-resolving lipid mediators: A randomized, double-blind, crossover study. Atherosclerosis. 2021 Jan;316:90-98. doi: 10.1016/j.atherosclerosis.2020.11.018.
7. Harris WS et al. Blood n-3 fatty acid levels and total and cause-specific mortality from 17 prospective studies. Nat Commun. 2021 Apr 22;12(1):2329. doi: 10.1038/s41467-021-22370-2.
8. Burhani MD et al. Fish oil and depression: The skinny on fats. J Integr Neurosci. 2017;16(s1):S115-S124. doi: 10.3233/JIN-170072.
9. Gertsik L et al. Omega-3 Fatty Acid Augmentation of Citalopram Treatment for Patients With Major Depressive Disorder. J Clin Psychopharmacol. 2012 Feb;32(1):61-4. doi: 10.1097/JCP.0b013e31823f3b5f.
10. Jensen TK et al. Associations of fish oil supplement use with testicular function in young men. JAMA Netw Open. 2020 Jan 3;3(1):e1919462. doi: 10.1001/jamanetworkopen.2019.19462.
11. Mischoulon D et al. Omega-3 fatty acids for mood disorders. Harvard Health Publishing. August 03, 2018.
12. Philpott JD et al. Applications of omega-3 polyunsaturated fatty acid supplementation for sport performance. Res Sports Med. Apr-Jun 2019;27(2):219-237. doi: 10.1080/15438627.2018.1550401.

PhGABA

1. Lapin I. Phenibut: a tranquilizer and nootropic drug. CNS Drug Rev. 2001 Winter;7(4):471-81.
2. Blackshaw LA. Receptors and transmission in the brain-gut axis: potential for novel therapies. IV. GABA(B) receptors in the brain-esophageal axis. Am J Physiol Gastrointest Liver Physiol. 2001 Aug;281(2):G311-5.
3. Lapin IP. Differences and similarity in the interaction of fenibut, baclofen, and diazepam with phenylethylamine. Farmakol Toksikol. 1985 Jul-Aug;48(4).
4. Shulgina GI. On neurotransmitter mechanisms of reinforcement and internal inhibition. Pavlov J Biol Sci. Oct-Dec 1986;21(4):129-40. doi: 10.1007/BF02734511.

Phosphatidic Acid

1. Hoffman JR et al. Efficacy of phosphatidic acid ingestion on lean body mass, muscle thickness and strength gains in resistance-trained men. Journal of the International Society of Sports Nutrition. 9:47 2012. ABSTRACT: Background: Phosphatidic acid (PA) has been reported to activate the mammalian target of rapamycin (mTOR) signaling pathway and is thought to enhance the anabolic effects of resistance training. The purpose of this pilot study was to examine if oral phosphatidic acid administration can enhance strength, muscle thickness and lean tissue accruement during an 8-week resistance training program. Methods: Sixteen resistance-trained men were randomly assigned to a group that either consumed 750 mg of PA (n = 7, 23.1 ± 4.4 y; 176.7 ± 6.7 cm; 86.5 ± 21.2 kg) or a placebo (PL, n = 9, 22.5 ± 2.0 y; 179.8 ± 5.4 cm; 89.4 ± 13.6 kg) group. During each testing session subjects were assessed for strength (one repetition maximum [1-RM] bench press and squat) and body composition. Muscle thickness and pennation angle were also measured in the vastus lateralis of the subject’s dominant leg. Results: Subjects ingesting PA demonstrated a 12.7% increase in squat strength and a 2.6% increase in LBM, while subjects consuming PL showed a 9.3% improvement in squat strength and a 0.1% change in LBM. Although parametric analysis was unable to demonstrate significant differences, magnitude based inferences indicated that the Δ change in 1-RM squat showed a likely benefit from PA on increasing lower body strength and a very likely benefit for increasing lean body mass (LBM). Conclusions: Results of this study suggest that a combination of a daily 750 mg PA ingestion, combined with a 4-day per week resistance training program for 8-weeks appears to have a likely benefit on strength improvement, and a very likely benefit on lean tissue accruement in young, resistance trained individuals.
2. Joy JM et al. Phosphatidic acid enhances mTOR signaling and resistance exercise induced hypertrophy. Nutrition & Metabolism 2014, 11:29. ABSTRACT: Introduction: The lipid messenger phosphatidic acid (PA) plays a critical role in the stimulation of mTOR signaling. However, the mechanism by which PA stimulates mTOR is currently unknown. Therefore, the purpose of this study was to compare the effects of various PA precursors and phospholipids on their ability to stimulate mTOR signaling and its ability to augment resistance training-induced changes in body composition and performance. Methods: In phase one, C2C12 myoblasts cells were stimulated with different phospholipids and phospholipid precursors derived from soy and egg sources. The ratio of phosphorylated p70 (P-p70-389) to total p70 was then used as readout for mTOR signaling. In phase two, resistance trained subjects (n = 28, 21 ± 3 years, 77 ± 4 kg, 176 ± 9 cm) consumed either 750 mg PA daily or placebo and each took part in an 8 week periodized resistance training program. Results: In phase one, soy-phosphatidylserine, soy-Lyso-PA, egg-PA, and soy-PA stimulated mTOR signaling, and the effects of soy-PA (+636%) were significantly greater than egg-PA (+221%). In phase two, PA significantly increased lean body mass (+2.4 kg), cross sectional area (+1.0 cm), and leg press strength (+51.9 kg) over placebo. Conclusions: PA significantly activates mTOR and significantly improved responses in skeletal muscle hypertrophy, lean body mass, and maximal strength to resistance exercise.
3. Joy JM et al. Phosphatidic Acid Supplementation Increases Skeletal Muscle Hypertrophy and Strength. Poster presented at the 2013 annual ISSN conference, and manuscript now in review for submission to publication. ABSTRACT: Introduction: The accretion of skeletal muscle tissue can be critical for a varied population including athletes and elderly. Skeletal muscle hypertrophy is largely mediated through increased muscle protein synthesis. The mammalian target of rapamycin (mTOR) has been shown to regulate rates of muscle protein synthesis and a mechanical stimulus (resistance exercise) has been shown to activate mTOR with the phospholipid Phosphatidic Acid (PA) playing a key role. A first pilot study found that oral supplementation with soy-derived PA in athletes undergoing progressive resistance training very likely resulted in greater increases in squat strength and lean mass over the placebo. However, this pilot study was likely underpowered, the workout was not supervised and no direct measures of skeletal muscle hypertrophy were taken. Therefore, the purpose of this study was to investigate the effects of PA on body composition, strength, power and muscular hypertrophy. Methods: Twenty-eight resistance trained, male subjects (21 ± 3 years of age, bodyweight of 76 ± 9 kg, and height of 176 cm ± 9 cm) participated in this study. Subjects were equally divided into experimental and control conditions, and each subject took part in an 8 week periodized resistance training program. The resistance training program consisted of two hypertrophy oriented workouts per week and one strength oriented workout per week. The experimental condition (EXP) received 750 mg of soy-derived PA (Mediator™, Chemi Nutra, White Bear Lake, MN), while the control condition (CON) received a visually identical placebo (rice flour). Measurements of DEXA-determined body composition, rectus femoris CSA, 1RM strength, and anaerobic power were taken prior to and following the 8 week training intervention. A 2x2 repeated measures ANOVA was used to determine group, time, and group x time interactions. A Tukey post-hoc was used to locate differences. Results: There was a significant group x time effect (p=0.02) for CSA, in which the EXP group increased (+1.01 cm2, ES = 0.92) to a greater extent than the CON group (+0.61 cm2, ES = 0.52). There was a significant group x time effect (p=0.01) for LBM, in which the EXP group (+2.4 kg, ES = 0.42) doubled the effects of resistance training alone (CON +1.2 kg, ES = 0.26). There was a significant group x time effect (p=0.04) for leg press 1RM, in which the EXP group increased to a greater extent (+52.0 kg, ES = 1.2) than the CON group (+32.5 kg, ES = 0.78). There was a trend group x time effect (p=0.06) for fat loss, in which the EXP group decreased body fat to a greater extent than the CON group (-1.3kg vs. -0.5kg). Conclusions: Supplementation with soy-derived PA can improve responses in skeletal muscle hypertrophy, lean body mass, and maximal strength.
4. Joy JM et al. The Effects of 8 Weeks of Phosphatidic Acid Supplementation on Cardiovascular, Kidney, and Liver Safety in Health Young Males. Poster presented at the 2013 annual ISSN conference. ABSTRACT: Background: The mammalian target of rapamycin (mTOR) has been shown to regulate rates of muscle protein synthesis, and one novel nutritional activator of mTOR is the phospholipid Phosphatidic Acid (PA). We have recently found that PA supplementation over 8 weeks of resistance training augmented responses in skeletal muscle hypertrophy and strength. However, we are unaware of research investigating the safety of PA in human subjects. Therefore the purpose of this study was to investigate the effects of 8 weeks of 750 mg per day of PA supplementation on safety parameters in healthy college aged males. Methods: Twenty-eight healthy, college aged male subjects (21 ± 3 years of age, bodyweight of 76 ± 9 kg, and height of 176 cm ± 9 cm) participated in this study. Subjects were equally divided into experimental and control conditions. The experimental condition (EXP) received 750 mg of soy-derived PA (Mediator™, Chemi Nutra, White Bear Lake, MN), while the control condition (CON) received a visually identical placebo (rice flour). Measures of cardiovascular, kidney, and liver function were analyzed with a full CMP and CBC prior to and 8 weeks following supplementation. This analysis included: total, high density, and low density lipoproteins, blood glucose, blood urea nitrogen, creatinine, eGFR, Na, K, Cl, CO2, Ca, protein, albumin, globulin, albumin:globulin ratio, total bilirubin, alkaline phosphatase, aspartate aminotransferase, and alanine aminotransferase. In addition a sample of urine was submitted for analysis of urine specific gravity and pH. A 2x2 repeated measures ANOVA was used to determine group, time, and group x time interactions. A Tukey post-hoc was used to locate differences. Results: There were no differences at baseline in blood chemistry and hematology between the CON and EXP supplemented groups. Additionally no differences were observed in urinalysis values between the groups. Moreover no group, or group X time effects were found following 8 weeks of supplementation. Conclusions: Soy-derived PA is a safe nutritional supplement for healthy college aged subjects if taken up to a dosage of 750 mg over an eight week period.
5. Fang Y et al. Phosphatidic Acid-Mediated Mitogenic Activation of mTOR Signaling. Science. 30 Nov 2001: Vol. 294, Issue 5548, pp. 1942-1945. DOI: 10.1126/science.1066015. ABSTRACT: The mammalian target of rapamycin (mTOR) governs cell growth and proliferation by mediating the mitogen- and nutrient-dependent signal transduction that regulates messenger RNA translation. We identified phosphatidic acid (PA) as a critical component of mTOR signaling. In our study, mitogenic stimulation of mammalian cells led to a phospholipase D–dependent accumulation of cellular PA, which was required for activation of mTOR downstream effectors. PA directly interacted with the domain in mTOR that is targeted by rapamycin, and this interaction was positively correlated with mTOR's ability to activate downstream effectors. The involvement of PA in mTOR signaling reveals an important function of this lipid in signal transduction and protein synthesis, as well as a direct link between mTOR and mitogens. Furthermore, these studies suggest a potential mechanism for the in vivo actions of the immunosuppressant rapamycin.
6. Gundermann D et al. Soy-derived Phosphatidic Acid, Lysophosphatidic Acid and Phosphatidylserine are Sufficient to Induce an Increase in mTOR Signaling. Poster presented at the 2013 annual ISSN conference, and manuscript now in review for submission to publication. ABSTRACT: Background: A protein kinase called the mechanistic target of rapamycin (mTOR) is a well-known regulator of cellular growth. In fact, several studies have indicated that the kinase activity of mTOR is required for mechanically-induced increases in skeletal muscle protein synthesis and hypertrophy. Previous studies have also determined that the lipid messenger phosphatidic acid (PA) plays a critical role in the stimulation of mTOR signaling and, an increase in PA concentration is sufficient for the activation of mTOR signaling. However, the mechanism by which PA stimulates mTOR is currently unknown. A primary target of mTOR includes the phosphorylation of p70 on the threonine 389 residue (P-p70-389), and thus, is a commonly accepted readout for the activation of mTOR. PA can be synthesized from a variety of reactions via multiple reactants. Therefore, the purpose of this study was to compare the effects of various PA precursors on their ability to stimulate mTOR signaling and determine if any other phospholipid species are also capable of stimulating mTOR signaling. Methods: C2C12 myoblasts were plated at approximately 30% confluence and grown for 24 hours in 10% FBS High Glucose DMEM. Cells were switched to 2mL/well serum free high glucose DMEM (no antibiotics) for 16 hours prior to the experiment. Cells were approximately 70% confluent at the time of the experiment. Cells were then stimulated for 20 minutes with vehicle (Control) or 10, 30 or 100µM of soy-derived phosphatidylserine (S-PS, SerinAid, Chemi Nutra, White Bear Lake, MN), phosphatidylinositol (S-PI), phosphatidylethanolamine (S-PE), phosphatidylcholine (S-PC), PA (S-PA, Mediator, Chemi Nutra, White Bear Lake, MN), lysophosphatidic acid (S-LPA), diacylglycerol (DAG), glycerol-3-phosphate (G3P), and egg-derived PA (E-PA). Cells were harvested in lysis buffer and subjected to immunoblotting. The ratio of P-p70-389 to total p70 was used as readout for mTOR signaling. Results: S-PI, S-PE, S-PC, DAG, and G3P elicited no increase in the ratio of P-p70-389 to total p70 compared to vehicle stimulated cells. In contrast, elevated mTOR signaling was observed at all tested concentrations of S-PS (529, 588, and 457%), S-LPA (649, 866, and 1,132%), and S-PA (679, 746, and 957%; P < 0.05). Egg-PA induced an 873% increase in mTOR signaling with the 100µM dose (P < 0.05), whereas no significant increase was observed with the 10 or 30µM doses. Conclusions: S-PA, S-LPA and S-PS are each sufficient to induce an increase in mTOR signaling. Therefore, they may be capable of enhancing the anabolic effects of resistance training and contributing to muscle accretion over time. Furthermore, S-PA is a more potent stimulator of mTOR signaling than PA derived from egg.
7. Purpura M et al. Effect of Oral Administration of Soy-Derived Phosphatidic Acid on Concentrations of Phosphatidic Acid and lyso-Phosphatidic Acid Molecular Species In Human Plasma. Poster presented at the 2013 annual ISSN conference. ABSTRACT: Background: The glycerophospholipid Phosphatidic acid (PA) has been identified as a potential nutritional treatment for gastrointestinal disorders. Dietary food sources rich in PA include cabbage and radish leaves as well as Mallotus japonicas, a Japanese edible herb historically used for the treatment of stomach ulcers. The mammalian target of rapamycin (mTOR) has been shown to regulate rates of muscle protein synthesis and a mechanical stimulus (resistance exercise) has been shown to activate mTOR with PA playing a key role. Supplementation with soy-derived PA significantly increases responses in skeletal muscle hypertrophy, lean body mass, and maximal strength to resistance exercise. PA accounts for less than 0.1% of the total glycerophospholipid concentration of 201 mg/dl in the human plasma. 15 of the more than 600 distinct molecular lipid species quantified in human plasma are PA, 6 are lysophosphatidic acid (LPA). Orally administered PA can be metabolized to LPA and glycerophosphate by pancreatic phospholipases A1 and A2, which hydrolyze the fatty acid at the sn-1 position and the sn-2 position, respectively. Lysophospholipids are absorbed by the mucosal cells of the gastrointestinal tract and are rapidly re-acylated with fatty acids of the body pool resulting in a newly-formed phospholipid-molecule whose fatty acid composition is determined by the physiological and nutritional status and not by its source. This study sought to assess the effect of soy-derived PA supplementation on concentrations LPA and PA molecular species in human plasma. Methods: After a 12 hour overnight fast one subject (20 years of age, bodyweight of 82 kg, and height of 178 cm) was assigned to receive 1.5 grams of soy-derived PA (Mediator, Chemi Nutra, White Bear Lake, MN). Blood draws were taken immediately prior to, and at 30 min, 1, 2, 3, and 7 hours following supplementation. The samples were analyzed by an ultra-performance liquid chromatograph with triple quadrupole mass spectrometry (LC/MS/MS) using 17:1-LPA and 37:4-PA as internal standards to determine the concentration of LPA and PA molecular species in human plasma. Results: At baseline, 19 PA (highest concentrations: C34:2 (15%), C40:4 (11%), and C36:4 (10%)) and 5 LPA (16:0 (45%), 18:2 (19%), 20:4 (17%), 14:0 (11%) and 18:1 (8%)) molecular species could be quantified with total concentrations of PA of 2.66 nmol/ml, and LPA of 0.11 nmol/ml. Plasma concentrations of PA peaked at 3 hours (+32%) after ingestion and stayed elevated even after 7 hours (+18%). LPA showed a bimodal absorption kinetic with peaks after 1 hour (+500%) and 3 hours (+264%), after almost dropping back to baseline levels after 2 hours. On an individual fatty acid level, most prominent was a 23-fold increase in 20:4-LPA after 1 hour compared to baseline. The increase in 20:4-LPA does not result from the administration of PA, since soy-derived PA does not contain any arachidonic acid (fatty acids distribution of soy-PA: 18:2 (66.1%), 18:1 (12.6%), 16:0 (11.7%), 18:3 (6.1%) and 18:0 (3.4%)). Absorption of soy-derived PA must yield glycerophosphate which is re-acylated with arachidonic acid. Conclusion: LPA and PA can be molecularly identified and measured. LPA, PA and LPA+PA plasma levels increase 30 min after ingestions, plateau at 1-3 hours and remain above baseline levels after 7 hours. This is the first case study showing that orally administered PA is bioavailable. Future research should repeat this case study with a larger n-size and include the analysis of omega 3 fatty acid-LPA molecular species.
8. Hornberger TA et al. The role of phospholipase D and phosphatidic acid in the mechanical activation of mTOR signaling in skeletal muscle. PNAS 103(12): 4741-4746, 2006. ABSTRACT: Signaling by the mammalian target of rapamycin (mTOR) has been reported to be necessary for mechanical load-induced growth of skeletal muscle. The mechanisms involved in the mechanical activation of mTOR signaling are not known, but several studies indicate that a unique [phosphotidylinositol-3-kinase (PI3K)- and nutrient-independent] mechanism is involved. In this study, we have demonstrated that a regulatory pathway for mTOR signaling that involves phospholipase D (PLD) and the lipid second messenger phosphatidic acid (PA) plays a critical role in the mechanical activation of mTOR signaling. First, an elevation in PA concentration was sufficient for the activation of mTOR signaling. Second, the isozymes of PLD (PLD1 and PLD2) are localized to the z-band in skeletal muscle (a critical site of mechanical force transmission). Third, mechanical stimulation of skeletal muscle with intermittent passive stretch ex vivo induced PLD activation, PA accumulation, and mTOR signaling. Finally, pharmacological inhibition of PLD blocked the mechanically induced increase in PA and the activation of mTOR signaling. Combined, these results indicate that mechanical stimuli activate mTOR signaling through a PLD-dependent increase in PA. Furthermore, we showed that mTOR signaling was partially resistant to rapamycin in muscles subjected to mechanical stimulation. Because rapamycin and PA compete for binding to the FRB domain on mTOR, these results suggest that mechanical stimuli activate mTOR signaling through an enhanced binding of PA to the FRB domain on mTOR.
9. O’Neil TK et al. The role of phosphoinositide 3-kinase and phosphatidic acid in the regulation of mammalian target of rapamycin following eccentric contractions. J Physiol 581.14: 3691-3701, 2009. ABSTRACT: Resistance exercise induces a hypertrophic response in skeletal muscle and recent studies have begun to shed light on the molecular mechanisms involved in this process. For example, several studies indicate that signalling by the mammalian target of rapamycin (mTOR) is necessary for a hypertrophic response. Furthermore, resistance exercise has been proposed to activate mTOR signalling through an upstream pathway involving the phosphoinositide 3-kinase (PI3K) and protein kinase B (PKB); however, this hypothesis has not been thoroughly tested. To test this hypothesis, we first evaluated the temporal pattern of signalling through PI3K-PKB and mTOR following a bout of resistance exercise with eccentric contractions (EC). Our results indicated that the activation of signalling through PI3K-PKB is a transient event (< 15 min), while the activation of mTOR is sustained for a long duration (>12 h). Furthermore, inhibition of PI3K-PKB activity did not prevent the activation of mTOR signalling by ECs, indicating that PI3K-PKB is not part of the upstream regulatory pathway. These observations led us to investigate an alternative pathway for the activation of mTOR signalling involving the synthesis of phosphatidic acid (PA) by phospholipase D (PLD). Our results demonstrate that ECs induce a sustained elevation in [PA] and inhibiting the synthesis of PA by PLD prevented the activation of mTOR. Furthermore, we determined that similar to ECs, PA activates mTOR signalling through a PI3K-PKB-independent mechanism. Combined, the results of this study indicate that the activation of mTOR following eccentric contractions occurs through a PI3K-PKB-independent mechanism that requires PLD and PA.
10. Winter JN et al. Phosphatidic acid mediates activation of mTORC1 through the ERK signaling pathway. Am J Physiol Cell Physiol. 299: C335-C344, 2010. ABSTRACT: The mammalian target of rapamycin (mTOR) assembles into two distinct multiprotein complexes known as mTORC1 and mTORC2. Of the two complexes, mTORC1 acts to integrate a variety of positive and negative signals to downstream targets that regulate cell growth. The lipid second messenger, phosphatidic acid (PA), represents one positive input to mTORC1, and it is thought to act by binding directly to mTOR, thereby enhancing the protein kinase activity of mTORC1. Support for this model includes findings that PA binds directly to mTOR and addition of PA to the medium of cells in culture results in activation of mTORC1. In contrast, the results of the present study do not support a model in which PA activates mTORC1 through direct interaction with the protein kinase but, instead, show that the lipid promotes mTORC1 signaling through activation of the ERK pathway. Moreover, rather than acting directly on mTORC1, the results suggest that exogenous PA must be metabolized to lysophosphatidic acid (LPA), which subsequently activates the LPA receptor endothelial differentiation gene (EDG-2). Finally, in contrast to previous studies, the results of the present study demonstrate that leucine does not act through phospholipase D and PA to activate mTORC1 and, instead, show that the two mediators act through parallel upstream signaling pathways to activate mTORC1. Overall, the results demonstrate that leucine and PA signal through parallel pathways to activate mTORC1 and that PA mediates its effect through the ERK pathway, rather than through direct binding to mTOR.

Phosphatidylcholine

1. Chung SY. Administration of phosphatidylcholine increases brain acetylcholine concentration and improves memory in mice with dementia. J Nutr. 1995 Jun;125(6):1484-9. doi: 10.1093/jn/125.6.1484.
2. Wiedeman AM et al. Dietary Choline Intake: Current State of Knowledge Across the Life Cycle. Nutrients. 2018 Oct 16;10(10):1513. doi: 10.3390/nu10101513.
3. van der Veen JN et al. The critical role of phosphatidylcholine and phosphatidylethanolamine metabolism in health and disease. Biochim Biophys Acta Biomembr. 2017 Sep;1859(9 Pt B):1558-1572. doi: 10.1016/j.bbamem.2017.04.006.

Piperine

1. Park UH et al. Piperine, a component of black pepper, inhibits adipogenesis by antagonizing PPARγ activity in 3T3-L1 cells. J Agric Food Chem. 2012 Apr 18;60(15):3853-60. doi: 10.1021/jf204514a. Epub 2012 Apr 6.
2. Kim J et al. Piperine enhances carbohydrate/fat metabolism in skeletal muscle during acute exercise in mice. Nutr Metab (Lond). 2017; 14: 43. Published online 2017 Jul 4. doi: 10.1186/s12986-017-0194-2.

Punicalagins (Pomegranate)

1. Sharma P et al. Pomegranate for Prevention and Treatment of Cancer: An Update. Molecules. 24 January 2017. Summary: This review summarizes preclinical and clinical studies highlighting the role of pomegranate in prevention and treatment of skin, breast, prostate, lung, and colon cancers.
2. Vicinanza R et al. Pomegranate Juice Metabolites, Ellagic Acid and Urolithin A, Synergistically Inhibit Androgen-Independent Prostate Cancer Cell Growth via Distinct Effects on Cell Cycle Control and Apoptosis. Evidence Based Complementary and Alternative Medicine. 19 February 2013. Summary: This study describes how ellagitannins (ETs) from pomegranate juice (PJ) are bioactive polyphenols with chemopreventive potential against prostate cancer (PCa).
3. Wang L et al. Cellular and molecular mechanisms of pomegranate juice-induced anti-metastatic effect on prostate cancer cells. Integr Biol. 2011 May 19. Summary: This study demonstrates how, in addition to causing cell death of hormone-refractory prostate cancer cells, pomegranate also increases cell adhesion and decreases cell migration of the cells that do not die.
4. Chaves FM et al. Pomegranate Juice and Peel Extracts are Able to Inhibit Proliferation, Migration and Colony Formation of Prostate Cancer Cell Lines and Modulate the Akt/mTOR/S6K Signaling Pathway. Plant Foods Hum Nutr. 2020 March;75(1):54-62. Summary: This study presents evidence that both juice and isolated peel extracts from pomegranate fruit have important anti-cancer effects against prostate cancer cells, modulating the mTOR/S6K signaling pathway.
5. Kroeger N et al. Pomegranate Extracts in the Management of Men's Urologic Health: Scientific Rationale and Preclinical and Clinical Data. Evid Based Complement Alternat Med. 2013 Mar 26. Abstract: Numerous micronutrients and polyphenols found in soy, green tea, and many fruits and vegetables have been described to impact diseases including erectile dysfunction, benign prostatic hyperplasia, and prostate cancer. However, oftentimes these reports lack both a scientific rationale and supportive evidence base. The efficacy of pomegranate, on the other hand, in the modulation of central biological processes like inflammation, hypoxia, and oxidative stress that are important in the pathogenesis of urological maladies has been robustly demonstrated in preclinical in vitro and in vivo studies. Moreover, clinical trials have further supported its use in the treatment of several diseases, in particular in the management of prostate cancer.
6. Forest CP et al. Efficacy and safety of pomegranate juice on improvement of erectile dysfunction in male patients with mild to moderate erectile dysfunction: a randomized, placebo-controlled, double-blind, crossover study. Int J Impot Res. Nov-Dec 2007;19(6):564-7. Abstract: This randomized-controlled trial examined the efficacy of wonderful variety pomegranate juice versus placebo in improving erections in 53 completed subjects with mild to moderate erectile dysfunction. The crossover design consisted of two 4-week treatment periods separated by a 2-week washout. Efficacy was assessed using International Index of Erectile Function (IIEF) and Global Assessment Questionnaires (GAQ). Of the 42 subjects who demonstrated improvement in GAQ scores after beverage consumption, 25 reported improvement after drinking pomegranate juice. Further, 17 subjects showed preference of one beverage to the other. Subjects were more likely to have improved scores when pomegranate juice was consumed (P=0.058).
7. Jeranka JS. Therapeutic Applications of Pomegranate (Punica granatum L.): A Review. Alternative Medicine Review: A Journal of Clinical Therapeutic, July 2008,13(2):128-44. Abstract: In addition to its ancient historical uses, pomegranate is used in several systems of medicine for a variety of ailments. In the past decade, numerous studies on the antioxidant, anticarcinogenic, and anti-inflammatory properties of pomegranate constituents have been published, focusing on treatment and prevention of cancer, cardiovascular disease, diabetes, dental conditions, erectile dysfunction, bacterial infections and antibiotic resistance, and ultraviolet radiation-induced skin damage. Other potential applications include infant brain ischemia, male infertility, Alzheimer's disease, arthritis, and obesity.
8. Gur S et al. Characterisation of pomegranate juice effects on human corpus cavernosum. Andrologia 2017 Oct;49(8). Abstract: This study evaluates the molecular characterisation and confirmation of POM's action on human corpus cavernosum (HCC) obtained from patients (n = 16) undergoing penile prosthesis implantation. We conclude that POM induced marked relaxation of HCC via: (i) nNOS stimulation, and (ii) downstream relaxation stimulated by nNOS and cGMP and bypassing the NO and PDE5. This action provides a rationale for the therapeutic or preventative use of POM in men with erectile dysfunction who do not respond well to PDE5 inhibitors.
9. Azadzoi K et al. Oxidative stress in arteriogenic erectile dysfunction: prophylactic role of antioxidants. J Urol. 2005 Jul;174(1):368-93. Abstract: Antioxidant activity of known antioxidant beverages, such as pomegranate juice (PJ), red wine, blueberry juice, cranberry juice, orange juice and green tea, was examined spectrophotometrically. PJ demonstrated the highest free radical scavenging capacity. Antioxidant therapy may be a useful prophylactic tool for preventing smooth muscle dysfunction and fibrosis in ED.
10. Deng Y et al. The extract from Punica granatum (pomegranate) peel induces apoptosis and impairs metastasis in prostate cancer cells. Biomed Pharmacother. 2017 Se9;93:976-984. Abstract: This study aimed to investigate the effects of pomegranate peel extract (PoPx) on the apoptosis and metastasis of prostate cancer cells and the related mechanism. They found that PoPx showed growth inhibition on prostate cancer cells. Nuclei morphological and flow cytometer (FCM) analysis indicated that PoPx could induce prostate cancer apoptosis. Wound healing assay and transwell migration and invasion assay implied that PoPx has the potential to inhibit migration and invasion, two critical steps in prostate cancer metastasis. Downregulation of MMP2/MMP9 and upregulation of TIMP2 showed accordance with the inhibition of migration and invasion. In summary, the present data showed that PoPx could be a promising drug candidate to treat prostate cancer, showing us a better way to develop novel drugs from natural compounds.
11. Bassiri-Jahromi S. Punica granatum (Pomegranate) activity in health promotion and cancer prevention. Oncol Res. 2018 Jan 30;12(1):345. Abstract: The available data suggest that Punica granatum (pomegranate) might be used in the control and potential therapeutic for some disease conditions and benefits human health status. This review summarizes in vitro, in vivo and clinical trial studies highlighting the pomegranate role in prevent and treatment of breast, prostate, lung, colon, skin and hepatocellular cell cancers.
12. Chrubasik-Hausman S et al. Pomegranate juice and prostate cancer: importance of the characterisation of the active principle. Phytoter Res. 2014 Nov;28(11):1676-8. Abstract: Two exploratory clinical studies investigating proprietary pomegranate products showed a trend of effectiveness in increasing prostate-specific antigen doubling time in patients with prostate cancer. A recent clinical study did not support these results. We therefore analysed a lot of the marketed pomegranate blend for co-active pomegranate compounds. The results show that the co-active compounds in the daily dose of the pomegranate blend were far below those previously tested and that the photometric assessment is not reliable for the standardisation of study medications. Not pomegranate but the low amount of co-active compounds in the proprietary pomegranate blend was responsible for its clinical ineffectiveness.
13. Wang L et al. Pomegranate and Its Components as Alternative Treatment for Prostate Cancer Int J Mol Sci. 2014 Sep; 15 (9):14949-14966. Abstract: Recent research has shown that pomegranate juice (PJ) and/or pomegranate extracts (PE) significantly inhibit the growth of prostate cancer cells in culture. In preclinical murine models, PJ and/or PE inhibit growth and angiogenesis of prostate tumors. More recently, it has been shown that three components of PJ, luteolin, ellagic acid and punicic acid together, have similar inhibitory effects on prostate cancer growth, angiogenesis and metastasis. Results from clinical trials are also promising. PJ and/or PE significantly prolonged the prostate specific antigen (PSA) doubling time in patients with prostate cancer.
14. Amri Z et al. Growth Inhibitory and Pro-Apoptotic Effects of Ornamental Pomegranate Extracts in Du145 Human Prostate Cancer Cells. Nutrition and Cancer. Volume 72. 2020. Issue 6. Abstract: This study was aimed to investigate the influence of dwarf pomegranate extracts (peel, juice, and seeds oil) on the proliferation and apoptosis of human prostate androgen-independent cell line DU145. The three tested extracts exhibited a dose-response cytotoxic effect and antiproliferative action on DU145 cell line and induce morphological changes. The three extracts could also induce prostate cancer cell apoptosis by an increase of DNA fragmentation, PARP cleavage, and inhibition of the COX-2 expression. The strongest pro-apoptotic effect was shown after peel treatment.

Protein

1. Yasudea J et al. Evenly Distributed Protein Intake over 3 Meals Augments Resistance Exercise-Induced Muscle Hypertrophy in Healthy Young Men. The Journal of Nutrition. 2020, April 22.
2. James P et al. Efficacy and tolerability of an undenatured type II collagen supplement in modulating knee osteoarthritis symptoms: a multicenter randomized, double-blind, placebo-controlled study. Nutrition Journal. 29 January 2016.
3. Reidy P. Muscle or Nothing! Where is the Excess Protein Going in Men with High Protein Intakes Engaged in Strength Training? The Journal of Nutrition. Volume 150, Issue 3, March 2020, pages 421-422.
4. Marinangeli et al. Potential impact of the digestible indispensable amino acid score as a measure of protein quality on dietary regulations and health. Nutr Rev. 2017 Aug; 75(8): 658-667.
5. Mathai JK et al. Values for digestible indispensable amino acid scores (DIAAS) for some dairy and plant proteins may better describe protein quality than values calculated using the concept for protein digestibility-corrected amino acid scores (PDCAAS). Br J Nutr. 2017 Feb;117(4):490-499.
6. Mettler S et al. Increased protein intake reduces lean body mass loss during weight loss in athletes. Med Sci Sports Exerc. 2010 Feb;42(2):326-37.
7. Morton R et al. A systematic review, meta-analysis and meta-regression of the effect of protein supplementation on resistance training-induced gains in muscle mass and strength in healthy adults. British Journal of Sports Medicine. 2017, July 11th.
8. Schoenfeld B et al. How much protein can the body use in a single meal for muscle-building? Implications for daily protein distribution. International Journal of Sports Nutrition. 2018, 15:10.
9. Wolfe R et al. Protein quality as determined by the Digestible Indispensable Amino Acid Score: evaluation of factors underlying the calculation. Nutr Rev. 2016 Sep; 74(9): 584-599.
10. Glenna J et al. Protein Digestibility-Corrected Amino Acid Scores (PDCAAS) for Soy Protein Isolate and Concentrate: Criteria for Evaluation. Journal of Agricultural and Food Chemistry. 2011, 59, 23, 12707-12712.
11. Schaafsma G. The Protein Digestibility-Corrected Amino Acid Score. The Journal of Nutrition. Volume 130, Issue 7, July 2000, Pages 1865S-1867S.

Raspberry Ketone

1. Park KS. Raspberry ketone, a naturally occurring phenolic compound, inhibits adipogenic and lipogenic gene expression in 3T3-L1 adipocytes. Pharm Biol 2015 Jun;53(6):870-5.

Resveratrol

1. Juan ME et al. Trans-Resveratrol, a natural antioxidant from grapes, Increases sperm output in healthy rats. J Nutr. 2005 Apr;135(4):757-60.
2. Bhat KP et al. Estrogenic and antiestrogenic properties of resveratrol in mammary tumor models. Cancer Res. 2001 Oct 15;61(20):7456-63.
3. Henry LA et al. Resveratrol: phytoestrogen effects on reproductive physiology and behavior in female rats. Horm Behav. 2002 Mar;41(2):220-8.
4. Matsumura A et al. Comparative study of oestrogenic properties of eight phytoestrogens in MCF7 human breast cancer cells. J Steroid Biochem Mol Biol. 2005 Apr;94(5):431-43.
5. Bowers JL et al. Resveratrol acts as a mixed agonist/antagonist for estrogen receptors alpha and beta. Endocrinology. 2000 Oct;141(10):3657-67.
6. Lu R et al. Resveratrol, a natural product derived from grape, exhibits antiestrogenic activity and inhibits the growth of human breast cancer cells. J Cell Physiol. 1999 Jun;179(3):297-304.
7. Turner RT et al. Is resveratrol an estrogen agonist in growing rats? Endocrinology. 1999 Jan;140(1):50-4.
8. Bhat KP et al. Resveratrol exhibits cytostatic and antiestrogenic properties with human endometrial adenocarcinoma (Ishikawa) cells. Cancer Res. 2001 Aug 15;61(16):6137-44.
9. Wang Y et al. The Red Wine Polyphenol Resveratrol Displays BI-Level Inhibition on Aromatase in Breast Cancer Cells. Toxicol Sci. 2006 Apr 11
10. Wallerath T et al. A blend of polyphenolic compounds explains the stimulatory effect of red wine on human endothelial NO synthase. Nitric Oxide. 2005 Mar;12(2):97-104.
11. Lekakis J et al. Polyphenolic compounds from red grapes acutely improve endothelial function in patients with coronary heart disease. Eur J Cardiovasc Prev Rehabil. 2005 Dec;12(6):596-600.
12. Buluc M et al. Resveratrol decreases calcium sensitivity of vascular smooth muscle and enhances cytosolic calcium increase in endothelium. Vascul Pharmacol. 2006 Apr;44(4):231-7.
13. Labinskyy N et al. Vascular dysfunction in aging: potential effects of resveratrol, an anti-inflammatory phytoestrogen. Curr Med Chem. 2006;13(9):989-96.
14. Bhat KPL et al. Biological effects of resveratrol. Antioxid Redox Signal. 2001 Dec;3(6):1041-64.
15. Bradamante S, et al. Cardiovascular protective effects of resveratrol. Cardiovasc Drug Rev. 2004 Fall;22(3):169-88.
16. de la Lastra CA et al. Resveratrol as an anti-inflammatory and anti-aging agent: mechanisms and clinical implications. Mol Nutr Food Res. 2005 May;49(5):405-30. doi: 10.1002/mnfr.200500022.
17. Delmas D et al. Resveratrol: preventing properties against vascular alterations and ageing. Mol Nutr Food Res. 2005 May;49(5):377-95.
18. Valenzano DR et al. Resveratrol prolongs lifespan and retards the onset of age-related markers in a short-lived vertebrate. Curr Biol. 2006 Feb 7;16(3):296-300.
19. Marambaud P et al. Resveratrol promotes clearance of Alzheimer's disease amyloid-beta peptides. J. Biol. Chem 2005 Nov;280(45): 37377-37382
20. Molnar V et al. Plant-derived anti-inflammatory compounds affect MIF tautomerase activity. Int Immunopharmacol. 2005 May;5(5):849-56.
21. Elmali N et al. Effect of resveratrol in experimental osteoarthritis in rabbits. Inflamm Res. 2005 Apr;54(4):158-62.
22. Kopp P. Resveratrol, a phytoestrogen found in red wine. A possible explanation for the conundrum of the 'French paradox'? Eur J Endocrinol. 1998 Jun;138(6):619-20.
23. Constant J. Alcohol, ischemic heart disease, and the French paradox. Coron. Artery Dis. 1997; 8:645 – 649.
24. Das D K et al. Cardioprotection of red wine: role of polyphenolic antioxidants. Drugs Exp Clin Res. 1999;25(2-3):115-20.
25. Soleas GJ, Diamandis EP, Goldberg DM. The world of resveratrol. Adv Exp Med Biol. 2001;492:159-82.
26. Wyke SM, Tisdale MJ. Induction of protein degradation in skeletal muscle by a phorbol ester involves upregulation of the ubiquitin-proteasome proteolytic pathway. 2006 May;78(25):2898-2910
27. Tisdale MJ. The ubiquitin-proteasome pathway as a therapeutic target for muscle wasting. J Support Oncol. 2005 May-Jun;3(3):209-17.
28. Wyke SM et al. Induction of proteasome expression in skeletal muscle is attenuated by inhibitors of NF-kappaB activation. Br J Cancer. 2004 Nov 1;91(9):1742-50.
29. Borra MT et al. Mechanism of human SIRT1 activation by resveratrol. J Biol Chem. 2005 Apr 29;280(17):17187-95.
30. Picard F, et al. Sirt1 promotes fat mobilization in white adipocytes by repressing PPAR-gamma. Nature. 2004 Jun 17;429(6993):771-6.
31. Wolf G. Calorie restriction increases life span: a molecular mechanism. Nutr Rev. 2006 Feb;64(2 Pt 1):89-92.
32. Ingram DK et al. Calorie restriction mimetics: an emerging research field. Aging Cell. 2006 Apr;5(2):97-108.
33. Roth GS et al. Caloric restriction mimetics: the next phase. Ann N Y Acad Sci. 2005 Dec;1057:365-71.
34. Tian WX. Inhibition of fatty acid synthase by polyphenols. Curr Med Chem. 2006;13(8):967-77.
35. Kasdallah-Grissa A, et al. Protective effect of resveratrol on ethanol-induced lipid peroxidation in rats. Alcohol Alcohol. 2006 May-Jun;41(3):236-9
36. Sener G et al. Protective effects of resveratrol against acetaminophen-induced toxicity in mice. Hepatol Res. 2006 Apr 1; E-Published Ahead of Print
37. Docherty JJ et al. Effect of resveratrol on herpes simplex virus vaginal infection in the mouse. Antiviral Res. 2005 Sep;67(3):155-62.
38. Jung HJ et al. Fungicidal effect of resveratrol on human infectious fungi. Arch Pharm Res. 2005 May;28(5):557-60.
39. Palamara AT et al. Inhibition of influenza A virus replication by resveratrol. J Infect Dis. 2005 May 15;191(10):1719-29.
40. Yoo KM et al. Potent Inhibitory Effects of Resveratrol Derivatives on Progression of Prostate Cancer Cells. Arch Pharm (Weinheim). 2006 Apr 18;339(5):238-241
41. Jones SB et al. Resveratrol-induced gene expression profiles in human prostate cancer cells. Cancer Epidemiol Biomarkers Prev. 2005 Mar;14(3):596-604.
42. Scifo C et al. Resveratrol and propolis as necrosis or apoptosis inducers in human prostate carcinoma cells. Oncol Res. 2004;14(9):415-26.
43. Kim YA et al. Antiproliferative effect of resveratrol in human prostate carcinoma cells. J Med Food. 2003 Winter;6(4):273-80.
44. Stewart JR et al. Resveratrol: a candidate nutritional substance for prostate cancer prevention. J Nutr. 2003 Jul;133(7 Suppl):2440S-2443S.
45. Ratan HL et al. Resveratrol – a prostate cancer chemopreventive agent? Urol Oncol. 2002 Nov-Dec;7(6):223-7.
46. Aggarwal BB, et al. Role of resveratrol in prevention and therapy of cancer: preclinical and clinical studies. Anticancer Res. 2004 Sep-Oct;24(5A):2783-840.
47. Aziz MH et al. Cancer chemoprevention by resveratrol: in vitro and in vivo studies and the underlying mechanisms (review). Int J Oncol. 2003 Jul;23(1):17-28.
48. Delmas D et al. Resveratrol as a chemopreventive agent: a promising molecule for fighting cancer. Curr Drug Targets. 2006 Apr;7(4):423-42.

Sleep

1. Bhaskar S et al. Prevalence of chronic insomnia in adult patients and its correlation with medical comorbidities. J Family Med Prim Care. Oct-Dec 2016;5(4):780-784. doi: 10.4103/2249-4863.201153. ABSTRACT: Introduction: Insomnia is one of the common but neglected conditions seen in family practice with long term and serious effects on health of a patient. Family physicians have the responsibility of diagnosing and adequately treating this. This study was done to find the prevalence of chronic insomnia in adult patients visiting a family medicine outpatient department (OPD) in a hospital and to assess the risk factors and co morbidities associated with it. Materials and methods: A cross-sectional study was done in the family medicine OPD at St. Philomena's Hospital, Bengaluru. All adult patients attending the OPD from September 1 to October 30, 2015 were enrolled in the study after obtaining written consent. Athens Insomnia Scale was used to diagnose insomnia and information regarding medical co morbidities was collected. Data was analyzed for the prevalence of insomnia and its association with co morbidities. Results: Chronic insomnia was seen in 33% of the adult population sampled. Increasing age and diabetes were significantly associated with insomnia, while other socioeconomic factors and co morbidities were not significantly associated. Twenty-seven percent of patients who had insomnia did not perceive the condition, which was statistically significant. Conclusion: Insomnia is a common sleep disorder which is many times missed by a primary care physician until/unless asked for. Since there is a higher incidence with increasing age and co morbidities such as diabetes, all patients, especially middle-aged and diabetics, should be screened for insomnia by the primary care physician with a self assessed questionnaire and counseled.

Superfood

1. Bahramsoltani R et al. The preventive and therapeutic potential of natural polyphenols on influenza. Expert Rev Anti Infect Ther. 2016;14(1):57-80. doi: 10.1586/14787210.2016.1120670.
2. Michael B et al. Human coronavirus OC43 3CL protease and the potential of ML188 as a broad-spectrum lead compound: Homology modelling and molecular dynamic studies. BMC Structural Biology. 2015;15:#8.
3. Vázquez-Calvo A et al. Antiviral Properties of the Natural Polyphenols Delphinidin and Epigallocatechin Gallate against the Flaviviruses West Nile Virus, Zika Virus, and Dengue Virus. Front Microbiol. 2017;(8):1314.
4. Cuevas et al. Modulation of Immune Function by Polyphenols: Possible Contribution of Epigenetic Factors. Nutrients. 2013 Jul; 5(7): 2314–2332.
5. Galland MD L. Doctor offers coronavirus protection advice. Fox 10 Phoenix. March 24, 2020.
6. Jagieta GC et al. 'Spicing up' of the immune system by curcumin. J Clin Immunology, 2006 Jan;27(1):19-35.
7. Le Sage V et al. Adapting the Stress Response: Viral Subversion of the mTOR Signaling Pathway. Viruses. 2016 Jun;8(6): 152.
8. Mathew D et al. Antiviral potential of curcumin. Journal of Functional Foods. 2018 Jan;40:692-699.
9. Mounce BC et al. Curcumin inhibits Zika and chikungunya virus infection by inhibiting cell binding. Antiviral Research. 2017 Jun;142:148-157.
10. Nakayama M et al. Inhibition of the Infectivity of Influenza Virus by Tea Polyphenols. Antiviral Res. 1993 Aug;21(4):289-299.
11. Teymouri M et al. Curcumin as a multifaceted compound against human papilloma virus infection and cervical cancers: A review of chemistry, cellular, molecular, and preclinical features. Biofactors. 2017 May6;43(3):331-346.
12. Ting D et al. Multisite Inhibitors for Enteric Coronavirus: Antiviral Cationic Carbon Dots Based on Curcumin. ACS Publications. 2018 Sep 12.
13. Jo S et al. Inhibition of SARS-CoV 3CL Protease by Flavonoids. J Enzyme Inhib Med Chem. 35 (1), 145-151, Dec 2020.
14. Zorofchian S et al. A Review of Antibacterial, Antiviral, and Antifungal Activity of Curcumin. Biomed Research International. Volume 2014, Article ID 186864.
15. Yang ZF et al. Comparison of in Vitro Antiviral Activity of Tea Polyphenols Against Influenza A and B Viruses and Structure-Activity Relationship Analysis. Fitoterapia. 93, 47-53, Mar 2014.
16. Rodriguez-Mateos A et al. Circulating anthocyanin metabolites mediate vascular benefits of blueberries: insights from randomized controlled trials, metabolomics, and nutrigenomics. The Journals of Gerontology: Series A, 2019.
17. Yu J et al. The Effect of Diet on Improved Endurance in Male C57BL/6 Mice. Nutrients 2018, 10(8), 1101.
18. Vendrame S et al. Wild blueberry consumption affects aortic vascular function in the obese Zucker rat. Applied Physiology, Nutrition, and Metabolism, 2013.
19. Blueberries may inhibit development of fat cells. Federation of American Societies for Experimental Biology. April 11, 2011.
20. Dastmalchi K et al. Edible Neotropical Blueberries: Antioxidant and Compositional Fingerprint Analysis. Journal of Agricultural and Food Chemistry, 2011.
21. Wang D et al. Fruit and Vegetable Intake and Mortality. Circulation. 2021;143:00-00.

Tribulus terrestris

1. Gamal El Din SF et al. Tribulus terrestris versus placebo in the treatment of erectile dysfunction and lower urinary tract symptoms in patients with late-onset hypogonadism: A placebo-controlled study. Urologia. 2019 May;86(2):74-78. doi: 10.1177/0391560318802160. Epub 2018 Sep 25. ABSTRACT: Aging is associated with a series of morphological and functional modifications that leads to reduced physiological efficiency and atrophy of various organs and systems. Tribulus terrestris induces its effect in fertility and sexual functions through the steroidal saponins, particularly the dominant saponins protodioscin. We aimed in this study to evaluate the efficacy and safety profiles of Tribulus terrestris in aging males with partial androgen deficiency who suffered from erectile dysfunction and lower urinary tract symptoms. A total of 70 randomized aging patients with erectile dysfunction and lower urinary tract symptoms were recruited from June 2017 to March 2018 from our andrology outpatient clinic. Thirty-five patients (group A) received Tribulus terrestris three times daily for 3 months and the other 35 patients (group B) received placebo. The mean of aspartate transaminase was elevated in group A after 3 months of receiving Tribulus terrestris (26.5 (before), 27.8 (after), respectively, p = 0.03). Moreover, there were significant elevations in the means of both total testosterone together with the score of the validated Arabic index of erectile function (5-item version of the International Index of Erectile Function) (2.2, 10.7 (before), 2.7, 16.1 (after), p < 0.001, p < 0.001, respectively). Finally, the mean of the total prostate-specific antigen was elevated in this group (1.4 (before), 1.7 (before), p = 0.007, respectively). Interestingly, there were no worsening of the lower urinary tract symptoms in group A as there was no change in the mean score of the international prostate symptom score, which was used to assess these symptoms before and after treatment (mean 14.4 (before), 14.6 (after), p = 0.67, respectively). In sum, this study replicates the findings of previous reports about the robust effect of this herbal medicine in elevating the testosterone level and improving the sexual function of patients who suffered from erectile dysfunction with partial androgen deficiency.
2. Salgado RM et al. Effect of oral administration of Tribulus terrestris extract on semen quality and body fat index of infertile men. Andrologia. 2017 Jun;49(5). doi: 10.1111/and.12655. Epub 2016 Jul 12. ABSTRACT: Male fertility can be evaluated through complete semen analysis. Plants belonging to the Tribulus genus are known for their role in enhancing sex hormone levels and semen quality. The aim of this study was to evaluate the effects of T. terrestris on semen quality and physiological parameters. Sixty-five men with abnormal semen evaluation were included in this study, in which they were prescribed with oral administration of Androsten® (250 mg of Tribulus terrestris dried extract per capsule). Body fat percentage, lean muscle mass gain, fluctuation in steroid hormone levels and all semen parameters were analysed during the period of treatment. The results demonstrated that decrease in the percentage of body fat and increase in lean mass were significant, as well as increase in dihydrotestosterone levels. Complete semen analysis evaluated at the end of treatment showed significant enhancement in sperm concentration, motility and liquefaction time. Protodioscin, the main phytochemical agent of the Tribulus genus, acts on sertoli cells, germ cell proliferation and growth of seminiferous tubules. This component is known to convert testosterone into dihydrotestosterone, which plays important roles in male attributes. Our results indicate the therapeutic use of Tribulus terrestris by men presenting altered semen parameters, and/or undergoing infertility treatment.
3. Wilk M et al. Endocrine Responses to Physical Training and Tribulus Terrestris Supplementation in Middle-Age Men. Central European Journal of Sports Sciences and Medicine, Vol. 13, No. 1/2016: 65-71. ABSTRACT: The aim of this study was to evaluate the effects of steroidal saponin supplementation on blood concentration of T, GH and IGF-1. The research involved 14 men between the age of 45 and 60 years. The duration of the experiment was 12 weeks. There were two series of laboratory tests. Independent tests were conducted at the beginning and after 12 weeks of the inter vention. A t wo-way repeated measures ANOVA revealed a statistically significant effect of the intervention on the following variables: T-Ch (η2 = 0.542), HDL-Ch (η2 = 0.522), LDL-Ch (η2 = 0.587), T (η2 = 0.603), IGF-1 (η2 = 0.512) and GH (η2 = 0.621). Thus, FFM significantly increased while TBF and BM decreased in comparison to pre-intervention levels. The analyzed results indicate that treatment or supplementation of individual hormone deficiencies can be a successful form of counteracting the aging process. Nevertheless, the effects of TT supplementation on the concentration of T as well as GH and IGF-1, requires further studies, especially in middle-aged and older subjects, along with different exercise programs. The analyzed results indicate that treatment or supplementation of individual hormone deficiencies can be a major form of counteracting the aging process
4. Haghmorad D et al. Improvement of fertility parameters with Tribulus Terrestris and Anacyclus Pyrethrum treatment in male rats. Int Braz J Urol. Sep-Oct 2019;45(5):1043-1054. doi: 10.1590/S1677-5538.IBJU.2018.0843. ABSTRACT: Objective: Anacyclus Pyrethrum (AP) and Tribulus Terrestris (TT) have been reported as male infertility treatment in several studies; however, in Iranian traditional medicine these two plants are prescribed simultaneously. In this study, we aimed to determine the effects of AP and TT extracts both separately and simultaneously on the male Wistar rat fertility parameters. Materials and methods: 32 male Wistar rats were divided into 4 groups: Control, TT, AP, and AT treated groups. Treatment continued for 25 days and rats were weighed daily. Their testes were dissected for histological studies. Sperm analysis including sperm count, viability and motility were performed. Serum was obtained to evaluate testosterone, LH and FSH levels. Histological studies were conducted to study Leydig, and Sertoli cells, spermatogonia and spermatid cell numbers, and to measure seminiferous diameter and epithelium thickness. Results: Sperm count increased in all the treatment groups. Sperm viability and motility in AT and AP groups were elevated. TT and AT groups showed signifi cantly increased testosterone level compared to control group (P=004, P=0.000, respectively) and TT, AP and AT treatment groups showed increased LH level (P=0.002, P=0.03 and P=0.000, respectively) compared to control, while only AT group showed increased FSH (p=0.006) compared to control. Histological studies showed signifi cant increase of spermatogonia, Leydig and Sertoli cell numbers and epithelial thickness in AT group compared to other groups. All the treatment groups had higher number of Leydig, spermatogonia and spermatid cells. Conclusion: TT and AP improved sexual parameters; however, their simultaneous administration had higher improving effects on studied parameters.
5. Adaikan G et al. History of herbal medicines with an insight on the pharmacological properties of Tribulus terrestris September 2001. The Aging Male 4(3):163-169. DOI:10.1080/tam.4.3.163.169. ABSTRACT: Phytochemicals have played a vital role in the past and will continue to do so in the future. Although synthetic drugs can produce dramatic results in most cases, the side-effects associated with them are a major concern. The source of many compounds used in modern medicine today can be traced down to plant origin. Whether or not scientific justification is available for the use of most plant products, the continued use of these compounds is due to their safety profile, ease of availability and also economic reasons. Each medicinal plant that has been used in the traditional system of medicine must be scientifically tested in order to bring forth its active principle that might be effectively used as a phytomedicine. In this vast resource of phytoproducts there are various plants that are claimed to improve the sexual deficiency in man. Tribulus terrestris Linn. (TT) is one such plant that has been used for a long time in both the Indian and Chinese systems of medicine. The plant is also said to possess various other pharmacological properties. The extract obtained from the air-dried aerial parts of this plant contains mainly steroidal glycosides, the major saponin being protodioscin (PTN). In our study on this plant product it was observed that PTN produced a moderate increase in testosterone, dihydrotestosterone and dehydroepiandrosterone sulfate levels in primates following bolus intravenous administration of the TT extract at doses of 7.5, 15 and 30 mg/kg body weight. It also improved libido, sexual activity and intracavernous pressure in rats following TT extract administration orally (for 8 weeks at doses of 2.5, 5 and 10 mg/kg body weight) and had a proerectile effect on the corpus cavernosum smooth muscle of rabbits (orally for 8 weeks at doses of 2.5, 5 and 10 mg/kg body weight). In this article, the various pharmacological effects of TT have been reviewed and our studies based on TT extract in relation to male erectile dysfunction have been summarized.

Vitamin D3

1. Aranow C. Vitamin D and the Immune System. J Investigative Medicine, 2011 Aug;59(6):881-886. ABSTRACT: Quote: "It is now clear that vitamin D has important roles in addition to its classic effects on calcium and bone homeostasis. As the vitamin D receptor is expressed on immune cells (B cells, T cells and antigen presenting cells) and these immunologic cells are all are capable of synthesizing the active vitamin D metabolite, vitamin D has the capability of acting in an autocrine manner in a local immunologic milieu. Vitamin D can modulate the innate and adaptive immune responses. Deficiency in vitamin D is associated with increased autoimmunity as well as an increased susceptibility to infection. As immune cells in autoimmune diseases are responsive to the ameliorative effects of vitamin D, the beneficial effects of supplementing vitamin D deficient individuals with autoimmune disease may extend beyond the effects on bone and calcium homeostasis."
2. Autier P, Gandini S. Vitamin D Supplementation and Total Mortality: A Meta-analysis of Randomized Controlled Trials. Arch Intern Med. Sept. 10, 2007;167(16):1730-1737. ABSTRACT: This study identified 18 independent randomized controlled trials, including 57 311 participants. A total of 4777 deaths from any cause occurred during a trial size-adjusted mean of 5.7 years. Daily doses of vitamin D supplements varied from 300 to 2000 IU. The trial size-adjusted mean daily vitamin D dose was 528 IU. In 9 trials, there was a 1.4- to 5.2-fold difference in serum 25-hydroxyvitamin D between the intervention and control groups. Intake of ordinary doses of vitamin D supplements seems to be associated with decreases in total mortality rates.
3. Adit A et al. Association Between Serum 25-Hydroxyvitamin D Level and Upper Respiratory Tract Infection in the Third National Health and Nutrition Examination Survey. Arch Intern Med. 2009;169(4):384-390. ABSTRACT: Study performed a secondary analysis of the Third National Health and Nutrition Examination Survey, a probability survey of the US population conducted between 1988 and 1994. They examined the association between 25(OH)D level and recent URTI in 18 883 participants 12 years and older. The analysis adjusted for demographics and clinical factors (season, body mass index, smoking history, asthma, and chronic obstructive pulmonary disease). Serum 25(OH)D levels were inversely associated with recent upper respiratory tract infections.
4. Marineua AR et al. Vitamin D supplementation to prevent acute respiratory tract infections: systematic review and meta-analysis of individual participant data. BMJ, 15 February 2017. ABSTRACT: 25 eligible randomized control trials totaling 11321 participants aged 0 to 95 years were identified. Vitamin D supplementation reduced the risk of acute respiratory tract infection among all participants.
5. Urshima M et al. Randomized trial of vitamin D supplementation to prevent seasonal influenza A in schoolchildren. Am J Clin Nutr. 2010, May;91(5):1255-60. ABSTRACT: This randomized, double-blind, placebo-controlled trial compared vitamin D(3) supplements (1200 IU/d) with placebo in schoolchildren. The results suggested that vitamin D(3) supplementation during the winter may reduce the incidence of influenza A, especially in specific subgroups of schoolchildren.
6. Simoliunas E et al. Bioavailability of Different Vitamin D Oral Supplements in Laboratory Animal Model. Medicina, 2019, 55(6), 265. 2597 C. ABSTRACT: The results of this study suggest that the oral vitamin D supplement vehicle has an impact on its bioavailability, thus it is important to take into account how much of the supplied vitamin D will be absorbed. To maximize the full exploit of supplement, the best delivery strategy should be employed. In this study, the microencapsulated form of vitamin D was the most bioavailable.

Yohimbine

1. Galitzky J et al. Alpha 2-antagonist compounds and lipid mobilization: evidence for a lipid mobilizing effect of oral yohimbine in healthy male volunteers. Eur J Clin Invest. 1988 Dec;18(6):587-94.
2. Lafontan m et al. Alpha-2 adrenoceptors in lipolysis: alpha 2 antagonists and lipid-mobilizing strategies. Am J Clin Nutr. 1992 Jan;55(1 Suppl):219S-227S. doi: 10.1093/ajcn/55.1.219s.
3. Kucio C et al. Does yohimbine act as a slimming drug? Isr J Med Sci. 1991 Oct;27(10):550-6.
4. Ostojic SM Yohimbine: The effects on body composition and exercise performance in soccer players. Res Sports Med. 2006, Oct-Dec;14(4):289-99.

Zinc

1. Hunt CD et al. Effects of dietary zinc depletion on seminal volume and zinc loss, serum testosterone concentrations, and sperm morphology in young men. Am J Clin Nutr. 1992 Jul;56(1):148-57. doi: 10.1093/ajcn/56.1.148.
2. Farahzadi R et al. Zinc sulfate contributes to promote telomere length extension via increasing telomerase gene expression, telomerase activity and change in the TERT gene promoter CpG island methylation status of human adipose-derived mesenchymal stem cells. PLoS One. 2017 Nov 16;12(11):e0188052. doi: 10.1371/journal.pone.0188052. eCollection 2017.
3. Nemoto K. et al. Modulation of telomerase activity by zinc in human prostatic and renal cancer cells. Biochem Pharmacol. 2000 Feb 15;59(4):401-5. doi: 10.1016/s0006-2952(99)00334-2.
4. Prasad AS et al. Zinc status and serum testosterone levels of healthy adults. Nutrition. 1996 May;12(5):344-8. doi: 10.1016/s0899-9007(96)80058-x.
5. Dissanayake DMAB et al. Effects of zinc supplementation on sexual behavior of male rats. J Hum Reprod Sci. 2009 Jul-Dec; 2(2): 57–61. doi: 10.4103/0974-1208.57223.

ZMA

1. Brilla L et al. Effects of a Novel Zinc-Magnesium Formulation on Hormones and Strength. Journal of Exercise Physiology (online). 3(4): 26-36, 2000.
2. Wilborn CD et al. Effects of Zinc Magnesium Aspartate (ZMA) Supplementation on Training Adaptations and Markers of Anabolism and Catabolism. J Int Soc Sports Nutr. 2004; 1(2): 12–20. doi: 10.1186/1550-2783-1-2-12