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PUREHEALTH RESEARCH TESTOSTERONE BOOSTER
CLINICAL STUDIES ON THE FOLLOWING INGREDIENTS:

Tribulus Terrestris (whole plant) Extract
Facts:
Tribulus terrestris is a Mediterranean plant/ plant from Ayurveda where it’s roots and fruits are used to enhance male virility and vitality. It’s primary functions are leaning more towards cardiovascular and urogenital health. Various researches had been done and established as to it’s functions and relevance.
Source:
http://examine.com/supplements/Tribulus+terrestris
http://www.webmd.com/vitamins-and-supplements/tribulus-terrestris-uses-and-risks

Study 1:
Clinical study of Tribulus terrestris Linn. in Oligozoospermia: A double blind study
After detailed discussion on observations made and results achieved, this present study shows significant remission in the signs and symptoms of Kshina Shukra, vis-à-vis oligozoospermia, corroborated with definite improvement in the total sperm count. In toto, from this study it can be concluded that the Alternate Hypothesis of this study is accepted, that is, Tribulus terrestris. Linn is effective in the management of Kshina Shukra, with lifestyle modification. Tribulus terrestris. Linn Granules have shown superior results in the management of Kshina Shukra, as compared to placebo granules.
Source:
Thirunavukkarasu M. Sellandi, Anup B. Thakar, and Madhav Singh Baghel. Clinical study of Tribulus terrestris Linn. in Oligozoospermia: A double blind study. Ayu. 2012 Jul-Sep; 33(3): 356–364. doi: 10.4103/0974-8520.108822.

Study 2:
Identification of fruits of Tribulus terrestris Linn. and Pedalium murex Linn.: A pharmacognostical approach
Gokshura is a well-known Ayurvedic drug that is used in many preparations. Botonically it is identified as Tribulus terrestris Linn., especially the roots and fruits of the plant. But instead the fruits of another plant Pedalium murex Linn. are commonly used and the drug is frequently substituted. Pharmacognostical study has been carried out to identify the distinguishing features, both morphological and microscopic, of the fruits of Tribulus terrestris Linn. and Pedalium murex Linn. This knowledge should help reduce the problem of substitution of the genuine drug.
Source:
Kevalia J, Patel B.Identification of fruits of Tribulus terrestris Linn. and Pedalium murex Linn.: A pharmacognostical approach.Ayu. 2011 Oct;32(4):550-3. doi: 10.4103/0974-8520.96132.

Study 3:
Short term impact of Tribulus terrestris intake on doping control analysis of endogenous steroids
Tribulus terrestris is a nutritional supplement highly debated regarding its physiological and actual effects on the organism. The main claimed effect is an increase of testosterone anabolic and androgenic action through the activation of endogenous testosterone production. Even if this biological pathway is not entirely proven, T. terrestris is regularly used by athletes. Recently, the analysis of two female urine samples by GC/C/IRMS (gas chromatography/combustion/isotope-ratio-mass-spectrometry) conclusively revealed the administration of exogenous testosterone or its precursors, even if the testosterone glucuronide/epitestosterone glucuronide (T/E) ratio and steroid marker concentrations were below the cut-off values defined by World Anti-Doping Agency (WADA). Hence, the short-term treatment with T. terrestris showed no impact on the endogenous testosterone metabolism of the two subjects.
Source:
Saudan C1, Baume N, Emery C, Strahm E, Saugy M.Short term impact of Tribulus terrestris intake on doping control analysis of endogenous steroids.Forensic Sci Int. 2008 Jun 10;178(1):e7-10. doi: 10.1016/j.forsciint.2008.01.003. Epub 2008 Feb 20.

Study 4:
Evaluation of the aphrodisiac activity of Tribulus terrestris Linn. in sexually sluggish male albino rats
A dose-dependent improvement in sexual behavior was observed with the Tribulus terrestris (LAET) treatment as characterized by an increase in mount frequency, intromission frequency, and penile erection index, as well as a decrease in mount latency, intromission latency, and ejaculatory latency. The enhancement of sexual behavior was more prominent on chronic administration of LAET. Chronic administration of LAET produced a significant increase in serum testosterone levels with no significant effect on the sperm count. No overt body system dysfunctions were observed in 28-day oral toxicity study.
Source:
Surender Singh, Vinod Nair, and Yogendra K. Gupta. Evaluation of the aphrodisiac activity of Tribulus terrestris Linn. in sexually sluggish male albino rats.J Pharmacol Pharmacother. 2012 Jan-Mar; 3(1): 43–47. doi: 10.4103/0976-500X.92512

Study 5:
A systematic review on the herbal extract Tribulus terrestris and the roots of its putative aphrodisiac and performance
Randomized control trials, which included healthy human subjects ingesting TT as sole or combined supplement, along with animal studies with TT as a sole treatment across a number of species were included. A limited number of animal studies displayed a significant increase in serum testosterone levels after TT administration, but this effect was only noted in humans when TT was part of a combined supplement administration. Literature available for the effectiveness of TT on enhancing testosterone concentrations is limited. Evidence to date suggests that TT is ineffective for increasing testosterone levels in humans, thus marketing claims are unsubstantiated. The nitric oxide release effect of TT may offer a plausible explanation for the observed physiological responses to TT supplementation, independent of the testosterone level.
Source:
Qureshi A, Naughton DP, Petroczi A. A systematic review on the herbal extract Tribulus terrestris and the roots of its putative aphrodisiac and performance enhancing effect. J Diet Suppl. 2014 Mar;11(1):64-79. doi: 10.3109/19390211.2014.887602.

Study 6:
Phytopharmacological overview of Tribulus terrestris
Tribulus terrestris (family Zygophyllaceae), commonly known as Gokshur or Gokharu or puncture vine, has been used for a long time in both the Indian and Chinese systems of medicine for treatment of various kinds of diseases. It has diuretic, aphrodisiac, antiurolithic, immunomodulatory, antidiabetic, absorption enhancing, hypolipidemic, cardiotonic, central nervous system, hepatoprotective, anti-inflammatory, analgesic, antispasmodic, anticancer, antibacterial, anthelmintic, larvicidal, and anticariogenic activities. The aim of this review is to create a database for further investigations of the discovered phytochemical and pharmacological properties of this plant to promote research.
Source:
Chhatre S, Nesari T, Somani G, Kanchan D, Sathaye S.Phytopharmacological overview of Tribulus terrestris.Pharmacogn Rev. 2014 Jan;8(15):45-51. doi: 10.4103/0973-7847.125530.

Study 7:
Effect of Tribulus terrestris on nicotinamide adenine dinucleotide phosphate-diaphorase activity and androgen receptors in rat brain
Administration of Tribulus terrestris extract (TT) increased sexual behaviour and intracavernous pressure both in normal and castrated rats and these effects were probably due to the androgen increasing property of TT. The objective of the present study is to evaluate the effect of TT on nicotinamide adenine dinucleotide phosphate-diaphorase (NADPH-d) activity and androgen receptor (AR) immunoreactivity in rat brain. There was an increase in both NADPH-d (67%) and AR immunoreactivity (58%) in TT treated group and these results were statistically significant compared to the control. Chronic treatment of TT in rats increases the NADPH-d positive neurons and AR immunoreactivity in the PVN region. The mechanism for the observed increase in AR and NADPH-d positive neurons in the present study is probably due to the androgen increasing property of TT.
Source:
Gauthaman K, Adaikan PG. Effect of Tribulus terrestris on nicotinamide adenine dinucleotide phosphate-diaphorase activity and androgen receptors in rat brain. J Ethnopharmacol. 2005 Jan 4;96(1-2):127-32.

Study 8:
The Effect of Oral Feeding of Tribulus terrestris L. on Sex Hormone and Gonadotropin Levels in Addicted Male Rats
Opioids can exert adverse effects on the body. Morphine, an opioid drug, reduces hormone levels and fertility, and causes sexual activity disorders. Tribulus terrestris (TT) is a traditional herbal medicine used to enhance sexual activities. This study investigates the possible role of TT on sex hormones and gonadotropins with the intent to show its usefulness in treating fertility disorders in opioid users. Oral consumption of TT could markedly antagonize the reduction of sex hormones and gonadotropins (except for FSH) due to morphine addiction.
Source:
Ghosian Moghaddam MH, Khalili M, Maleki M, Ahmad Abadi ME. The Effect of Oral Feeding of Tribulus terrestris L. on Sex Hormone and Gonadotropin Levels in Addicted Male Rats. Int J Fertil Steril. 2013 Apr;7(1):57-62. Epub 2013 Mar 6.

Study 9:
Effects and Mechanism of Action of a Tribulus terrestris Extract on Penile Erection
Concentration-dependent relaxation effects of the extract on the corpus cavernosum (CC) were detected in the organ bath study. Relaxation of the CC by the T. terrestris extract was inhibited in both an endothelium-removed group and an L-arginen methyl ester pretreatment group. The intracavernous pressure (ICP) measured after oral administration of the T. terrestris extract for 1 month was higher than that measured in the control group, and a significant increase in cAMP was observed in the T. terrestris extract group. The T. terrestris extract induced concentration-dependent relaxation of the CC in an organ bath. The mechanism included a reaction involving the nitric oxide/nitric oxide synthase pathway and endothelium of the CC. Moreover, in an in vivo study, the T. terrestris extract showed a significant concentration-dependent increase in ICP. Accordingly, the T. terrestris extract may improve erectile function.
Source:
Jungmo Do, Seemin Choi, Jaehwi Choi, and Jae Seog Hyun. Effects and Mechanism of Action of a Tribulus terrestris Extract on Penile Erection. Korean J Urol. 2013 Mar; 54(3): 183–188. Published online 2013 Mar 15. doi: 10.4111/kju.2013.54.3.183

Study 10:
The effects of Tribulus terrestris on body composition and exercise performance in resistance-trained males
There were no changes in body weight, percentage fat, total body water, dietary intake, or mood states in either group. Muscle endurance (determined by the maximal number of repetitions at 100-200% of body weight) increased for the bench and leg press exercises in the placebo group (p <.05; bench press +/-28.4%, leg press +/-28.6%), while the tribulus group experienced an increase in leg press strength only (bench press +/-3.1%, not significant; leg press +/-28.6%, p <.05). Supplementation with tribulus does not enhance body composition or exercise performance in resistance-trained males.
Source:
Antonio J, Uelmen J, Rodriguez R, Earnest C. The effects of Tribulus terrestris on body composition and exercise performance in resistance-trained males. Int J Sport Nutr Exerc Metab. 2000 Jun;10(2):208-15.

Study 11:
Chemical constituents from Tribulus terrestris and screening of their antioxidant activity
Two oligosaccharides (1,2) and a stereoisomer of di-p-coumaroylquinic acid (3) were isolated from the aerial parts of Tribulus terrestris along with five known compounds (4-8). This is the first report for the complete NMR spectral data of the known 4,5-di-p-trans-coumaroylquinic acid (4). The antioxidant activity represented as DPPH free radical scavenging activity was investigated revealing that the di-p-coumaroylquinic acid derivatives possess potent antioxidant activity so considered the major constituents contributing to the antioxidant effect of the plant.
Source:
Hammoda HM, Ghazy NM, Harraz FM, Radwan MM, ElSohly MA, Abdallah II.Chemical constituents from Tribulus terrestris and screening of their antioxidant activity. Phytochemistry. 2013 Aug;92:153-9. doi: 10.1016/j.phytochem.2013.04.005. Epub 2013 May 2.

Study 12:
Sexual effects of puncturevine (Tribulus terrestris) extract (protodioscin): an evaluation using a rat model
The weight gain and improvement in sexual behavior parameters observed in rats could be secondary to the androgen increasing property of TT (PTN) that was observed in our earlier study on primates. The increase in ICP which confirms the proerectile aphrodisiac property of TT could possibly be the result of an increase in androgen and subsequent release of nitric oxide from the nerve endings innervating the corpus cavernosum.
Source:
Gauthaman K, Ganesan AP, Prasad RN. Sexual effects of puncturevine (Tribulus terrestris) extract (protodioscin): an evaluation using a rat model. J Altern Complement Med. 2003 Apr;9(2):257-65.

Study 13:
The hormonal effects of Tribulus terrestris and its role in the management of male erectile dysfunction–an evaluation using primates, rabbit and rat
TT extract was administered intravenously, as a bolus dose of 7.5, 15 and 30 mg/kg, in primates for acute study. Rabbits and normal rats were treated with 2.5, 5 and 10mg/kg of TT extract orally for 8 weeks, for chronic study. In addition, castrated rats were treated either with testosterone cypionate (10mg/kg, subcutaneously; biweekly for 8 weeks) or TT orally (5mg/kg daily for 8 weeks). In primates, the increases in T (52%), DHT (31%) and DHEAS (29%) at 7.5mg/kg were statistically significant. In castrated rats, increases in T levels by 51% and 25% were observed with T and TT extract respectively that were statistically significant.
Source:
Gauthaman K, Ganesan AP.The hormonal effects of Tribulus terrestris and its role in the management of male erectile dysfunction–an evaluation using primates, rabbit and rat.Phytomedicine. 2008 Jan;15(1-2):44-54.

Study 14:
The aphrodisiac herb Tribulus terrestris does not influence the androgen production in young men
There was no significant difference between Tribulus terrestris supplemented groups and controls in the serum testosterone. . All results were within the normal range. The findings in the current study anticipate that Tribulus terrestris steroid saponins possess neither direct nor indirect androgen-increasing properties
Source:
Neychev VK, Mitev VI. The aphrodisiac herb Tribulus terrestris does not influence the androgen production in young men.J Ethnopharmacol. 2005 Oct 3;101(1-3):319-23.

Study 15:
In vivo and in vitro animal investigation of the effect of a mixture of herbal extracts from Tribulus terrestris and Cornus officinalis on penile erection

  1. terrestris extract, C. officinalis extract, and the mixture of both extracts showed concentration-dependent relaxation effects of the CC. In both the endothelium-removed group and N(G)-nitro-L-arginine methyl ester pretreatment group, T. terrestris extract inhibited relaxation. ICP measured after oral administration of the extract mixture for a month was higher than that measured in the control group, and a significant increase in cAMP was observed in the mixture group. T. terrestris extract and C. officinalis extract exhibited concentration-dependent relaxation in an organ bath. In the in vivo study of the extract mixture, ICP and cAMP was significantly potentiated. Accordingly, the mixture of T. terrestris extract and C. officinalis extract may improve erectile function.

Source:
Kam SC, Do JM, Choi JH, Jeon BT, Roh GS, Hyun JS.In vivo and in vitro animal investigation of the effect of a mixture of herbal extracts from Tribulus terrestris and Cornus officinalis on penile erection.J Sex Med. 2012 Oct;9(10):2544-51. doi: 10.1111/j.1743-6109.2012.02889.x. Epub 2012 Aug 20.

Study 16:
Proerectile pharmacological effects of Tribulus terrestris extract on the rabbit corpus cavernosum
The relaxant responses to acetylcholine, nitroglycerin and EFS by more than 10%, 24% and 10% respectively compared to their control values and the lack of such effect on the contractile response to noradrenaline and histamine indicate that PTN has a proerectile activity. The enhanced relaxant effect observed is probably due to increase in the release of nitric oxide from the endothelium and nitrergic nerve endings, which may account for its claims as an aphrodisiac. However, further study is needed to clarify the precise mechanism of its action.
Source:
Adaikan PG, Gauthaman K, Prasad RN, Ng SC. Proerectile pharmacological effects of Tribulus terrestris extract on the rabbit corpus cavernosum. Ann Acad Med Singapore. 2000 Jan;29(1):22-6.

Chrysin (99.6%)
Facts:
Chrysin is a member of a chemical class known as flavanoids. Chrysin is known to have relevance in body building, treating anxiety, inflammation gout, erectile dysfunction and is known to have a significant role in preventing cancer. It is known to boost testosterone when directly injected into the testicle for maximum reabsorption and potentiating its results.
Source:
http://examine.com/supplements/Chrysin/
http://www.webmd.com/vitamins-supplements/ingredientmono-1047-chrysin.aspx?activeingredientid=1047&activeingredientname=chrysin

Study 1:
Beneficial effects of chrysin on the reproductive system of adult male rats
Tissue thiobarbituric acid reactive substances (TBARS) and glutathione (GSH) levels, antioxidant enzyme activity (CAT, SOD and GSH-Px), sperm parameters (motility, concentration and abnormal sperm rate), reproductive organ weight (testes, epididymis, vesicula seminalis, prostate) and serum testosterone levels were determined in the rats. Our results indicated that chrysin significantly increased GSH, CAT, GSH-Px and CuZn-SOD levels, but did not change the formation of TBARS significantly. In addition, sperm motility, sperm concentration and serum testosterone levels significantly increased, whereas abnormal sperm rate significantly decreased with chrysin treatment.
Source:

  1. Ciftci, İ. Ozdemir, M. Aydin andA. Beytur. Beneficial effects of chrysin on the reproductive system of adult male rats. Andrologia. Volume 44, Issue 3, pages 181–186, June 2012. Article first published online: 7 MAR 2011 DOI:

10.1111/j.1439-0272.2010.01127.x

Study 2:
Chrysin alleviates testicular dysfunction in adjuvant arthritic rats via suppression of inflammation and apoptosis: comparison with celecoxib
Long standing rheumatoid arthritis (RA) is associated with testicular dysfunction and subfertility. Few studies have addressed the pathogenesis of testicular injury in RA and its modulation by effective agents. Thus, the current study aimed at evaluating the effects of two testosterone boosting agents; chrysin, a natural flavone and celecoxib, a selective COX-2 inhibitor, in testicular impairment in rats with adjuvant arthritis, an experimental model of RA. Chrysin suppressed paw edema with comparable efficacy to celecoxib. Both agents upregulated steroidogenic acute regulatory (StAR) mRNA expression and serum testosterone with concomitant restoration of LH and FSH. Alleviation of the testicular impairment was accompanied with suppression of oxidative stress via lowering testicular lipid peroxides and nitric oxide. Generally, the 50mg/kg dose of chrysin exerted comparable protective actions to celecoxib.
Source:
Darwish HA, Arab HH, Abdelsalam RM. Chrysin alleviates testicular dysfunction in adjuvant arthritic rats via suppression of inflammation and apoptosis: comparison with celecoxib.Toxicol Appl Pharmacol. 2014 Sep 1;279(2):129-40. doi: 10.1016/j.taap.2014.05.018. Epub 2014 Jun 14.

Study 3:
Chrysin modulates ethanol metabolism in Wistar rats: a promising role against organ toxicities
Ethanol administration significantly induced CYP 2E1, ADH and XO in liver and kidneys, respectively, along with an enhancement in levels of malondialdehyde and serum alanine aminotransferase, aspartate aminotransferase, blood urea nitrogen, creatinine and lactate dehydrogenase when compared with the control group and this enhancement is significantly normalized with chrysin administration. Oxidative stress markers: reduced glutathione, glutathione peroxidase, catalase and glutathione reductase were significantly (P < 0.001) depleted in the ethanol-treated group, while chrysin administration significantly restored all of these. Only chrysin administration did not show any adverse effect. Results demonstrate that chrysin administration prevents the liver and kidney of Wistar rats against oxidative damage during chronic ethanol consumption by inhibiting the activities of ADH, CYP 2E1, XO and catalase.
Source:
Tahir M, Sultana S. Chrysin modulates ethanol metabolism in Wistar rats: a promising role against organ toxicities.Alcohol Alcohol. 2011 Jul-Aug;46(4):383-92. doi: 10.1093/alcalc/agr038. Epub 2011 Apr 29.

Study 4:
Testosterone and dihydrotestosterone inhibit gallbladder motility through multiple signalling pathways
T or DHT induced a concentration-dependent relaxation of cholecystokinin octapeptide (CCK)-induced tension. Pretreatment of the strips with PKA inhibitor 14-22 amide myristolated had no significant effect on the relaxation induced by either T or DHT. Pretreatment of strips with 2-APB, an inhibitor of IP(3) induced Ca(2+) release, produced a significant (p<0.001) reduction in the T- or DHT-induced relaxation. The flavone chrysin, an aromatase inhibitor, and genistein, an isoflavone, each produced a significant (p<0.01) reduction in CCK-induced tension. Chrysin significantly (p<0.05) increased T-induced relaxation; however, genistein had no effect on T-induced relaxation. It is concluded that T and DHT inhibits gallbladder motility rapidly by nongenomic actions of the hormones.
Source:
Kline LW, Karpinski E. Testosterone and dihydrotestosterone inhibit gallbladder motility through multiple signalling pathways.Steroids. 2008 Oct;73(11):1174-80. doi: 10.1016/j.steroids.2008.05.006. Epub 2008 May 23.

Study 5:
Effect of chrysin on nociception in formalin test and serum levels of noradrenalin and corticosterone in rats
Chrysin treatment can significantly decrease formalin-induced pain in rat in a dose-dependent manner. Chrysin (150 mg/kg) significantly inhibit the first phase (P < 0.01), whereas, the all concentration of chrysin were affected on the later phase of formalin-induced pain (P < 0.05).Chrysin could significantly attenuate the content of corticosterone and noradrenalin in the serum versus to the control rats (P < 0.01). The current study confirms that the chrysin decreased the nociceptive behaviors in the formalin test and indicate a correlation with decrease in serum corticosterone and noradrenalin levels.
Source:
Farkhondeh T, Samarghandian S, Azimin-Nezhad M, Samini F. Effect of chrysin on nociception in formalin test and serum levels of noradrenalin and corticosterone in rats.Int J Clin Exp Med. 2015 Feb 15;8(2):2465-70. eCollection 2015.

Study 6:
Flavonoid Chrysin prevents age-related cognitive decline via attenuation of oxidative stress and modulation of BDNF levels in aged mouse brain
In this study, the effect of Chrysin (5,7-dihydroxyflavone), an important member of the flavonoid family, on memory impairment, oxidative stress and BDNF reduction generated by aging in mice were investigated. Mice were trained and tested in Morris Water Maze task. Results demonstrated that the age-related memory decline was partially protected by Chrysin at a dose of 1mg/kg, and normalized at the dose of 10mg/kg (p<0.001). Treatment with Chrysin significantly attenuated the increase of RS levels and the inhibition of SOD, CAT and GPx activities of aged mice. Inhibition of Na+, K+-ATPase activity in PFC and HP of aged mice was also attenuated by Chrysin treatment. Moreover, Chrysin marked mitigated the decrease of BDNF levels in the PFC and HC of aged mice. These results demonstrated that flavonoid Chrysin, an antioxidant compound, was able to prevent age-associated memory probably by their free radical scavenger action and modulation of BDNF production. Thus, this study indicates that Chrysin may represent a new pharmacological approach to alleviate the age-related declines during normal age, acting as an anti-aging agent.
Source:
Souza LC, Antunes MS, Filho CB, Del Fabbro L, de Gomes MG, Goes AT, Donato F1, Prigol M, Boeira SP, Jesse CR. Flavonoid Chrysin prevents age-related cognitive decline via attenuation of oxidative stress and modulation of BDNF levels in aged mouse brain.Pharmacol Biochem Behav. 2015 Apr 28;134:22-30. doi: 10.1016/j.pbb.2015.04.010. [Epub ahead of print]

Study 7:
Chrysin-organogermanium (IV) complex induced Colo205 cell apoptosis-associated mitochondrial function and anti-angiogenesis
We found that Chry-Ge could induce apoptosis in Colo205 cells in mitochondrial-dependent pathway, cause the reorganization of cytoskeleton and induce the damage of nucleus in Colo205 cells. Besides, Chry-Ge was also found to induce membrane ultrastructural changes in Colo205 cells by AFM. Further, we found that Chry-Ge can inhibit tube formation of human umbilical vascular endothelial cell in vitro. Chry-Ge was also tested in vivo in the chicken chorioallantoic membrane (CAM) assay and found to inhibit bFGF-treated CAMs development. These results suggested that Chry-Ge could induce Colo205 cell apoptosis by mitochondrial pathway and anti-angiogenesis, highlighting the use of organic germanium agents for the treatment of colorectal cancer
Source:
Yang F, Gong L, Jin H, Pi J, Bai H, Wang H, Cai H, Yang P, Cai J. Chrysin-organogermanium (IV) complex induced Colo205 cell apoptosis-associated mitochondrial function and anti-angiogenesis.Scanning. 2015 Apr 23. doi: 10.1002/sca.21205.

Study 8:
Chrysin and its emerging role in cancer drug resistance
This letter illustrates the significant chemosensitizing effects of chrysin to resistance cancer cells and refers to the article on “Combination ofchrysin and cisplatin promotes the apoptosis of Hep G2 cells by up-regulating p53” by Li et al., published in your journal recently. Recent studies have demonstrated that chrysin is able to sensitize or kill cancer cells which are resistant to chemotherapeutic drugs such as cisplatin, doxorubicin and adriamycin. Owing to its potential anti-cancer effects and devoid of toxicity to non-transformed cells, further research is required to completely explore its chemosensitizing effects in other cancers and also assess and evaluate its safety, before going for possible human application.
Source:
Kasala ER, Bodduluru LN, Barua CC, Gogoi R. Chrysin and its emerging role in cancer drug resistance.Chem Biol Interact. 2015 Apr 24;236:7-8. doi: 10.1016/j.cbi.2015.04.017.

Study 9:
Beneficial effects of chrysin on the reproductive system of adult male rats
In this study, the beneficial effect of chrysin, a natural flavonoid currently under investigation due to its important biological activities, on reproductive system of rats was investigated. Rats (n = 16) were divided randomly into two equal groups. Rats in control group were given corn oil as carrier.Chrysin was orally administered at the dose of 50 mg kg(-1) per day by gavages, and it was dissolved in corn oil for 60 days. Tissue thiobarbituric acid reactive substances (TBARS) and glutathione (GSH) levels, antioxidant enzyme activity (CAT, SOD and GSH-Px), sperm parameters (motility, concentration and abnormal sperm rate), reproductive organ weight (testes, epididymis, vesicula seminalis, prostate) and serum testosterone levels were determined in the rats. Our results indicated that chrysin significantly increased GSH, CAT, GSH-Px and CuZn-SOD levels, but did not change the formation of TBARS significantly. In addition, sperm motility, sperm concentration and serum testosterone levels significantly increased, whereas abnormal sperm rate significantly decreased with chrysin treatment. In conclusion, it is suggested that treatment with chrysin can positively affect the reproductive system in rats, and it can be used for the treatment of male infertility.
Source:
Ciftci O, Ozdemir I, Aydin M, Beytur A. Beneficial effects of chrysin on the reproductive system of adult male rats. Andrologia. 2012 Jun;44(3):181-6. doi: 10.1111/j.1439-0272.2010.01127.x. Epub 2011 Mar 7.

Study 10:
Chrysin, a natural flavonoid enhances steroidogenesis and steroidogenic acute regulatory protein gene expression in mouse Leydig cells
To explore the possibility of delaying the decline using food supplements, we have studied steroidogenic effects of a natural flavonoid,chrysin, in mouse Leydig cells. Chrysin dramatically increased cyclic AMP (cAMP)-induced steroidogenesis in MA-10 mouse Leydig tumor cells. This result was confirmed using Leydig cells isolated from mouse testes. Also, chrysin significantly increased StAR promoter activity and StAR mRNA level. In the absence of cAMP, chrysin did not increase steroidogenesis. However, when a sub-threshold level of cAMP was used, StAR protein and steroid hormone were increased by chrysin to the levels seen with maximal stimulation of cAMP. These results suggest that while chrysin itself is unable to induce StAR gene expression and steroidogenesis, it appears to function by increasing the sensitivity of Leydig cells to cAMP stimulation.
Source:
Jana K, Yin X, Schiffer RB, Chen JJ, Pandey AK, Stocco DM, Grammas P, Wang X. Chrysin, a natural flavonoid enhances steroidogenesis and steroidogenic acute regulatory protein gene expression in mouse Leydig cells. J Endocrinol. 2008 May;197(2):315-23. doi: 10.1677/JOE-07-0282.

Study 11:
Effects of chrysin on urinary testosterone levels in human males
The equilibrium of sexual hormones in both sexes is controlled in vertebrates by the enzyme aromatase, a member of the cytochrome P450 superfamily, which catalyzes the conversion of androstenedione and testosterone into estrone and estradiol, respectively. Flavonoids are diphenolic compounds present in whole grains, legumes, fruits, and vegetables that are strongly implicated as protective in coronary heart disease, stroke, and cancer. The aim of this study was to verify if daily treatment for 21 days with propolis and honey, containing chrysin, would modify urinary concentrations of testosterone in volunteer male subjects. In fact, aromatase inhibition by chrysin could block the conversion of androgens into estrogens with a consequent increase oftestosterone, eventually measurable in urine samples.
Source:
Gambelunghe C, Rossi R, Sommavilla M, Ferranti C, Rossi R, Ciculi C, Gizzi S, Micheletti A, Rufini S. Effects of chrysin on urinary testosterone levels in human males.J Med Food. 2003 Winter;6(4):387-90.

Study 12:
Ameliorative effect of chrysin on adenine-induced chronic kidney disease in rats
Using several indices in plasma, urine and kidney homogenates, adenine was found to impair kidney function as it lowered creatinine clearance and increased plasma concentrations of creatinine, urea, neutrophil gelatinase-associated lipocalin and N-Acetyl-beta-D-glucosaminidase activity. Furthermore, it raised plasma concentrations of the uremic toxin indoxyl sulfate, some inflammatory cytokines and urinary albumin concentration. In renal homogenates, antioxidant indices, including superoxide dismutase and catalase activities, total antioxidant capacity and reduced glutathione were all adversely affected. Most of these adenine – induced actions were moderately and dose -dependently mitigated by chrysin, especially at the highest dose. Chrysin did not cause any overt adverse effect on the treated rats.
Source:
Ali BH, Adham SA, Al Za’abi M, Waly MI, Yasin J, Nemmar A, Schupp N. Ameliorative effect of chrysin on adenine-induced chronic kidney disease in rats.PLoS One. 2015 Apr 24;10(4):e0125285. doi: 10.1371/journal.pone.0125285. eCollection 2015.

Study 13:
Selenium-Containing Chrysin and Quercetin Derivatives: Attractive Scaffolds for Cancer Therapy
Selenium-containing chrysin (SeChry) and 3,7,3′,4′-tetramethylquercetin (SePQue) derivatives were synthesized by a microwave-based methodology. In addition to their improvement in terms of DPPH scavenging and potential GPx-like activities, when tested in a panel of cancer cell lines both selenium-derivatives revealed consistently to be more cytotoxic when compared with their oxo and thio-analogues, evidencing the key role of selenocabonyl moiety for these activities. In particular, SeChry elicited a noteworthy cytotoxic activity with mean IC50 values 18- and 3-fold lower than those observed for chrysin and cisplatin, respectively. Additionally, these seleno-derivatives evidenced an ability to overcome cisplatin and multidrug resistance. Notably, a differential behavior toward malignant and nonmalignant cells was observed for SeChry and SePQue, exhibiting higher selectivity indexes when compared with the chalcogen-derivatives and cisplatin. Our preliminary investigation on the mechanism of cytotoxicity of SeChry and SePQue in MCF-7 human mammary cancer cells demonstrated their capacity to efficiently suppress the clonal expansion along with their ability to hamper TrxR activity leading to apoptotic cell death.
Source:
Martins IL, Charneira C, Gandin V, Ferreira da Silva JL, Justino GC, Telo JP, Vieira AJ, Marzano C, Antunes AM. Selenium-Containing Chrysin and Quercetin Derivatives: Attractive Scaffolds for Cancer Therapy.J Med Chem. 2015 May 7.

Study 14:
Chrysin Inhibits Tumor Promoter-Induced MMP-9 Expression by Blocking AP-1 via Suppression of ERK and JNK Pathways in Gastric Cancer Cells
Cell invasion is a crucial mechanism of cancer metastasis and malignancy. High expression levels of MMP-9 in gastric cancer positively correlate with tumor aggressiveness and have a significant negative correlation with patients’ survival times. In the present study, we tested the effects of chrysin on MMP-9 expression in gastric cancer cells, and determined its underlying mechanism. We examined the effects of chrysin on MMP-9 expression and activity via RT-PCR, zymography, promoter study, and western blotting in human gastric cancer AGS cells. Chrysin inhibited phorbol-12-myristate 13-acetate (PMA)-induced MMP-9 expression in a dose-dependent manner. Using AP-1 decoy oligodeoxynucleotides, we confirmed that AP-1 was the crucial transcriptional factor for MMP-9 expression. Furthermore, AGS cells pretreated with PMA showed markedly enhanced invasiveness, which was partially abrogated by chrysin and MMP-9 antibody.
Source:
Xia Y, Lian S, Khoi PN, Yoon HJ, Joo YE1, Chay KO, Kim KK, Do Jung Y. Chrysin Inhibits Tumor Promoter-Induced MMP-9 Expression by Blocking AP-1 via Suppression of ERK and JNK Pathways in Gastric Cancer Cells. PLoS One. 2015 Apr 15;10(4):e0124007. doi: 10.1371/journal.pone.0124007. eCollection 2015.

Study 15:
Chrysin inhibits foam cell formation through promoting cholesterol efflux from RAW264.7 macrophages
Chrysin dose dependently inhibited the formation of foam cells and prevented the enhanced cholesterol accumulation by ox-LDL. Treatment with chrysin (10 μM) significantly enhanced cholesterol efflux and substantially inhibited cholesterol influx. Simultaneously, chrysin significantly increased the mRNA levels of PPARγ, liver X receptor alpha (LXRα), ATP-binding cassette, sub-family A1 (ABCA1), and sub-family G1 (ABCG1), decreased scavenger receptor A1 (SR-A1) and SR-A2, and increased the transcriptional activity of PPARγ.Chrysin is a new inhibitor of foam cell formation that may stimulate cholesterol flow. Up-regulation of the classical PPARγ-LXRα-ABCA1/ABCG1 pathway and down-regulation of SR-A1 and SR-A2 may participate in its suppressive effect on intracellular cholesterol accumulation.
Source:
Wang S, Zhang X, Liu M, Luan H, Ji Y, Guo P, Wu C. Chrysin inhibits foam cell formation through promoting cholesterol efflux from RAW264.7 macrophages.Pharm Biol. 2015 Apr 10:1-7.

Study 16:
Protective effects of the flavonoid chrysin against methylmercury-induced genotoxicity and alterations of antioxidant status, in vivo.
One of dietary flavonoids is chrysin (CR), found mainly in passion fruit, honey, and propolis. Methylmercury (MeHg) is a toxic metal whose main toxic mechanism is oxidative damage. Thus, the study aimed to evaluate the antioxidant effects of CR against oxidative damage induced by MeHg in Wistar rats. Animals were treated with MeHg (30 µg/kg/bw) in presence and absence of CR (0.10, 1.0, and 10 mg/kg/bw) by gavage for 45 days. Moreover, exposure to MeHg decreased the levels of GSH and GSH levels were restored in the animals that received CR plus MeHg.
Source:
Manzolli ES, Serpeloni JM, Grotto D, Bastos JK, Antunes LM, Barbosa Junior F, Barcelos GR. Protective effects of the flavonoid chrysin against methylmercury-induced genotoxicity and alterations of antioxidant status, in vivo.Oxid Med Cell Longev. 2015;2015:602360. doi: 10.1155/2015/602360.

Diindolylmethane (DIM)
Facts:
Diindolylmethane (DIM) is a molecule commonly found in broccoli known to prevent breast, uterine and colorectal cancer. It affects the metabolism of estrogen and is able to maintain the body’s balance.
Source:
http://examine.com/supplements/Diindolylmethane
http://www.webmd.com/vitamins-supplements/ingredientmono-1049-diindolylmethane.aspx?activeingredientid=1049&activeingredientname=diindolylmethane

Study 1:
Safety and tolerability of DIM-based therapy designed as personalized approach to reverse prostatic intraepithelial neoplasia (PIN).
The trial revealed that Infemin treatment is associated with minimal toxicity and no serious adverse events when administered orally for 3 months. We noted three adverse events including nausea and diarrhea in two patients (14%). Combined 95% confidence interval (CI) was 1.8%-42.8%. Therapy was continued in all cases of adverse events. Good tolerability of DIM-based formulation allows us to recommend it for further clinical trials among men diagnosed with PIN for its efficacy and long-term safety parameters.
Source:
Paltsev M, Kiselev V, Muyzhnek E, Drukh V, Kuznetsov I, Pchelintseva O. Safety and tolerability of DIM-based therapy designed as personalized approach to reverse prostatic intraepithelial neoplasia (PIN). EPMA J. 2014 Oct 8;5(1):18. doi: 10.11
86/1878-5085-5-18. eCollection 2014.

Study 2:
FOXM1-mediated downregulation of uPA and MMP9 by 3,3′-diindolylmethane inhibits migration and invasion of human colorectal cancer cells.
DIM significantly inhibited the migration and invasion of colorectal cancer cells as assessed by wound healing and Matrigel invasion assays. The migratory ability of the DLD-1 and HCT116 cells was significantly reduced by DIM at 24 and 48 h. DIM also significantly inhibited the invasion rate of the DLD-1 and HCT116 cells in a dose-dependent manner. DIM also decreased the protein levels of uPA and MMP9, yet significantly increased E-cadherin protein expression. In addition, DIM significantly reduced the mRNA and protein levels of FOXM1 in the DLD-1 and HCT116 cells.
Source:
Jin H, Li XJ, Park MH, Kim SM. FOXM1-mediated downregulation of uPA and MMP9 by 3,3′-diindolylmethane inhibits migration and invasion of human colorectal cancer cells. Oncol Rep. 2015 Jun;33(6):3171-7. doi: 10.3892/or.2015.3938. Epub 2015 Apr 28.

Study 3:
3,3′-Diindolylmethane inhibits VEGF expression through the HIF-1α and NF-κB pathways in human retinal pigment epithelial cells under chemical hypoxic conditions.
Hypoxia induces oxidative damage in retinal pigment epithelial cells (RPE cells). Human RPE cells (ARPE-19 cells) were treated with cobalt chloride (CoCl2, 200 µM) and/or DIM (10 and 20 µM). The production of VEGF was measured by enzyme-linked immunosorbent assay. The translocation of hypoxia-inducible factor-1α (HIF-1α) and nuclear factor-κB (NF-κB) was determined by western blot analysis. The results revealed that DIM significantly attenuated the CoCl2-induced expression of VEGF in the ARPE-19 cells. The CoCl2-induced translocation and activation of HIF-1α and NF-κB were also attenuated by treatment with DIM. Taken together, the findings of our study demonstrate that DIM inhibits the CoCl2-induced production of VEGF by suppressing mitochondrial ROS production, thus attenuating the activation of HIF-1α and p38 MAPK/NF-κB.
Source:
Park H, Lee DS, Yim MJ, Choi YH, Park S, Seo SK, Choi JS, Jang WH, Yea SS, Park WS, Lee CM, Jung WK, Choi IW.3,3′-Diindolylmethane inhibits VEGF expression through the HIF-1α and NF-κB pathways in human retinal pigment epithelial cells under chemical hypoxic conditions.Int J Mol Med. 2015 May 5. doi: 10.3892/ijmm.2015.2202.

Study 4:
Synergistic anticancer activity of capsaicin and 3,3′-diindolylmethane in human colorectal cancer.
A promising area of cancer research is focused on chemoprevention by nutritional compounds. Although individual active compounds have demonstrated significant anticancer activity, an emerging area of research is focusing on the combination of multiple dietary compounds that act synergistically on cancer to exert greater effects. The current study evaluated the potential synergistic effects of capsaicin, an active compound from red chili peppers, in combination with 3,3′-diindolylmethane (DIM), from cruciferous vegetables. The present study suggests capsaicin and DIM work synergistically to inhibit cell proliferation and induce apoptosis in colorectal cancer through modulating transcriptional activity of NF-κB, p53, and target genes associated with apoptosis.
Source:
Clark R, Lee J, Lee SH.Synergistic anticancer activity of capsaicin and 3,3′-diindolylmethane in human colorectal cancer. J Agric Food Chem. 2015 May 6;63(17):4297-304. doi: 10.1021/jf506098s. Epub 2015 Apr 22.

Study 5:
A therapeutically relevant, 3,3′-diindolylmethane derivative NGD16 attenuates angiogenesis by targeting glucose regulated protein, 78kDa (GRP78).
Angiogenesis remain a critical procedure for tumor progression and malignancy. NGD16 suppressed the viability of prostate cancer (PC-3), pancreatic adenocarcinoma (MiaPaca-2), colorectal cancer (COLO-205) and human umbilical vein endothelial cells (HUVECs) effectively with IC50 values 0.8μM, 2.8μM, 5.3μM and 2.5μM respectively. At the molecular level, NGD16 inhibited the expression of glucose regulated protein, 78kDa (GRP78), vascular endothelial growth factor receptor-2 (VEGFR2) and matrix metalloproteinase-9 (MMP-9) expression, the main mediators of angiogenesis and neovessel formation. Antibody blocking of GRP78 further potentiated NGD16 in attenuating angiogenesis through inhibition of MMP-9. NGD16 depicted its promising biodistribution profile in a pharmacokinetic study with 46.9% intraperitoneal bioavailability.
Source:
Nayak D, Amin H, Rah B, Ur Rasool R, Sharma D, Gupta AP, Kushwaha M, Mukherjee D, Goswami A.A therapeutically relevant, 3,3′-diindolylmethane derivative NGD16 attenuates angiogenesis by targeting glucose regulated protein, 78kDa (GRP78).Chem Biol Interact. 2015 May 5;232:58-67. doi: 10.1016/j.cbi.2015.03.008. Epub 2015 Mar 17.

Study 6:
Natural indoles, indole-3-carbinol (I3C) and 3,3′-diindolylmethane (DIM), attenuate staphylococcal enterotoxin B-mediated liver injury by downregulating miR-31 expression and promoting caspase-2-mediated apoptosis.
Staphylococcal enterotoxin B (SEB) is a potent superantigen capable of inducing inflammation characterized by robust immune cell activation and proinflammatory cytokine release. In the current study, we investigated the effect of natural indoles including indole-3-carbinol (I3C) and 3,3′-diindolylmethane (DIM) on SEB-mediated liver injury. Administration of I3C and DIM (40 mg/kg), by intraperitonal injection, attenuated SEB-induced acute liver injury, as evidenced by decrease in AST levels, inflammatory cytokines and cellular infiltration in the liver. I3C and DIM triggered apoptosis in SEB-activated T cells primarily through activation of the intrinsic mitochondrial pathway. Both compounds also down-regulated miR-31, which directly targets caspase-2 and influences apoptosis in SEB-activated cells.
Source:
Busbee PB, Nagarkatti M, Nagarkatti PS.Natural indoles, indole-3-carbinol (I3C) and 3,3′-diindolylmethane (DIM), attenuate staphylococcal enterotoxin B-mediated liver injury by downregulating miR-31 expression and promoting caspase-2-mediated apoptosis.PLoS One. 2015 Feb 23;10(2):e0118506. doi: 10.1371/journal.pone.0118506. eCollection 2015.

Study 7:
3,3′-Diindolylmethane increases bone mass by suppressing osteoclastic bone resorption in mice.
This study evaluated the effect of DIM on bone mass in mice under physiological and pathological conditions. Further, Bone histomorphometric analyses showed that this treatment significantly reduced bone resorption parameters, but did not increase bone formation parameters. Furthermore, we use ovariectomized (OVX)-induced osteoporotic mouse model, and explore function of DIM in skeletal pathological processes. Bone phenotype analyses revealed that the administration of DIM in this study effectively prevented OVX-induced bone loss resulting from increased bone resorption. Our results demonstrated that DIM increased bone mass by suppressing osteoclastic bone resorption in bone metabolism under both physiological and pathological conditions.
Source:
Yu TY, Pang WJ, Yang GS.3,3′-Diindolylmethane increases bone mass by suppressing osteoclastic bone resorption in mice.J Pharmacol Sci. 2015 Jan;127(1):75-82. doi: 10.1016/j.jphs.2014.11.006. Epub 2014 Nov 20.

Study 8:
3,3′-Diindolylmethane potentiates paclitaxel-induced antitumor effects on gastric cancer cells through the Akt/FOXM1 signaling cascade.
Forkhead box M1 (FOXM1) is overexpressed in gastric cancer, suggesting that it is important in gastric cancer oncogenesis. Here, we report for the first time that DIM effectively downregulated Akt/FOXM1 in gastric cancer cells. Combination treatment with DIM and paclitaxel significantly and dose-dependently inhibited the proliferation of SNU638 cells when compared to treatment with DIM or paclitaxel alone. DIM dose-dependently sensitized gastric cancer cells through downregulation of FOXM1 and potentiated the effects of paclitaxel. FOXM1 effector genes such as CDK4, p53 and cyclin D1 were downregulated in gastric cancer cells by combination treatment with DIM and paclitaxel. In addition, DIM significantly and dose-dependently inhibited phosphorylation of Akt and potentiated paclitaxel-induced inhibition of Akt function in gastric cancer cells. Our findings suggest that DIM enhances the therapeutic efficacy of paclitaxel in gastric cancer and is a potential clinical anticancer agent for the prevention and/or treatment of gastric cancer.
Source:
Jin H, Park MH, Kim SM.3,3′-Diindolylmethane potentiates paclitaxel-induced antitumor effects on gastric cancer cells through the Akt/FOXM1 signaling cascade. Oncol Rep. 2015 Apr;33(4):2031-6. doi: 10.3892/or.2015.3758. Epub 2015 Jan 28.

Study 9:
Androgen receptor splice variants contribute to prostate cancer aggressiveness through induction of EMT and expression of stem cell marker genes.
Overexpression of AR led to the induction of EMT phenotype, while overexpression of AR3 not only induced EMT but also led to the expression of stem cell signature genes. More importantly, ADT enhanced the expression of AR and AR3 concomitant with up-regulated expression of EMT and stem cell marker genes. Dihydrotestosterone (DHT) treatment decreased the expression of AR and AR3, and reversed the expression of these EMT and stem cell marker genes. BR-DIM administered to PCa patients prior to radical prostatectomy inhibited the expression of cancer stem cell markers consistent with inhibition of self-renewal of PCa cells after BR-DIM treatment.AR variants could contribute to PCa progression through induction of EMT and acquisition of stem cell characteristics, which could be attenuated by BR-DIM, suggesting that BR-DIM could become a promising agent for the prevention of CRPC and/or for the treatment of PCa.
Source:
Kong D, Sethi S, Li Y, Chen W, Sakr WA, Heath E, Sarkar FH.Androgen receptor splice variants contribute to prostate cancer aggressiveness through induction of EMT and expression of stem cell marker genes. Prostate. 2015 Feb;75(2):161-74. doi: 10.1002/pros.22901. Epub 2014 Oct 13.

Study 10:
Preclinical antitumor activity of the diindolylmethane formulation in xenograft mouse model of prostate cancer.
Statistically significant tumor xenograft regressions have been shown in group which received the DIM, cod liver oil, polysorbate 80 and α-tocopherol acetate (vitamin E) on the 37(th) day of observation post inoculation. The highest antitumor activity was achieved on the 39(th) day (T/C = 16,8%). Therapeutic effect lasts for 6 days after the end of therapy period.Our findings demonstrate inhibitory effect of the formulation on tumor development in the xenograft animal model due to the tumor growth rate reduction.
Source:
Kiselev VI, Drukh VM, Muyzhnek EL, Kuznetsov IN, Pchelintseva OI, Paltsev MA.Preclinical antitumor activity of the diindolylmethane formulation in xenograft mouse model of prostate cancer.Exp Oncol. 2014 Jun;36(2):90-3.

Study 11:
Beyond androgen deprivation: ancillary integrative strategies for targeting the androgen receptor addiction of prostate cancer.
The large majority of clinical prostate cancers remain dependent on androgen receptor (AR) activity for proliferation even as they lose their responsiveness to androgen deprivation or antagonism. AR activity can be maintained in these circumstances by increased AR synthesis–often reflecting increased NF-κB activation; upregulation of signaling pathways that promote AR activity in the absence of androgens; and by emergence of AR mutations or splice variants lacking the ligand-binding domain, which render the AR constitutively active. Concurrent measures that suppress AR synthesis or boost AR turnover could be expected to complement the efficacy of such drugs. The proteasomal turnover of the AR is abetted by diets with a high ratio of long-chain omega-3 to omega-6 fatty acids, which are beneficial in prostate cancer xenograft models; berberine and sulforaphane, by inhibiting AR’s interaction with its chaperone Hsp90, likewise promote AR proteasomal degradation and retard growth of human prostate cancer in nude mice.
Source:
McCarty MF, Hejazi J, Rastmanesh R. Beyond androgen deprivation: ancillary integrative strategies for targeting the androgen receptor addiction of prostate cancer.Integr Cancer Ther. 2014 Sep;13(5):386-95. doi: 10.1177/1534735414534728. Epub 2014 May 26.

Study 12:
Effects of sulforaphane and 3,3′-diindolylmethane on genome-wide promoter methylation in normal prostate epithelial cells and prostate cancer cells.
The goal of this study was to determine the genome-wide effects of SFN and DIM on promoter methylation in normal prostate epithelial cells and prostate cancer cells. Both SFN and DIM treatment decreased DNA methyltransferase expression in normal prostate epithelial cells (PrEC), and androgen-dependent (LnCAP) and androgen-independent (PC3) prostate cancer cells. The effects of SFN and DIM on promoter methylation profiles in normal PrEC, LnCAP and PC3 prostate cancer cells were determined using methyl-DNA immunoprecipitation followed by genome-wide DNA methylation array. In particular, SFN and DIM altered promoter methylation in distinct sets of genes in PrEC, LnCAP, and PC3 cells, but shared similar gene targets within a single cell line.
Source:
Wong CP, Hsu A, Buchanan A, Palomera-Sanchez Z, Beaver LM, Houseman EA, Williams DE, Dashwood RH, Ho E.Effects of sulforaphane and 3,3′-diindolylmethane on genome-wide promoter methylation in normal prostate epithelial cells and prostate cancer cells.PLoS One. 2014 Jan 22;9(1):e86787. doi: 10.1371/journal.pone.0086787. eCollection 2014.

Study 13:
Indole-3-carbinol and 3′,3′-diindolylmethane modulate androgen’s effect on C-C chemokine ligand 2 and monocyte attraction to prostate cancer cells.
We hypothesized that androgen could modulate CCL2 expression in hormone-responsive prostate cancer cells and thereby promote recruitment of monocytes. Dihydrotestosterone was found to induce a time-dependent (0-72 hours) and concentration-dependent (0-1 nmol/L) increase in CCL2 mRNA levels in androgen-responsive human prostate cancer cells (LNCaP). This increase in CCL2 mRNA corresponded with increased secretion of CCL2 protein. The effect of dihydrotestosterone was mediated through an androgen receptor (AR)-dependent pathway as small inhibitor RNA against AR negated the induction of CCL2. Both I3C and DIM inhibited promotional effects of dihydrotestosterone on CCL2 and migration.
Source:
Kim EK, Kim YS, Milner JA, Wang TT. Indole-3-carbinol and 3′,3′-diindolylmethane modulate androgen’s effect on C-C chemokine ligand 2 and monocyte attraction to prostate cancer cells.Cancer Prev Res (Phila). 2013 Jun;6(6):519-29. doi: 10.1158/1940-6207.CAPR-12-0419. Epub 2013 Apr 12.

Study 14:
Indole-3-carbinol and 3′,3′-diindolylmethane modulate androgen’s effect on C-C chemokine ligand 2 and monocyte attraction to prostate cancer cells.
We hypothesized that androgen could modulate CCL2 expression in hormone-responsive prostate cancer cells and thereby promote recruitment of monocytes. Dihydrotestosterone was found to induce a time-dependent (0-72 hours) and concentration-dependent (0-1 nmol/L) increase in CCL2 mRNA levels in androgen-responsive human prostate cancer cells (LNCaP). This increase in CCL2 mRNA corresponded with increased secretion of CCL2 protein. The effect of dihydrotestosterone was mediated through an androgen receptor (AR)-dependent pathway as small inhibitor RNA against AR negated the induction of CCL2. Both I3C and DIM inhibited promotional effects of dihydrotestosterone on CCL2 and migration.
Source:
Kim EK, Kim YS, Milner JA, Wang TT. Indole-3-carbinol and 3′,3′-diindolylmethane modulate androgen’s effect on C-C chemokine ligand 2 and monocyte attraction to prostate cancer cells.Cancer Prev Res (Phila). 2013 Jun;6(6):519-29. doi: 10.1158/1940-6207.CAPR-12-0419. Epub 2013 Apr 12.

Longjack Eurycoma Longifolia (root)
Facts:
Eurycoma Longifolia is an Asian herb known to boost men’s sexual ability and virility. It is known to be an Aphrodisiac which has anti-estrogen characteristics. A handful clinical studies has been conducted to support these functions.
Sources:
http://examine.com/supplements/Eurycoma%20Longifolia%20Jack.
http://www.webmd.com/vitamins-supplements/ingredientmono-1132-eurycoma%20longifolia.aspx?activeingredientid=1132&activeingredientname=eurycoma%20longifolia

Study 1:
The In Vitro and In Vivo Anti-Cancer Activities of a Standardized Quassinoids Composition from Eurycoma longifolia on LNCaP Human Prostate Cancer Cells.
We hypothesized that androgen could modulate CCL2 expression in hormone-responsive prostate cancer cells and thereby promote recruitment of monocytes. Dihydrotestosterone was found to induce a time-dependent (0-72 hours) and concentration-dependent (0-1 nmol/L) increase in CCL2 mRNA levels in androgen-responsive human prostate cancer cells (LNCaP). This increase in CCL2 mRNA corresponded with increased secretion of CCL2 protein. The effect of dihydrotestosterone was mediated through an androgen receptor (AR)-dependent pathway as small inhibitor RNA against AR negated the induction of CCL2. Both I3C and DIM inhibited promotional effects of dihydrotestosterone on CCL2 and migration.
Source:
Tong KL, Chan KL, AbuBakar S, Low BS, Ma HQ, Wong PF. The In Vitro and In Vivo Anti-Cancer Activities of a Standardized Quassinoids Composition from Eurycoma longifolia on LNCaP Human Prostate Cancer Cells. LoS One. 2015 Mar 31;10(3):e0121752. doi: 10.1371/journal.pone.0121752. eCollection 2015.

Study 2:
Production of biomass and bioactive compounds from adventitious roots by optimization of culturing conditions of Eurycoma longifolia in balloon-type bubble bioreactor system.
The effects of the type and concentration of auxin on root growth were studied, as well as the effects of the NH4(+):NO3(-) ratio on adventitious root growth and the production of phenolics and flavonoids. The adventitious roots were thin, numerous, and elongated in 3/4 MS medium supplemented with 5.0 and 7.0 mg L(-1) IBA, whereas the lateral roots were shorter and thicker with 5.0 mg L(-1) NAA compared with IBA treatment. High phenolic and flavonoid productions (38.59 and 11.27 mg L(-1) medium, respectively) were also obtained with a ratio of 15:30. These results suggest that balloon-type bubble bioreactor cultures are suitable for the large-scale commercial production of E. longifolia adventitious roots which contain high yield of bioactive compounds.
Source:
Lulu T, Park SY, Ibrahim R, Paek KY.Production of biomass and bioactive compounds from adventitious roots by optimization of culturing conditions of Eurycoma longifolia in balloon-type bubble bioreactor system.J Biosci Bioeng. 2015 Jun;119(6):712-7. doi: 10.1016/j.jbiosc.2014.11.010. Epub 2014 Dec 12.

Study 3:
Eurycomanone and eurycomanol from Eurycoma longifolia Jack as regulators of signaling pathways involved in proliferation, cell death and inflammation.
Eurycomanone and eurycomanol are two quassinoids from the roots of Eurycoma longifolia Jack. The aim of this study was to assess the bioactivity of these compounds in Jurkat and K562 human leukemia cell models compared to peripheral blood mononuclear cells from healthy donors. Both eurycomanone and eurycomanol inhibited Jurkat and K562 cell viability and proliferation without affecting healthy cells. Interestingly, eurycomanone inhibited NF-κB signaling through inhibition of IκBα phosphorylation and upstream mitogen activated protein kinase (MAPK) signaling, but not eurycomanol. In conclusion, both quassinoids present differential toxicity towards leukemia cells, and the presence of the α,β-unsaturated ketone in eurycomanone could be prerequisite for the NF-κB inhibition.
Source:
Hajjouli S, Chateauvieux S, Teiten MH, Orlikova B, Schumacher M, Dicato M, Choo CY, Diederich M.Eurycomanone and eurycomanol from Eurycoma longifolia Jack as regulators of signaling pathways involved in proliferation, cell death and inflammation. Molecules. 2014 Sep 16;19(9):14649-66. doi: 10.3390/molecules190914649.

Study 4:
Supplementation of Eurycoma longifolia Jack Extract for 6 Weeks Does Not Affect Urinary Testosterone: Epitestosterone Ratio, Liver and Renal Functions in Male Recreational Athletes.
Supplementation of ElJ i.e. Physta(®) at a dosage of 400 mg/day for 6 weeks did not affect the urinary T:E ratio and hence will not breach any doping policies of the International Olympic Committee for administration of exogenous testosterone or its precursor. In addition, the supplementation of ElJ at this dosage and duration was safe as it did adversely affect the liver and renal functions.
Source:
Chen CK, Mohamad WM, Ooi FK, Ismail SB, Abdullah MR, George A.Supplementation of Eurycoma longifolia Jack Extract for 6 Weeks Does Not Affect Urinary Testosterone: Epitestosterone Ratio, Liver and Renal Functions in Male Recreational Athletes.Int J Prev Med. 2014 Jun;5(6):728-33.

Study 5:
Effects of a Proprietary Freeze-Dried Water Extract of Eurycoma longifolia (Physta) and Polygonum minus on Sexual Performance and Well-Being in Men: A Randomized, Double-Blind, Placebo-Controlled Study.
Background. Methods. Outcome measures included validated questionnaires that aimed to evaluate erectile function, satisfaction with intervention, sexual intercourse performance, erectile hardness, mood, and overall quality of life. Results. 12 subjects in the active group and 14 in the placebo group completed the study. Significant improvements were noted in scores for the Sexual Intercourse Attempt diary, Erection Hardness Scale, Sexual Health Inventory of Men, and Aging Male Symptom scale (P < 0.05 for all). Conclusion. Supplementation for twelve weeks with Polygonum minus and the proprietary Eurycoma longifolia extract, Physta, was well tolerated and more effective than placebo in enhancing sexual performance in healthy volunteers.
Source:
Udani JK, George AA, Musthapa M, Pakdaman MN, Abas A.Effects of a Proprietary Freeze-Dried Water Extract of Eurycoma longifolia (Physta) and Polygonum minus on Sexual Performance and Well-Being in Men: A Randomized, Double-Blind, Placebo-Controlled Study.Evid Based Complement Alternat Med. 2014;2014:179529. doi: 10.1155/2014/179529. Epub 2014 Jan 12.

Study 6:
Four new quassinoids from the roots of Eurycoma longifolia Jack.
Seven compounds were isolated from the roots of Eurycoma longifolia, and characterized by comprehensive analysis of 1D and 2D NMR experiments along with single crystal X-ray diffraction. Among them, four new quassinoids were identified and three of them were diastereomers for each other. Compounds 1-7 were evaluated for cytotoxicities against HT-29, MCF-7, LOVO, BGC-823, MGC-803, HepG2, HeLa, and A549 cancer cell lines. Compounds 2 and 5 exhibited the lowest IC50 values of 24.9 μM, 11.8 μM, and 44.1 μM, 14.1 μM towards MCF-7, MGC-803 cancer cell lines, respectively, while compound 6 exhibited moderate cytotoxicity towards all the selected cancer cell lines.
Source:
Meng D, Li X, Han L, Zhang L, An W, Li X.Four new quassinoids from the roots of Eurycoma longifolia Jack. Fitoterapia. 2014 Jan;92:105-10.

Study 7:
Eurycoma longifolia in Radix for the treatment of ethanol-induced gastric lesion in rats.
The effect of treatment with Radix on ethanol-induced gastric lesions was investigated. Three groups were given 0.5 mL 100% ethanol orally. One group that was administered with ethanol was only given distilled water orally (no treatment). (Radix) and oral ranitidine 21.4 mg kg(-1) b.wt. (Ranitidine), respectively. There was no difference in ulcer index between the Radix and ranitidine group. The gastric MDA content was significantly higher in all the groups that were induced with ethanol compared to the control group but no difference between all the ethanol-induced groups.
Source:
Qodriyah HM, Asmadi AY.Eurycoma longifolia in Radix for the treatment of ethanol-induced gastric lesion in rats.Pak J Biol Sci. 2013 Dec 1;16(23):1815-8.

Study 8:
Phytoandrogenic properties of Eurycoma longifolia as natural alternative to testosterone replacement therapy.
This significant decline in testosterone levels is further closely linked with medical conditions such as obesity, metabolic syndrome, diabetes or hypertension. Apart from the beneficial effects of TRT, significant adverse side effects have been described, and prostate cancer (PCa) as absolute contraindication is debated. Eurycoma longifolia (Tongkat Ali; TA) is natural alternative to TRT and has been shown to restore serum testosterone levels, thus significantly improving sexual health. This includes significant positive effects on bone health and physical condition of patients.
Source:
George A, Henkel R.Phytoandrogenic properties of Eurycoma longifolia as natural alternative to testosterone replacement therapy.Andrologia. 2014 Sep;46(7):708-21. doi: 10.1111/and.12214. Epub 2014 Jan 6. Review.

Study 9:
Acute, reproductive toxicity and two-generation teratology studies of a standardized quassinoid-rich extract of Eurycoma longifolia Jack in Sprague-Dawley rats.
The roots of Eurycoma longifolia Jack are popularly sought as herbal medicinal supplements to improve libido and general health amongst the local ethnic population. The major quassinoids of E. longifolia improved spermatogenesis and fertility but toxicity studies have not been well documented. The reproductive toxicity, two generation of foetus teratology and the up-and-down acute toxicity were investigated in Sprague-Dawley rats orally treated with quassinoid-rich E. longifolia extract (TAF273). The results showed that the median lethal dose (LD50 ) of TAF273 for female and male rats was 1293 and >2000 mg/kg, respectively. Fertility index and litter size of the TAF273 treated were significantly increased when compared with those of the non-treated animals. The TAF273-treated dams decreased in percentage of pre-implantation loss, post-implantation loss and late resorption. hus, any human dose derived from converting the rat doses of 100 mg/kg and below may be considered as safe for further clinical studies.
Source:
Low BS, Das PK, Chan KL.Acute, reproductive toxicity and two-generation teratology studies of a standardized quassinoid-rich extract of Eurycoma longifolia Jack in Sprague-Dawley rats. Phytother Res. 2014 Jul;28(7):1022-9. doi: 10.1002/ptr.5094. Epub 2013 Dec 6.

Study 10:
Evaluation of Acute 13-Week Subchronic Toxicity and Genotoxicity of the Powdered Root of Tongkat Ali (Eurycoma longifolia Jack).
Tongkat Ali (Eurycoma longifolia) is an indigenous traditional herb in Southern Asia. Its powdered root has been processed to produce health supplements, but no detailed toxicology report is available. In this study, neither mutagenicity nor clastogenicity was noted, and acute oral LD50 was more than 6 g/kg b.w. After 4-week subacute and 13-week subchronic exposure paradigms (0, 0.6, 1.2, and 2 g/kg b.w./day), adverse effects attributable to test compound were not observed with respect to body weight, hematology, serum biochemistry, urinalysis, macropathology, or histopathology. However, the treatment significantly reduced prothrombin time, partial thromboplastin time, blood urea nitrogen, creatinine, aspartate aminotransferase, creatine phosphate kinase, lactate dehydrogenase, and cholesterol levels, especially in males (P < 0.05). These changes were judged as pharmacological effects, and they are beneficial to health. The calculated acceptable daily intake (ADI) was up to 1.2 g/adult/day. This information will be useful for product development and safety management.
Source:
Li CH, Liao JW, Liao PL, Huang WK, Tse LS, Lin CH, Kang JJ, Cheng YW. Evaluation of Acute 13-Week Subchronic Toxicity and Genotoxicity of the Powdered Root of Tongkat Ali (Eurycoma longifolia Jack).Evid Based Complement Alternat Med. 2013;2013:102987. doi: 10.1155/2013/102987. Epub 2013 Aug 25.

Study 11:
Anti-angiogenic quassinoid-rich fraction from Eurycoma longifolia modulates endothelial cell function.
In the present study, antiangiogenic potential of partially purified quassinoid-rich fraction (TAF273) of E. longifolia root extract was evaluated using ex vivo and in vivo angiogenesis models and the anti-angiogenic efficacy of TAF273 was investigated in human umbilical vein endothelial cells (HUVEC). TAF273 caused significant suppression in sprouting of microvessels in rat aorta with IC50 11.5μg/ml. TAF273 (50μg/ml) showed remarkable inhibition (63.13%) of neovascularization in chorioallantoic membrane of chick embryo. In vitro, TAF273 significantly inhibited the major angiogenesis steps such as proliferation, migration and differentiation of HUVECs. Phytochemical analysis revealed high content of quassinoids in TAF273.
Source:
Al-Salahi OS, Kit-Lam C, Majid AM, Al-Suede FS, Mohammed Saghir SA, Abdullah WZ, Ahamed MB, Yusoff NM.Anti-angiogenic quassinoid-rich fraction from Eurycoma longifolia modulates endothelial cell function.Microvasc Res. 2013 Nov;90:30-9. doi: 10.1016/j.mvr.2013.07.007. Epub 2013 Jul 27.

Study 12:
Eurycomanone, the major quassinoid in Eurycoma longifolia root extract increases spermatogenesis by inhibiting the activity of phosphodiesterase and aromatase in steroidogenesis.
Eurycomanone enhanced testosterone steroidogenesis at the Leydig cells by inhibiting aromatase conversion of testosterone to oestrogen, and at a high concentration may also involve phosphodiesterase inhibition. The quassinoid may be worthy for further development as a phytomedicine to treat testosterone-deficient idiopathic male infertility and sterility.
Source:
Low BS, Choi SB, Abdul Wahab H, Das PK, Chan KL.Eurycomanone, the major quassinoid in Eurycoma longifolia root extract increases spermatogenesis by inhibiting the activity of phosphodiesterase and aromatase in steroidogenesis. J Ethnopharmacol. 2013 Aug 26;149(1):201-7. doi: 10.1016/j.jep.2013.06.023. Epub 2013 Jun 27.

Study 13:
Tongkat Ali as a potential herbal supplement for physically active male and female seniors–a pilot study.
Thirteen physically active male and 12 physically active female seniors (57-72 years) were supplemented with 400-mg TA extract daily for 5 weeks. After treatment, hemoglobin, testosterone, and dihydroepiandrosterone concentrations, and the ratio of total testosterone/cortisol and muscle force remained significantly lower in female seniors than in male seniors. Treatment resulted in significant increases in total and free testosterone concentrations and muscular force in men and women. The study affirms the ergogenic benefit of TA through enhanced muscle strength.
Source:
Henkel RR, Wang R, Bassett SH, Chen T, Liu N, Zhu Y, Tambi MI.Tongkat Ali as a potential herbal supplement for physically active male and female seniors–a pilot study.Phytother Res. 2014 Apr;28(4):544-50. doi: 10.1002/ptr.5017. Epub 2013 Jun 11.

Study 14:
In vivo effects of Eurycoma longifolia Jack (Tongkat Ali) extract on reproductive functions in the rat.
An aqueous extract of Eurycoma longifolia (Tongkat Ali; TA) roots is traditionally used to enhance male sexuality. 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). 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. Testosterone concentration increased by 30.2% (P = 0.0544). Muscle weight also increased, yet not significantly.
Source:
Solomon MC, Erasmus N, Henkel RR.In vivo effects of Eurycoma longifolia Jack (Tongkat Ali) extract on reproductive functions in the rat. Andrologia. 2014 May;46(4):339-48. doi: 10.1111/and.12082. Epub 2013 Mar 6.

Study 15:
Standardized quassinoid-rich Eurycoma longifolia extract improved spermatogenesis and fertility in male rats via the hypothalamic-pituitary-gonadal axis.
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. 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). 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). 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.
Source:
Low BS, Das PK, Chan KL.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. doi: 10.1016/j.jep.2012.11.013. Epub 2012 Dec 20.

Study 16:
Randomized Clinical Trial on the Use of PHYSTA Freeze-Dried Water Extract of Eurycoma longifolia for the Improvement of Quality of Life and Sexual Well-Being in Men.
A randomized, double-blind, placebo controlled, parallel group study was carried out to investigate the clinical evidence of E. longifolia in men. Primary endpoints were the Quality of Life investigated by SF-36 questionnaire and Sexual Well-Being investigated by International Index of Erectile Function (IIEF) and Sexual Health Questionnaires (SHQ); Seminal Fluid Analysis (SFA), fat mass and safety profiles. The E. longifolia (EL) group significantly improved in the domain Physical Functioning of SF-36, from baseline to week 12 compared to placebo (P = 0.006) and in between group at week 12 (P = 0.028). All safety parameters were comparable to placebo.
Source:
Ismail SB, Wan Mohammad WM, George A, Nik Hussain NH, Musthapa Kamal ZM, Liske E.Randomized Clinical Trial on the Use of PHYSTA Freeze-Dried Water Extract of Eurycoma longifolia for the Improvement of Quality of Life and Sexual Well-Being in Men.Evid Based Complement Alternat Med. 2012;2012:429268. doi: 10.1155/2012/429268. Epub 2012 Nov 1.

Study 17:
Eurycoma longifolia upregulates osteoprotegerin gene expression in androgen- deficient osteoporosis rat model.
Supplementation with Eurycoma longifolia (EL) extract elevated the testosterone levels, reduced the bone resorption marker and upregulated OPG gene expression of the orchidectomised rats. These actions may be responsible for the protective effects of EL extract against bone resorption due to androgen deficiency.
Source:
Shuid AN, El-arabi E, Effendy NM, Razak HS, Muhammad N, Mohamed N, Soelaiman IN.Eurycoma longifolia upregulates osteoprotegerin gene expression in androgen- deficient osteoporosis rat model.BMC Complement Altern Med. 2012 Sep 12;12:152. doi: 10.1186/1472-6882-12-152.

Study 18:
Effects of Eurycoma longifolia on Testosterone Level and Bone Structure in an Aged Orchidectomised Rat Model.
Testosterone replacement is the choice of treatment in androgen-deficient osteoporosis. Thirty-six male Sprague-Dawley rats aged 12 months were divided into normal control, normal rat supplemented with EL, sham-operated, orchidectomised-control, orchidectomised with testosterone replacement, and orchidectomised with EL supplementation groups. Testosterone replacement was able to raise the testosterone level and restore the bone volume of orchidectomised rats. EL supplementation failed to emulate both these testosterone actions.
Source:
Tajul Ariff AS, Soelaiman IN, Pramanik J, Shuid AN.Effects of Eurycoma longifolia on Testosterone Level and Bone Structure in an Aged Orchidectomised Rat Model.Evid Based Complement Alternat Med. 2012;2012:818072. doi: 10.1155/2012/818072. Epub 2012 Aug 26.

Study 19:
Combined Effects of Eurycoma longifolia and Testosterone on Androgen-Deficient Osteoporosis in a Male Rat Model.
Forty male Sprague-Dawley rats were divided into: sham-operated (SHAM), orchidectomized-control (ORX), orchidectomized with testosterone (ORX + T), orchidectomized with EL (ORX + EL), and orchidectomized with combined T and EL therapy (ORX + T + EL). T was injected intramuscularly at 8 mg/kg and 4 mg/kg for the ORX + T and ORX + T + EL groups, respectively. Biomechanically, the strain parameter of the ORX + T + EL group was significantly higher than the ORX group (P < 0.05). Thus, the combination therapy of EL and low-dose T has potential for treatment of androgen-deficient osteoporosis.
Source:
Saadiah Abdul Razak H, Shuid AN, Naina Mohamed I.Combined Effects of Eurycoma longifolia and Testosterone on Androgen-Deficient Osteoporosis in a Male Rat Model.Evid Based Complement Alternat Med. 2012;2012:872406. doi: 10.1155/2012/872406. Epub 2012 Aug 9.

Study 20:
Eurycoma longifolia: Medicinal Plant in the Prevention and Treatment of Male Osteoporosis due to Androgen Deficiency.
Osteoporosis in elderly men is now becoming an alarming health issue due to its relation with a higher mortality rate compared to osteoporosis in women. Androgen deficiency (hypogonadism) is one of the major factors of male osteoporosis and it can be treated with testosterone replacement therapy (TRT). However, one medicinal plant, Eurycoma longifolia Jack (EL), can be used as an alternative treatment to prevent and treat male osteoporosis without causing the side effects associated with TRT. EL exerts proandrogenic effects that enhance testosterone level, as well as stimulate osteoblast proliferation and osteoclast apoptosis. This will maintain bone remodelling activity and reduce bone loss. Phytochemical components of EL may also prevent osteoporosis via its antioxidative property. Hence, EL has the potential as a complementary treatment for male osteoporosis.
Source:
Mohd Effendy N, Mohamed N, Muhammad N, Naina Mohamad I, Shuid AN.Eurycoma longifolia: Medicinal Plant in the Prevention and Treatment of Male Osteoporosis due to Androgen Deficiency.Evid Based Complement Alternat Med. 2012;2012:125761. doi: 10.1155/2012/125761. Epub 2012 Jul 15.

Study 21:
Effect of Eurycoma longifolia Jack (Tongkat ali) extract on human spermatozoa in vitro.
A sample without addition of TA served as control. For washed spermatozoa, significant dose-dependent trends were found for vitality, total motility, acrosome reaction and reactive oxygen species-positive spermatozoa. Contrary, the increase in the percentage of acrosome-reacted spermatozoa with increasing TA concentrations is very significant (P < 0.0001), and a significant difference (P = 0.0069) to the control could even be recorded at 20 μg TA per ml. Results indicate that the TA extract has no deleterious effects on sperm functions at therapeutically used concentrations (<2.5 μg ml(-1) ). However, at very high concentrations, TA may have harmful effects in vitro.
Source:
Erasmus N, Solomon MC, Fortuin KA, Henkel RR.Effect of Eurycoma longifolia Jack (Tongkat ali) extract on human spermatozoa in vitro. Andrologia. 2012 Oct;44(5):308-14. doi: 10.1111/j.1439-0272.2012.01282.x. Epub 2012 Feb 15.

Study 22:
Standardised water-soluble extract of Eurycoma longifolia, Tongkat ali, as testosterone booster for managing men with late-onset hypogonadism?
Considering that human studies are not available, 76 of 320 patients suffering from late-onset hypogonadism (LOH) were given 200 mg of a standardised water-soluble extract of Tongkat ali for 1 month. The Ageing Males’ Symptoms (AMS) according to the standardised rating scale and the serum testosterone concentration were taken. Results show that treatment of LOH patients with this Tongkat ali extract significantly (P < 0.0001) improved the AMS score as well as the serum testosterone concentration.
Source:
Tambi MI, Imran MK, Henkel RR.Standardised water-soluble extract of Eurycoma longifolia, Tongkat ali, as testosterone booster for managing men with late-onset hypogonadism?Andrologia. 2012 May;44 Suppl 1:226-30. doi: 10.1111/j.1439-0272.2011.01168.x. Epub 2011 Jun 15.

Study 23:
The effect of Eurycoma longifolia on sperm quality of male rats.
The standardized MeOH extract at doses of 50, 100 and 200 mg/kg, the EtOAc fraction (70 mg/kg), and standardized MeOH extract at 200 mg/kg co-administered with the EtOAc fraction of A. paniculata at 70 mg/kg were each given orally to male Sprague-Dawley albino rats for 48 consecutive days. The spermatozoa count, morphology, motility, plasma testosterone level and Leydig cell count of the animals were statistically analyzed by ANOVA with a post-hoc Tukey HSD test. The plasma testosterone level of the rats treated with the standardized MeOH extract at 200 mg/kg was significantly increased (p < 0.01) when compared with that of the control and infertile animals.
Source:
Chan KL, Low BS, Teh CH, Das PK.The effect of Eurycoma longifolia on sperm quality of male rats.Nat Prod Commun. 2009 Oct;4(10):1331-6.

Study 24:
Influence of Eurycoma longifolia on the copulatory activity of sexually sluggish and impotent male rats.
Concerning the copulatory activity of sexually sluggish rats, both acute (dosed at 500 and 1000 mg/kg) and subacute treatments with the root powder significantly reduced ejaculation latencies, increasing also the percentage of mounting and ejaculating animals; in addition the subacute administration reduced post-ejaculatory interval. In impotent rats both subacute and subchronic treatments increased the percentage of mounting and ejaculating rats. The motivational behavior of sluggish rats during the partner preference test was not affected by the treatments. Testosterone serum levels were increased in rats subacutely treated in comparison with controls.
Eurycoma longifolia root improved sexual performance but not motivation in sluggish rats after acute or subacute administration. The effect could be mainly ascribed to increased testosterone levels.
Source:
Zanoli P, Zavatti M, Montanari C, Baraldi M.Influence of Eurycoma longifolia on the copulatory activity of sexually sluggish and impotent male rats. J Ethnopharmacol. 2009 Nov 12;126(2):308-13. doi: 10.1016/j.jep.2009.08.021. Epub 2009 Aug 22.

Gamma Oryzanol
Facts:
Gamma oryzanol is a mixture of compounds usually found in bran and in other fruits and vegetables. It is known to have anti-cholesterol characteristics and is also used to increase the level of testosterone and human growth hormone. It also possess strengthening potentials during resistance exercise training.
Sources:
http://examine.com/supplements/Gamma%20Oryzanol
http://www.webmd.com/vitamins-supplements/ingredientmono-770-gamma%20oryzanol.aspx?activeingredientid=770&activeingredientname=gamma%20oryzanol

Study 1:
Gamma oryzanol-plant sterol supplementation: metabolic, endocrine, and physiologic effects.
The use of gamma-oryzanol and phytosterols is gaining popularity among various athletic populations. These compounds are being consumed in the belief that they elicit anabolic effects ranging from increased testosterone production and release to stimulating human growth hormone release. However, published scientific studies suggest that these compounds are poorly absorbed. Furthermore, animal studies indicate that when these compounds are injected subcutaneously or intravenously, they induce antianabolic or catabolic activity. Normally, less than 5% of orally consumed phytosterols are absorbed from the intestinal tract, with the majority being excreted in the feces. Intravenous or subcutaneous injections of gamma-oryzanol in rats have been shown to suppress luteinizing hormone release, reduce growth hormone synthesis and release, and increase release of the catecholamines, dopamine and norepinephrine, in the brain. Although it hasn’t been directly measured, this metabolic milieu, if accurate, may actually reduce testosterone production.
Source:
Wheeler KB, Garleb KA.Gamma oryzanol-plant sterol supplementation: metabolic, endocrine, and physiologic effects. Int J Sport Nutr. 1991 Jun;1(2):170-7. Review.

Study 2:
The effects of gamma-oryzanol supplementation during resistance exercise training.
The effects of the oral administration of trans-ferulic acid and gamma-oryzanol (mixture of steryl ferulates) with ethanol (5.0 g per kg) for 30 days to c57BL mice on ethanol-induced liver injury were investigated. Preventions of ethanol-induced liver injury by trans-ferulic acid and gamma-oryzanol were reflected by markedly decreased serum activities of plasma aspartate aminotransferase, alanine aminotransferase and significant decreases in hepatic lipid hydroperoxide and TBARS levels. Furthermore, the trans-ferulic acid- and gamma-oryzanol-treated mice recovered ethanol-induced decrease in hepatic glutathione level together with enhancing superoxide dismutase activity. These results demonstrate that both trans-ferulic acid and gamma-oryzanol exert a protective action on liver injury induced by chronic ethanol ingestion.
Source:
Fry AC, Bonner E, Lewis DL, Johnson RL, Stone MH, Kraemer WJ.The effects of gamma-oryzanol supplementation during resistance exercise training.Int J Sport Nutr. 1997 Dec;7(4):318-29.

Study 3:
γ-Oryzanol protects pancreatic β-cells against endoplasmic reticulum stress in male mice.
In islets from high-fat diet-fed diabetic mice, oral administration of γ-oryzanol improved glucose-stimulated insulin secretion on following reduction of exaggerated ER stress and apoptosis. Furthermore, we examined the impact of γ-oryzanol on low-dose streptozotocin-induced diabetic mice, where exaggerated ER stress and resultant apoptosis in β-cells were observed. Taken together, our findings demonstrate that γ-oryzanol directly ameliorates ER stress-induced β-cell dysfunction and subsequent apoptosis, highlighting usefulness of γ-oryzanol for the treatment of diabetes mellitus.
Source:
Kozuka C, Sunagawa S, Ueda R, Higa M, Tanaka H, Shimizu-Okabe C, Ishiuchi S, Takayama C, Matsushita M, Tsutsui M, Miyazaki J, Oyadomari S, Shimabukuro M, Masuzaki H.γ-Oryzanol protects pancreatic β-cells against endoplasmic reticulum stress in male mice.Endocrinology. 2015 Apr;156(4):1242-50. doi: 10.1210/en.2014-1748. Epub 2015 Jan 16.

Study 4:
Effects of gamma oryzanol supplementation on anthropometric measurements &amp; muscular strength in healthy males following chronic resistance training.
Our findings indicated that 600 mg/day gamma oryzanol supplementation during the 9-week resistance training did not change anthropometric and body measurements, but it increased muscular strength in young healthy males. Further, studies need to be done in trained athletes, women, and in patients who suffer from muscular fatigue.
Source:
Eslami S, Esa NM, Marandi SM, Ghasemi G, Eslami S.Effects of gamma oryzanol supplementation on anthropometric measurements &amp; muscular strength in healthy males following chronic resistance training. Indian J Med Res. 2014 Jun;139(6):857-63.

Study 5:
Rice bran enzymatic extract-supplemented diets modulate adipose tissue inflammation markers in Zucker rats.
Chronic administration of a novel water-soluble RBEE, rich in polyphenols, tocotrienols and γ-oryzanol, could be a suitable treatment to ameliorate the obesity-associated proinflammatory response.
Source:
Candiracci M, Justo ML, Castaño A, Rodriguez-Rodriguez R, Herrera MD.Rice bran enzymatic extract-supplemented diets modulate adipose tissue inflammation markers in Zucker rats.Nutrition. 2014 Apr;30(4):466-72. doi: 10.1016/j.nut.2013.09.016. Epub 2013 Oct 16.

Study 6:
Antioxidant effects of gamma-oryzanol on human prostate cancer cells.
This study highlighted effects of gamma-oryzanol via the down-regulation of antioxidant genes, catalase and GPX, not cytotoxic roles. This might be interesting for adjuvant chemotherapy to make prostate cancer cells more sensitive to free radicals. It might be useful for the reduction of cytotoxic agents and cancer chemoprevention.
Source:
Klongpityapong P, Supabphol R, Supabphol A.Antioxidant effects of gamma-oryzanol on human prostate cancer cells. Asian Pac J Cancer Prev. 2013;14(9):5421-5.

Study 7:
Effect of gamma-oryzanol-enriched rice bran oil on quality of cryopreserved boar semen.
The semen was divided into three portions in which lactose-egg yolk (LEY) extender used to resuspend the centrifuged sperm pellet was supplemented with 2 types of rice bran oils, at a gamma-oryzanol concentration of 0 mg/ml of lactose egg yolk (LEY) freezing extender (group A, control), 0.1 mg/ml(0.16 mMol) of freezing extender (group B) and 0.1 mg/ml of freezing extender (group C). In conclusion, addition of gamma-oryzanol-enriched rice bran oil to LEY freezing extender is appropriated for improving the quality of frozen-thawed boar semen.
Source:
Kaeoket K, Donto S, Nualnoy P, Noiphinit J, Chanapiwat P.Effect of gamma-oryzanol-enriched rice bran oil on quality of cryopreserved boar semen.J Vet Med Sci. 2012 Sep;74(9):1149-53. Epub 2012 May 18.

Study 8:
Gamma-oryzanol ameliorates insulin resistance and hyperlipidemia in rats with streptozotocin/nicotinamide-induced type 2 diabetes.
Gamma-oryzanol is a component of rice bran oil (RBO) with purported health benefits. This study evaluated the effects of gamma-oryzanol on insulin resistance and lipid metabolism in Wistar rats with type 2 diabetes (T2DM). Adding gamma-oryzanol to PO improved the negative influence of PO on lipid metabolism in T2DM rats. In addition, gamma-oryzanol tended to increase insulin sensitivity in T2DM rats compared to control and PO groups. In addition, gamma-oryzanol tended to increase insulin sensitivity in T2DM rats compared to control and PO groups. Longer-term studies are needed to evaluate these effects further.
Source:
Cheng HH, Ma CY, Chou TW, Chen YY, Lai MH.Gamma-oryzanol ameliorates insulin resistance and hyperlipidemia in rats with streptozotocin/nicotinamide-induced type 2 diabetes.Int J Vitam Nutr Res. 2010 Jan;80(1):45-53. doi: 10.1024/0300-9831/a000005.

Study 9:
Optimization, in vitro release and bioavailability of gamma-oryzanol-loaded calcium pectinate microparticles reinforced with chitosan.
Response surface methodology was used to optimize coating conditions, including chitosan concentration (X(1)) and coating time (X(2)), for sustained release of chitosan-coated Ca-pectinate (CP) microparticles containing oryzanol (OZ). The optimized values of X(1) and X(2) were found to be 1.48% and 69.92 min, respectively. These optimized values agreed favorably with the predicted results, indicating the utility of predictive models for the release of OZ in simulated intestinal fluid. In vitro release studies revealed that the chitosan-coated CP microparticles were quite stable under acidic conditions, but swell and disintegrate under alkaline conditions. In vivo release study of OZ, physically entrapped within chitosan-coated CP microcapsules, demonstrated the sustained release of OZ and could be used to improve the bioavailability of OZ following oral administration.
Source:
Kim JS, Lee JS, Chang PS, Lee HG.Optimization, in vitro release and bioavailability of gamma-oryzanol-loaded calcium pectinate microparticles reinforced with chitosan. N Biotechnol. 2010 Sep 30;27(4):368-73. doi: 10.1016/j.nbt.2010.02.018. Epub 2010 Mar 1.

Study 10:
Cycloartenyl ferulate, a component of rice bran oil-derived gamma-oryzanol, attenuates mast cell degranulation.
IgE-targeting therapy could provide significant progress in the treatment of allergic inflammation. In this study, we examined the effect of cycloartenyl ferulate (cycloartenol ferulic acid ester; CAF), a natural product from rice bran oil-derived gamma-oryzanol, on allergic reaction. When CAF and gamma-oryzanol were injected intradermally with anti-DNP IgE into the dorsal skin of rats, the passive cutaneous anaphylaxis reaction induced by DNP-HSA was attenuated. CAF and gamma-oryzanol also inhibited the degranulation of DNP-IgE sensitized RBL-2H3 mast cells stimulated with anti-DNP-HSA. IgE conjugated with CAF could not be detected by anti-IgE antibody in the ELISA analysis. Although incubation of IgE with CAF did not decrease the amount of IgE, it was possible to precipitate IgE by centrifugation. These results demonstrate that CAF captures IgE, prevents it from binding to FcepsilonRI, and attenuates mast cell degranulation.
Source:
Oka T, Fujimoto M, Nagasaka R, Ushio H, Hori M, Ozaki H.Cycloartenyl ferulate, a component of rice bran oil-derived gamma-oryzanol, attenuates mast cell degranulation.Phytomedicine. 2010 Feb;17(2):152-6. doi: 10.1016/j.phymed.2009.05.013. Epub 2009 Jul 3.
Oka T, Fujimoto M, Nagasaka R, Ushio H, Hori M, Ozaki H.Cycloartenyl ferulate, a component of rice bran oil-derived gamma-oryzanol, attenuates mast cell degranulation.Phytomedicine. 2010 Feb;17(2):152-6. doi: 10.1016/j.phymed.2009.05.013. Epub 2009 Jul 3.