Help Interpreting This Study On Limonene

[Captain_Coconut]

I am interested on the health effects of limonene supplementation. I found this study which I think would help shed some light on what exactly it does in the body. However I am not learned enough to interpret the results.

“We conducted metabolite profiling in 39 paired (pre/postintervention) plasma samples from early-stage breast cancer patients receiving limonene treatment (2 g QD) before surgical resection of their tumor. ”

Plasma Metabolomic Profiles of Breast Cancer Patients after Short-term Limonene Intervention

Table 1 has a lot of valuable data. It looks clear it helps with bile production. I am interested in what the more biologically astute have to say about this, particularly what the results relating to androgen metabolites would be indicating. Do these results indicate anything remarkable to you?

 

[Captain_Coconut]

Big stretch here, but I found this with a small sample size, and not much credibility. But it shows blue spruce essential oil increasing free testosterone and total testosterone... the main constituent of the oil is limonene and pinene...

US9675653B2 - Composition containing an essential oil product and method for using such to maintain normal levels of testosterone - Google Patents

 

[Captain_Coconut]

Bump

 

[ecstatichamster]

good idea to get someone's input on it. These "metabolanomics" studies (or whatever they call it) are very hard to decipher. It is a form of laziness academically to just provide a bunch of numbers with no real interpretation and no hypothesis or conclusion.

 

[Captain_Coconut]

good idea to get someone's input on it. These "metabolanomics" studies (or whatever they call it) are very hard to decipher. It is a form of laziness academically to just provide a bunch of numbers with no real interpretation and no hypothesis or conclusion.

Yea it is pretty hard for me to make much from it though I’m sure someone on this forum must know how to interpret the results. The main things in the paper which worry me is the potential androgen effects, however the circulation of the metabolites mentioned in relation to e.g. levels of free testosterone or dhea or estrogen could be totally counterintuitive... also there is this:

“Pyruvate, a terminal product of glycolysis, was significantly elevated in plasma following limonene intervention, as was fructose of the sorbitol pathway, suggesting restricted entry or usage of glucose-derived carbons by the TCA cycle in the mitochondria. A shift in energy metabolism to preferential use of fatty acid incorporation into the TCA cycle is also supported by a small but significant increase in β-hydroxybutyrate which is increased during oxidation of fatty acids ”


However I am not clear how extreme the effect is. Also my understanding is limonene is protective against lipid peroxidation, so maybe it is not all that bad that it causes that shift...

 

[ecstatichamster]

also it lowered sulfation of prohormones. That is the a concerning part of this to me in addition to what you pointed out.

 

[Whichway?]

I’ve had a look at it. If you have any specific questions fire away.

They were looking for specific avenues by which limonene produces it anti-cancer effects. Looks like it acts systemically on a large number of metabolic systems. Androgen and gonadal steroids were down, bile synthesis was up, as was collagen re-modeling, and it seems to inhibit the burning of glucose in mitochondria and instead promotes gluconeogenesis.

What are looking to use it for?

 

[Captain_Coconut]

I’ve had a look at it. If you have any specific questions fire away.

They were looking for specific avenues by which limonene produces it anti-cancer effects. Looks like it acts systemically on a large number of metabolic systems. Androgen and gonadal steroids were down, bile synthesis was up, as was collagen re-modeling, and it seems to inhibit the burning of glucose in mitochondria and instead promotes gluconeogenesis.

What are looking to use it for?

Thanks. I am interested in table 1 more so than the correlations to cancer. I take around a gram a day and find I feel better on it. It is strongly anxiolytic and offers better digestion and some pain relief as well. I am curious which steroids it specifically is shown to reduce and to what degree, and to what degree it promotes gluconeogenesis. I have read before that it assists the liver in clearing estrogen, do these results indicate that? It is a big component of citrus juice, so I’m guessing it must be somewhat Peaty, what do you think? Thanks!

 

[Inaut]

Citrus Oils and Breast Health
by Robert Tisserand, with thanks to Christine Carson PhD and Linda Halcon PhD for their input.

In the United States, breast cancer is the most common cancer in women, and it is the second most common cause of death from any cancer. According to the National Cancer Institute 12.3% of women will be diagnosed with breast cancer at some point during their lifetime, based on 2010-2012 data. In 2012, there were an estimated 2.9 million women living with breast cancer in the United States. Risk factors, and common ways to avoid them, are elaborated here. Types of breast cancer are listed here.

There has been much research on citrus fruits, citrus oils, and limonene in relation to breast cancer, and recent developments seem to suggest at least a preventive effect. Treatment of any cancer is a significant challenge, and I want to be clear that I am not advocating an alternative breast cancer treatment regime. This is about prevention. That doesn’t mean this regime will be useless in combination with other therapies. It may have some effect, but is not likely to be effective as a stand-alone treatment.

Citrus x limon

Limonene
Limonene is the major constituent of citrus fruit oils, and is found in orange, tangerine and grapefruit oils at about 90%, and in lemon and mandarin oils at about 70%. Research in the 80s showed that limonene (or in one case orange oil) could either inhibit the development of breast cancer in female rodents, or could cause regression of existing tumors (6,7,8,9,12,19). In the mid 1990s, 11 patients with advanced breast cancer were included in one clinical trial using high oral doses of limonene. There was regression of disease in only one patient (22). Perillyl alcohol, a metabolite of limonene, was similarly trialed in a small group of patients with advanced breast cancer, also without success (2). However, while the use of oral limonene as a cancer treatment option has stalled, it is now showing great promise in breast cancer prevention and for this purpose, topical application appears to be as effective as oral dosing.


[It is also worth noting that two minor constituents of many citrus oils, bergapten and citral, have shown in vitro activity against breast cancer cells (3,4).]
Prevention
Prevention is difficult to prove, but a scientific team around Jessica Miller at the University of Arizona has been focusing their energies on limonene and breast health for the past 7 years. So far they have published five papers. First, the team established a method to measure limonene in body fat (13), and then they reported that limonene is preferentially absorbed into buttock fat (14). This was found in 7 healthy adults (one male, 6 females) who drank lemonade made from whole lemons every day for four weeks. After the first 6 hours, there was 8 times more limonene in body fat than in the blood; after four weeks, the difference increased to 34 times, showing that limonene accumulates in fatty tissue. So at this point we know that limonene is drawn to fat cells in the body. In fact, as far as we know, it stays there for longer than most other essential oil constituents.

Data from Miller et al 2013

The team then found that either oral or topical administration of orange oil to female mice resulted in preferential absorption of limonene into breast tissue, with similar amounts being present from oral as from topical (15). In the same study, healthy women were recruited to apply orange oil, diluted to 10% or 20% in fractionated coconut oil, to their breasts. This was determined to be both safe and feasible.

Following this, 43 women with newly diagnosed, operable breast cancer were recruited to ingest two grams of limonene daily for 2-6 weeks (the time varied with scheduled surgery). In this clinical trial, limonene again preferentially concentrated in breast tissue (see illustration). In addition, there was a 22% reduction in cyclin D expression. This is interesting, since cyclin D is an important factor in rates of cell growth, and is overexpressed in hyperplasia and intraductal carcinoma of the breast, suggesting this plays a role in the earliest stages of breast cancer development (16).

Cancer prevention with natural compounds is not just about direct action against any cancer cells that may arise, and also involves effects such as immune-enhancing, antimutagenic and antioxidant (21). Interestingly, these indirect cancer protective effects have all been established for limonene (1,5,10,18). In their latest research paper, Jessica Miller and colleagues studied the effect of limonene on hundreds of substances in the body, such as adrenal hormones, amino acids and glucose metabolites. These were blood tests that were conducted during the previously described clinical trial (17). An important conclusion was that limonene counteracts many of the metabolic changes that happen in people with breast cancer, strongly suggesting a preventive effect.

Dr. Jessica Miller

Citrus fruit consumption
The above studies on limonene suggest breast cancer preventive effects, and it is worth noting that there is also epidemiological evidence from the consumption of citrus fruit. Several such reports looked at the relationship between citrus fruit consumption and breast cancer incidence. A meta-analysis of five studies found an overall 10% reduction in breast cancer incidence, associated with “high intake” of citrus fruits. A Japanese report not included in this review concluded that daily citrus consumption was correlated with an 11% reduced incidence of all cancers in both sexes. The effect was even greater (17%) in those who also drank green tea every day (11). An association with citrus fruits does not necessarily mean that the essential oil, or limonene, is solely responsible, in fact this is unlikely as citrus fruits contain many anticancer substances, not all found in the essential oil.

Conclusions
There is growing evidence that citrus fruit, and limonene-rich citrus essential oils could play a role in the prevention of breast cancer. It is not implausible that combination of fresh oranges, green tea and a topical essential oil formulation could be preventive for many people. However, we need to know more about the safety and efficacy of long-term topical use.

References

1. Bacanlı, M., Başaran, A. A., & Başaran, N. (2015). The antioxidant and antigenotoxic properties of citrus phenolics limonene and naringin. Food and Chemical Toxicology, 81, 160–170
   The antioxidant and antigenotoxic properties of citrus phenolics limonene and naringin. - PubMed - NCBI
2. Bailey, H. H., Attia, S., Love, R. R. et al (2008). Phase II trial of daily oral perillyl alcohol (NSC 641066) in treatment-refractory metastatic breast cancer. Cancer Chemotherapy and Pharmacology, 62(1), 149–157
   Phase II trial of daily oral perillyl alcohol (NSC 641066) in treatment-refractory metastatic breast cancer. - PubMed - NCBI
3. Chaouki, W., Leger, D. Y., Liagre, B. et al (2009). Citral inhibits cell proliferation and induces apoptosis and cell cycle arrest in MCF-7 cells. Fundamental & Clinical Pharmacology, 23(5), 549–556
   Citral inhibits cell proliferation and induces apoptosis and cell cycle arrest in MCF-7 cells. - PubMed - NCBI
4. De Amicis, F., Aquila, S., Morelli, C. et al (2015). Bergapten drives autophagy through the up-regulation of PTEN expression in breast cancer cells. Molecular Cancer, 14(1), 130
   http://www.molecular-cancer.com/content/pdf/s12943-015-0403-4.pdf
5. Del Toro-Arreola, S., Flores-Torales, E., Torres-Lozano, C. et al (2005). Effect of d-limonene on immune response in BALB/c mice with lymphoma. International Immunopharmacology, 5(5), 829–838
   Effect of D-limonene on immune response in BALB/c mice with lymphoma. - PubMed - NCBI
6. Elegbede, J. A., Elson, C. E., Qureshi, A. et al (1984). Inhibition of DMBA-induced mammary cancer by the monoterpene d-limonene. Carcinogenesis, 5(5), 661–664
   Inhibition of DMBA-induced mammary cancer by the monoterpene d-limonene. - PubMed - NCBI
7. Elegbede, J. A., Elson, C. E., Tanner, M. A. et al (1986). Regression of rat primary mammary tumors following dietary d-limonene. Journal of the National Cancer Institute, 76(2), 323–325
   Regression of rat primary mammary tumors following dietary d-limonene. - PubMed - NCBI
8. Elson, C. E., Maltzman, T. H., Boston, et al (1988). Anti-carcinogenic activity of d-limonene during the initiation and promotion/progression stages of DMBA-induced rat mammary carcinogenesis. Carcinogenesis, 9(2), 331–332
   Anti-carcinogenic activity of d-limonene during the initiation and promotion/progression stages of DMBA-induced rat mammary carcinogenesis. - PubMed - NCBI
9. Haag, J., Lindstrom, M., & Gould, M. (1992). Limonene-induced regression of mammary carcinomas. Cancer Research, 4021–4026
   Limonene-induced Regression of Mammary Carcinomas
10. Hamada, M., Uezu, K., Matsushita, J. et al (2002). Distribution and immune responses resulting from oral administration of d-limonene in rats. Journal of Nutritional Science and Vitaminology, 48(2), 155–160
    Distribution and immune responses resulting from oral administration of D-limonene in rats. - PubMed - NCBI
11. Li, W. Q., Kuriyama, S., Li, Q. et al (2010). Citrus consumption and cancer incidence: The Ohsaki cohort study. International Journal of Cancer, 127(8), 1913–1922
    http://onlinelibrary.wiley.com/doi/10.1002/ijc.25203/epdf
12. Maltzman, T. H., Hurt, L. M., Elson, C. E. et al (1989). The prevention of nitrosomethylurea-induced mammary tumors by d-limonene and orange oil. Carcinogenesis, 10(4), 781–783
    The prevention of nitrosomethylurea-induced mammary tumors by d-limonene and orange oil. - PubMed - NCBI
13. Miller, J. A., Hakim, I. A,, Thomson, C. et al (2008). Determination of d-limonene in adipose tissue by gas chromatography-mass spectrometry. Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences, 870(1), 68–73
    Determination of d-limonene in adipose tissue by gas chromatography-mass spectrometry
14. Miller, J. A., Hakim, I. A., Chew W. et al (2010). Adipose tissue accumulation of d-limonene with the consumption of a lemonade preparation rich in d-limonene content. Nutrition & Cancer, 62(6), 783–788
    Adipose tissue accumulation of d-limonene with the consumption of a lemonade preparation rich in d-limonene content. - PubMed - NCBI
15. Miller, J. A., Thompson, P., Hakim, I. A. et al (2012). Safety and feasibility of topical application of limonene as a massage oil to the breast. Journal of Cancer Therapy, 3, 1–12
    http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3824622/
16. Miller, J. A., Lang, J., Ley, M. et al (2013). Human breast tissue disposition and bioactivity of limonene in women with early stage breast cancer. Cancer Prevention Research 6(6), 577–584
    http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3692564/
17. Miller, J. A., Pappan, K., Thompson, P. A. et al (2015). Plasma metabolomic profiles of breast cancer patients after short-term limonene intervention. Cancer Prevention Research, 8(1), 86–93
    http://www.ncbi.nlm.nih.gov/pubmed/25388013
18. Raphael, T. J., & Kuttan, G. (2003). Immunomodulatory activity of naturally occurring monoterpenes carvone, limonene, and perillic acid. Immunopharmacology and Immunotoxicology, 25(2), 285–294
    http://www.ncbi.nlm.nih.gov/pubmed/12784919
19. Russin, W. A., Hoesly, J. D., Elson, C. E. et al (1989). Inhibition of rat mammary carcinogenesis by monoterpenoids. Carcinogenesis, 10(11), 2161–2164
    http://www.ncbi.nlm.nih.gov/pubmed/2509095
20. Song, J. K., & Bae, J. M. (2013). Citrus fruit intake and breast cancer risk: A quantitative systematic review. Journal of Breast Cancer, 16(1), 72–76
    http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3625773/pdf/jbc-16-72.pdf
21. Tsuda, H., Ohshima, Y., Nomoto, H. et al (2004). Cancer prevention by natural compounds. Drug Metabolism and Pharmacokinetics, 19(4), 245–263
    https://www.jstage.jst.go.jp/article/dmpk/19/4/19_4_245/_pdf
22. Vigushin, D. M., Poon, G. K., Boddy, A. et al (1998). Phase I and pharmacokinetic study of d-limonene in patients with advanced cancer. Cancer Research Campaign Phase I/II Clinical Trials Committee. Cancer Chemotherapy and Pharmacology, 42(2), 111–117
    http://www.ncbi.nlm.nih.gov/pubmed/9654110

https://tisserandinstitute.org/citrus-oils-and-breast-health/

I think the accumulation in fat cells is part of the bigger benefit with limonene

 

[ecstatichamster]

Thanks much !

 

[Inaut]

Honestly d-limonene is becoming one of my top supplements. I'm even applying it on my eczema and it seems to be drying it up/reducing redness quite well.

 

[Inaut]

Use of d-limonenes as testosterone-5-alpha-reductase inhibitor and as hair grower.
European Patent Application EP0467660
Kind Code:
A2


Abstract:
A 5 alpha -reductase inhibitor according to the invention comprises d-limonene, which has an excellent activity inhibitory effect and is highly safe without any drawback as noted by side effects and thus is a suitable component for a hair grower.



Inventors:
Miyauchi, Yutaka (JP)
Application Number:
EP19910306490
Publication Date:
01/22/1992
Filing Date:
07/17/1991
Export Citation:
Click for automatic bibliography generation
Assignee:
Miyauchi, Yutaka (JP)
International Classes:
A61K8/00; A61K8/31; A61K8/97; A61K31/015; A61P43/00; A61Q5/00; A61Q7/00; A61Q7/02; (IPC1-7): A61K7/06; A61K31/035
European Classes:
A61K8/31; A61K8/97; A61K31/015; A61Q7/02
View Patent Images:
Download PDF EP0467660 PDF help



Foreign References:
WO1991006278A1 1991-05-16

COMPOSITION FOR HAIR GROWTH STIMULATION
FR2338035A1 1977-08-12


Other References:
PATENT ABSTRACTS OF JAPAN, vol. 11, no. 163 (C-424)Ä2610Ü, 26th May 1987; & JP-A-61 291 508 (YUTAKA MIYAUCHI) 22-12-1986
PATENT ABSTRACTS OF JAPAN, vol. 8, no. 145 (C-232)Ä1582Ü, 6th July 1984; & JP-A-59 53 419 (SHISEIDO K.K.) 28-03-1984
PATENT ABSTRACTS OF JAPAN, vol. 8, no. 145 (C-232)Ä1582Ü, 6th July 1984; & JP-A-59 53 417 (SHISEIDO K.K.) 28-03-1984
PATENT ABSTRACTS OF JAPAN, vol. 13, no. 536 (C-660)Ä3884Ü, 29th November 1989; & JP-A-1 221 309 (SANYO CHEM. IND. LTD) 04-09-1989
Claims:
1. Use of d-limonene as a testosterone 5 alpha -reductase inhibitor.

2. Use of d-limonene as a hair grower.

3. Use of d-limonene for the manufacture of a medicament for use as a hair grower.

4. Use according to any preceding claim wherein the d-limonene is as collected from a refined oil from the skin of a citrus fruit.

5. Use according to claim 4, wherein the citrus fruit is selected from a navel orange [citrus sinensis L. Osbeck (Rutaceae)], an iyokan [citrus iyo Hort. et Tanaka (Rutaceae)], a hassaku [citrus hassaku Hort. et Tanaka (Rutaceae)], a sweet summer orange [citrus Natsudaidai var. (Rutaceae)], a lemon and a mandarin orange.

Description:
Background of the Invention 1. Field of Industrial Application

The present invention relates to a testosterone 5 alpha -reductase (hereinafter designated simply as "5 alpha -reductase") inhibitor and a hair grower comprising the same. 2. Description of the Prior Art

Recently, increased numbers of studies have been made on a 5 alpha -reductase inhibitor as a hair grower, and there have been many patent applications for hair growers featured by the effect of preventing loss of hair by inhibition of 5 alpha -reductase (for example, Japanese Patent Laid-open Nos. 53,417/1984 and 53,419/1984).

According to a certain view, it is considered that physiological symptoms of, for example, male pattern baldness, psilosis, or seborrhea are caused by an increased hormonal stimulus of androgen, which results from excessive accumulation of the androgenic hormone (5 alpha -dihydro- testosterone) in a metabolic system. It is considered that this androgenic hormone is produced from testosterone by a reducing action of 5 alpha -reductase, and, in particular, the androgenic hormone is produced and accumulated in some organs such as hair roots and sebaceous glands due to the action of 5 alpha -reductase. As a result, hair-matrix cell division may be suppressed, leading to involution of the hair roots, and thereby symptoms of alopecia or baldness may be induced. It is therefore possible to prevent epilation or to promote hair-growth by inhibitory actions on the 5 alpha -reductase.

This is the reason why the 5 alpha -reductase inhibitor is used for hair growers.

From this point of view, some compounds used for this purpose have been found. However, though they have a high 5 alpha -reductase activity inhibitory ratio, they have drawbacks of producing side effects or causing high irritation to the skin. Summary of the Invention

Accordingly, an object of the present invention is to provide a 5 alpha -reductase inhibitor which has no drawback as noted by the side effects and is highly safe.

Another object of the present invention is to provide a hair grower having an excellent hair-growth effect upon application to the human scalp.

Other objects, features and advantages of the invention will hereinafter become more readily apparent from the following description. Brief Description of the Drawings

Fig. 1 is a graph for explaining an enzymatic activity inhibitory ratio of sodium beta -glycyrrhetinic acid.

Fig. 2 is a graph for explaining an enzymatic activity inhibitory ratio of Duke extract.

Fig. 3 is a graph for explaining an enzymatic activity inhibitory ratio of the hair grower of the present invention.

Fig. 4 is a graph for explaining an enzymatic activity inhibitory ratio of d-limonene.

Fig. 5 is a graph for explaining an enzymatic activity inhibitory ratio of swertiamarin. Description of the preferred Embodiment Example 1

Various effects of the hair grower of the present invention were studied, and the results are explained in the followings.

The present inventor has already disclosed a novel toilet lotion containing components from citrus fruits and other components (Japanese Patent Publication No. 20,123/1989). This toilet lotion is, of course, provided with the ordinary functions of a toilet lotion, e.g., astringent and cleansing effects, dampness, and lubrication of the skin. It has been confirmed that the toilet lotion gives, besides the above ordinary functions, anti-dandruff and antiprustic effects and an effect of preventing the loss of hair when rubbed into the pores of the scalp after shampooing. It has been also confirmed that some people have enjoyed the benefits of the toilet lotion, which not merely restrained the loss of their hair but also positively promoted the growth of their hair.

The toilet lotion consists of each extract from the skin of a navel orange [Citrus sinensis L. Osbeck (Rutaceae)], the skin of an iyokan [Citrus iyo Hort. et Tanaka (Rutaceae)], the skin of a hassaku [Citrus hassaku Hort. et Tanaka (Rutaceae)], the skin of a sweet summer orange [Citrus Natsudaidai var. (Rutaceae)], the skin of a lemon, the skin of a mandarin orange, and aloe, all of said extracts being prepared by extracting with sake. The refined oil from the citrus fruits contains d-limonene in the amount of not less than 90% by weight. It is therefore clear that the d-limonene functions as the 5 alpha -reductase inhibitor and thereby contributes to prevention of the loss of hair and to promotion of the growth of hair.

The following table shows a formulation of the hair grower (toilet lotion) in a preferred form of the present invention. EMI4.1 EMI5.1

The above extracts are all those extracted with sake (first class refined sake) from the skin of each of the above citrus fruits or other materials. A slight variation in the proportion of each of the above components is expected in the stages of manufacturing. It was, however, confirmed that some +/-5% of the proportion out of the defined range can be tolerated to ensure the same performance and safety of the composition. Application to the Scalp

As the symptoms identified by the loss of hair in the pilose regions of the body, especially the head, many sorts are known, such as androgenic alopecia, alopecia areata, and alopecia seborrheica. In this example, however, the hair grower containing as a major component d-limonene, which is the 5 alpha -reductase inhibitor according to the present invention, was applied to the scalp of dozens of persons irrespective of the sort of alopecia. The results are shown in the following tables. EMI6.1 EMI6.2 EMI7.1

As evidenced from the above results, the hair grower of the present invention is excellently effective or effective on the anti-dandruff, prevention of depilation, and hair-growth.

Such excellent effect of the hair grower of the present invention may be due to some action of amino acid in the hair grower of the present invention (contained in the amount of 0.2% by weight in the citrus skin and the refined sake). The following is an explanation for the kinds of amino acid contained in 100 ml of the hair grower of the present invention. EMI7.2 EMI8.1 Example 2

A more detailed experiment was performed on the inhibitory actions of d-limonene on the 5 alpha -reductase activity. As previously mentioned, it is known that testosterone is converted into 5 alpha -dihydrotestosterone (5 alpha DHT) having an androgenic action and thereby expresses physiological activities. 5 alpha -Reductase is present in living bodies and functions as an enzyme which converts testosterone into 5 alpha DHT. Therefore substances capable of impeding the activities of the enzyme will probably be prophylactics for androgenic alopecia.

From the above point of view, a system capable of measuring a testosterone 5 alpha -reductase activity was formed, and, using the system, studies were performed on the 5 alpha -reductase inhibitory activities of the hair grower of the present invention and of the limonene that is a constituent of the hair grower.

Two hair growers of the present invention, one aged and another fresh, and d-limonene were subjected to a test. For control groups, Duke extract and sodium beta -glycyrrhetinic acid were selected as positive control groups and swertiamarin as a negative control group.

The test was performed according to the following methods. Preparation of a crude enzyme solution

A Wristar male rat weighing 250-300 g was sacrificed by bleeding, and its prostata was excised. The prostata was cut into fragments under ice-cooling. A 0.1 M HEPES buffer solution (pH: 7.4) containing 0.25 M sucrose (the buffer solution is hereinafter referred to as buffer solution A) was added to the fragments in the amount three times the wet weight of the fragments. Using a Potter-type homogenizer made of Teflon, the mixture was homogenized. The homogenate was filtered using a stainless mesh of a 200 mu size, and then the filtrate was centrifuged at 3000 rpm (900 xg) for ten minutes. The precipitate was suspended in 2.5 parts of buffer solution A, followed by centrifuging at 3000 rpm for five minutes. The residue recovered was suspended in buffer solution A weighing 0.8 times the wet weight of the fragments to produce a crude enzyme solution.

The crude enzyme solution was stored at -70 DEG C. Enzymatic reaction

The test sample (less than 10 mu l) previously prepared was added to [4-<1><4>C]-testosterone (1.5 nmol, 2.85 kBq), NADPH (0.5 mu mol), and the enzyme solution (30 mu l) cited above. To this was added the buffer solution A to the extent that the total volume became 100 mu l, to prepare a reaction liquid. An enzymatic reaction was performed using the reaction liquid at 37 DEG C for two hours. The enzymatic reaction was initiated by removing the reaction liquid from an ice-water-cooled bath to a 37 DEG C warm bath. The enzymatic reaction was terminated by cooling the reaction liquid in ice-water. After completion of the reaction, 400 mu l of n-hexane was added to the liquid, followed by vigorously stirring the mixture to uniformly mix.

The uniformly blended mixture was centrifuged at 3000 rpm and a temperature of 4 DEG C for five minutes to obtain a supernatant liquid. 200 mu l of the supernatant liquid partially collected was evaporated under reduced pressure to a solid, to which was added 30 mu l of n-hexane to redissolve. 10 mu l of the liquid mixture collected in part was applied to a thin layer plate of silica gel. After drying, the deposited substance was developed using cyclohexane-ethyl acetate (1:1, v/v). Quantitative determination of reaction product and Enzymatic activity inhibitory ratio measuring method

A radiation activity of the developed material on the TLC plate cited above was measured by means of an AMBIS image analyzer.

Two spots shown in a chromatogram were subjected to readings of the radiation activities identified by their positions corresponding to testosterone (T) and dihydrotestosterone (DHT) respectively. The rate of conversion of testosterone into dihydrotestosterone was given by: DHT/(T + DHT) x 100% Defining the conversion rate in the presence of the negative control as 100, a percentage of the conversion rate in the presence of the sample was calculated to determine the degree of the enzymatic activity inhibitory ratio of the sample. Results

First, various organic solvents were investigated to seek an eluent that could clearly isolate testosterone from dihydrotestosterone on the TLC plate, and as a result it was confirmed that both materials could be clearly separated independently by the use of the aforementioned developing solvent mixture, cyclohexane-ethyl acetate (1:1, v/v). Specifically, on the TLC plate there were the two spots representing respective radiation activities. One spot with a lower Rf value was for testosterone and another spot with a higher Rf value for dihydrotestosterone. The two spots could be thus distinguished.

Second, conditions of the enzymatic reaction were studied, and as a result it was confirmed that the reaction at 37 DEG C for two hours was relatively effective for the conversion of testosterone into dihydrotestosterone. In this case, because the liposoluble sample was barely soluble in water, it was mixed with water by shaking at 120 rpm.

Further studies were carried out on the kinds of extracting solvents and thin layer plates, and the like.

The reaction conditions thus established are as follows: [4-<1><4>C]-testosterone 1.9 mu l NADPH 10 mu l Enzyme solution 30 mu l Sample 3-10 mu l

The process was carried out by heating at 37 DEG C for two hours a reaction liquid made up to 100 ml by the addition of a buffer solution, extracting with hexane, and developing by means of TLC.

Next, studies were carried out on the inhibitory activities of sodium beta -glycyrrhetinic acid and Duke extract, which were used as the positive controls. As shown in Fig. 1 and Fig. 2, the amounts of sodium beta -glycyrrhetinic acid and Duke extract exhibiting a inhibitory ratio of 50% are 0.5 mM and 0.75 mu l respectively. Their remarkably high inhibitory activities are thus confirmed.

The inhibitory actions on the testosterone 5 alpha -reductase activity of the hair growers of the present invention and of d-limonene, which is a constituent of the hair grower, were studied. As shown in Figs. 3 and 4, either of the test samples can give a considerable high inhibitory effect. Among the hair growers the aged sample gives a higher inhibitory effect than the fresh sample. On the other hand, the negative control swertiamarin has no inhibitory effect as shown in Fig. 5.

As explained above, the hair grower containing as a major component the 5 alpha -reductase inhibitor according to the present invention provides an excellent hair-growth effect upon application to the human scalp. The hair grower is very safe in contact with the skin. Also, it can be readily produced from the skin of citrus fruits and hence has the advantages of easy preparation and reduced cost.

Use of d-limonenes as testosterone-5-alpha-reductase inhibitor and as hair grower. - Miyauchi, Yutaka


does this mean it's good for hair but bad for androgens

 

[danishispsychic]

Stanley Burroughs writes about it in his yellow Master Cleanser book-

 

[Inaut]

what does he say @danishispsychic if you care to summarize (as best you can)

 

[danishispsychic]

He talks about its benefits and being something that breaks down toxins and he believed that " toxemia " is and was the only disease. ( I am inclined to agree these days ) Lemon with pith and rind and well as juice is part of his Lemon/Maple syrup/ Cayenne drink that one fasts on doing his cleanse. A lot of people do not take this cleanse seriously as it seems so simple etc but I have done it several times. He talks about the minerals and vitamins in the B grade syrup fortifying and mixed with the fresh lemon ( rind or no rind ) to break down and " melt " toxins in the color , liver , kidneys ( hence the Master Cleanse ) . The Cayenne added then helps the vascular system and also additionally heals the gut . He talks about this formula being healing and treated cancer patients with it as well. Some more successfully than others I am sure. Basically by drinking the specifically formulated drink, you are dissolving and cleaning. The salt water flush each morning after taking a laxative tea at night then adds the salt ( very Peaty ) back into the system and does a " flush". That salt flush is taken from all old Ayurvedic cure . I talk about it all the time and basically because the salt, limonine , maple syrup, cayenne and of course the superstar water are a powerful detoxer. It is to be done for 10 days. I have never made it past 6 but I have done it about 10 times.

 

[danishispsychic]

Quotations from Stanley Burroughs
On germs, viruses and epidemics:

“In recent times it has been believed that these many diseases are contagious and that germs have spread them….

“Disease, old age, and death are the result of accumulated poisons and congestion throughout the entire body. These toxins become crystallized and hardened, settling around the joints, in the muscles, and throughout the billions of cells all through the body … Lumps and growths are formed all over the body as storage spots for unusable and accumulated wast products … These growths and lumps appear to us as forms of fungi….

“When we stop feeding this fungi and cleanse our system … they dissolve or break up and pass from the body…. Germs and viruses do not and cannot cause any of our diseases … Germs and viruses are in the body to help break down waste material and can do no harm to healthy tissues….

“Basically, all of our diseases are created by ourselves because we have never taken the time to discover the true foods meant for man’s use.

“Since germs do not cause our disorders, there must be another logical reason for the triggering of an epidemic. This is a matter of simple ‘vibration.’ The better the physical and mental condition a person is in, the higher becomes his vibration, but as he steadily becomes clogged with more and more waste matter, his vibration goes constantly downward….”

 

[Inaut]

Thanks @danishispsychic. Very useful information :)

Since I started drinking more lemon juice, I’ve noticed a definite effect on my guts and colon. I eat a lot of ginger also so I can see that working similarly to the cayenne. Pretty much a detoxing lemonade.

Your second post resonates with my own thoughts on disease in general. I think limonene is powerful and cheap for busting up the crude in the lymph and blood

 

[danishispsychic]

I agree and I would even go as far as to say that it aligns with Dr Morse obsession with clean lymph ( I am not a total fan of his tho ) and actually I think that the whole orange juice thing with Dr Peat probably does some of the same citrus orange oil " stripping " thing if it is fresh. The further I go down the rabbit hole the more I think that detoxing is more important than what you put in really. :) and P.S if Cancer is a fungi like a lot of health gurus think - no wonder there is no " cure for cancer ". I love Burroughs, he was jailed for practicing medicine without a license. My kind of guy.

 

[Inaut]

I agree and I would even go as far as to say that it aligns with Dr Morse obsession with clean lymph ( I am not a total fan of his tho ) and actually I think that the whole orange juice thing with Dr Peat probably does some of the same citrus orange oil " stripping " thing if it is fresh. The further I go down the rabbit hole the more I think that detoxing is more important than what you put in really. :)

Once again I’m in full agreement to pretty much everything you just said! Even to your point about Morse... He made me aware of the importance of the lymphatic movement and cleansing but I don’t subscribe to most of his ideas.

 

[Inaut]

Monoterpenes: Essence of a Cancer Cure

MAY 22, 2014 BY CINDY JONES 1 COMMENT

by Cindy L.A. Jones, Ph.D.

Essential oils, the highly concentrated volatile, aromatic essences of plants, are a mainstay of aromatherapy but are also used in flavoring, perfumes and even as solvents. Researchers now think that two components of orange oil and lavender oil are a good bet to prevent and treat cancer.

Most essential oils contain monoterpenes, compounds that contain 10 carbon molecules often arranged in a ring. Monoterpenes are formed in the mevalonic acid pathway in plants. This is the same pathway that makes cholesterol in animals and humans. Early on, cancer researchers realized that some aspects of cholesterol metabolism were involved in cancer growth. They then discovered that plant monoterpenes interfered with animal cholesterol synthesis, thereby reducing cholesterol levels and reducing tumor formation in animals.

Limonene and Perillyl Alcohol
Two widely studied monoterpenes are being evaluated for their anticancer activity, limonene from orange peel (Citrus sinensis) and perillyl alcohol from lavender (Lavandula angustifolia). Because limonene and perillyl affect the pathway that produces cholesterol, they can inhibit cholesterol synthesis, thereby eliminating a minor contributor to cancer formation. Monoterpenes also increase the levels of liver enzymes involved in detoxifying carcinogens, an effect that decreases the possibility carcinogens will cause cellular damage. In addition, monoterpenes stimulate apoptosis, a cellular self-destruction mechanism triggered when a cell’s DNA is badly damaged. This safety feature is generally activated before a cell becomes cancerous. Finally, monoterpenes inhibit protein isoprenylation. The cell uses this process to help a protein, in this case the ras protein involved in cell growth, find its proper location within the cell. If ras is not in the right place, it becomes overactive and can spur cancerous cell growth.

Where Do They Come From? Most plant matter contains a wide variety of monoterpenes. Rich sources include: herbs, spices, wine, essential oils, eggs, olive & palm oil, rice bran oil, barley oil, and dairy products.
Laboratory animal studies demonstrate that these two monoterpenes inhibit the formation of chemically induced breast, colon, liver, skin and pancreatic tumors. For example, animals fed a diet containing 5 percent orange peel oil had a significantly reduced risk of developing mammary tumors when treated with the chemical tumor inducer DMBA. Similarly, animals fed a 5-percent limonene diet had less chance of mammary tumor growth. Researchers noticed that in this experiment rat tumors also regressed, suggesting limonene may treat existing cancer as well as prevent it.

New drugs typically undergo three phases of clinical tests, each more rigorous than the previous: Phase I trials establish a toxic human dose, phase II trials determine a therapeutic dose and how it is metabolized, and phase III trials determine drug effectiveness. Extensive animal studies are done before phase I trials begin.

Phase I trials have so far shown that limonene is well tolerated by cancer patients and has little toxicity. Phase II trials, to test how well limonene actually works to reduce cancer, are under way at several institutions including the National Cancer Institute (NCI) in Bethesda, Md. In other research, perillyl alcohol, a related compound, was found to be five times as active as limonene in regressing tumors. A diet of 2.5 percent perillyl alcohol caused 75 percent of chemically induced rat mammary tumors to regress. Perillyl alcohol is now being tested in NCI-sponsored phase I clinical trials as a treatment for advanced breast, ovarian and prostatic cancers at the University of Wisconsin, Madison. Researchers speculate that perillyl alcohol may also be effective against pancreatic cancer, which is extremely difficult to treat.

The amount of monoterpenes needed to prevent cancer in humans is not established. Toxicity studies are incomplete, but the high doses required for chemotherapy may cause kidney damage and gastrointestinal problems. Both orange and lavender essential oils are safe to ingest; in fact, orange oil is a common food additive used for flavoring.

Few drugs have been developed that effectively treat cancer, so NCI is constantly searching for potential drug candidates. Many of these candidates, like limonene and perillyl alcohol, are natural products from herbs. For now though, these two concentrated substances remain in the realm of the laboratory, the doses being used in clinical trials are intended for treating cancer and must be monitored by a physician.

References
1. Elson CE, Yu SG. The chemoprevention of cancer by mevalonate-derived constituents of fruits and vegetables. J Nutr 1994;124:607-14.
2. Gould MN. Cancer chemoprevention and therapy by monoterpenes. Environ Health Perspect 1997;105:S977-9.
3. Mills JJ, et al. Induction of apoptosis in liver tumors by the monoterpene perillyl alcohol. Cancer Res 1995; 55:979-83.
4. Hohl RJ. Monoterpenes as regulators of malignant cell proliferation. In: American Institute for Cancer Research. Dietary Phytochemicals in Cancer Prevention and Treatment. New York: Plenum Press;1996.
5. Elson CE. Suppression of mevalonate pathway activities by dietary isoprenoids: protective roles in cancer and cardiovascular disease. J Nutr 1995;125:1666S-72S.
6. [Anonymous]. Clinical Development Plan: l-Perillyl Alcohol, J Cellular Biochem 1996;26S:137-48.
7. Crowell PL, et al. Antitumor effects of limonene and perillyl alcohol against pancreatic and breast cancer. In: American Institute for Cancer Research. Dietary Phytochemicals in Cancer Prevention and Treatment. New York: Plenum Press;1996.
8. Vigushin DM, et al. Phase I and pharmacokinetic study of d-limonene in patients with advanced cancer. Cancer Research Campaign Phase I/II Clinical Trials Committee. Cancer Chemother & Pharmacol 1998;42:111-17.
9. Ziegler J. Raloxifen, retinoids and lavender: “me too” tamoxifen alternatives under study. J Natl Canc Inst 1996;88:1100-1.
10. Stark MJ, et al. Chemotherapy of pancreatic cancer with the monoterpene perillyl alcohol. Cancer Letters 1995; 96:15-21.

Originally printed April 1999 Nutrition Science News




Monoterpenes: Essence of a Cancer Cure