This one is really long but this study deserved a very detailed look.  I explained my feelings on it at the end.  I started this at 5:30 this morning and am posting it at 2:30 in the afternoon. 

Alfonso, Mara de O Silva, Carvalho, Rivelli, Barros, Rogero, Lottenberg, Torres, and Mancini-Filho (2013) compared two different forms of rosemary extract and their effect on high cholesterol. 

One was a water extract, 20 grams of herb to 100 milliliters of water at room temp for 1 hour.  It was then centrifuged, filtered then 100 milliliters of water added.

The non-estrified phenolic fraction (NEPF)was powdered, defatted leaves. They did this with tetrahydrofuran, 6 one-minute periods of 20ml, at room temperature.  They filtered it with sodium sulfate anhydrous.  They used vacuum evaporation at 30 degrees Celsius. The dried matter was then placed in 5 milliliters of methanol then filtered then the flask was filled to 20 milliliters.  

This study was conducted using 48 mice which were divided into six groups. All groups were fed a high cholesterol (HC) diet except the control group.  The HC group received distilled water. 

There were two water extract groups. One was treated with 70 milligrams of the extract per kilogram of body weight per day, the other with 140 (AQ 70 and 140).  The other extract was given to the last two groups at 7 milligrams per kilogram of body weight per day and 14. 

When they evaluated the phenolic content of the extracts, there was twice as much phenolic compound in the water extract, 15. 67 to 8.59 in the NEPF.  The water extract had 1.87 grams of rosmarinic acid per 100 grams of dry weight.  The NEPF extract had 5.71 grams of carnosic acid per 100 grams of weight. 

The antioxidant activity was ten times higher in the NEPF extract.  The total cholesterol was 39.8% lower in the AQ70 group and the non-high density lipoprotein cholesterol (non-HDL) was 44.4 % lower.  The higher dosage did not have any significant result in comparison.  The triglyceride levels were higher and the HDL were lower but not significantly.  In the AQ70, the thiobarbituric acid reactive substances (TBARS) were reduced from 1.34 to .83 micromoles/milligram.

With NEPF, at 7 and 14 milligrams per kilogram, TBARS in the heart dropped by 24.32% and 16.05 respectively. In renal tissues, levels dropped by 33.61% and 27.29% respectively.  Both rosemary groups showed significant drops in TBARS levels in the brain and liver in comparison with the high cholesterol group. 

In both rosemary extracts, the catalase activity was significantly increased, even higher in the higher dosage groups. For glutathione, the higher dosage of water extract performed the best.  In the kidneys, the NEPF increased catalase and superoxide dismutase activities.  Neither extract helped the catalase and superoxide dismutase activities in the cardiac tissue.

An important note on the catalase activity: The mice fed a high cholesterol diet and no rosemary had poor catalase activity.  Adding the rosemary treatment improved this activity seeming to reverse that negative effect from the diet. This means it seemed to reduce the cardiovascular disease caused by the poor diet by reducing the lipid profile.

            NOTES: I know this is a long one but this is an interesting study that is missing some things.  I want to be fair to it so I’m sharing as much of this data as I can.  The authors say only the water extract significantly reduced total cholesterol and HDL and attributed this to overall phenolic compound levels being more than twice as high in the water extract. 

My issue with this is they don’t break this out in this study, just the rosmarinic acid for the water extract and the carnosic acid for the NEPF extract. This leaves the question of what were the other compounds and how much of each was in each extract.  I also don’t like how they provide numbers in some comparisons and percentages in others which doesn’t provide a “fair” comparison between these two groups unless you can see the graphs they include in the study.  As an analyst, I start questioning why the numbers are not presented in the same form when doing comparisons. 

I do think this form of study is vital, in that the readers need to understand there is a reason to use different types of extraction methods.  When you can see the differences in effectiveness for different issues, it helps bring this concept home.  It helps to understand why we use teas, infusions, decoctions, tinctures and other methods of use, often different methods for each herb. 
 
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Alfonso, M. S., Mara de O Silva, A., Carvalho, E. B. T., Rivelli, D. P., Barros, S. B. M., Rogero, M. M., Lottenberg, A. M., Torres, R. P., and Mancini-Filho, J. (2013). Phenolic compounds from Rosemary (Rosmarinus officinalis L.) attenuate oxidative stress and reduce blood cholesterol concentrations in diet-induced hypercholesterolemic rats. Nutrition and Metabolism, 10:19.

I confess, when I first opened this article, I thought there was NO way this herb would be effective here and they killed these poor rats for nothing.  I am now humbled after fully reading this article.  Such a detailed study!

Ashtiyani, Zohrabi, Hassanpoor, Hosseini, and Hajihashemi (2013) tested a water-extract of rosemary in a study using 40 rats, 10 per group to investigate the effectiveness of rosemary in treating kidney failure.  The authors said the most common cause of kidney failure is renal ischemia which can be caused by vasoconstriction. Decline in kidney function results in the buildup of nitrogen waste and the increase of blood urea and creatinine.

Two of the groups were given rosemary for 1 week, one of these groups underwent renal failure by an induced ischema-reperfusion. One group was given reperfusion with saline over the same period. 

The water extract was created by coarsely grinding fresh herb and putting 8 grams of herb in 100 milliliters of water.  It was boiled for 5 minutes and infused for 10.  The rats were given 10 milliliters per kilogram of weight per day through a feeding tube.

In the rosemary group, there was a 86% decrease in creatinine clearance, 34% decrease in urine osmolarity, 39% decrease in erythrocytes inside the glomeruli, and 35% decrease in absolute urinary excretion of potassium. There were increases in levels of plasma creatinine, BUN, absolute levels of sodium urinary excretion, fractional sodium urinary excretion, and fractional potassium urinary excretion.  This is in comparison to the sham group that was given rosemary but not the reperfusion. 

Compared to the other group that was given the reperfusion, the rosemary group had a 71% decrease in plasma creatinine, decrease in BUN by 48%, decrease in absolute urinary sodium excretion by 74%, decrease in fractional sodium excretion by 94%, decrease in fractional potassium excretion by 56% and a significant decrease in malondialdehyde.  There were significant increases in erythrocytes in glomeruli, absolute urinary potassium excretion, creatinine clearance and ferric-reducing antioxidant capability. 

In the degradation of the kidneys comparison, the rosemary group did incredibly well.  The authors used a grading system with zero being not present and 5 being 80-100% present.  The reperfusion group got a 3 in increase in Bowman space, a 4 in vascular congestion, cell exfoliation got a 4, and a 4 in proteinaceous casts.  In the rosemary group, Bowman space got a 1, vascular congestion scored a 2, cell exfoliation got a 2, and proteinaceous casts got a 1.

The results indicate the rosemary group had better blood flow than the reperfusion group.  This demonstrates the anti-inflammatory and antispasmodic abilities of the herb. The rosemary group also had significant decrease in cellular damage and functionality.
 
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Ashtiyani, S. C., Zohrabi, M., Hassanpoor, A., Hosseini, N., and Hajihashemi, S. (2013). Oral Administration of the Aqueous Extract of Rosmarinus Officinalis in Rats Before Renal Reperfusion Injury. Iranian Journal of Kidney Diseases, 7(5).

Gauch, Silveira-Gomes, Esteves, Pedrosa, Gurgel, Arruda, and Marques-de-Silva (2013) collected the dentures of 10 people and retrieved samples of Candida albicans from them. The intent was to see if rosemary essential oil would have an effect against this fungus.  Four sets of dentures had Type I and four had Type II.  The other two didn’t have any. 

No matter the type of C. albicans tested, when 4% rosemary oil was added, germ tube formation was completely inhibited.
 
Gauch, L. M. R., Silveira-Gomes, F., Esteves, R. A., Pedrosa, S. S., Gurgel, E. S. C., Arruda, A. C., and Marques-de-Silva, S. H. (2013). Effects of Rosmarinus offi cinalis essential oil on germ tube formation by Candida albicans isolated from denture wearers. Revista da Sociedade Brasileira de Medicina Tropical 47(3):389-391. doi: 10.1590/0037-8682-0137-2013

Shin, Choi, Ryu, Lee, Kim, Choi, Chang, Lee, Jang, and Inn (2013) conducted a study testing the effectiveness of carnosic acid in treating human respiratory syncytial virus (hRSV).  At the time of the study, there was no vaccine and no effective study.  They tested three herbs, the other two were not very effective against hRSV so they just used rosemary in this study.  They extracted carnosic acid, rosmarinic acid, and butalinic acid but focused the study on carnosic acid.  They also determined carnosol has no effect on hRSV.  The authors also evaluated cytotoxicity and determined there was minimal cell degradation due to toxicity. 

The results using carnosic acid against hRSV were effective.  After two days at a dose of 20 micrograms per milliliter, the virus had reduced by 26-fold.  At a dose of 50 midcr50 micrograms the number had risen to 1160-fold.  After four days at the 50 dose, there was no virus remaining. It was effective against both a and b type hRSV viruses.  However effective this was against hRSV, they tested it against the flu and had no effect suggesting it has a very specific way of working against some viruses.  Treatment 8 and 12 hours after infection were still effective at 70% and 50%.  

They also tested using pretreatment and the results suggested carnosic acid may inhibit hRSV infection.  While they still do not fully understand how it works, they think because it was ineffective against influenza A, it must attack the virus itself rather than protect the host cells.  For this same reason, they believe it does not attack the RNA of a virus.  The current thinking it is works against proteins which inhibits RNA synthesis.
 
Shin, H., Choi, M. Ryu, B., Lee, N., Kim, H., Choi, H., Chang, J., Lee, K. T., Jang, D. S., and Inn, K. (2013). Antiviral activity of carnosic acid against respiratory syncytial virus.  Virology Journal, 10:303.

Zhao, Lee, Ma, Huang, Sun, Li, Chen, and Niu (2012) studied the effects of carnosol, an antioxidant on estrogen receptors. This is an attempt to find a solution to postmenopausal cardiovascular symptoms that do not bear a cancer-causing risk such as hormone replacement therapy.

Estrogen receptor a played a bigger role in this study that estrogen receptor b but it was clear that carnosol was effective overall in this study.  Carnasol played a significant role in reducing induced oxidative stress in a vascoprotective way.  Meaning, it prevented cell death caused by oxidative stress.  It displayed phytoestrogenic activity in BAECs. 

The estrogen receptors have also demonstrated antioxidant properties, especially estrogen receptor a which inhibited some of the carnosol activity. This study showed carnosol used the estrogen receptor pathway to exert its antioxidative properties.  However, the binding ability is different due to the structure of the phytoestrogen cells.  When the cells were pretreated with carnosol 24 hours prior to induced oxidative stress, the cardiovascular cells were not damaged by the oxidative stress. 

The authors of the study say there are existing studies which indicate daily intake if carnasol in animals is well tolerated.
 
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Zhao, P., Lee, D. Y., Ma, Z., Huang, L., Sun, L., Li, Y., Chen, J., and Niu, J. (2012). The Antioxidant Effect of Carnosol in Bovine Aortic Endothelial Cells Is Mainly Mediated via Estrogen Receptor α Pathway. Biological Pharmacological Bulletin, 35(11):1947–1955

This constituent isn't really "new" but it was not listed in any of the books I used as sources so I do not have it defined in my constituent list nor is it listed in my rosemary herbarium yet.

Miyazawa, Sugie, and Shindo (2002) looked at the biotransformation of the constituent verbenone which is found is the essential oil of rosemary.  In the study they were able to determine it transforms into a metabolite called 10-hydroxyverbenone.  The authors conclude the study with indicating further studies were underway on this topic.

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Miyazawa, M., Sugie, A., and Shindo, M. (2002). Biotransformation of Verbenone by Human Liver Microsomes. Bioscience, Biotechnology and Biochemistry, 66(11): 2458-2460.

Vicente, Martin, Garcia-Risco, Fornari, and Reglero (2012) examined the extraction of rosemary’s antioxidant constituents using carbon dioxide and combing them with sunflower oil, linseed oil and grapeseed oil to determine if there was any effect on the antioxidant ability of the rosemary constituents. 

They determined if ethanol is not used with the carbon dioxide extraction process, the antioxidant ability is greatly reduced. No matter which method, with or without ethanol, the linseed oil had the most carnasol and carnosic acid, increasing as the dosage increased, The grapeseed oil had the least.

The interesting part the authors noted was the linseed oil without the rosemary was the most likely to oxidize. There was a direct relationship to the amount of PUFA (polyunsaturated fatty acids) to the likelihood of susceptibility of oxidation. 

The sunflower oil had the least PUFA and was in between the other two oils in terms of absorption of the rosemary constituents. Linseed oil had the most PUFA and absorbed the most. Grapeseed oil was the middle on PUFA but the last on absorption.

The authors are now wondering if the composition of the PUFA is the cause of oxidation.  Linseed oil was 43% linolenic acid (Omega-3) and 19% linoleic acid (Omega-6). Grapeseed oil had 68% linoleic acid, and Sunflower oil had 44% linoleic acid. They also are wondering if the position of the double bonds has an effect. Constituents which are a natural part of the original oils may also play a role.
 
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Vicente, G., Martin, D., Garcia-Risco, M. R., Fornari, T., and Reglero, G. (2012). Supercritical Carbon Dioxide Extraction of Antioxidants from Rosemary (Rosmarinus officinalis) Leaves for Use in Edible Vegetable Oils. Journal of Oleo Science, 61(12):689-697.

Kayashima and Matsubara (2012) studies carnosic acid and carnosol themselves and determined carnosic acid is neuroprotective and is dose dependent where carnosol is not dose defendant with the same activity.  In mice, they fed these constituents then looked at the inside of the aorta of each mouse to determine the effectiveness.  The images in this study speak volumes though they only show the comparison between carnosic acid and the control, not one for carnosol.  For carnosol, at a dose of 10 micrometers, microvessel growth was inhibited at 32% (these support the growth of tumors).  At a dose of 50, that percentage only went up to 37%.  The authors felt the carnosic acid was converting a small amount to carnosol to support this activity. The effect of carnosol and carnosic acid on endothelial cells were different which may have resulted on the carnosol being weaker. Carnosic acid seems to work by activating the Keap1/Nrf2 transcription pathway to express antioxidant enzymes and protect against oxidative stress. They end their study mentioning there are studies underway to determine the exact pathways that these 2 constituents use to be effective in protecting these cells.

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Kayashima, Y., and Matsubara, K. (2012). Antiangiogenic Effects of Carnosic Acid and Carnosol, Neuroprotective Compounds in Rosemary Leaves. Bioscience, Biotechnology and Biochemistry, 76(1): 115-119. Doi: 10.1271/bbb.110584

Wada, Nagano, Kido, Ikeda, Kuroda, and Nakashima (2011) evaluated rosemary extract in both water and isopropanol to evaluate antioxidative effects using the thiobarbituric acid (TBA) method. 

The water extract was 65% oxidative and the isopropanol extract was 31%.  The control was 8% oxidative. This was indicated by the amount of rosemary extract in the blood plasma tested, which was taken from volunteers. 

They tested the extracts at different concentrations: 10, 20, 30, 40, and 50 milligrams per milliliter.  With each increase in concentration, there was in increase in antioxidative activity.  In each, the water extract was more than the other, with 50 being more than twice as effective.

Constituents were not determined using this method therefore it was not determined what was causing the effect.

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Wada, M., Nagano, M., Kido, H., Ikeda, R., Kuroda, N., and Nakashima, K. (2011). Suitability of TBA Method for the evaluation of the Oxidative Effect of Non-water-soluble and water-soluble Rosemary Extracts. Journal of Oleo Science 60(11): 579-584

While the picture above is of my rosemary bush and the title says this article is about cinnamon, I came across this article while researching rosemary and the plant was included in the study.  While it was effective, it was the LEAST effective of those studied with cinnamon being the most.  This was quite an interesting article.  I hope you enjoy the summary I wrote!

Taguchi, Takizawa, Ishibashi, Sagawa, Arai, Inoue, Yamaguchu, Abe, (2010) conducted a study to determine herbal effectiveness against candida albicans. Herbs tested were lemongrass, lemon balm, rosemary, roselle, spearmint, green tea, cinnamon (Cinnamomum cassia), and thyme.  Lemongrass, green tea and cinnamon were the strongest when tested against clove (which had a numbing effect in previous studies making some results difficult to read). Cinnamon was the strongest. 

All herbs were extracted using water at 90 degrees Celsius for 5 minutes. This methodology was to test the viability of a tea used medicinally. This extract was then diluted to determine what amount was needed for effectiveness. 

Of the herbs tested, rosemary performed the worst with having to use 25-50% of the extract to get a 50% kill ratio of the candida.  Spearmint was 12.5-25%. Lemon balm and thyme were 6.25-12.5%.  Lemon grass and green tea were less than 6.25% of the extract to achieve a 50% kill ratio.  Cinnamon was put directly in the mouth of the mice instead of in the water and it was 1-5% of the extract.  It took twice as much for each herb to achieve an 80% kill ratio except with cinnamon and clove which was the control. The clove numbers were 5-25% of the extract for both a 50% kill ratio and an 80% kill ratio. 

Since cinnamon doesn’t dissolve completely in water, they wanted to make sure the undissolved particles were included in the treatment, hence the variation. They did a symptom comparison between the control and cinnamon at different ratios.  Only the 100% cinnamon treatment had a significant result in comparison. 

There was about 200 milligrams of solid matter per milliliter in the cinnamon extract. In looking at cinnamon closer, the original extract had 19.5 milligrams per milliliter of cinnamaldehyde and 3.09 milligrams per milliliter of coumarin.  They put it in a centrifuge and tested the top layer (supernatant) and bottom layer (precipitate). The supernatant portion had 5.38 milligrams per milliliter of cinnamaldehyde and .99 milligrams of coumarin while precipitate portion was 3 or 4 times that for each.

In testing effectiveness, the supernatant needed 20-100% dilution while the precipitate was 256-1024%. The dry matter in the precipitate was ten times that of the supernatant.
 
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Taguchi, Y., Takizawa, T., Ishibashi, H., Sagawa, T., Arai, R., Inoue, S., Yamaguchu, H., Abe, S. (2010). Therapeutic Effects on Murine Oral Candidiasis by Oral Administration of Cassia (Cinnamomum cassia) Preparation.  Jpn. Med. Mycol. 51: 13-21.