Cranberry’s medicinal properties have been recognized for centuries for a number of ailments and Native Americans even used raw cranberries as a wound dressing. Clinical studies have been conducted to determine how Cranberries help support Urinary Tract Health. The most recent studies have shown constituents found naturally in Cranberries can help reduce the ability of bacteria like E. coli to adhere to the lining of the bladder and urethra, potentially reducing the susceptibility for and advancement of urinary tract infections (UTIs). Cranberries are also high in antioxidants and contain a potent vasodilator, which help reduce free radicals in the body. However, recent studies show that the Cranberry extract itself did not display the ability to kill the bacteria like E. Coli. Therefore, Nature’s Essentials formulation team worked to find the best ingredients found in nature that are known to have the ability to combat bacteria such as E. Coli. After extensive searches Oregano Oil Extract and South African Buchu Extract were found to have the most scientific evidence supporting them as potent anti-microbial agents with specific results in Urinary Tract Health. First we found the best Wildcrafted Organic Oregano Oil Extract with potent bacteria fighting agents Carvacrol and Thymol (standardized to 75-85% carvacrol). Next, we added Buchu, which originated in South Africa that contains active microbial fighting ingredient diosphenol among others. Both of these ingredients possess powerful anti-microbial activities and attack harmful microbes with breathtaking results but through different mechanisms from conventional antimicrobial products. Therefore, Nature’s Essentials Cranberry-Rx™ with Oregano Oil Extract and Buchu Extract combined with our potent Cranberry 50:1 Extract is the ultimate synergistic blend for optimal Urinary Tract Health. †

[Buchu. Review of Natural Products. factsandcomparisons4.0 [online]. 2005. Available from Wolters Kluwer Health, Inc. Accessed April 16, 2007.]

As for the Cranberry Extract, the high level of antioxidants in cranberry fruit is partly from substances called proanthocyanidins. Antioxidants scavenge damaging particles in the body known as free radicals. Environmental toxins (including ultraviolet light, radiation, cigarette smoking, and air pollution) can increase the number of free radicals in the body, which are believed to contribute to the aging process as well as the development of a number of health problems such as heart disease, cancer, and infections. Antioxidants can neutralize free radicals and may reduce or even help prevent some of the damage they cause. Research is underway to determine if the antioxidant ability of cranberries will translate into supporting the body in protection from certain precursors that lead to heart disease. Adding to cranberry’s potential health benefits, a recent study found that an extract of cranberry inhibited an enzyme that has been associated with supporting a reduction in cancer risk.

Combatting UTIs

Urinary tract infections (UTIs) are a serious health problem affecting millions of people each year. Infections of the urinary tract are so common that only respiratory infections occur more often. There are approximately 157 million women in the United States and each year, UTIs result in about 9.6 million health care visits and one out of every five woman develops a UTI during her lifetime. UTIs in men are less common, but still occur. Nearly 20% of women who have a UTI will have more than one and 30% of those will have more than two. Cranberry appears to be more effective than some probiotics in supporting the body in preventing recurrent UTIs.

NIH-funded research suggests that one factor behind recurrent UTIs may be the ability of bacteria to attach to cells lining the urinary tract. Current belief is that the prevention of UTI is achieved by inhibiting the infecting bacteria, E. coli, from adhering to uroepithelial cells (Sobota 1984; Schmidt and Sobota 1988; Zafriri 1989; Ofek 1991; Howell 1998). Bacterial adherence to these cells is a critical step in the development of infection, without which the causative bacteria are flushed, preventing their colonization of the urinary tract.

A research group found that a nondialyzable polymeric compound isolated from cranberry juice had the most potent effect on urinary tract health.

Cranberry appears to work by inhibiting the adhesion of type I and P-fimbriated uropathogens (e.g. uropathogenic E. coli) to the uroepithelium, thus impairing colonization and subsequent infection…Reasonable evidence suggests that the anthocyanidin/proanthocyanidin moieties are potent antiadhesion compounds.
[Drugs. 2009;69(7):775-807. doi: 10.2165/00003495-200969070-00002.
Cranberry and urinary tract infections.
Guay DR1. Department of Experimental and Clinical Pharmacology, College of Pharmacy, University of Minnesota, Minneapolis, Minnesota 55455, USA.]

Cranberry Capsules for Urinary Tract Infection Study

Spinal Cord. 2008 Sep;46(9):622-6. doi: 10.1038/sc.2008.25. Epub 2008 Apr 8.
Evaluation of cranberry tablets for the prevention of urinary tract infections in spinal cord injured patients with neurogenic bladder.
Hess MJ1, Hess PE, Sullivan MR, Nee M, Yalla SV.
Author information
Abstract
STUDY DESIGN:
Randomized, double blind, placebo-controlled trial with a crossover design.
OBJECTIVE:
To evaluate cranberry tablets for the prevention of urinary tract infection (UTI) in spinal cord injured (SCI) patients.
SETTING:
Spinal Cord Injury Unit of a Veterans Administration Hospital, MA, USA.
METHODS:
Subjects with spinal cord injury and documentation of neurogenic bladder were randomized to receive 6 months of cranberry extract tablet or placebo, followed by the alternate preparation for an additional 6 months. The primary outcome was the incidence of UTI.
RESULTS:
Forty-seven subjects completed the trial. We found a reduction in the likelihood of UTI and symptoms for any month while receiving the cranberry tablet (P CONCLUSION:
Cranberry extract tablets should be considered for the prevention of UTI in SCI patients with neurogenic bladder. Patients with a high GFR may receive the most benefit.
SPONSORSHIP:
Spinal Cord Research Foundation, sponsored by the Paralyzed Veterans of America.

Cranberry Extract for Urinary Tract Infection Study

Eur Rev Med Pharmacol Sci. 2015 Jan;19(1):77-80.
Cranberry supplementation in the prevention of non-severe lower urinary tract infections: a pilot study.
Ledda A1, Bottari A, Luzzi R, Belcaro G, Hu S, Dugall M, Hosoi M, Ippolito E, Corsi M, Gizzi G, Morazzoni P, Riva A, Giacomelli L, Togni S.

Abstract
OBJECTIVE:
Cranberry extracts have been tested as a nutritional supplementation in the prevention of recurrent lower-urinary tract infections (R-UTIs), with mixed results. This pilot, registry study evaluates the prophylactic effects of oral supplementation with a new well-standardized cranberry extract in patients with R-UTI, over a 2-month follow-up.
PATIENTS AND METHODS:
All subjects were suggested to take one capsule containing a cranberry extract (36mg of proanthocyanidins) for 60 days and were also given lifestyle advice. Clinical outcomes were compared between patients on cranberry extracts and those who don’t take this supplementation.
RESULTS:
In total, 22 subjects completed the study in each of the two groups. In the cranberry group, the reduction in the frequency of UTI episodes during the study period compared with the two months before the inclusion was 73.3% (p < 0.05). This figure was 15.4% in the control group (p < 0.05; p = 0.012 vs cranberry group). Seven (31.8%) subjects in the cranberry group were symptom-free; no patient was symptom-free in the control group (p < 0.05). The mean duration of UTI episodes was 2.5 ± 1.3 days in the cranberry group, compared with 3.6 ± 1.7 days in subjects not on cranberry (p < 0.05). Three subjects (13.6%) in the cranberry group and 8 (36.3%) in the control group required medical consultation for UTI symptoms (p < 0.05). Urine evaluation was completely negative in 20/22 subjects in the Cranberry group (90.9%) and in 11 control subjects (50.0%; p < 0.005). No adverse events were observed. CONCLUSIONS: These preliminary results, obtained in a field-practice setting, indicates the effectiveness and safety of a well-standardized cranberry extract in the prevention of R-UTI. Cranberry proanthocyanidin Urinary Tract Infection Study Cranberry products represent a non-antimicrobial method for prevention of urinary tract infection (UTI). Cranberry proanthocyanidin (PAC), a type of condensed tannin, is the active ingredient in cranberry that inhibits adherence of P-fimbriated Escherichia coli (E.coli) to uroepithelial cells.1, 2 Previous cranberry studies for UTI prevention yielded conflicting results, likely because of variability of PAC dose and clinical populations studied. (3, 4) In a clinical trial of 300ml of cranberry juice beverage daily (36mg PAC), older women (mean age 78.5 years) had a 58% reduction in the odds of having bacteriuria plus pyuria compared to controls. (5) However, nursing home residents have difficulty ingesting the volume of juice that is necessary to prevent bacteriuria. Cranberry capsules are feasible to administer to nursing home residents, but their efficacy has not been demonstrated. (6) In vitro, 36 to 108mg of PAC is efficacious at inhibiting bacterial adherence to uroepithelial cells, (7) but the most efficacious dose among older nursing home residents has not been identified. The goal of this study was to identify the optimal dose of cranberry capsules that reduced the incidence of bacteriuria plus pyuria over one month. Go to: METHODS This study was a pilot double-blind, randomized, placebo-controlled trial of 3 cranberry capsules daily (108mg PAC), 2 cranberry plus one placebo capsule daily (72mg PAC), 1 cranberry plus two placebo capsules daily (36mg PAC), and 3 placebo capsules daily for 30 days. The primary outcome was episodes of bacteriuria plus pyuria at 7, 14, 21, and 28 days of cranberry capsule treatment. Participants were stratified by presence or absence of baseline bacteriuria with 20 participants randomized by strata to each arm of the study. Cranberry and placebo capsules were donated by the manufacturer. Urine cultures and urinalyses were collected at baseline and then on a weekly basis for 4 weeks (total = 5 specimens). No additional follow-up was performed after the completion of cranberry capsule treatment. Bacteriuria was defined as >100,000 colony forming units per milliliter (cfu/ml) of any bacteria. Pyuria was defined as any white blood cells seen on microscopic urinalysis. (5) Inclusion criteria were: 1)female; 2)history of UTI; 3)age≥65 years; 4)long term residence; 5)English speaking. Exclusion criteria included: 1)total incontinence; 2)warfarin therapy;8 3)

RESULTS
In 11 homes, 1929 residents were screened; 1381 residents did not meet inclusion criteria (no history of UTI n=665[48.2%], male n=479[34.7%], short-term rehabilitation n=163[11.8%], non-English speaking n=38[2.8%], age < 65 years n=36[2.6%]). Of 548 remaining residents, 308 residents met exclusion criteria (totally incontinent n=104[33.8%], warfarin use n=72[23.4%], residence < 4 weeks n=31[10.1%], chronic indwelling catheter n=23[7.5%], discharged n=20[6.5%], terminal n=16[5.2%], antibiotic therapy n=14[4.5%], kidney stones n=9[2.9%], cranberry therapy n=7[2.3%], administrative decision n=6[1.9%], dialysis n=5[1.6%], cranberry allergy n=1[0.3%]) and 240 residents were eligible; 90 residents consented (37.5% consent rate), and 80 residents enrolled (10 participants met an exclusion criterion prior to enrollment). Demographics of the 80 participants included: 98% white(n=78), mean age 89.2 years(S.D., 7 years), and mean number of comorbidities 4.1(S.D., 1.7). Most participants were totally dependent in bathing(54%) and had some bowel(67%) and bladder(76%) incontinence. Of 80 baseline urine cultures, 1 had no growth, 8 had ≤100,000 cfu/ml, 41 had >100,000 cfu/ml, and 30 had mixed flora (3 or more organisms). The placebo, one, and three capsule groups each had 10 of 20 participants with >100,000 cfu/ml; the two capsule group had 11 of 20 participants with >100,000 cfu/ml on baseline urine culture. Of 80 baseline urinalyses requested, 73 were obtained: 11 had no pyuria and 62 had pyuria. Of 320 urine specimens that should have been collected, 302(94%) urine cultures and 294(92%) urinalyses were obtained. Results of bacteriuria plus pyuria by cranberry capsule group are provided in Table 1. These results represent 4 weekly follow-up urines obtained per participant while consuming cranberry capsules.

Cross-classification of cranberry capsule dose and presence of bacteriuria plus pyuria
Go to:
DISCUSSION
This study showed a dose-dependent trend toward decrease in bacteriuria plus pyuria, particularly with E.coli, among female nursing home residents ingesting cranberry capsules over one month. Previous studies in older patients were conducted using cranberry juice, (5, 9) and studies of cranberry capsules are lacking. In this study, E.coli bacteriuria was reduced and is consistent with the mechanism of PAC, (7) but bacteriuria with other pathogens did not show this same pattern of results. E.coli accounts for about 50% of uropathogens in nursing home residents, (10) and reduction in bacteriuria may reduce treatment for UTI. Since the effect of two and three capsules was comparable and to reduce capsule burden, further investigation of two cranberry capsules daily in nursing home residents is warranted to determine if the reduction of E.coli bacteriuria is sustained over a longer period of time and whether it impacts clinical outcomes related to UTI (e.g., hospitalization, antibiotic therapy for UTI).

References
1. Gupta K, Chou MY, Howell A, et al. Cranberry products inhibit adherence of p-fimbriated Escherichia coli to primary cultured bladder and vaginal epithelial cells. J Urol. 2007;177:2357–2360. [PMC free article] [PubMed]
2. Howell AB, Vorsa N, Der Marderosian A, et al. Inhibition of the adherence of P-fimbriated Escherichia coli to uroepithelial-cell surfaces by proanthocyanidin extracts from cranberries. N Engl J Med. 1998;339:1085–1086. [PubMed]
3. Raz R, Chazan B, Dan M. Cranberry juice and urinary tract infection. Clin Infect Dis. 2004;38:1413–1419. [PubMed]
4. Barbosa-Cesnik C, Brown MB, Buxton M, et al. Cranberry juice fails to prevent recurrent urinary tract infection: Results from a randomized placebo-controlled trial. Clin Infect Dis. 2011;52:23–30. [PMC free article] [PubMed]
5. Avorn J, Monane M, Gurwitz JH, et al. Reduction of bacteriuria and pyuria after ingestion of cranberry juice. JAMA. 1994;271:751–754. [PubMed]
6. Juthani-Mehta M, Perley L, Chen S, et al. Feasibility of cranberry capsule administration and clean-catch urine collection in long-term care residents. J Am Geriatr Soc. 2010;58:2028–2030. [PMC free article] [PubMed]
7. Lavigne JP, Bourg G, Combescure C, et al. In-vitro and in-vivo evidence of dose-dependent decrease of uropathogenic Escherichia coli virulence after consumption of commercial Vaccinium macrocarpon (cranberry) capsules. Clin Microbiol Infect. 2008;14:350–355. [PMC free article] [PubMed]
8. Suvarna R, Pirmohamed M, Henderson L. Possible interaction between warfarin and cranberry juice. BMJ. 2003;327:1454. [PMC free article] [PubMed]
9. McMurdo ME, Bissett LY, Price RJ, et al. Does ingestion of cranberry juice reduce symptomatic urinary tract infections in older people in hospital? A double-blind, placebo-controlled trial. Age Ageing. 2005;34:256–261. [PubMed]
10. Das R, Perrelli E, Towle V, et al. Antimicrobial susceptibility of bacteria isolated from urine samples obtained from nursing home residents. Infect Control Hosp Epidemiol. 2009;30:1116–1169. [PMC free article] [PubMed]

Cranberry RISKS: The use of supplemental cranberry products are not recommended for individuals who are taking warfarin.

Cranberry & Oregano Extracts on Urinary Tract Infections and Lifespan

J Gerontol A Biol Sci Med Sci. 2010 Jan; 65A(1): 41–50.
Published online 2009 Nov 11. doi: 10.1093/gerona/glp176
PMCID: PMC2796885
Prolongevity Effects of an Oregano and Cranberry Extract are Diet Dependent in the Mexican Fruit Fly (Anastrepha ludens)
Sige Zou,corresponding author1 James R. Carey,2 Pablo Liedo,3 Donald K. Ingram,4 Binbing Yu,5 and Reza Ghaedian6

Go to:
Abstract
Botanicals have numerous health benefits. Here, we used the Mexican fruit fly to screen 14 compounds and botanicals for their prolongevity effects and found an oregano and cranberry mixture (OC) improved survival. We then evaluated prolongevity effects of OC within the context of diet composition. Individual flies were fed 0%, 1%, or 2% OC in one of the three diets containing sugar and yeast extract (SY) at a ratio of 3:1, 9:1, or 24:1. We found that prolongevity effects of OC depended upon dose, gender, and diet composition. The greatest increase in longevity was observed in females fed the SY24:1 diet with 2% OC compared to the non-supplemented diet. OC did not reduce egg laying and, hence, did not compromise fecundity under any dietary condition tested here. This study reveals the prolongevity effects of OC and supports the emerging view that benefits of botanicals on aging depend on diet composition and gender.

THE health benefits of diets high in fruit and vegetable content have been well established. Extracts from various plants, fruits, and herbs have diverse biological activities, including antimicrobial infection, anti-inflammation, anticancer, and anticardiovascular diseases as well as prevention of several other types of diseases, such as diabetes and neurodegenerative disorders (1). Many of these properties are attributable to the presence of a variety of phytochemicals, including polyphenolics and flavonoids (2). These phytochemicals often exert their influence through multiple mechanisms, acting as antioxidants, ligands for receptors and activators or inhibitors of enzymes, and transcription factors linked to disease pathways (3–5). Although botanical extract can affect a broad range of biological signatures of aging, few have been directly demonstrated to provide prolongevity effects.

Oregano (Origanum vulgare) and cranberry (Vaccimium macrocarpon) are prominent examples of botanicals offering multiple health benefits (6,7). Oregano is widely used as a food and drink additive and contains high levels of phytochemicals, especially carvacrol and thymol (6,8). Extracts from oregano exhibit antimicrobial activity against a variety of pathogens (6). Oregano oil, for example, has been found to be more effective than benomyl, a commercial fungicide, at inhibiting mycelial growth of a number of phytopathogenic fungi (9). Anticancer effects of oregano have been demonstrated in several cell-based and animal models as well as in clinical investigations and appear to be at least partly attributable to antioxidant activity (10,11). Oregano extracts also exhibit selective proapoptotic and cytotoxic effects on colon cancer cells, suggesting that the wide use of oregano as a spice in the Mediterranean region contributes to the relatively low regional risk for colon cancer (10). In a 3-month supplement study of patients with mild hyperlipidemia, oregano has been shown to improve lipid profiles, antioxidant status, and endothelial function in patients (12). Biochemical studies indicate that oregano extracts potently inhibit both diabetes-related alpha-glucosidase and hypertension-related angiotensin-converting enzyme-1 (ACE-1) (13). These findings suggest that oregano supplementation is potentially effective for management of diabetes and hypertension, two common age-related conditions (14). Together with evidence that oregano can reduce mortality and improve reproductive success in a commercial farm animal setting (15), the range of potential applications of this dietary intervention appears quite broad.

Cranberry provides a rich source of phytochemicals (16,17). Consumption of cranberry juice is popularly regarded as effective in the treatment and prevention of urinary tract infection (UTI) in humans (18). Indeed, recent clinical studies have demonstrated that cranberry consumption for a 12-month period significantly decreases the incidence of symptomatic UTIs, especially among women with recurrent infections (19,20). Randomized control trials in older women with recurrent UTI further indicate that cranberry is equally effective as the commonly prescribed antibiotic, Trimethoprim, but has less adverse effects, especially reduced risk of developing antimicrobial resistance (21). Cranberry constituents, especially proanthocyanidins with A-type linkages, are thought to exert antimicrobial activity by inhibiting adhesion of P-fimbriated uropathogenic Escherichia coli to uroepithelial cells, thereby suppressing bacterial growth and improving urinary tract health (22). Alongside this antimicrobial action, other studies indicate that cranberry has the potential to exert multiple biological effects through its antioxidant properties and by engaging many signaling pathways, such as Jun kinase (JNK), mitogen-activated protein kinase, and NFκB signaling pathways (7,18,23).

Consistent with the proposal that cranberry may offer widespread health benefits, dietary supplementation of cranberry reduces the levels of total and low density lipid cholesterol as well as the total: high density lipid cholesterol ratio without affecting the glycemic control in type 2 diabetic patients on glucose-lowering agents, suggesting that persons with impaired glucose tolerance can benefit from cranberry consumption (24). In addition, a number of in vitro and in vivo experiments have revealed a chemopreventive action of cranberry. For instance, cranberry extracts can inhibit the proliferation and invasion of several types of tumor cells in vitro and tumor formation in vivo, including Rev-2-T-6 murine lymphoma cells and tumors, SGC-7901 cancer cells, and human tumor xenografts in mice (25). Cranberry also stimulates the generation of anti-lymphoma antibodies, suggesting the potential for protective effects against malignant lymphoma in immune competent hosts (26). Cranberry appears to exert these effects by inducing apoptosis in tumor cells and by reducing the expression of tumor metastasis–related matrix metalloproteinases and promoting anti-inflammatory activity (23). Finally, current evidence also suggests that cranberry reduces the risk of cardiovascular disease and blunts the toxicity of Alzheimer’s disease–related A-beta peptide (17,27).

Different botanical extracts have unique phytochemical profiles comprising various polyphenolics and flavonoids. A number of experiments have demonstrated synergistic or additive effects between different phytochemicals in exerting their biological functions (2,28). For instance, synergistic interactions among anthocyanins, proanthocyanidins, and flavonol glycosides are reportedly coupled to the inhibitory effects of whole cranberry extract on the proliferation of cancer cells (29). The synergy is also evident in investigations combining extracts of different botanical sources. Combining water-soluble cranberry extract with extracts of oregano, rosemary, or Rhodiola rosea, for example, enhances inhibition of markers linked to diabetes and hypertension, including alpha-glucosidase and pancreatic alpha-amylase and ACE-I, respectively (28). Cranberry and oregano together, compared with either alone, also augments the inhibition of Helicobacter pylori, a bacterium linked to ulcers (30), and Listeria monocytogenes, a virulent food-borne pathogen causing Listeriosis, presumably by suppressing microbial urease and proline dehydrogenase activity (28). These studies suggest that developing maximally beneficial botanical extracts may require bringing together phytochemicals from different sources to enhance functional activities.

In the current study using the Mexican fruit fly (mexfly) Anastrepha ludens (Loew) as a model organism, we first screened for potential prolongevity effects from a number of candidate compounds and botanical extracts selected on the basis of a variety of hypothesized actions in aging. Mexfly provides several advantages for use as a model system for screening compounds. First, extensive demographic studies of aging have been conducted in this species (40). In particular, the effects of dietary composition on life span and reproduction are well documented (41). Second, millions of mexflies are readily available on a daily basis from the site of the present study (Moscafrut facility, Chiapas, Mexico) (41), which enables usage of a large number of flies reared in identical conditions for screening. Third, unlike the commonly used labarotory fly, Drosophila melanogaster, mexfly females lay their eggs away from the food, allowing a direct assessment of egg laying or fecundity independent of the influence of food availability and composition on egg-laying behavior (38,41). Capitalizing on this model, we screened 14 compounds and botanical extracts and identified a botanical product with a mixture of oregano and cranberry extract (OC) that exhibited robust prolongevity effects. With this lead, we explored this benefit on individually housed flies in the context of diet composition. The results demonstrate that the prolongevity effect of OC in mexfly is both diet and gender specific.

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Materials and Methods
Reagents

Compounds were purchased from Sigma-Aldrich Inc (St Louis, MO). Botanical extracts were obtained from Decas Cranberry Product Inc (Carver, MA) and Barrington Chemicals Inc (Harrison, NY).

Life-Span Assay Using Population Cages

To screen compounds or botanicals for prolongevity effects, approximately 2,000 adult mexflies of both sexes were housed in a large population cage (size: 0.5′ × 1′ × 2′ [W × L × H]) as described previously (42). Briefly, to get these adult flies, pupae were obtained from the mass rearing facility and placed in Petri dishes inside the population cages 1 day before the maximum emergence. After 24 hours, the Petri dishes with the empty pupal cases and nonemerged pupae were removed. Therefore, the age of all individual flies inside each cage was the same (within 24 hours). The amount of pupae placed in each cage was estimated by weight and the possible percentage of emergence so that there were approximately 2,000 flies per cage. The flies were then fed a standard solid diet with sugar: yeast at a ratio of 9:1, 1.5% agar, and a specified concentration of a dietary supplement in each cage. Vivaria were maintained at 24°C ± 2°C, 65% ± 9% relative humidity, and 12:12 hours light–dark cycle. Each treatment was repeated with two cages of flies. Approximately 20 ml of fresh food was provided in a Petri dish to each cage once a week. Dead flies were collected, sorted by sex, and recorded every day for life-span measurements. The screening experiments were conducted in three batches, which were terminated after flies reached 107, 71, or 54 days, respectively. The control was set up and replicated in four separate population cages for each batch of experiments. The number of surviving flies was counted after the termination date. For each treatment, median life-span and survival curves were derived by combining 2,000–2,400 of each sex from two population cages. Median life span of the control was calculated by combining data from four replicates.

Life-Span Assay Using Condominium Cages

Mass-reared virgin male and female flies were individually housed in clear Plexiglass cages (4 × 4 × 10 cm per unit), termed as condominium cages as described previously (41). Environmental conditions were the same as described previously for the population assay. Flies were individually provided 20 μl of one of the following nine diets and a 20-μl droplet of water was provided daily to each fly on a glass slide through an opening in each cage. The nine diets were designed by combination of one of the three different sugar: yeast extracts at a ratio of 3:1, 9:1, and 24:1 with one of the three concentrations of OC: 0%, 1%, and 2% (weight/volume). For each diet, sugar and yeast extract (SY) were first dissolved in double the amount of water by weight to make the sugar: yeast mother stock solution. The OC was then added to each mother stock solution to obtain the final concentration of 0%, 1%, and 2% OC in weight/volume. Eighty-two males and 82 females were randomly assigned into the 164-unit cages with the stipulation that females and males occupied alternate units to avoid mixing eggs from two females. Each unit contained a black silicon membrane attached to the rear of the cage in which females could insert their ovipositor to lay eggs. Sex-specific survival of males and females and egg laying of females were recorded daily.

Data Analysis

Event life history graphs and response surfaces were generated using DeltaGraph 5 software (Red Rock Software, Inc, Salt Lake City, UT). LogRank analyses were performed for the data obtained with the population cages using the SAS PROC LIFETEST program (Cary, NC). The logistic model analysis for the fertility rate data was performed using the R Project for Statistical Computing program (http://www.r-project.org/). Analyses of variance (ANOVA) were conducted to assess the statistical significance of life-span and egg-laying data from the assays with individual flies using Statview Version 5.0 (SAS, Cary, NC). p < .0014 after Bonferroni adjustment for multiple comparisons was considered statistically significant for the experiments using individual flies.

Go to:
Results
We initially surveyed 14 compounds and botanical extracts for their potential prolongevity activity using population cages with approximately 2,000 mixed males and females per cage fed a standard diet with sugar: yeast at the 9:1 ratio. Median life span of flies under these treatments is shown in the Supplementary Tables 1 and 2. A treatment was considered to induce a significant life-span extension if the p value is less than .00083 after Bonferroni adjustment and median life span of both males and females under that condition is 2 SDs more than median life span of the corresponding control. Using these criteria, OC, an oregano and cranberry mixture, was found to extend median life span of both males and females at the highest concentration tested (1.6% OC in food) in this screening (Figure 1). No significant extension was observed with other supplements (online Supplementary Tables 1 and 2). OC has been described previously as a potent antimicrobial botanical extract containing 75% water-soluble oregano extract and 25% water-soluble cranberry extract (43). OC contains 15%–19% total phenolics as measured by the Folin–Ciocalteu assay and has antioxidant activity of 3,250 μmol Trolox per gram based on the oxygen radical absorbance capacity assay (44,45).

Figure 1.
Figure 1.
Life-span curves of mexflies fed the standard 9:1 sugar and yeast extract diet in population cages. Concentrations of oregano and cranberry (OC) mixture in the food as percentages and the life expectancy of flies in days from each treatment are depicted …
To further investigate aging-related effects of OC, we utilized virgin males and females housed individually in plexiglass chambers, which we refer to as “condominium” cages. Flies were fed a standard laboratory diet containing SY at a ratio of 9:1. Under this condition, the OC-supplemented diet significantly extended mean life span in females, but not males, in a dose-dependent manner (Figure 2). Specifically, the mean life span of females was increased by 20.5% by supplementation of 2% OC compared with the control, whereas 1% supplementation had no statistically significant effect (Table 1, see sugar: yeast 9:1 ratio). Maximum life span was not significantly affected in any treatment condition (Table 1). These results indicate that OC extends mean life span in a dose- and gender-specific fashion in mexflies under a standard diet condition.

Table 1.
Table 1.
Life span and Fecundity of Individual Mexflies Treated With Oregano and Cranberry (OC) Mixture
Figure 2.
Figure 2.
Life-span curves of female and male mexflies individually fed the standard 9:1 sugar and yeast extract diet supplemented with 0%, 1%, or 2% oregano and cranberry (OC) mixture. Concentrations of OC in the food as percentages and mean life span of flies …
Dietary customs vary dramatically across geographical regions and ethnic backgrounds (39), and accordingly, it would be important to consider the potential modulatory influence of diet when evaluating the prolongevity effects of any promising supplement. To this end, we assessed whether diet composition, defined by the relative balance of carbohydrate and protein, affects the prolongevity effect of OC. Separate groups of mexflies were provided different compositions of SY, representing carbohydrate and protein, respectively, at one of three ratios: 24:1, 9:1, or 3:1. This evaluation was conducted using the condominium cages, in which individual flies received fresh food daily. Results from the standard diet (SY 9:1) are described previously (Figure 2). In comparison with those findings, increasing the ratio of yeast or decreasing the ratio of sugar (SY 3:1) completely blocked the prolongevity effect of 2% OC relative to nonsupplemented controls maintaining on the same diet (Figure 3A and B and Table 1). In the lower yeast or higher sugar condition (SY 24:1), however, 2% OC significantly increased both mean and maximum life span of females and males (p < .0001; Figure 3C and D and Table 1). Although a numerical benefit of 1% OC was also apparent (Figure 3C and D and Table 1), this effect did not reach statistical significance after adjustment for multiple comparisons. These findings suggest that, besides gender, the prolongevity activity of OC is influenced by the diet composition and is potentiated by a low protein or high sugar level in the food.

Figure 3.
Figure 3.
Life-span curves of female and male mexflies individually fed a 3:1 sugar and yeast extract (SY) diet (A and B) or a 24:1 diet (C and D) supplemented with 0%, 1%, and 2% oregano and cranberry (OC) mixture. SY ratios, concentrations of OC in the food as …
The interactive influence of diet composition and botanical supplementation on life span is further illustrated in Figure 4 using surface response maps, providing a two-dimensional representation of longevity as a function of dietary yeast and OC concentrations. The interaction between OC and yeast extract on life span was also analyzed (Supplementary Table 3). Mean life span of females in general increased with higher concentrations of OC and reached a peak when yeast represented roughly 10% of the available diet (Figure 4A), whereas mean life span of males increased with increasing concentration of OC or yeast under the conditions tested in this study (Figure 4B). The effect of OC on mean life span of both males and females depended on diet composition represented by the percentage yeast as revealed by two-way ANOVA (Supplementary Table 3). The overall pattern of maximum life span in response to yeast and the OC supplement was similar to that for mean life span of females with some differences (Figure 4A and C). There is no significant interaction between OC and yeast concentrations for maximum life span of females. Maximum life span of males in general increased with increasing concentration of OC, but not with the concentration of yeast extract (Figures 4D and Supplementary Table 3). Surface response mapping thereby complements the preceding analyses, consistent with the interpretation that the prolongevity effect of OC is gender and diet dependent.

Figure 4.
Figure 4.
Life-span response surfaces showing the effects of diet composition and oregano and cranberry (OC) mixture concentrations in the diet on mean (A and B) and maximum (C and D) life span of females and males. x-axis represents OC concentration and y-axis …
Reproduction has profound effects on life span, and under certain conditions, life-span extension is associated with reduced reproduction (46). To evaluate whether the prolongevity effect of OC is coupled with suppressed reproduction, daily egg laying was recorded for females maintained in condominium cages. Event life history maps are shown in Figure 5, representing daily egg laying and life span for individual flies. Consistent with previously published results (41), fertility rate and egg laying were significantly reduced in flies as a function of relative dietary protein content (SY 24:1 < SY 9:1 < SY 3:1; Figure 5A, Table 1, and Supplementary Table 4). Within dietary conditions, however, fecundity of fertile females was similar across OC-supplemented and control subjects (Figure 5A and B). Of particular note, OC supplementation did not decrease lifetime egg laying per fertile fly under any concentration tested here (Figure 5A and Table 1). Indeed, the data trended in the opposite direction, and in one condition (SY 9:1, 1% OC), supplementation was associated with increased egg production relative to values for nonsupplemented controls. Fertility rate was significantly increased by 2% OC supplementation (Supplementary Table 4). Together, these findings indicate that OC can have prolongevity effects without compromising reproductive output, thus suggesting this effect is largely independent of processes regulating the reproductive pathway. Figure 5. Figure 5. The effect of dietary composition and oregano and cranberry (OC) mixture on fecundity. (A) Lifetime egg laying per fertile female in relation to diet and OC concentration. (B) Life event history depicting the relationship among life span, egg laying, … Go to: Discussion After screening 14 compounds and botanicals for prolongevity effects using large groups of mexflies housed in population cages, we have found one botanical product that has robust effects on life span. This product is a blend of OC extracts, which increases median life span of both males and females in this initial screening with mixed population. The prolongevity effect of OC has been further examined within the context of diet composition in individually housed virgin flies. Using the standard 9:1 SY diet, we note a significant effect of OC on the mean life span of females. No effect is detected on the mean life span of males or on the maximum life span of either gender. With the higher protein or lower sugar diet (SY 3:1), OC supplementation produces no significant effect on life span. However, with the lower protein or higher sugar diet (SY 24:1), OC significantly increases mean and maximum life span of both males and females. This indicates that the higher sugar or lower protein content in food, the greater the effect of OC on life span, and the prolongevity effect of OC is sex dependent. Using egg laying as a measure of healthspan, it is clear that OC has no detrimental effects on this measure, further supporting the health benefits of the OC supplementation. Diet has an enormous impact on health and life span in almost all organisms including humans (47–49). Dietary restriction without causing malnutrition has been shown to extend life span and delay onset of age-related functional decline and diseases in diverse model organisms (46,49,50). Diet composition is also critical and can significantly influence health and aging. Nutritional geometry studies related to aging in D melanogaster and A ludens have indicated that optimal life span depends on a balanced level of protein and carbohydrates (41,51,52). How a supplement interacts with protein and carbohydrate in food to modulate life span has not been well addressed. The effects of dietary supplements on life span may vary significantly depending on gender and diet composition. Indeed, we have found that the prolongevity effect of OC is more prominent when the mexflies are on diets with relatively higher carbohydrate or lower protein content. OC does not extend life span of mexflies fed a high protein or low carbohydrate diet. Although the exact role of carbohydrate and protein in affecting the prolongevity effect of OC remains to be determined, results from this study suggest that one should take into consideration the physiological difference of gender and dietary composition when evaluating candidate prolongevity agents. In addition, we have manipulated only dietary carbohydrate and protein in this study. The standard fly diet contains only a small percentage of fat from the yeast extract. Given the importance of fat in human diets, it would be necessary in the future to evaluate the interaction of dietary fat and prolongevity supplements using modified fly diets. The observation that females have a more robust response to OC treatment than males when they were maintained in condominium cages but not when they were housed in population cages suggests that the sex-specific effects of OC are affected by mating status as well as by density. Both of these factors have age- and sex-specific effects on the Mediterranean fruit fly (53–55) and thus it is likely they have similar effects on the mexfly. For example, mated flies tend to have shorter life span compared with virgins but the effects differ between the sexes. The cost of reproduction in laying females can also explain the difference in life span of males and females because their reproductive responses differ qualitatively but both depend on dietary, mating, and/or density conditions. Both sexes maintained in population cages tend to live shorter than those individually housed in condominium cages due to heightened stress from high densities, competition for food and mates, and exposure to pheromones that elicit different behaviors. Most or all of these stress factors are avoided when flies are maintained in solitary confinement. In general, the finding that the sex-specific outcome of OC-supplemented diet is conditional on cage and mating conditions underscores the complex life table dynamics involved in the longevity outcomes due to different dietary restriction treatments. Together, our findings point to the challenges of identifying effective aging interventions and the importance of including comprehensive dietary manipulations when evaluating potential prolongevity supplements. What are the potential mechanisms of life-span extension by OC? Egg laying is not reduced by OC and is even increased by one concentration of OC, suggesting that OC does not extend life span through reproduction-related pathways. Cranberry and oregano are rich in polyphenolics (8,16), and a number of polyphenolics can act as antioxidants or modulators of signaling pathways, such as the JNK and NFκB pathways (3–5). Shetty and colleagues found in their studies on antimicrobial activities of OC that proline could suppress the inhibitory activities of OC on microbial growth (30,43). Based on these observations, they proposed that phenolics in OC can disrupt the flow of the electrons along the bacterial membrane by either quenching free electrons or by inhibiting proline dehydrogenase in bacteria, which, in turn, would affect ATP production and consequently microbial growth (56,57). Considering the prokaryotic origin of mitochondria and the fact that a reduction of expression of mitochondrial genes, such as those involved in oxidative phosphorylation, can modulate life span in C elegans (58–60), polyphenolics in OC may modulate life span by influencing mitochondrial function, especially ATP production in eukaryotes. Consistent with this hypothesis, we have found that OC is more effective for life-span extension in mexflies under a higher glucose or a lower protein diet. OC may suppress mitochondrial respiration induced by sugar through the inhibitory effects of its polyphenolics on the electron flow and/or proline dehydrogenase. This suppression may result in lower production of ATP and/or oxidants and lead to longer life span in animals fed OC (61,62). Further experiments need to be conducted to confirm this hypothesis and dissect potential molecular mechanisms responsible for the prolongevity effect of OC in genetically tractable organisms, such as D melanogaster. In many cell-based and whole animal experiments, the beneficial effects of supplementation of one fruit or herb become apparent only at fairly high concentrations, which is either hard to achieve or has unpleasant side effects when applied to humans (26,27,43). Failure to detect robust prolongevity effects for compounds or botanicals other than OC in our study may be due to the possibility that the concentrations of supplements are not optimal. In clinical trials to evaluate the effects of cranberry juice on preventing UTIs, a high percentage of dropouts occurred, mostly due to the fact that cranberry often causes too much acidity in the digestive system and upsets the stomach (19,20). Therefore, regimens with a lower dose of botanical extracts are desirable to achieve both effective concentrations and to reduce side effects. In this study, we have found that a mixture of oregano and cranberry at 2% in food has a prolongevity effect. This mixture is more potent in terms of antimicrobial activity compared with either cranberry or oregano extract alone (30,43). Our findings suggest that the prolongevity effect of OC may be at least partially due to the synergy of two botanical extracts. However, it remains to be determined whether an oregano or cranberry extract alone at the concentrations tested in this study produces the prolongevity effect. Although it is hard to estimate effective dosages for human consumption based on our current study due to physiological differences between flies and humans, this study supports health benefits of consuming fruit-enriched diets. Go to: Acknowledgments We would like to thank A. Oropeza, R. Bustamente, E. de Leon, S. Salgado, S. Rodriguez, R. Rincon, and G. Rodas for dedicated technical work and the Moscamed-Moscafrut facility in Tapachula, Mexico, for flies and laboratory space. We also want to thank Dr Peter Rapp, Edwards Spangler, and Dr Luc Poirier for critical reading of the manuscript and valuable suggestions. Go to: References 1. Son TG, Camandola S, Mattson MP. Hormetic dietary phytochemicals. Neuromolecular Med. 2008;10:236–246. [PMC free article] [PubMed] 2. Liu RH. Potential synergy of phytochemicals in cancer prevention: mechanism of action. J Nutr. 2004;134:3479S–3485S. [PubMed] 3. Ferguson LR, Philpott M. Cancer prevention by dietary bioactive components that target the immune response. Curr Cancer Drug Targets. 2007;7:459–464. [PubMed] 4. Spencer JP. Flavonoids: modulators of brain function? Br J Nutr. 2008;99(E suppl 1):ES60–ES77. [PubMed] 5. Stevenson DE, Hurst RD. Polyphenolic phytochemicals—just antioxidants or much more? Cell Mol Life Sci. 2007;64:2900–2916. [PubMed] 6. Baser KH. Biological and pharmacological activities of carvacrol and carvacrol bearing essential oils. Curr Pharm Des. 2008;14:3106–3119. [PubMed] 7. Neto CC. Cranberry and its phytochemicals: a review of in vitro anticancer studies. J Nutr. 2007;137:186S–193S. [PubMed] 8. Lukas B, Schmiderer C, Franz C, Novak J. Composition of essential oil compounds from different Syrian populations of Origanum syriacum L. (Lamiaceae) J Agric Food Chem. 2009;57:1362–1365. [PubMed] 9. Kordali S, Cakir A, Ozer H, Cakmakci R, Kesdek M, Mete E. Antifungal, phytotoxic and insecticidal properties of essential oil isolated from Turkish Origanum acutidens and its three components, carvacrol, thymol and p-cymene. Bioresour Technol. 2008;99:8788–8795. [PubMed] 10. Savini I, Arnone R, Catani MV, Avigliano L. Origanum vulgare induces apoptosis in human colon cancer caco2 cells. Nutr Cancer. 2009;61:381–389. [PubMed] 11. Srihari T, Sengottuvelan M, Nalini N. Dose-dependent effect of oregano (Origanum vulgare L.) on lipid peroxidation and antioxidant status in 1,2-dimethylhydrazine-induced rat colon carcinogenesis. J Pharm Pharmacol. 2008;60:787–794. [PubMed] 12. Ozdemir B, Ekbul A, Topal NB, et al. Effects of Origanum onites on endothelial function and serum biochemical markers in hyperlipidaemic patients. J Int Med Res. 2008;36:1326–1334. [PubMed] 13. Kwon YI, Vattem DA, Shetty K. Evaluation of clonal herbs of Lamiaceae species for management of diabetes and hypertension. Asia Pac J Clin Nutr. 2006;15:107–118. [PubMed] 14. Camici GG, Sudano I, Noll G, Tanner FC, Luscher TF. Molecular pathways of aging and hypertension. Curr Opin Nephrol Hypertens. 2009;18:134–137. [PubMed] 15. Mauch C, Bilkei G. Strategic application of oregano feed supplements reduces sow mortality and improves reproductive performance—a case study. J Vet Pharmacol Ther. 2004;27:61–63. [PubMed] 16. He X, Liu RH. Cranberry phytochemicals: isolation, structure elucidation, and their antiproliferative and antioxidant activities. J Agric Food Chem. 2006;54:7069–7074. [PubMed] 17. McKay DL, Blumberg JB. Cranberries (Vaccinium macrocarpon) and cardiovascular disease risk factors. Nutr Rev. 2007;65:490–502. [PubMed] 18. Howell AB. Bioactive compounds in cranberries and their role in prevention of urinary tract infections. Mol Nutr Food Res. 2007;51:732–737. [PubMed] 19. Jepson RG, Craig JC. A systematic review of the evidence for cranberries and blueberries in UTI prevention. Mol Nutr Food Res. 2007;51:738–745. [PubMed] 20. Jepson RG, Craig JC. Cranberries for preventing urinary tract infections. Cochrane Database Syst Rev. 2008 CD001321. [PubMed] 21. McMurdo ME, Argo I, Phillips G, Daly F, Davey P. Cranberry or trimethoprim for the prevention of recurrent urinary tract infections? A randomized controlled trial in older women. J Antimicrob Chemother. 2009;63:389–395. [PMC free article] [PubMed] 22. Howell AB, Reed JD, Krueger CG, Winterbottom R, Cunningham DG, Leahy M. A-type cranberry proanthocyanidins and uropathogenic bacterial anti-adhesion activity. Phytochemistry. 2005;66:2281–2291. [PubMed] 23. Neto CC, Amoroso JW, Liberty AM. Anticancer activities of cranberry phytochemicals: an update. Mol Nutr Food Res. 2008;52(suppl 1):S18–S27. [PubMed] 24. Lee IT, Chan YC, Lin CW, Lee WJ, Sheu WH. Effect of cranberry extracts on lipid profiles in subjects with Type 2 diabetes. Diabet Med. 2008;25:1473–1477. [PubMed] 25. Hochman N, Houri-Haddad Y, Koblinski J, et al. Cranberry juice constituents impair lymphoma growth and augment the generation of antilymphoma antibodies in syngeneic mice. Nutr Cancer. 2008;60:511–517. [PubMed] 26. Liu M, Lin LQ, Song BB, et al. Cranberry phytochemical extract inhibits SGC-7901 cell growth and human tumor xenografts in Balb/c nu/nu mice. J Agric Food Chem. 2009;57:762–768. [PubMed] 27. Joseph JA, Fisher DR, Carey AN. Fruit extracts antagonize Abeta- or DA-induced deficits in Ca2+ flux in M1-transfected COS-7 cells. J Alzheimers Dis. 2004;6:403–411. discussion 443–409. [PubMed] 28. Apostolidis E, Kwon YI, Shetty K. Potential of cranberry-based herbal synergies for diabetes and hypertension management. Asia Pac J Clin Nutr. 2006;15:433–441. [PubMed] 29. Seeram NP, Adams LS, Hardy ML, Heber D. Total cranberry extract versus its phytochemical constituents: antiproliferative and synergistic effects against human tumor cell lines. J Agric Food Chem. 2004;52:2512–2517. [PubMed] 30. Lin YT, Kwon YI, Labbe RG, Shetty K. Inhibition of Helicobacter pylori and associated urease by oregano and cranberry phytochemical synergies. Appl Environ Microbiol. 2005;71:8558–8564. [PMC free article] [PubMed] 31. Wilson MA, Shukitt-Hale B, Kalt W, Ingram DK, Joseph JA, Wolkow CA. Blueberry polyphenols increase lifespan and thermotolerance in Caenorhabditis elegans. Aging Cell. 2006;5:59–68. [PMC free article] [PubMed] 32. Allard JS, Perez E, Zou S, de Cabo R. Dietary activators of Sirt1. Mol Cell Endocrinol. 2009;299:58–63. [PMC free article] [PubMed] 33. Baur JA, Pearson KJ, Price NL, et al. Resveratrol improves health and survival of mice on a high-calorie diet. Nature. 2006;444:337–342. [PubMed] 34. Baur JA, Sinclair DA. Therapeutic potential of resveratrol: the in vivo evidence. Nat Rev Drug Discov. 2006;5:493–506. [PubMed] 35. Pearson KJ, Baur JA, Lewis KN, et al. Resveratrol delays age-related deterioration and mimics transcriptional aspects of dietary restriction without extending life span. Cell Metab. 2008;8:157–168. [PMC free article] [PubMed] 36. Bass TM, Weinkove D, Houthoofd K, Gems D, Partridge L. Effects of resveratrol on lifespan in Drosophila melanogaster and Caenorhabditis elegans. Mech Ageing Dev. 2007;128:546–552. [PubMed] 37. Kaeberlein M, Kennedy BK. Does resveratrol activate yeast Sir2 in vivo? Aging Cell. 2007;6:415–416. [PubMed] 38. Zou S, Carey JR, Liedo P, et al. The prolongevity effect of resveratrol depends on dietary composition and calorie intake in a tephritid fruit fly. Exp Gerontol. 2009;44:472–476. [PMC free article] [PubMed] 39. Harrison GG. Proceedings of the workshop on food-consumption surveys in developing countries: methodologic considerations in descriptive food-consumption surveys in developing countries. Food Nutr Bull. 2004;25:415–419. [PubMed] 40. Carey JR, Liedo P, Muller HG, Wang JL, Senturk D, Harshman L. Biodemography of a long-lived tephritid: reproduction and longevity in a large cohort of female Mexican fruit flies, Anastrepha ludens. Exp Gerontol. 2005;40:793–800. [PMC free article] [PubMed] 41. Carey JR, Harshman LG, Liedo P, Muller HG, Wang JL, Zhang Z. Longevity-fertility trade-offs in the tephritid fruit fly, Anastrepha ludens, across dietary-restriction gradients. Aging Cell. 2008;7:470–477. [PMC free article] [PubMed] 42. Zou S, Sinclair J, Wilson MA, et al. Comparative approaches to facilitate the discovery of prolongevity interventions: effects of tocopherols on lifespan of three invertebrate species. Mech Ageing Dev. 2007;128:222–226. [PMC free article] [PubMed] 43. Lin YT, Labbe RG, Shetty K. Inhibition of Listeria monocytogenes in fish and meat systems by use of oregano and cranberry phytochemical synergies. Appl Environ Microbiol. 2004;70:5672–5678. [PMC free article] [PubMed] 44. Cao G, Alessio HM, Cutler RG. Oxygen-radical absorbance capacity assay for antioxidants. Free Radic Biol Med. 1993;14:303–311. [PubMed] 45. Slinkard K, Singleton VL. Total phenol analysis—automation and comparison with manual methods. Am J Enol Vitic. 1977;28:49–55. 46. Partridge L, Pletcher SD, Mair W. Dietary restriction, mortality trajectories, risk and damage. Mech Ageing Dev. 2005;126:35–41. [PubMed] 47. Everitt AV, Hilmer SN, Brand-Miller JC, et al. Dietary approaches that delay age-related diseases. Clin Interv Aging. 2006;1:11–31. [PMC free article] [PubMed] 48. Fernandes G. Progress in nutritional immunology. Immunol Res. 2008;40:244–261. [PubMed] 49. Masoro EJ. Dietary restriction-induced life extension: a broadly based biological phenomenon. Biogerontology. 2006;7:153–155. [PubMed] 50. Mattison JA, Roth GS, Lane MA, Ingram DK. Dietary restriction in aging nonhuman primates. Interdiscip Top Gerontol. 2007;35:137–158. [PubMed] 51. Lee KP, Simpson SJ, Clissold FJ, et al. Lifespan and reproduction in Drosophila: new insights from nutritional geometry. Proc Natl Acad Sci U S A. 2008;105:2498–2503. [PMC free article] [PubMed] 52. Skorupa DA, Dervisefendic A, Zwiener J, Pletcher SD. Dietary composition specifies consumption, obesity, and lifespan in Drosophila melanogaster. Aging Cell. 2008;7:478–490. [PMC free article] [PubMed] 53. Carey JR, Liedo P, Harshman L, et al. A mortality cost of virginity at older ages in female Mediterranean fruit flies. Exp Gerontol. 2002;37:507–512. [PubMed] 54. Carey JR, Liedo P, Muller HG, et al. Female sensitivity to diet and irradiation treatments underlies sex-mortality differentials in the Mediterranean fruit fly. J Gerontol A Biol Sci Med Sci. 2001;56:B89–B93. [PubMed] 55. Carey JR, Liedo P, Vaupel JW. Mortality dynamics of density in the Mediterranean fruit fly. Exp Gerontol. 1995;30:605–629. [PubMed] 56. Phang JM, Donald SP, Pandhare J, Liu Y. The metabolism of proline, a stress substrate, modulates carcinogenic pathways. Amino Acids. 2008;35:681–690. [PubMed] 57. Wood JM. Membrane association of proline dehydrogenase in Escherichia coli is redox dependent. Proc Natl Acad Sci U S A. 1987;84:373–377. [PMC free article] [PubMed] 58. Dillin A, Hsu AL, Arantes-Oliveira N, et al. Rates of behavior and aging specified by mitochondrial function during development. Science. 2002;298:2398–2401. [PubMed] 59. Gray MW. The evolutionary origins of organelles. Trends Genet. 1989;5:294–299. [PubMed] 60. Lee SS, Lee RY, Fraser AG, Kamath RS, Ahringer J, Ruvkun G. A systematic RNAi screen identifies a critical role for mitochondria in C. elegans longevity. Nat Genet. 2003;33:40–48. [PubMed] 61. McMillin JB, Pauly DF. Control of mitochondrial respiration in muscle. Mol Cell Biochem. 1988;81:121–129. [PubMed] 62. Ungvari Z, Krasnikov BF, Csiszar A, et al. Testing hypotheses of aging in long-lived mice of the genus Peromyscus: association between longevity and mitochondrial stress resistance, ROS detoxification pathways, and DNA repair efficiency. Age (Dordr) 2008;30:121–133. [PMC free article] [PubMed] E. Coli and other microbes are becoming increasingly resistant to the reliable active antibiotics available to fight them. These clinical results show that about 65% of the E. Coli isolates are multi-drug resistant (meaning they are resistant to 3 or more drugs). [Antibiotic resistance of E. coli isolates from urine samples of Urinary Tract Infection (UTI) patients in Pakistan Saghir Ahmad Jafri,1 Muhammad Qasim,2,* Muhammad S Masoud,2 Mahmood-ur- Rahman,2 Mateen Izhar,3 and Saqib Kazmi4 ] Carvacrol inhibits the growth of several bacteria strains, e.g. Escherichia coli[6] and Bacillus cereus. Its low toxicity together with its pleasant taste and smell suggests its use as a food additive to prevent bacterial contamination.[7] In Pseudomonas aeruginosa it causes damages to the cell membrane of these bacteria and, unlike other terpenes, inhibits their proliferation.[8] The cause of the antimicrobial properties is believed to be disruption of the bacteria membrane.[9][10] It is a potent activator of the human ion channels transient receptor potential V3 (TRPV3) and A1 (TRPA1).[11] Application of carvacrol on the human tongue, as well as activation of TRPV3, causes a sensation of warmth. In addition, carvacrol also activates, but then rapidly desensitizes, the pain receptor TRPA1; this explains its pungency.[11] It activates PPAR and suppresses COX-2 inflammation.[12] In rats, carvacrol is quickly metabolized and excreted. The main metabolic route is esterification of the phenolic group with sulfuric acid and glucuronic acid. A minor pathway is oxidation of the terminal methyl groups to primary alcohols. After 24 hours, only very small amounts of carvacrol or its metabolites could be found in urine, indicating an almost complete excretion within one day.[13] A study led by Supriya Bavadekar reports that carvacrol stimulates apoptosis in prostate cancer cells.[14] 1 “Carvacrol data sheet from Sigma-Aldrich”. 2 Jump up 
^ Lide, David R. (1998). Handbook of Chemistry and Physics (87 ed.). Boca Raton, FL: CRC Press. pp. 3–346. ISBN 0-8493-0594-2. 3 Jump up 
^ Ultee A, Slump RA, Steging G, Smid EJ (2000). “Antimicrobial activity of carvacrol toward Bacillus cereus on rice”. J. Food Prot. 63 (5): 620–4. PMID 10826719. 4 Jump up 
^ De Vincenzi M, Stammati A, De Vincenzi A, Silano M (2004). “Constituents of aromatic plants: carvacrol”. Fitoterapia 75 (7–8): 801–4. doi:10.1016/j.fitote.2004.05.002. PMID 15567271. 5 Jump up 
^ Characterization of volatile compounds from ethnic Agave alcoholic beverages by gas chromatography-mass spectrometry. León-Rodríguez, A. de, Escalante-Minakata, P., Jiménez-García, M. I., Ordoñez-Acevedo, L. G., Flores Flores, J. L. and Barba de la Rosa, A. P., Food Technology and Biotechnology, 2008, Volume 46, Number 4, pages 448-455 (abstract) 6 Jump up 
^ Du WX, Olsen C. E., Avena-Bustillos R. J., McHugh T. H., Levin C. E., Friedman M. (2008). “Storage Stability and Antibacterial Activity against Escherichia coli O157:H7 of Carvacrol in Edible Apple Films Made by Two Different Casting Methods”. J. Agric. Food Chem. 56 (9): 3082–8. doi:10.1021/jf703629s. PMID 18366181. 7 Jump up 
^ Ultee A., Smid E. J. (2001). “Influence of carvacrol on growth and toxin production by Bacillus cereus”. Int. J. Food Microbiol. 64 (3): 373–8. doi:10.1016/S0168-1605(00)00480-3. PMID 11294360. 8 Jump up 
^ Cox SD, Markham JL (2007). “Susceptibility and intrinsic tolerance of Pseudomonas aeruginosa to selected plant volatile compounds”. J. Appl. Microbiol. 103 (4): 930–6. doi:10.1111/j.1365-2672.2007.03353.x. PMID 17897196. 9 Jump up 
^ Di Pasqua R, Betts G, Hoskins N, Edwards M, Ercolini D, Mauriello G (2007). “Membrane toxicity of antimicrobial compounds from essential oils”. J. Agric. Food Chem. 55 (12): 4863–70. doi:10.1021/jf0636465. PMID 17497876. 10 Jump up 
^ Cristani M, D’Arrigo M, Mandalari G; et al. (2007). “Interaction of four monoterpenes contained in essential oils with model membranes: implications for their antibacterial activity”. J. Agric. Food Chem. 55 (15): 6300–8. doi:10.1021/jf070094x. PMID 17602646. 11 ^ Jump up to: 
a b Xu H, Delling M, Jun JC, Clapham DE (2006). “Oregano, thyme and clove-derived flavors and skin sensitizers activate specific TRP channels”. Nat. Neurosci. 9 (5): 628–35. doi:10.1038/nn1692. PMID 16617338. 12 Jump up 
^ Hotta, M.; Nakata, R.; Katsukawa, M.; Hori, K.; Takahashi, S.; Inoue, H. (2010). “Carvacrol, a Component of Thyme Oil, Activates PPAR and Suppresses COX-2 Expression”. Journal of Lipid Research 51: 132–9. doi:10.1194/jlr.M900255-JLR200. 13 Jump up 
^ Austgulen LT, Solheim E, Scheline RR (1987). “Metabolism in rats of p-cymene derivatives: carvacrol and thymol”. Pharmacol. Toxicol. 61 (2): 98–102. doi:10.1111/j.1600-0773.1987.tb01783.x. PMID 2959918. 14 Jump up 
^ http://www.liu.edu/Brooklyn/About/News/Press-Releases/2012/April/BK-PR-Apr25-2012.aspx 15 Jump up 
^ http://www.hort.purdue.edu/newcrop/proceedings1993../V2-628.html 16 Jump up 
^ http://www.herbapolonica.pl/magazines-files/2733672-art.3.pdf 17 ^ Jump up to: 
a b Bouchra, Chebli; Achouri, Mohamed; Idrissi Hassani, L.M; Hmamouchi, Mohamed; et al. (2003). “Chemical composition and antifungal activity of essential oils of seven Moroccan Labiatae against Botrytis cinerea Pers: Fr”. Journal of Ethnopharmacology 89 (1): 165–169. doi:10.1016/S0378-8741(03)00275-7. PMID 14522450. 18 Jump up 
^ Liolios, C.C.; Gortzi, O; Lalas, S; Tsaknis, J; Chinou, I; et al. (2009). “Liposomal incorporation of carvacrol and thymol isolated from the essential oil of Origanum dictamnus L. and in vitro antimicrobial activity”. Food Chemistry 112 (1): 77–83. doi:10.1016/j.foodchem.2008.05.060. 19 ^ Jump up to: 
a b Aligiannis, N.; Kalpoutzakis, E.; Mitaku, Sofia; Chinou, Ioanna B.; et al. (2001). “Composition and Antimicrobial Activity of the Essential Oils of Two Origanum Species”. J. Agric. Food Chem. 49 (9): 4168–4170. doi:10.1021/jf001494m. PMID 11559104. 20 Jump up 
^ Coskun, Sevki; Girisgin, O; Kürkcüoglu, M; Malyer, H; Girisgin, AO; Kirimer, N; Baser, KH; et al. (2008). “Acaricidal efficacy of Origanum onites L. essential oil against Rhipicephalus turanicus (Ixodidae)”. Parasitology Research 103 (2): 259–261. doi:10.1007/s00436-008-0956-x. PMID 18438729. 21 Jump up 
^ Ruberto, Giuseppe; Biondi, Daniela; Meli, Rosa; Piattelli, Mario; et al. (2006). “Volatile flavour components of Sicilian Origanum onites L”. Flavour and Fragrance Journal 8 (4): 197–200. doi:10.1002/ffj.2730080406. 22 Jump up 
^ Kanias, G. D.; Souleles, C.; Loukis, A.; Philotheou-Panou, E.; et al. (1998). “Trace elements and essential oil composition in chemotypes of the aromatic plant Origanum vulgare”. Journal of Radioanalytical and Nuclear Chemistry 227 (1 – 2): 23–31. doi:10.1007/BF02386426. 23 Jump up 
^ Figiel, Adam; Szumny, Antoni; Gutiérrez-Ortíz, Antonio; Carbonell-Barrachina, ÁNgel A.; et al. (2010). “Composition of oregano essential oil (Origanum vulgare) as affected by drying method”. Journal of Food Engineering 98 (2): 240–247. doi:10.1016/j.jfoodeng.2010.01.002. 24 Jump up 
^ Özkan, Aysun; Erdoğan, Ayşe (2010). “A comparative evaluation of antioxidant and anticancer activity of essential oil from Origanum onites (Lamiaceae) and its two major phenolic components”. Tübitak 35 (2011): 735–742. doi:10.3906/biy-1011-170. 25 ^ Jump up to: 
a b c d American College of Toxicology (2006). “Final Report on the Safety Assessment of Sodium p-Chloro-m-Cresol, p-Chloro-m-Cresol, Chlorothymol, Mixed Cresols, m-Cresol, o-Cresol, p-Cresol, Isopropyl Cresols, Thymol, o-Cymen-5-ol, and Carvacrol”. International Journal of Toxicology 25: 29–127. doi:10.1080/10915810600716653. PMID 16835130. Jump up 
^ The British Pharmacopoeia Secretariat (2009). “Index, BP 2009” (PDF). Retrieved 29 March 2010. “The recommended doses of cranberry products for the prevention of UTIs have been poorly defined, and beverage formulations vary widely. The most highly studied formulation has 25% pure juice (13). Clinical research suggests that daily dosages of 240–300 ml of cranberry juice cocktail can prevent 50% of the recurrences of UTIs and can reduce bacteriuria (11,15,16,30). An ex vivo study examining human urine following cranberry juice cocktail consumption suggests that twice-daily dosages of cranberries (36 mg of PAC) might offer additional protection during a 24 h period (25,28). Recommended doses of dried, concentrated juice extract range from 600 to >1,200 mg/day (56) divided into two or three daily doses. It is important to consider that dried cranberry extract can be broken down by exposure to light, heat or cold. However, the addition of vitamins C and E exert a stabilizing influence (57).”
[Clinics (Sao Paulo). 2012 Jun; 67(6): 661–667.
doi: 10.6061/clinics/2012(06)18
PMCID: PMC3370320
Cranberries and lower urinary tract infection prevention
Marcelo Hisano,I Homero Bruschini,I Antonio Carlos Nicodemo,II and Miguel SrougiI]

“A significant dose-dependent decrease in bacterial adherence in vitro was noted after the consumption of 108 and 36 mg of cranberry (p <0.001). The in-vivo model confirmed that E. coli strains had a reduced ability to kill C. elegans after growth in the urine of patients who consumed cranberry capsules. Overall, these in-vivo and in-vitro studies suggested that consumption of cranberry juice represents an interesting new strategy to prevent recurrent urinary tract infection.”
[Clin Microbiol Infect. 2008 Apr;14(4):350-5. doi: 10.1111/j.1469-0691.2007.01917.x. Epub 2008 Jan 7.
In-vitro and in-vivo evidence of dose-dependent decrease of uropathogenic Escherichia coli virulence after consumption of commercial Vaccinium macrocarpon (cranberry) capsules.
Lavigne JP1, Bourg G, Combescure C, Botto H, Sotto A.]

OREGANO OIL STUDIES

Oregano Oil and Detrimental Bacteria (Research)
Peer-Reviewed Professional Journals
1 Sivropolou, A., et al. Antimicrobial and cytotoxic activities of Origanum essential oils. J Agric Food Chem. 44:1202-1205, 1996.

Laypersons’ Publications
• Gormley, J. J. Oregano: more than a nice, Italian spice. Better Nutrition. 61(1):32-33, 1999.
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• Terhanian, H. Bactericidal oregano oil. Well Being Journal. 9(1):18-20, 2000.
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• When oregano oil was first tested in 1910 it was described as the most powerful plant-derived antiseptic known. It was demonstrated to be 26 times as potent as phenol.
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• Puotinen, C. J. Natural solutions to drug-resistant infections. Well Being Journal. 9(1):1-16, 2000.
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• Oregano oil is one of the most effective antiseptic essential oils for most types of infections.

Oregano Oil and Eschericia coli (Research)
Peer-Reviewed Professional Journals
• Burt, S. A., et al. Antibacterial activity of selected plant essential oils against Escherichia coli O157:H7. Lett Appl Microbiol. 36(3):162-167, 2003.
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• Department of Public Health and Food Safety, Faculty of Veterinary Medicine, University of Utrecht, Utrecht, The Netherlands.
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• The aim of this study was to quantify the antibacterial properties of five essential oils (EO) on a non-toxigenic strain of Escherichia coli O157:H7 in the presence and absence of a stabilizer and an emulsifier and at three different temperatures. Five EOs known to exhibit antibacterial properties were screened by disc diffusion assay and the most active were selected for further study in microdilution colorimetric assays. Oregano (Origanum vulgare) and thyme (Thymus vulgaris; light and red varieties) EO had the strongest bacteriostatic and bactericidal properties, followed by bay (Pimenta racemosa) and clove bud (Eugenia caryophyllata synonym: Syzygium aromaticum) EO. Oregano oil was colicidal at 625 &mgr;l l-1 at 10, 20 and 37 degrees C. The addition of 0.05% (w/v) agar as stabilizer reinforced the antibacterial properties, particularly at 10 degrees C, whereas 0.25% (w/v) lecithin reduced antibacterial activity. Scanning electron micrographs showed extensive morphological changes to treated cells. Oregano and thyme EO possess significant in vitro colicidal and colistatic properties, which are exhibited in a broad temperature range and substantially improved by the addition of agar as stabilizer. Bay and clove bud EO are less active. Lecithin diminished antibacterial properties. The bactericidal concentration of oregano EO irreversibly damaged E. coli O157:H7 cells within 1 min. Oregano and light thyme EO, particularly when enhanced by agar stabilizer, may be effective in reducing the number or preventing the growth of E. coli O157:H7 in foods.
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• Dorman, H. J. D., et al. Antimicrobial agents from plants: antibacterial activity of plant volatile oils. J Applied Microbiology. 88:308-316, 2000.
• Sivropolou, A., et al. Antimicrobial and cytotoxic activities of Origanum essential oils. J Agric Food Chem. 44:1202-1205, 1996.

Oregano Oil and Staphylococcus (Research)
Laypersons’ Publications
• Oregano oil – a promising antibacterial agent. Healthy Buzz. 5(1), 2002.
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• In an in vitro study, oregano oil was added to test tubes containing Staphylococcus bacteria and compared with the effects of streptomycin, penicillin and vancomycin. The oregano oil at relatively low doses was found to inhibit the growth of staphylococcus bacteria in the test tube as effectively as the standard antibiotics did. Another aspect of the study examined the efficacy of oregano oil and carvacrol, which is believed to be the major antibacterial component of oregano, in 18 mice infected with the Staphylococcus bacteria. Six mice received oregano oil for 30 days, and 50% of this group survived the 30-day treatment. Six received the carvacrol in olive oil, not in oregano oil, and none survived longer than 21 days. Six mice received olive oil alone with no active agents (the control group) and all died within three days. A repeat study corroborated these findings, which demonstrates that there are components of oregano oil other than carvacrol that have antibiotic properties.

Oregano Oil and Staphylococcus aureus (Research)
Peer-Reviewed Professional Journals
• Dorman, H. J. D., et al. Antimicrobial agents from plants: antibacterial activity of plant volatile oils. J Applied Microbiology. 88(2):308-316, 2000.
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• Aromatic and Medicinal Plant Group, Scottish Agricultural College, Auchincruive, South Ayrshire, UK.
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• The volatile oils of black pepper [Piper nigrum L. (Piperaceae)], clove [Syzygium aromaticum (L.) Merr. & Perry (Myrtaceae)], geranium [Pelargonium graveolens L’Herit (Geraniaceae)], nutmeg [Myristica fragrans Houtt. (Myristicaceae)], oregano [Origanum vulgare ssp. hirtum (Link) Letsw. (Lamiaceae)] and thyme [Thymus vulgaris L. (Lamiaceae)] were assessed for antibacterial activity against 25 different genera of bacteria. These included animal and plant pathogens, food poisoning and spoilage bacteria. The volatile oils exhibited considerable inhibitory effects against all the organisms under test while their major components demonstrated various degrees of growth inhibition.

Oregano Oil and Streptococcus pneumoniae (Research)
Peer-Reviewed Professional Journals
Horne, D. 98th general assembly of the American Society of Microbiology.
Streptococcus pneumoniae cells “fell apart” in the presence of oregano oil.

†These statements have not been evaluated by the FDA. This Product is not intended to diagnose, treat, cure or prevent any disease.