ACTIVE INGREDIENTS, THEIR BIOAVAILABILITY AND THE HEALTH BENEFITS OF THE PUNICA GRANATUM LINN (POMEGRANATE

)
A Research Review

Dirk Budka, M.Sc. Senior Biomedical Scientist, MSML Research Unit

© Front picture: Cleanfoods Ltd, Bangalore, India

CONTENT Preface The Claims Scientific Classification Nutritional Value Compact History Cultivars Pollination Climate Soil Propagation Culture Harvesting and Yield Keeping Quality & Storages Pest and Disease Food Uses Introduction to Health Benefits of Pomegranate Active Ingredients of the Pomegranate Antioxidants Phenolics TANNINS Tannin Table I Punicalagin Ellagic Acid Pelagonidin, Cyanidin Gallic Acid Quercetin Pomegranate and Cancer Pomegranate and Atherosclerosis page page page page page page page page page page page page page page page page page page page page page page page page page page page page page 01 02 04 04 05 08 12 14 14 15 15 15 12 16 18 18 20 22 23 25 27 29 29 30 32 33 33 34 45

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Chronic Obstructive Pulmonary Disease Osteoarthritis Increase in Sperm Quality Erectile Dysfunction Alzheimer’s Disease Menopausal Symptoms Cosmeceutical Antimicrobial

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Reference List List of Research Centers

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Correspondence Address:

MSML c/o Hale Clinic, 7 Park Crescent, London W1B 1PF Senior biomedical Scientist: Dirk Budka Senior Practitioner: Dr. Peter Gruenewald, MD dirk.budka@ntlworld.com

www.immuneclinic.com www.bacteriaclinic.com www.virusmedicalclinic.com www.parasiteclinic.com www.fungusandyeast.com www.nutritionlondon.net www.ageless-technologies.com www.ibsforum.co.uk www.stop-readymeals.com

Of course. Dirk Budka June 2008 . to treat ailments/diseases? Let’s start with the pomegranate. a lot has still to be done – a lot of research is necessary to understand the interactions between the many active ingredients and the interactions between pharmaceuticals and the active ingredients in pomegranate. According to the WHO (World Health Organization). Novel treatments are essential… and what’s wrong in using natural products. The Pomegranate was once named the “most medicinal fruit in the world” and you will read in this paper about the many health benefits of this fruit. inexpensive and available in the next shop. mycologist and parasitologist are just at the beginning of the understanding of the microbial world. medical plants would be the best source to obtain a variety of drugs.-1- PREFACE Plants are a valuable source of natural products for maintaining human health and the use of plant compounds for pharmaceutical purposes has increased. Virologist. bacteriologist.

It can protect/is protective against Prostate Cancer Lung Cancer Colon Cancer Skin Cancer Eosophageal Cancer Diabetes Osteoarthritis Hypercholesterolemia Atherosclerosis Obstructive Pulmonary Disease . This study is looking at the research. THEIR BIOAVAILABILITY AND THE HEALTH BENEFITS OF THE PUNICA GRANATUM LINN (POMEGRANATE) Many claims have been made over the past decade regarding the health benefits of the Pomegranate. scientific studies and the proven and non-proven claims. once named “the most medicinal fruit in the world”. .-2- ACTIVE INGREDIENTS. fungi and parasites. The claims: . viruses.Pomegranate has antimicrobial properties and can therefore be protective and/or fights off bacteria.

Pomegranate helps to stabilize/increase sperm quality . Scientific Classification Kingdom Division Class Subclass Order Family Genus Species Plantae Magnoliophyta Magnoliopsida Rosidae Myrtales Lythraceae Punica P.it has anti-malarial properties. it protects the neonatal brain against hypoxic-ischemic injury and is classified within the cosmetic industry as a Cosmeceutical (combining a feature of both cosmetic and pharmaceutical).-3- - Alzheimer’s Disease Tuberculosis Macular Degeneration and Vision Loss Erectile Dysfunction Chronic inflammation (arthritis and cystic fibrosis) Menopausal Symptoms Anti-histaminic . granatum .

dietary fibre Fat Protein Thiamin (B1) Riboflavin (B2) Niacin (B3) Pantothenic acid (B5) Vitamin B6 Folate (B9) Vitamin C Calcium Iron Magnesium Phosphorus Potassium Zinc 16.17 g 0.95 g 0.30 mg 3 mg 8 mg 259 mg 0.030 mg 0.-4- The Nutritional Value per 100 g of the Pomegranate (aril only) Carbohydrates -sugars .596 mg 0.063 mg 0.60 g 17.105 mg 6 µg 3 mg 3 mg 0.300 mg 0.30 g 0.12 mg Source: USDA Nutrient Database .57 g 0.

b) CORONARY HEART DISEASE: The effects of pomegranate have been studied in patients suffering from CHD and myocardial ischemia.-5- COMPACT a) ANTI ATHEROGENIC: Pomegranate significantly reduces oxidative stress by inhibiting the formation of oxidized LDL lipoproteins and macrophage lipid peroxidation. One group was given 240 ml pomegranate juice daily for three months. while the other group drank a beverage of similar caloric content. After three months. c) CAROTID ARTERY STENOSIS: Consumption for three years of pomegranate juice by atherosclerotic patients with carotid artery stenosis reduced blood pressure and LDSL oxidation. the extent of stress induced ischemia decreased in the pomegranate group and increased in the control-group.5 mmol of total polyphenols per day for two weeks) by hypertensive patients showed a 36% decrease in serum angiotensin converting enzyme (ACE) activity and a 5% reduction in systolic blood pressure. They were randomly assigned into two groups. The pomegranate juice decreases LDL susceptibility to aggregation and retention. . flavour and colour. and suppresses oxidized LDL degration and cholesterol biosynthesis in macrophages. increases the activity of serum paraoxinase (a HDL esterase). that can lead to reduced cellular cholesterol accumulation and foam cell formation. d) HYPERTENSION: Pomegranate juice consumption (50 ml/1. amount.

The effect of concentrated pomegranate juice consumption (40g/daily for eight weeks) on lipid profiles of type II diabetic patients (14 female/8male) with hyperlipidemia was also evaluated. g) CANCER. Luteolin and Punicic acid were tested in vitro as inhibitors on the invasion of human PC-3 prostate cancer cells. inhibiting the growth of the cells through induction of apoptosis. The interaction between pomegranate . Antioxidant compounds – present in the diet – are considered chemo-preventative and chemotherapeutic agents. pomegranate extract had significant cytotoxic and growth inhibition effects on human breast cancer cells. but resulted in antioxidative effects on serum and macrophages. and lipophilic fractions prepared from pomegranate seed promoted regeneration of epidermis in human skin cells in laboratory conditions. COLON: Pomegranate juice suppressed inflammatory cell signalling in the HT-29 human colon cancer cell line. LDLcholesterol. Caffeic acid. f) SKIN: Pomegranate aqueous extract promotes regeneration of the dermis. PROSTATE CANCER: Ellagic acid. h) ANTIMICROBIAL: Pomegranate seeds have potent antimicrobial activities against bacteria and fungi/yeasts. Pomegranate juice significantly reduced the level of total cholesterol.-6- e) DIABETES: Pomegranate juice consumption (50 ml/day for three months) by diabetic patients did not worsen the diabetic parameters such as serum glucose. All compounds significantly inhibited the cell-invasion when they were employed individually. BREAST CANCER: In laboratory conditions. cholesterol and triglyceride levels.

(Salbutamol inhaler brought on an immediate positive response). Nevertheless. tetracycline. 13 had sensitivity to pollen. One case was very interesting. Not only was bacterial growth tested. j) ADVERSE REACTION: One person developed a late-onset tongue-oedema due to pomegranate intake as proven by a double blind oral challenge test. demonstrates that pomegranate extract dramatically enhances the activity of all antibiotics tested. but also bacterial enterotoxin production and it was shown that staphylococcal enterotoxin production was inhibited. her mother had frequently eaten this fruit while breastfeeding the daughter. 10 to nuts and 8 to peaches. Pomegranate may increase the risk of rhabdomyolysis during rosuvastatin treatment for McArdle disease (serious muscle damage). In this case. Another case relates to a 7-year-old asthmatic child who showed clinical conditions of bronchospasm. maternal milk could have been the source of the child’s sensitization. ampicillin.-7- i) (methanolic extract) and the antibiotics chloramphenicol. gentamicin. . because the female patient stated that she never ate pomegranate before. moments after ingesting several pomegranate seeds. Pomegranate juice was also screened for inhibitory activity against HIV-1 IIIB using CD4 and CXCR4 as cell receptors. and oxacillin against 30 clinical isolates of methicillin-resistant and methicillin-sensitive Staphylococcus aureus. In a study of 15 patients allergic to pomegranate.

The Pomegranate has appeared throughout history in some of the greatest documents and art and architecture. to Shakespeare and Raphael. from the mysterious ‘Tree of Life’. Moving way back in time. and appearing in Greek mythology. The ancient Chinese believed the seeds symbolized longevity and immortality. pomegranates appear in many contexts. The pomegranate tree is said to have flourished in the Garden of Eden and is very likely the “apple” of the Adam and Eve story in Genesis. from Homer and Chaucer. cited in the Old Testament as ‘rimmon’. Well before the Christian era.-8HISTORY Photo: Courtesy of OZ Granate PTY Ltd. they were introduced into China from Samarkhand. in Roman history and in the Koran. regeneration. and to Cezanne in more modern times. cultural and religious. Tree of Life Symbolism of the Pomegranate The fruit is mentioned by various cultures and religions. and marriage. we find the Pomegranate celebrated in Egyptian papyri. In Judaism. . Greek and Persian mythology mentions the fruit as representing life.

A large. The cultivation of the pomegranate has a long history in Armenia. While the pomegranate originated generally from Persia (Iran) and has been cultivated in Central Asia. It has always been a symbol of many virtues. In fact. and as a decoration on the robes of priests. Armenia and the Mediterranean region for several millennia. In Georgia. Carbonized pips and pieces of the peel of the fruit have been identified in Early Bronze Age levels of Jericho.-9- The fruit is also a symbol of resurrection and life in Christianity. Georgia. and it is one of the three “blessed fruits” in Buddhism. as well as a likeness to female breasts. a thimble-sized. decayed remains of pomegranates dating back to 1000 BC have been found in the country. there are wild pomegranate groves outside of ancient abandoned settlements. ivory pomegranate bearing an ancient Hebrew inscription is the only relic ever recovered from the Solomon’s Temple. as a regal symbol for Kings and Queens. The juice has been compared to blood. The Pomegranate was used as a decoration in the Temple of Solomon. and Armenia to the east of the Black Sea. and a spiritual dimension as well. dry pomegranate was found in the tomb of Djehuty. and the obvious ‘crown’ has led to royal connections. including love and fertility in particular. as well as Late Bronze Age levels of Hala Sultan Tekke on Cyprus and Tiryns. The abundance of seeds suggested all these virtues. and health and abundance. Mesopotamian cuneiform records mention . the butler of Queen Hatshepsut.

even New England. “Plant it against the side of thy house. 1762. It succeeded in the South: Bartram received a barrel of pomegranates and oranges from a correspondent in Charleston. 1764.10 - pomegranates from the mid-Third millennium BC onwards. death. but in the English colonies it was less at home: “Don’t use the pomegranate inhospitably. South Carolina. The ancient city of Granada in Spain was renamed after the fruit during the Moorish period. it is a symbol of hope. and flowers beautifully. whether originally spread along the route of the Silk Road or brought by sea traders. by John Tradescant the elder. a stranger that has come so far to pay his respects to thee” the English Quaker Peter Collinson wrote to the botanizing John Bartram in Philadelphia. I have twenty-four on one tree… Doctor Fothergill says. and eternity and was an emblem of the Eleusinian Mysteries. the pomegranate is still prized both as a medication and as a symbol of beauty. and bears fruit this hot year. Thomas Jefferson planted pomegranates at Monticello in 1771: he had them from George Wythe of Williamsburg In the Mesopotamian region. the pomegranate came to symbolize fertility. but the disappointment that it did not set fruit there led to its repeated introduction to the American colonies. Spanish colonists later introduced the fruit to the Caribbean and Latin America. In Christian art. of all trees this is most salutiferous to mankind. In this manner it thrives wonderfully with us. fertility and wisdom worldwide.. . It is also extensively grown in South China and in Southeast Asia. longevity. nail it close to the wall.”The pomegranate had been introduced as an exotic to England the previous century. Because of its role in the Greek legend of Persephone.

It travelled to central and southern India from Iran about the first century A. Afghanistan. In California. Production declined from lack of demand in the 1930's but new plantings were made when demand increased in the 1960's. Iran. the East Indies and tropical Africa. Iraq. Africa and Europe. It is grown for its fruit mostly in the dry zones of that state and Arizona. commercial pomegranate cultivation is concentrated in Tulare. and it was carried by desert caravans for the sake of its thirstquenching juice. There were 2. The most important growing regions are Egypt. Many people grow it at cool altitudes in the interior of Honduras. The fruit was used in many ways as it is today and was featured in Egyptian mythology and art. with small plantings in Imperial and Riverside counties. Burma and Saudi Arabia. and was reported growing in Indonesia in 1416. It has been widely cultivated throughout India and drier parts of southeast Asia. praised in the Old Testament of the Bible and in the Babylonian Talmud. India. China. Pakistan.. West Indies and warm areas of South and Central America. but only occasionally seen in the Bahamas. Fresno and Kern counties. In Mexico it is frequently planted.D. Malaya. It is rather commonly planted and has become naturalized in Bermuda where it was first recorded in 1621.000 acres (810 ha) of hearing trees in these areas in the 1920's. and it is sometimes found in gardens in Hawaii.11 - The pomegranate tree is native from Iran to the Himalayas in northern India and has been cultivated since ancient times throughout the Mediterranean region of Asia. . The tree was introduced in California by Spanish settlers in 1769. Bangladesh. There are some commercial orchards in Israel on the coastal plain and in the Jordan Valley.

Types with relatively soft seeds are often classed as "seedless". 'Kandhari' is large. In India there are several named cultivars. non-suckering. subacid pulp. sweet. or all-over greenish-white. Among the best are 'Bedana' and 'Kandhari'. . very hard seeds. pulp. sweet. with brownish or whitish rind. Others include: 'Alandi' ('Vadki')–medium-sized. with fleshy red or pink. Preference is usually given those with fleshy. 'Kabul'–large. 'Aswad'. purplish-white or white. with deep-pink or blood-red. dark-red. while the sweeter. The plant is evergreen. 'Wonderful' and 'Red Loufani' are often grown in the Jewish sector of Israel. less tangy 'Malissi' and 'Ras el Baghl'.. are favored in the Arab sector. fleshy. with dark-red and pale-yellow rind. 'Halwa' are important in Iraq. 'Bedana' is medium to large.12 - Cultivars There is little information available on the types grown in the Near East. and 'Mangulati' in Saudi Arabia. deep-red. slightly bitter pulp. hard seeds. fleshy. juicy pulp around the seeds. subacid pulp and hard seeds. yellow-red. pulp pinkish-white. seeds soft. except that the cultivars 'Ahmar'. with patches of dark-pink and purple at base. 'Dholka'–large. thick rind. sweet. desirable for commercial purposes in Delhi.

Plant is vigorous and productive. medium-hard seeds. juicy. sweet pulp. with dark-red. with medium-thick rind. with thin or fairly thick rind. fleshy. reddish or pink. 'Muscat White'–large. creamcolored. bright-red. with very thin rind. juicy. thin rind. with thin rind. 'Spanish Ruby'–round. and small to medium. juicy pulp and mediumhard seeds. sweet. fleshy.13 - 'Muscat Red'–small to medium. and orange-red or pink-and-red pulp. winey pulp. seeds medium-hard. . fleshy. medium to large. The plant is a moderately prolific bearer. 'Paper Shell'–round. The fruit is oblate. soft or medium-hard seeds. gray or grayish-green rind. 'Vellodu'–medium to large. with medium-thick rind. very juicy pulp and soft seeds. mediumsweet pulp. fleshy. 'Poona'–large. Bears heavily. sweet. 'Wonderful'–originated as a cutting in Florida and propagated in California in 1896. Desirable for commercial planting in Delhi. Considered medium in quality. small to medium or large. aromatic pulp. very large. pale-yellow blushed with pink. sometimes spotted. dark purple-red. deep-red. fairly soft seeds. fleshy. creamy-white tinged with pink. Bears well.. rose-colored.

red. 6 to 20% of the pollen may be infertile. in male. scarlet bordered and streaked with yellowish-white. 'Spanish Ruby' and 'Sweet Fruited' were the leading cultivars in the past century. Among other ornamental cultivars are 'Multiplex' with double. 'Mme. orange-red. the fruit only 2 in (5 cm) wide but borne abundantly. is especially hardy and widely grown as an ornamental in pots. Self-pollination of bagged flowers has resulted in 45% fruit set. Cross-pollination has increased yield to 68%. The size and fertility of the pollen vary with the cultivar and season. . but certain types are grown in home dooryards in tropical areas. creamy white blooms. In hermaphrodite flowers. Climate The species is primarily mild-temperate to subtropical and naturally adapted to regions with cool winters and hot summers. The Japanese dwarf pomegranate.14 - In California. 'Pleniflora'. including 'Granada de China' and 'Granada Agria'. such as various islands of the Bahamas and West Indies. granatum var. There is very little wind dispersal of pollen. 'Rubra Plena'. The flowers are scarlet. 14 to 28%. double. double. nana. double. Legrelle' and 'Variegata'. Puebla. Many cultivars are grown. Pollination The pomegranate is both self-pollinated and cross-pollinated by insects. Mexicans take especial pride in the pomegranates of Tehuacan. 'Chico'.. P. but were superseded by 'Wonderful'. double. red. In recent years 'Wonderful' is losing ground to the more colorful 'Grenada'.

The plant favors a semi-arid climate and is extremely drought -tolerant. alkaline soil and on deep.15 - In southern Florida. the pomegranate can be grown outdoors as far north as Washington County. In northern India. it is spontaneous on rockstrewn gravel.11º C). However. and Washington.95% greatly enhances root development and survival.. fruit development is enhanced after a cold winter. Elsewhere in the United States. Propagation Pomegranate seeds germinate readily even when merely thrown onto the surface of loose soil and the seedlings spring up with vigor.5 m) apart. Culture Rooted cuttings or seedlings are set out in pre-fertilized pits 2 ft (60 cm) deep and wide and are spaced 12 to 18 ft (3. Utah. It can be severely injured by temperatures below 12º F (-11. D. indole-butyric acid and planting at a moisture level of 15. Treatment with 50 ppm.5-5. acidic loam and a wide range of soils in between these extremes. and then transplanted to the field. Grafting has never been successful but branches may be airlayered and suckers from a parent plant can be taken up and transplanted. The cuttings are set in beds with 1 or 2 buds above the soil for 1 year.. though it doesn't fruit in the latter locations. to avoid seedling variation.C. depending on the fertility of the . Soil The pomegranate thrives on calcareous. selected cultivars are usually reproduced by means of hardwood cuttings 10 to 20 in (25-50 cm) long.

In Israel.16 - soil. Harvesting and Yield The fruits ripen 6 to 7 months after flowering. After the 3rd year. For good fruit production. Initially. it is recommended that. irrigation water is supplied by overhead sprinklers which also provide frost protection during cold spells. the branches be judiciously shortened annually to encourage the maximum number of new shoots on all sides. only suckers and dead branches are removed. maturity has been equated with 1. and achieve a strong. Inasmuch as fruits are borne only at the tips of new growth.. the plant must be irrigated. for the first 3 years. The fruit must be picked before over maturity when it tends to crack open if rained upon or under certain conditions of atmospheric humidity. the plants are cut back to 24 to 30 in (60-75 cm) in height and after they branch out the lower branches are pruned to provide a clear main stem. prevent straggly development. well-framed plant. but 2 1/2 to 3 years is more common. Growers generally consider the fruit ready for harvest if it makes a metallic sound when tapped. cultivar 'Wonderful' is deemed ready for harvest when the soluble solids (SSC) reach 15%. The pomegranate may begin to bear in 1 year after planting out.8% titratable acidity (TA) and SSC of 17% or more. In Israel. brackish water is utilized with no adverse effect. Of course. one might assume that ultimate splitting is the natural means of seed release and dispersal. The fruit cannot be ripened off the tree even with ethylene treatment. In California. In California. or insufficient irrigation. dehydration by winds. .

for nearby markets. unwrapped but topped with shredded plastic. in baskets. Commercial California growers grade the fruits into 8 sizes. Appearance is important. Subsequent transfer to 68º F (20º C) dispels off-flavour from ethanol accumulation. precool rapidly. streaky pulp of flat flavor. become juicier and more flavorful. pack in layers. without shrinking or spoiling.17 - The fruits should not be pulled off but clipped close to the base so as to leave no stem to cause damage in handling and shipping.. the fruit can be kept only 2 months at 41º F (5º C). may be kept for a period of 7 months within this temperature range and at 80 to 85% relative humidity. After prolonged storage. for longer periods at 50º F (10º C). The fruit ships well. Control has been achieved by delaying harvest and storing in 2% O2 at 35. .6º F (2º C). stored in Israel for Christmas shipment to Europe. especially in the United States where pomegranates may be purchased primarily to enhance table arrangements and other fall (harvest-time) decorations. It is best maintained at a temperature of 32º to 41º F (0º-5º C). The fruits improve in storage. Too much sun exposure causes sunscald–brown. in wooden crates or. and ship in refrigerated trucks. 'Wonderful' pomegranates. in covered wood boxes. are subject to superficial browning ("husk scald"). At 95% relative humidity. russeted blemishes and roughening of the rind. internal breakdown is evidenced by faded. Keeping Quality and Storage The pomegranate is equal to the apple in having a long storage life. cushioned with paper or straw.

or Zythia versoniana may destroy as much as 80% of the crop unless these organisms are controlled by appropriate spraying measures. and defoliation by Euproctis spp.. The fruits may be sometimes disfigured by Sphaceloma punicae.. Discoloration of fruits and seeds results from infestation by Aspergillus castaneus. A post-harvest rot caused by Alternaria solani was observed in India in 1974. in a few days the caterpillars enter the fruit by way of the calyx. Gloeosporium and Pestalotia sp. A stem borer sometimes makes holes right through the branches. paper or plastic bags or other covers may be put over the fruits to protect them from borers. Virachola isocrates. Minor problems are leaf and fruit spot caused by Cercospora. Excessive rain during the ripening season may induce soft rot. lays eggs on flower-buds and the calyx of developing fruits. Dry rot from Phomopsis sp. mealybugs and scale insects. Food Uses For enjoying out-of-hand or at the table. It is particularly prevalent in cracked fruits. In India. Termites may infest the trunk. and Archyophora dentula. bats and squirrels. Twig dieback may be caused by either Pleuroplaconema or Ceuthospora Phyllosticta. These fruit borers may cause loss of an entire crop unless the flowers are sprayed 2 times 30 days apart. birds. the fruit is deeply scored several times vertically and then broken apart. also foliar damage by whitefly. then the clusters of juice sacs can be lifted out of the . thrips.18 - Pests and Diseases The pomegranate butterfly.

In northern India. family or group activity.19 - rind and eaten. or the whole fruits may be pressed and the juice strained out. Most simply. the cut-open fruits may be stomped by a person wearing special shoes in a clay tub and the juice runs through outlets into clay troughs. the juice can be easily extracted by reaming the halved fruits on an ordinary orange-juice squeezer. the juice is a very popular beverage. For beverage purposes. . it is usually sweetened. In Iran. Pomegranate juice is widely made into grenadine for use in mixed drinks.. Housewives in South Carolina make pomegranate jelly by adding 7 1/2 cups of sugar and 1 bottle of liquid pectin for every 4 cups of juice. Italians and other pomegranate fanciers consider this not a laborious handicap but a social. then strained and bottled. a major use of the wild fruits is for the preparation of "anardana"– the juice sacs being dried in the sun for 10 to 15 days and then sold as a spice.175%) and this is precipitated out by a gelatin process. the fruits are quartered and crushed. The juice from crushed whole fruits contains excess tannin from the rind (as much as . allowed to settle for 2 days. for future use. In Saudi Arabia. Hydraulic extraction of juice should be at a pressure of less than 100 psi to avoid undue yield of tannin. In the home kitchen. such as Iran. In the Asiatic countries it may be made into a thick sirup for use as a sauce. prolonging the pleasure of dining. Otherwise. the juice sacs are removed from the fruit and put through a basket press. After filtering. the juice may be preserved by adding sodium benzoate or it may be pasteurized for 30 minutes. the juice sacs may be frozen intact or the extracted juice may be concentrated and frozen. It is also often converted into wine. In some countries.

Juice. (3) peel.20 - INTRODUCTION TO HEALTH BENEFITS OF POMEGRANATE The last decades have seen an increase of interest in active ingredients and natural products with proven health benefits of plants.possesses a vast ethnomedical history and represents a phytochemical reservoir of heuristic medicinal value. The pomegranate tree . - Juice and peels have potent antioxidant properties. (4) leaf.especially its fruit . peel and oil are all weakly estrogenic and heuristically of interest for the treatment of menopausal symptoms and sequellae.. Juice. The tree/fruit can be divided into several anatomical compartments: (1) seed. peel and oil have also been shown to possess anticancer activities. including interference with tumour cell proliferation. (6) bark. The “most medicinal fruit of all”. (5) flower. the Punica granatim Linn (pomegranate) has been an important key to a rising interest of scientist in researching alternatives (or additions) to conventional pharmaceutical approaches. invasion and angiogenesis. and (7) roots. (2) juice. The phytochemistry and pharmacological actions of all Punica granatum components suggest a wide range of clinical applications for the treatment and prevention of . each of which has interesting pharmacologic activity. cell cycle.

. as well as other diseases where chronic inflammation is believed to play an essential etiologic role.21 - cancer. .

cyanidin. malvidin.22 - ACTIVE INGREDIENTS OF THE POMEGRANATE Punicalagin Ellacic Acid ellagitannins punicalagin A and punicalagin B potent tannins anthocyanins (delphinidin. petunidin) caffeic acid luteolin punicic acid gallic acid Punicotannic Acid Gallic Acid Mannite Pelletierine N-Methylisopelletierine Pelargonidin Punicalin Punicalagin syringic acid sinapic acid protocatechuic acid ferulic acid 3.4-dihydroxy-phenylacetic acid (PAA) . idin. pelargonidin..

such as fat. A free radical is a damaged molecule.. it reacts with any molecule from which it can take an electron. By taking an electron from certain key components in the cell. free radicals can damage cells. Eventually. Even though most free radical damage is repaired. The environment is also a source of free radicals caused by ultraviolet radiation or airborne pollutants. protein or DNA molecules. Free radicals are highly reactive chemicals which attack molecules by capturing electrons and thus modifying chemical structures. free radical damage may . a fraction may still remain. which are toxic molecules. Because the free-radical molecule needs its full complement of electrons. Antioxidants that occur naturally in the body and certain foods may block this damage by donating electrons to stabilize and neutralize the harmful effects of the free radicals. As part of their normal function. Oxygen damage (oxidation) to the cells is partly responsible for the effects of aging and certain diseases. cells produce free radicals.23 - Quercetin Kaempferol Antioxidants Any substance that reduces oxidative damage (damage due to oxygen) such as that caused by free radicals is called an antioxidant. because it is missing an electron. such as cigarette smoke.

interception. and . Despite the support for the health benefits of vitamin C.18 - overwhelm the body's natural defence. there are also studies which suggest that one should not take large supplemental doses. is organized at multiple levels. Antioxidants are also commonly added to food products like vegetable oils and prepared foods to prevent or delay their deterioration from the action of air. Antioxidants may possibly reduce the risks of cancer and age-related macular degeneration (AMD). As cell damage accumulates. Research has shown a relationship to a number of diseases. Antioxidants clearly slow the progression of AMD. The cellular protection against the deleterious effects of reactive oxidants generated in aerobic metabolism. vitamin E and carotenoids. Evidence from more than a hundred studies suggests that eating fruits and vegetables rich in vitamin C or carotenoids is linked with a reduced risk of many cancers.. Scientists theorize that low-density lipoprotein (LDL) cholesterol damages the lining of the arteries when it becomes oxidized. It is suspected that cataracts develop partly as a result of oxidation of proteins in the lens of the eye. called oxidative stress. Vitamin C. The defense strategies include three levels of protection: prevention. vitamin E and carotenoids may help protect against the oxidation of LDL cholesterol by neutralizing free radicals. and some studies have shown that antioxidants might be effective in reducing age-related macular degeneration and the resulting vision loss. More antioxidant vitamins from one's diet may help counter some of the damage. it may contribute to aging and certain diseases like cardiovascular disease and some cancers.

Commercial pomegranate juices showed an antioxidant . Short-term and long-term adaptation and cell specialization in these functions are new areas of interest. DMPD/TMPD are effective electron-transfer mediators for an enzyme. Four different methods (ABTS. Synthetic antioxidants mimic biological strategies. However. Control over the activity of pro-oxidant enzymes. FRAP ferric reducing antioxidant power..24 - repair. Antioxidants prevent free radical induced tissue damage by preventing the formation of radicals. or by promoting their decomposition. such as NADPH oxidase and NO synthases.1-diphenyl-2-picrylhydrazyl) is used for free radical scavenging assays. The antioxidant activity of the pomegranate juices were compared to those of red wine and a green tea infusion. DPPH (1. and FRAP) were used. Free radical production occurs continuously in all cells as part of normal cellular function. DMPD. scavenging them. A common use for it is in the enzyme-linked immunosorbent assay (ELISA) to detect for binding of molecules to each other. In one research. ABTS (2'-azino-bis(3-ethylbenzthiazoline-6-sulphonic acid) is chemical compound used to observe the reaction kinetics of specific enzymes. DPPH. the antioxidant activity of pomegranate juices was evaluated. is crucial. Regulation of the antioxidant capacity includes the maintenance of adequate levels of antioxidant and the localization of antioxidant compounds and enzymes. excess free radical production originating from endogenous or exogenous sources might play a role in many diseases.

seeds. Phenolics can also determine plant properties such as flavour and palatability. They naturally exist in grape stems. and pulp. The simplest of . Phenolics Phenolics are plant compounds (structurally characterized by an alcohol group on an aromatic ring) that impart a variety of functions to plants.25 - activity (18-20 TEAC) {Teac = trolox equivalent antioxidant capacity} three times higher than those of red wine and green tea (6-8 TEAC).. Phenols. including defence mechanisms and interactions with other organisms. are a class of chemical compounds consisting of a hydroxyl group (. In addition. ellagic acid derivatives. and hydrolyzable tannins were detected and quantified in the pomegranate juices. This shows that pomegranate industrial processing extracts some of the hydrolyzable tannins present in the fruit rind. anthocyanins. juice. The activity was higher in commercial juices extracted from whole pomegranates than in experimental juices obtained from the arils only (12-14 TEAC). sometimes called phenolics.O H) attached to an aromatic hydrocarbon group. HPLC-DAD and HPLC-MS analyses of the juices revealed that commercial juices contained the pomegranate tannin punicalagin (1500-1900 mg/L) while only traces of this compound were detected in the experimental juice obtained from arils in the laboratory. skins. This could account for the higher antioxidant activity of commercial juices compared to the experimental ones.

These studies have revealed a substantial and sometimes unexpected network of regulatory interactions. Phenols are similar to alcohol. . More recently.. Metabolic engineering of lignins and flavonoids has been deeply investigated. cardiovascular and neurodegenerative diseases or for use in anti-aging or cosmetic products. even in recent years. the demand and an increasing interest for practical applications has stimulated a wide range of biological and epidemiological studies aiming at characterizing the health promoting properties of specific phenolic compounds with antioxidant activities towards cancer. The extensive characterization of genes encoding the different enzymatic steps of flavonoid synthesis and cytochrome P450 genes have been among the most recent advances in this area.26 - the class is phenol (C6H5OH). Phenolic compounds represent the most studied phytochemicals and have been widely exploited as model systems in different areas of plant research. Example: the nature and functions of enzymes involved in lignin synthesis have been revisited several times. molecular biology and genomics have provided additional understanding of the mechanisms underlying the synthesis of these compounds with special emphasis on the regulation of gene expression by environmental factors. Some phenols are germicidal and are used in formulating disinfectants. Others possess estrogenic or endocrine disrupting activity. In the present time. The research is still active due to the complexity of the structures and the biosynthetic pathways. but they have unique properties and relatively higher acidities due to the aromatic ring's tight coupling with the oxygen and a relatively loose bond between the oxygen and hydrogen.

27 - TANNINS Since ancient times it is known that certain organic substances have tanning properties and are able to tan animal skins to form leather. they can be used in poisonings with these substances. precisely what happens to the skin during the tanning process was only elucidated during the twentieth century with the help of modern analytical techniques. and fruit juices. beer.. Other industrial uses of tannins include textile dyes. as antioxidants in the fruit juice. and as coagulants in rubber production. and haemostatic pharmaceuticals. In the food industry tannins are used to clarify wine. against diarrhoea. against stomach and duodenal tumours. as diuretics. In medicine. especially in Asian (Japanese and Chinese) natural healing. antiseptic. especially due to the increased incidence of deadly illnesses such as AIDS and various cancers. beer. As tannins can precipitate heavy metals and alkaloids (except morphine). Recently the tannins have attracted scientific interest. The search for new . Real tanning is understood as the cross-linking of the skin’s collagen chains. However. and also in the production of inks (iron gallate ink). It is also becoming clear that tannins often are the active principles of plant-based medicines. and wine industries. and as anti-inflammatory. Tannins are used in the dyestuff industry as caustics for cationic dyes (tannin dyes). Prehistoric tribes already knew about the tanning of protective animal hides with brain material and the fat of the killed animals. while false tanning entails the filling of hollow spaces between the skin’s collagen chains. the tannincontaining plant extracts are used as astringents.

28 - compounds for the development of novel pharmaceuticals has become increasingly important. especially. For example. . During the last twenty years many representatives of this class of compounds have been isolated and characterized. In extensive biological tests many representatives of this class were found to have antiviral. Tannins are polyphenolic secondary metabolites of higher plants. (Gallotannins. or from the animal kingdom. Complex Tannins. The polyphenolic structure of the secondary metabolites from higher plants is a necessary but not sufficient requirement for membership of the tannin class. certain tannins can selectively inhibit HIV replication. Ellagitannins. Corresponding polyphenolic natural products have not yet been isolated from lower plants such as algae. Condensed Tannins). Currently known tannins with unambiguously determined structures already number far more than 1000 natural products.. anti-tumour activity. antibacterial. especially as the biological action of tannin-containing plant extracts has been well documented. and.

Using preparative HSCCC about a 350 mg amount of the crude extract was separated. dellulose. is a high molecular weight polyphenolic compound. polyamides. the main ingredient of pomegranate (Punica granatum L. Diaion HP20). previous methods included labour intensive and expensive solid phase extractions by column chromatography (C-18. yielding 105 mg of punicalagin at a high-purity of over 92%.. Eighty milligrams of gallic acid was simultaneously separated as another product. High-speed countercurrent chromatography (HSCCC) was used for isolation and purification of punicalagin from pomegranate husk. Sephadex Lipophilic LH-20.) husk. at a purity of 75%. Punicalagin (PCG) .29 - Punicalagin Punicalagin. To isolate punicalagin. It has shown remarkable pharmacological activities attributed in the presence of dissociable OH groups.

The protective bioactivity of punicalagin. elagostasine. It is present in many red fruits and berries. and guanosine as a model for DNA has been investigated. strawberries. a high molecular weight polyphenol isolated from pomegranate fruit pith and carpellary membrane. amino acids constituting the proteins. Ellagic Acid (Benzoaric acid. gallogen) Ellagic acid is a fused four-ring polyphenol. against oxidative damages to lipids. including raspberries. In their study “The effects of dietary ellagic acid on rat hepatic and oesophageal mucosal cytochromes P450 and . Ellagic acid prevents the destruction of P53 gene by cancer cells. cranberries. based on its inhibitory action on the activation of the nuclear factor of activated T cells (NFAT). pomegranate and some nuts including pecans and walnuts. thereby making them inactive. PCG downregulated the mRNA and soluble protein expression of interleukin-2 from anti-CD3/anti-CD28-stimulated murine splenic CD4+ T cells and suppressed mixed leukocytes reaction (MLR) without exhibiting cytotoxicity to the cells. blackberries.. Pure ellagic acid is a cream to light yellow crystalline solid. It can bind with cancer causing molecules. eleagic acid.30 - isolated from the fruit of Punica granatum was identified as a potent immune suppressant.

thus preventing its carcinogenic alteration. Ellagic acid has also antiviral and antibacterial activities. One study confirmed that Ellagic acid effectively protects cells from damaging free radicals. With an increased concentration of these enzymes. thereby enhancing the ability of the target tissues to detoxify the reactive intermediates. a Nottingham. various tissues ability to detoxify harmful . n 1996. Additional phenolic compounds found in pomegranate known as anthocynadins (also well known scavengers of free radicals) combine synergistically with Ellagic acid to greatly augment pomegranate's potency as an antioxidant. Ellagic acid has several mechanisms of actions by which it exhibits its chemopreventive properties.” Ahn et al showed that ellagic acid causes a decrease in total hepatic mucosal cytochromes and an increase in some hepatic phase II enzyme activities. Ellagic acid showed also a chemo-protective effect against various chemically induced cancers. The discovery of this anti-viral activity instigated further experimentation and clinical trials. Initial experimentation by Stoner and Mukhtar showed that Ellagic acid decreased the number of chemically induced lung tumours.31 - phase II enzymes. Additionally. Mukhtar further illustrated that topical application of Ellagic acid provided protection against chemically induced skin tumours. PA University research team learned that pomegranate extract could destroy several viruses nearly on contact. Research by Barch et al demonstrated that Ellagic acid could actually bind to DNA. ellagic acid has been shown to induce the production of phase 11 detoxification enzymes through its manipulation of gene expression..

Cyanidin Pelargonidin and Cyanidin are both types of anthocyanidins. Pelagonidin. Anthocyanidins are found in blue. These are antioxidants that have been found to help improve blood vessel function in humans and animals. Finally.32 - compounds is augmented.. a molecule whose activity has been associated with the ability of certain viruses to transform normal cells into cancerous cells. More acidic cell sap gives a red colour. The colour of the pigment is altered by the pH of the cell sap. purple and red fruits and vegetables such as:        blueberries blackberries plums cranberries raspberries strawberries pomegranate Anthocyanidins are found in the cell's cell sap (unlike chlorophyll and carotene that are attached to cell membranes). Ellagic acid was found to be a potent inhibitor of tyrosine protein kinase. Less acidic cell sap express a more .

Quercetin … is also a powerful anti-histamine.. Anthocyanins are used in processed foods (drinks and confectionery) to avoid using synthetic additives. Gallic Acid (3. and anticancer properties. It is a bioflavonoid that gives pigment to many plants and herbs. and emphysema. The study found that histamine release was reduced 46 to 96 percent by the nutraceutical. It has anti-fungal and anti-viral properties and shows cytotoxicity against cancer cells without harming healthy cells. quercetin was effective against symptom-causing histamine activity in mast cells. antihistamine. The substance has been shown to exert anti-inflammatory. In a Japanese study published in the Journal of Allergy and Clinical Immunology. Gallic acid acts as a antioxidant and helps to protect our cells against oxidative damage.33 - purple colour. It also inhibits histamine release and pro-inflammatory cytokine production in mast cells and can therefore be of importance in future treatment for allergies.5-Trihydroxybenzoic acid) … is a free molecule or occurs a part of the tannin molecule. bronchitis. Additional studies have found high intakes quercetin (and of course other flavonoids) lower the risk of certain respiratory diseases such as asthma. . antioxidant.4.

34 - POMEGRANATE AND CANCER There is currently a shifting focus towards finding natural compounds that may prevent or treat cancer. It is important to develop non-cytotoxic therapies for solid malignancies such as prostate and breast cancer. and the ensuing aberrant proliferation leads to tumour growth.. Flavonoid-rich polyphenol fractions from the pomegranate fruit exert anti-proliferative. anti-invasive. Flavonoids are a group of differentiation-inducing chemicals with a potentially lower toxicology profile than retinoids. due to the problems that exist with current chemotherapeutic regimens. and proapoptotic actions in breast and prostate cancer cells and anti-angiogenic activities in vitro and in vivo. The fruit of the pomegranate contains hundreds of phytochemicals and pomegranate has been shown to exhibit antioxidant properties. total pomegranate tannin extract (TPT) 55% . One study examined the effects of PJ on inflammatory cell signaling proteins in the HT-29 human colon cancer cell line. At a concentration of 50 mg/L PJ significantly suppressed TNFalpha-induced COX-2 protein expression by 79% (SE = 0.042). Pomegranate juice (PJ) and its ellagitannins inhibited proliferation and induced apoptosis in HT-29 colon cancer cells. Cancer develops when the balance between cell proliferation and cell death is disrupted. anti-eicosanoid.

whereas ellagic acid (EA) (another pomegranate polyphenol) was ineffective.35 - (SE = 0. Pomegranate fruit extracts (PFE) inhibits the cell-growth and induces apoptosis of the highly aggressive human prostate carcinoma PC3 cells.4-fold.049). PJ reduced phosphorylation of the p65 subunit and binding to the NFkappaB response element 6. which are cancer cells derived from a bone marrow metastasis. Pomegranates possess strong antioxidant and anti-inflammatory agents. Therefore. TPT suppressed NFkappaB binding 10-fold. and thus even a modest delay in disease progression . punicalagin 3. This is done through modulations in the cyclin kinase inhibitor/cyclin kinase dependent machinery. PJ also abolished TNFalpha-induced AKT activation.regulate the cell cycle by adding phosphate groups to a variety of protein substrates that control processes in the cycle.achieved through pharmacological or nutritional intervention . because it is typically diagnosed in men over 50 years of age.. Prostate Cancer (CaP) is one of the leading causes of cancer-related deaths among males in most Western countries. The cyclin dependent kinase (cdk) is a family of kinases that . Additionally. and punicalagin 48% (SE = 0.022). CDKs are considered a potential target . the polyphenolic phytochemicals in the pomegranate can play an important role in the modulation of inflammatory cell signalling in colon cancer cells.6-fold. needed for NFkappaB activity.could significantly impact the quality of life of these patients. CaP is an ideal candidate disease for chemoprevention. Dietary antioxidants are proven chemo-preventative agents for CaP patients.once activated by cyclin .

The mitochondrial apoptotic pathway is largely mediated through Bcl-2 family proteins. before developing new drugs. Shifting research towards the effects for the use of natural products like active ingredients in pomegranate. . The balance between cell proliferation and apoptosis is crucial for the healthy functioning of organisms. If it is possible to selectively interrupt the cell cycleregulation in cancer cells by interfering with CDK action. is an area of extensive study because of the importance of maintaining the homeostatic balance in response to pro.. Much more research is necessary. including cancer. the cell will die. The biochemical mechanism of apoptosis. and BNIP3 that promote mitochondrial permeability. or programmed cell death (PCD). and anti-apoptotic members such as Bcl-2 and Bcl-xL that inhibit their effects. is one way forward. neurodegenerative disorders.36 - for anti-cancer medication. Dysregulation of apoptosis is implicated in many degenerative and autoimmune diseases. and viral and bacterial infections. The event of modulation of CDKs is associated with alterations of Bax and Bcl-2 – shifting the Bax:Bcl-2 ratio in favour of apoptosis. acquired immune deficiency syndrome. which include both pro-apoptotic members such as Bax. because the disruption of the CDK pathways can also lead to serious consequences/side-effects. The answers maybe found in more intensified studies of active ingredients in natural products. or inhibit the mitochondrial release of cyt c . which inhibit cell growth and induce apoptosis. Bak.or anti-apoptotic stimuli.

p21 acts as an inhibitor of apoptosis in a number of systems. P21 is often responsible for stress-induced p53-dependent and p53-independent cell cycle arrest. . the capacity of p21 to induce cell cycle arrest after stress can protect cells from stressinduced apoptosis.37 - Another important component is the tumor suppressor protein p53. leading to DNA degradation or fragmentation. p21 is poised to play an important role in preventing tumour development. and induction of p21 may cause cell cycle arrest.. Anti-apoptotic activity of p21 may contribute to its potential to act as an oncogene. On the other hand. Cell cycle arrest permits cells to pause and to repair damage and then to continue cell division. which activates the caspase-9 molecules (upon cleavage of the bound zymogen procaspases-9). and this may counteract its tumor-suppressive functions as a growth inhibitor. As a proliferation inhibitor. a number of recent studies have pointed out that in addition to being an inhibitor of cell proliferation. The cyclin-dependent kinase inhibitor p21 is induced by both p53-dependent and independent mechanisms following stress. which in turn activate caspase-3. whereas the inhibitor of apoptosis (IAP) inhibits both caspase-3 and caspase-9 activities. On one hand. Cyt c leakage supports the formation of an apoptosome complex by binding to apoptotic protease activating factor-1 (Apaf-1). Caspase-3 cleaves the inhibitor of caspase activated DNase (ICAD). which simultaneously suppresses Bcl-2 and activates Bax. the function of p21 to inhibit cell proliferation may contribute to its ability to act as tumor suppressor.

luteolin (L) and punicic acid (P). small molecules that eliminate p21 expression may improve the action of anticancer drugs. Here some examples of studies and the results: (detailed information regarding the studies please see reference list) A) Four pure chemicals. Therefore. All compounds significantly inhibited invasion when employed individually. functional p21 may suppress tumour growth in the organism. and each belonging to different representative chemical classes and showing known anticancer activities. but at the same time elimination of p21 may be beneficial during chemotherapy. and p21 protects cells from anticancer drug-induced apoptosis. The role of natural products/anti-oxidants of influencing/changing the activity of p53 and p21 should be studied intensively. P.38 - Anticancer drugs kill cancer cells by inducing p53-dependent and p53-independent apoptosis. a supradditive inhibition . and L were equally combined at the same gross dosage (4 microg/ml) as when the compounds were tested individually. Because loss of p21 usually increases sensitivity of tumor cells to apoptosis induced by different chemotherapeutic agents. all important components of the aqueous compartments or oily compartment of pomegranate fruit. were tested as potential inhibitors of in vitro invasion of human PC-3 prostate cancer cells in an assay employing Matrigel artificial membranes. When C. ellagic acid (E). caffeic acid (C)..

if the null hypothesis is true then what is the chance of obtaining a Kruskal-Wallis statistic as high (or higher) as observed in this experiment. then they both get the average of the two ranks for which they tie. Prism first ranks all the values from low to high. Prism displays a progress dialog and you can .. The largest number gets a rank of N. To perform the Kruskal-Wallis test. Prism then sums the ranks in each group. Prism can take a long time to calculate the exact P value. disregarding which group each value belongs. it approximates the P value from the chi-square distribution. and reports the sums. The P value answers this question: If the populations really have the same median.39 - of invasion was observed. what is the chance that random sampling would result in sums of ranks as far apart (or more so) as observed in this experiment? More precisely. measured by the Kruskal-Wallis non-parametric test. This test compares three or more unpaired groups. the P value will be small. If your samples are large or if there are ties. Prism calculates an exact P value. While it does the calculations. If the sums of the ranks are very different. If two values are the same. With medium size samples. If your samples are small and no two values are identical (no ties). The approximation is quite accurate with large samples. where N is the total number of values in all the groups. The discrepancies among the rank sums are combined to create a single value called the Kruskal-Wallis statistic (some books refer to this value as H). A larger KruskalWallis statistic corresponds to a larger discrepancy among rank sums. The smallest number gets a rank of 1.

mechanism of the cancer preventive and suppressive potential of pomegranate (here: fermented juice and pericarp.40 - press Cancel to interrupt the calculations if an approximate P value is good enough for your purposes. B) Another study tested flavonoid-rich fractions from fresh (J) and fermented (W) pomegranate juice and from an aqueous extraction of pomegranate pericarps (P) <the fruit wall> as potential differentiation-promoting agents of human HL-60 promyelocytic leukaemia cells. In addition.on its own showed important anti-cancer activities.ellagic acid (E). The four chemicals . The results highlight an important.. Extracts W and P were strong promoters of differentiation in all settings. the effect of these extracts on HL-60 proliferation was evaluated. . luteolin (L) and punicic acid (P). specific esterase activity. caffeic acid (C).increased the activity even more. and phagocytic activity. and DU 145 human cancer cell lines. previously unknown. Four assays were used to assess differentiation: nitro blue tetrazolium reducing activity. C) Another study showed the inhibition of the proliferation of LNCaP. PC-3. In this research scientists focussed on the quantities of . but the combination – the synergism of all four . The extracts had proportional inhibitory effects on HL-60 cell proliferation. nonspecific esterase activity. with extract J showing only a relatively mild differentiation-promoting effect.

wt/vol) to . (ii) inhibition of PI3K. Oral administration of PFE (0. Researchers further found that PFE treatment also resulted in (i) induction of WAF1/p21 and KIP1/p27. PFE treatment of A549 cells also resulted in dose-dependent arrest of cells in G0-G1 phase of the cell cycle (as assessed by DNA cell cycle analysis). (v) degradation and phosphorylation of IkappaBalpha.2%. cdk4 and cdk6 expression. scientists first compared the growth inhibitory effects of pomegranate fruit extract (PFE). (iii) phosphorylation of Akt at Thr308. The scientists also found that PFE treatment to A549 cells resulted in inhibition of NFkappaB DNA-binding activity. (iv) NF-kappaB and IKKalpha. D) Developing novel mechanism-based chemo-preventive approaches for lung cancer through the use of dietary substances which humans can accept has become an important goal. and (vi) Ki-67 and PCNA.1 and 0. In one study.41 - used active ingredients to induce apoptosis. and (iii) decrease in cyclin-dependent kinase (cdk) 2. employing normal human bronchial epithelial cells (NHBE) and human lung carcinoma A549 cells. Overall. The treatment of cells with PFE inhibited (i) phosphorylation of MAPK proteins. Treatment of PFE (50-150 microg/ml) for 72 h was found to result in a decrease in the viability of A549 cells but had only minimal effects on NHBE cells as assessed by the MTT and Trypan blue assays. this study demonstrates significant anti-tumour activity of pomegranate-derived materials against human prostate cancer. (ii) decrease in the protein expressions of cyclins D1. D2 and E..

metastatic potential. The results suggest vertical as well as the usual horizontal strategies for discovering pharmacological actives in plants. complementary and synergistic effects were proven in all models (again) by the Kruskal-Wallis non-parametric H test at p < 0. Supradditive. syringic acid.4-dihydroxyphenylacetic acid (PAA). Again. at concentrations more or less similar to those expected from normal consumption of foods.01 for invasion.05 for PLA2 expression.42 - athymic nude mice implanted with A549 cells resulted in a significant inhibition in tumour growth. E) One study investigated whether dissimilar biochemical fractions originating in anatomically discrete sections of the pomegranate fruit might act synergistically against proliferation. Proliferation of DU 145 human prostate cancer cells was measured following treatment with a range of therapeutically active doses of pomegranate. fermented pomegranate juice polyphenols (W) and sub-therapeutic doses of either pomegranate pericarp (peel) polyphenols (P) or pomegranate seed oil (Oil). 3. and phosholipase A2 (PLA2) expression of human prostate cancer cells. F) Another study focussed on the interaction/anti-proliferative activities of caffeic acid. sinapic acid. The results provide a suggestion that PFE can be a useful chemopreventive/chemotherapeutic agent against human lung cancer. the . and p < 0.001 for the proliferation tests.. protocatechuic acid and ferulic acid on the human breast cancer T47D cell line. p < 0.

which inhibited aromatase activity by 60-80%. were assessed in vitro for possible chemopreventive or adjuvant therapeutic potential in human breast cancer. in vitro. producing estrogens. F (aromatase are a group of enzymes of the cytochrome P450 superfamily. at concentrations ranging from 100 to 1. The ability to affect a blockade of endogenous active oestrogen biosynthesis was shown by polyphenols from fermented juice.) Fermented juice and pericarp polyphenols. G) In another study. pericarp. and of the crude whole oil and crude fermented and unfermented juice concentrate.. three components of the pomegranate were used to study the effects on breast cancer: fermented juice. it is an important factor in sexual development. inhibited 17-betahydroxysteroid dehydrogenase Type 1 from 34 to 79%. The activities of these ingredients.43 - results indicate that phenolic acids produce growth inhibition of cancer cells. . whose function is to aromatize androgens (that is. and whole seed oil.000 microg/ml according to seed oil. to selectively increase their aromaticity).) As such. indicating an additional protective effect on hormone-dependent breast tumours. aqueous pericarp extract and cold-pressed or supercritical CO2-extracted seed oil. and oil.

and 54% apoptosis in MDA-MB-435 oestrogen receptor negative metastatic human breast cancer cells at 50 microg/ml.12-dimethylbenz[a]anthracene (DMBA). . fermented pomegranate juice polyphenols consistently showed about twice the antiproliferative effect as fresh pomegranate juice polyphenols. In a murine mammary gland organ culture. Inhibition of cell lines by fermented juice and pericarp polyphenols was according to oestrogen-dependent (MCF-7) > estrogen-independent (MB-MDA-231) > normal human breast epithelial cells (MCF-10A). whereas the lyophilized juice by itself displayed only minimal estrogenic action. the fresh juice activities were also high enough to focus in further studies on the chemo-preventive and therapeutic use of pomegranate juice in human breast cancer. 75% inhibition of invasion of MCF-7 across a Matrigel membrane at 10 microg/ml. Pomegranate seed oil effected 90% inhibition of proliferation of MCF-7 at 100 microg/ml medium. In both MCF-7 and MB-MDA-231 cells. Although a higher inhibition through seed oil and fermented juice was proven..44 - In a yeast oestrogen screen (YES) lyophilized fresh pomegranate juice effected a 55% inhibition of the estrogenic activity of 17-beta-estradiol. fermented juice polyphenols effected 47% inhibition of cancerous lesion formation induced by the carcinogen 7.

cholesterol. high systolic/diastolic blood pressure) Atherosclerosis (or coronary artery disease) occurs.58 g/L polyphenols.37 g/L punicalagin isomers) for 5 days. stroke. . The most potent in vitro antioxidant polyphenol from this juice is the ellagitannin punicalagin. The antiatherogenic activity of pomegranate juice has been attributed to its antioxidant polyphenols. heart attack. In one experiment. cellular waste products and other clotting materials like calcium and fibrin is building up in the inner lining of an artery. diabetes.45 - POMEGRANATE AND ATHEROSCLEROSIS (… and implications re. In vitro antioxidant activity of plasma (ABTS and FRAP assays) and urine (ABTS and DPPH) were determined. including 4. six healthy subjects (four men and two women) consumed 1 L of pomegranate juice daily (5. and fatty substances. Its bioavailability and metabolism was studied to assess the effect on several blood parameters (including cardiovascular risk disease markers) and to compare the antioxidant activity of punicalagin with that of the in vivo generated metabolites. Fourteen haematological and twenty serobiochemical parameters including LDL.. HDL and VLDL as well as cholesterol and triglycerides in each lipoprotein were evaluated. when the normal lining of the arteries deteriorates the walls of the arteries thicken/harden. The polyphenols and the in vivo generated metabolites were measured by HPLC-DAD-MS-MS.

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RESULTS: Neither punicalagin nor ellagic acid present in the juice were detected in both plasma and urine. Three microbial ellagitannin-derived metabolites were detected: 3,8-dihydroxy-6H-dibenzo[b,d]pyran-6-one glucuronide, an unidentified aglycone (tentatively, trihydroxy-6H-dibenzo[b,d]pyran-6-one) and hydroxy-6-Hdibenzo[b,d]pyran-6-one glucuronide. These metabolites could reach up to 18.6 microM in plasma, although a large inter-individual variability was observed. In urine, the same metabolites and their corresponding aglycones became evident after 1 day of juice consumption. Total urine excretion of metabolites ranged from 0.7 to 52.7% regarding the ingested punicalagin. No relevant effect was observed on any blood parameter. The metabolites did not show significant antioxidant activity compared to punicalagin from pomegranate juice.

Although other studies have clearly shown the anti-oxidant activity of the active ingredients in pomegranate juice, this study shows that the potential systemic biological effects of pomegranate juice ingestion should be attributed to the colonic microflora metabolites rather than to the polyphenols present in the juice (more in chapter ‘microbes’.)

Polyphenolic antioxidants are associated with the inhibition of low density lipoproteins (LDL, the so called bad cholesterol), the macrophage foam cell formation and attenuation of atherosclerosis development. One study investigated ‘the effects of pomegranate juice (PJ, which contains potent tannins and anthocyanins) consumption by atherosclerotic patients with carotid artery stenosis (CAS) on the progression of

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carotid lesions and changes in oxidative stress and blood pressure. Ten patients were supplemented with PJ for 1 year and five of them continued for up to 3 years. Blood samples were collected before treatment and during PJ consumption. In the control group that did not consume PJ, common carotid intima-media thickness (IMT) increased by 9% during 1 year, whereas, PJ consumption resulted in a significant IMT reduction, by up to 30%, after 1 year. The patients' serum paraoxonase 1 (PON 1) activity was increased by 83%, whereas serum LDL basal oxidative state and LDL susceptibility to copper ion-induced oxidation were both significantly reduced, by 90% and 59%, respectively, after 12 months of PJ consumption, compared to values obtained before PJ consumption. Furthermore, serum levels of antibodies against oxidized LDL were decreased by 19%, and in parallel serum total antioxidant status (TAS) was increased by 130% after 1 year of PJ consumption. Systolic blood pressure was reduced after 1 year of PJ consumption by 21% and was not further reduced along 3 years of PJ consumption. For all studied parameters, the maximal effects were observed after 1 year of PJ consumption. Further consumption of PJ, for up to 3 years, had no additional beneficial effects on IMT and serum PON1 activity, whereas serum lipid peroxidation was further reduced by up to 16% after 3 years of PJ consumption. The results of the present study thus suggest that PJ consumption by patients with CAS decreases carotid IMT and systolic blood pressure and these effects could be related to the potent antioxidant characteristics of PJ polyphenols.

One study investigated whether daily consumption of pomegranate juice for 3 months would affect myocardial perfusion in 45 patients who had Coronary Heart Disease

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and myocardial ischemia in a randomized, placebo-controlled, double-blind study. Patients were randomly assigned into 1 of 2 groups: a pomegranate juice group (240 ml/day) or a placebo group that drank a beverage of similar caloric content, amount, flavour, and color. Participants underwent electrocardiographic-gated myocardial perfusion single-photon emission computed tomographic technetium-99m tetrofosmin scintigraphy at rest and during stress at baseline and 3 months. Visual scoring of images using standardized segmentation and nomenclature (17 segments, scale 0 to 4) was performed by a blinded independent nuclear cardiologist. To assess the amount of inducible ischemia, the summed difference score (SDS) was calculated by subtracting the summed score at rest from the summed stress score. The experimental and control groups showed similar levels of stress-induced ischemia (SDS) at baseline (p >0.05). After 3 months, the extent of stress-induced ischemia decreased in the pomegranate group (SDS -0.8 +/- 2.7) but increased in the control group (SDS 1.2 +/- 3.1, p <0.05). This benefit was observed without changes in cardiac medications, blood sugar, hemoglobin A1c, weight, or blood pressure in either group. In conclusion, daily consumption of pomegranate juice may improve stress-induced myocardial ischemia in patients who have Coronary Heart Disease.

Diabetes is associated with increased oxidative stress and atherosclerosis development. One study investigated the effects of pomegranate juice (PJ; which

the researchers observed increased level of cellular peroxides (by 36%). whereas serum SH groups content and paraoxonase 1 (PON1) activity. and decreased glutathione content (by 64%). were both decreased (by 23%). but it resulted in a significant reduction in serum lipid peroxides and TBARS levels by 56% and 28%. In the patients versus controls serum levels of lipid peroxides and thiobarbituric acid reactive substances (TBARS) were both increased. whereas serum SH groups and PON1 activity significantly increased by 12% and 24%.. by 350% and 51%.49 - contains sugars (!) and potent anti-oxidants) consumption by diabetic patients on blood diabetic parameters. which could contribute to attenuation of atherosclerosis development in these patients. Ten healthy test-persons (controls) and 10 non-insulin dependent diabetes mellitus (NIDDM) patients who consumed PJ (50ml per day for 3 months) participated in the study. and increased glutathione levels (by 141%) in the patients' HMDM. The patients' versus control HMDM took up oxidized LDL (Ox-LDL) at enhanced rate (by 37%) and PJ consumption significantly decreased the extent of Ox-LDL cellular uptake (by 39%). In the patients versus controls monocytes-derived macrophages (HMDM). . It was concluded that PJ consumption by diabetic patients did not worsen the diabetic parameters. but rather resulted in anti-oxidative effects on serum and macrophages. PJ consumption did not affect serum glucose. and on oxidative stress in their serum and macrophages. respectively. respectively. cholesterol and triglyceride levels. PJ consumption significantly reduced cellular peroxides (by 71%).

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Paraoxonase 2 (PON2), a member of the paraoxonase gene family, was shown to protect macrophages against oxidative stress. . Pomegranate juice (PJ), which contains potent polyphenol anti-oxidants, exhibits similar effects.

{(Additionally to PON2, PON1 and PON3 also hydrolyze and thereby inactivate Nacyl-homoserine lactones, which are quorum-sensing signals of pathogenic bacteria (more in chapter Microbes)}.

One study showed the association of pomegranate juice polyphenolics, macrophage oxidative stress, and cellular PON2 expression, in relation to the activation of specific PON2 transcription factors. Incubation of J774A.1 macrophages with PJ (0-50 microM of total polyphenols) dose-dependently increased expression (mRNA, protein) and activity and reduced macrophage oxidative status. These effects could be attributed to the PJ unique polyphenols, punicalagin and gallic acid. PJ polyphenolinduced up-regulation of PON2 was inhibited by 40% upon using the PPAR gamma inhibitor GW9662 (50 microM). Accordingly, the PPAR gamma ligand, rosiglitazone, dose-dependently stimulated macrophage PON2 expression, by up to 80%. Inhibition of AP-1 activation with SP600125, attenuated PJ-induced up-regulation of PON2 by 40%. Similarly, incubation of macrophages with PJ polyphenols in the presence of GW9662 or SP600125, significantly reduced their capacity to protect the cells against oxidative stress. It can be concluded that the anti-oxidative characteristics of PJ unique phenolics punicalagin and gallic acid could be related, at least in part, to their

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stimulatory effect on macrophage PON2 expression, a phenomenon which was shown to be associated with activation of the transcription factors PAPR gamma and AP-1.

The anti-atherosclerotic properties in pomegranate juice showed in one study a clear reduction in blood pressure. The effects of pomegranate juice consumption (50 ml, 1.5mmol of total polyphenols per day, for 2 weeks) by hypertensive patients on their blood pressure and on serum angiotensin converting enzyme (ACE) activity were tested in this research. A 36% decrement in serum ACE activity and a 5% reduction in systolic blood pressure were noted. Similar dose-dependent inhibitory effect (31%) of pomegranate juice on serum ACE activity was observed also in vitro. As reduction in serum ACE activity, even with no decrement in blood pressure, was previously shown to attenuate atherosclerosis, pomegranate juice can offer a wide protection against cardiovascular diseases which could be related to its inhibitory effect on oxidative stress and on serum ACE activity.

Pomegranate juice (PJ) can also revert the potent downregulation of the expression of endothelial nitric-oxide synthase (NOSIII) induced by oxidized low-density liporotein (oxLDL) in human coronary endothelial cells. analyses showed a significant decrease of NOSIII expression after a 24-h treatment with oxLDL. A significant dosedependent reduction in nitric oxide bioactivity represented by both basal and bradykinin-stimulated cellular cGMP accumulation was observed. These phenomena were corrected significantly by the concomitant treatment with PJ. The data of this study, done by the Department of General Pathology and Excellence Research Center

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on Cardiovascular Disease, University of Naples, Italy, suggest that PJ can exert beneficial effects on the evolution of clinical vascular complications, coronary heart disease, and atherogenesis in humans by enhancing the NOSIII bioactivity.

A study done by Department of Human Nutrition, National Nutrition and Food Technology Research Institute, Tehran, Iran was undertaken to assess the effect of concentrated pomegranate juice consumption on lipid profiles of type II diabetic patients with hyperlipidemia (total cholesterol or triglycerides > or = 200 mg/dL). In this pilot study 22 diabetic patients were recruited from the Iranian Diabetes Society. They were free of any other chronic diseases. The patients were followed for eight weeks to obtain more detailed data about their diet before concentrated pomegranate juice (CPJ) consumption period began. In this pre-study period a 24-hour food recall and a food record (containing flavonoid-rich foodstuffs) were completed every ten days. At the end of the eighth week, anthropometric and biochemical assessments were done. Thereafter the patients consumed 40 g CPJ for eight weeks. During this period, dietary assessment was continued. After completion of the study anthropometric and blood indices were evaluated again. The Wilcoxon signed-rank test was used for statistical analysis. P-value was considered significant at p < 0.05. There were 14 women (63.6%) and 8 men (36.4%) in this survey. Mean (+/- SD) of age, weight, and duration of diabetes were 52.5 (+/- 5.2) years, 71.5 (+/- 10.3) kg, and 7.9 (+/- 6.6) years, respectively. After consumption of concentrated pomegranate juice

and total cholesterol/HDL-c (p < 0. Therefore. .006). types and doses of oral hypoglycemic agents.53 - significant reductions were seen in total cholesterol (p < 0.006). and the intake of nutrients and flavonoid-rich foodstuffs did not change during the CPJ consumption period. its inclusion in their diets may be beneficial. It is concluded that CPJ consumption could modify heart disease risk factors in these hyperlipidemic patients..001). LDL-c/high-density lipoproteincholesterol (HDL-c) (p < 0. physical activity level. Anthropometric indices. However there were no significant changes in serum triacylglycerol and HDL-c concentrations. low-density lipoprotein-cholesterol (LDL-c) (p < 0.001).

coal-dust or congenital conditions (e. CEBAS-CSIC. like lung fibrosis. and different other lung disorders. The decrease in the expiratory flow rates leads to an increase in the total lung capacity. The Department of Food Science and Technology. 5-week randomized study with the use of pomegranate juice in COPD-patients. have shown.54 - CHRONIC OBSTRUCTIVE PULMONARY DISEASE (COPD) … also known as Chronic Obstructive Airway Disease is a pathological limitation of airflow in the airway. . The high TEAC of PJ cannot be extrapolated in vivo probably due to the metabolism of its polyphenols by the colonic microflora. In the Spanish study it was concluded that Pomegranate Juice (PJ) supplementation does not add benefits to the current standard therapy in patients with stable COPD. Spain. It includes chronic bronchitis.g. solvents. that antioxidants can be effective in attenuating fibroproliferative responses in the lung of animals and humans. It can include ‘classical’ asthma. placebo-controlled. This – and other research looking into interstitial lung diseases. emphysema. Murcia. Patients with this condition – triggered by smoking or other airborne irritants such as asbestos. The understanding of the different bioavailability of dietary polyphenols is critical before claiming any antioxidant-related health benefit. which is irreversible.. Research Group on Quality. alpha-1-antitrypsin deficiency) – are very prone to acute respiratory failure from infections. Safety and Bioactivity of Plant Foods. did a double blind. but more often it is an asthma-related breathing problem.

55 - It it/was proven that polyphenols in PJ can be very beneficial.. but the colonic microflora of each individual plays a major role in bioavailability. . More research is necessary.

induces the expression of matrix metalloproteinases (MMPs) implicated in cartilage resorption and joint degradation in osteoarthritis (OA). We will discuss the latter also in the ‘Microbe/Bacteria’ section. The expression of . no studies have been undertaken to investigate whether PFE constituents protect articular cartilage. A case study done by the Western Reserve University School of Medicine showed the significant effects of Pomegranate fruit extract (PFE). Here is a study extract: Interleukin (IL)-1ß .a pro-inflammatory protein molecule that plays a key role in cartilage degradation in osteoarthritis .namely cartilage protection -. However. researchers added a water extract of pomegranate fruit to the culture using a well-established in vitro model. OA chondrocytes or cartilage explants were pretreated with PFE and then stimulated with IL-1ß at different time points in vitro. The amounts of proteoglycan released were measured by a colorimetric assay. Using tissue samples of human cartilage affected by osteoarthritis. Pomegranate fruit extract (PFE) was recently shown to exert antiinflammatory effects in different disease models.56 - OSTEROARTHRITIS The pomegranate has antioxidant AND anti-inflammatory properties. The findings showed a new activity for pomegranate fruit extract -. In the present studies..in addition to its previously discovered antioxidant and antiinflammatory properties. Arthritis is one of the foremost diseases for which patients seek herbal or traditional medicine treatments.

05) and this was associated with the inhibition of mRNA expression.B (NF. MAPK enzyme activity was assayed by in vitro kinase assay. Expression of mRNA was quantified by real-time PCR. IL-1ß– induced phosphorylation of p38-MAPK. these novel results indicate that PFE or compounds derived from it may inhibit cartilage degradation in OA and may also be a useful nutritive supplement for maintaining joint integrity and function. At the cellular level. -3. phosphorylation of the inhibitor of B (I B ) and mitogen-activated protein kinases (MAPKs) was determined by Western immuno-blotting.B in OA chondrocytes. was most susceptible to inhibition by low doses of PFE. PFE also inhibited the IL-1ß–induced phosphorylation of I B and the DNA binding activity of the transcription factor NF. Taken together. and -13 protein in the medium (P < 0. . Activation of nuclear factor. but not that of c-Jun-N-terminal kinase or extracellular regulated kinase. PFE inhibited the IL-1ß–induced proteoglycan breakdown in cartilage explants in vitro.B) was determined by electrophoretic mobility shift assay. PFE (6. and the addition of PFE blocked the activity of p38-MAPK in a kinase activity assay..25–25 mg/L) inhibited the IL-1ß–induced expression of MMP-1.57 - MMPs.

PJ consumption provided an increase in epididymal sperm concentration.50 mL distilled water and 1 mL PJ were given daily for seven weeks by gavage to rats in the first.58 - INCREASE IN SPERM-QUALITY Regarding sperm-quality. 0. CONCLUSION: The results suggest that PJ consumption improves sperm quality and antioxidant activity of rats.. spermatogenic cell density and diameter of seminiferous tubules and germinal cell layer thickness. each group containing seven rats. sperm motility. Data were compared by analysis of variance (ANOVA) and the degree of significance was set at P<0. and it decreased abnormal sperm rate when compared to the control group. spermatogenic cell density. One milliliter distilled water. Pomegranate fruit is inescapably linked with fertility. and lipid peroxidation and. levels of antioxidant vitamins. antioxidant enzyme activities were investigated. only one study was done so far… with rats. testosterone. Body and reproductive organ weights. METHODS: Twenty-eight healthy adult male Wistar rats were divided into four groups. glutathione peroxidase (GSH-Px) and catalase (CAT) activities. birth and eternal life because of its many seeds. and vitamin C level were observed in rats treated with different doses of PJ.50 mL PJ plus 0. RESULTS: A significant decrease in malondialdehyde (MDA) level and marked increases in glutathione (GSH). antioxidant activity and testosterone level of male healthy rats.25 mL PJ plus 0. spermatogenic cell density.75 mL distilled water. second. third and fourth groups.05. sperm characteristics. respectively. The aim of this study was to investigate the effects of pomegranate juice (PJ) consumption on sperm quality. All analyses were done only once at the end of the seven week study period. . 0.

47% percent of the subjects reported that their erections improved with the pomegranate juice. be in a stable. and be willing to attempt sexual intercourse on at least one occasion per week during each study period. they were assigned to drink 8 oz. while only 32% reported improved erections with the placebo (p=0. For the first four weeks of the study. double-blind. At the end of each four week period. To qualify. crossover pilot study.. The GAQ elicits the patient's self-evaluation of the study beverages' effect on erectile activity. published by the International Journal of Impotence Research (Nature Group) examined the efficacy of pomegranate juice versus placebo in improving erections in 61 male subjects. placebo-controlled. participants had to experience mild to moderate ED for at least 3 months. efficacy was assessed using the International Index of Erectile Function (IIEF) and Global Assessment Questionnaires (GAQ).59 - ERECTILE DYSFUNCTION A randomized. These results compare favourably to a recent 24-week study using a PDE5 . After a two-week washout period during which the subjects did not consume any study beverage nor utilize any erectile dysfunction treatment. the subjects were assigned to drink either 8 oz. monogamous relationship with a consenting female partner.058). of Pomegranate Juice (used juice: POM Wonderful) daily with their evening meal or shortly after. The IIEF is a validated questionnaire that has been demonstrated to correlate with ED intensity. of the opposite study beverage every evening for another four weeks.

arterial plague. Although the study did not achieve overall statistical significance. experiencing improvement while on the placebo). in which roughly 73% of subjects reported a benefit from the PDE5 inhibitor and 26% reported a "placebo effect" (i. Looking at several causes of ED (diabetes. .e. high blood pressure. the authors conclude that additional studies with more patients and longer treatment periods may in fact reach statistical significance..60 - inhibitor (such as Cialis). heart disease) the anti-atherosclerosis activity of the active pomegranate ingredients can also help with ED conditions.

Loma Linda University. (all: Washington University School of Medicine. the Department of Molecular Biology and Pharmacology. California. that polyphenols are neuro-protective. that that further studies to validate and determine the mechanism of polyphenolic effects.. MO. University of California School of Medicine. Louis. Although there are no proven ways to delay onset or slow progression of Alzheimer's disease (AD). St.61 - ALZHEIMER’S DISEASE It is proven. . the Hope Center for Neurological Disorders. California. showed. as well as whether substances in PJ may be useful in Alzheimer’s Diseases. USA) and the David Geffen School of Medicine. Washington University School of Medicine. studies suggest that diet can affect the risk. should be considered. A study with mice done by the Department of Psychology. the Department of Neurology.

especially when they occur imultaneously. The onset of menopause triggers profound changes in cardiac health. While declining levels of oestrogen are what gives rise to the difficulties of menopause. osteoporosis. Menopause is a complex health condition. short-term symptomatic relief as well as longer-term benefits that support women’s health as their body chemistry changes. and cell proliferation. The ideal approach to addressing menopause would be comprehensive and holistic. and certain cancers.62 - MENOPAUSAL SYMPTOMS Active ingredients in pomegranate are helpful once we look at the complete picture of menopause. These health issues are not easy to correct. all of which combine to greatly elevate a woman’s risk for contracting heart disease.. both nutritional and hormonal support are required to address and ameliorate the physiological symptoms and other changes that accompany menopause. providing fast-acting. bone strength. Addressing menopause requires a diverse approach that both restores normal hormone balance and protects against the multitude of diseases that can arise during this period in a woman’s life. mental states. .

which help to reduce swelling and ease muscular aches and pains. Without moisture. and protects the skin. but had no growth-supporting effect on keratinocytes. was shown to stimulate keratinocyte proliferation in monolayer culture. The conjugated fatty acids give it strong anti-inflammatory properties. In parallel. Pomegranate seed oil adds moisture. Pomegranate seed oil. It provides relief from minor skin irritations and inflammation. a mild thickening of the epidermis (without the loss of ordered differentiation) was observed in skin organ culture. interstitial collagenase) production by dermal fibroblasts. eczema. improves skin elasticity. These results suggest heuristic potential of pomegranate fractions for facilitating skin repair in a . and to soothe minor skin irritations.. anti-microbial. the skin looks tight and lacks luster. both the fermented juice and seed cake extracts) stimulated type I procollagen synthesis and inhibited matrix metalloproteinase-1 (MMP-1. peel or seed cake. pomegranate peel extract (and to a lesser extent. psoriasis and sunburned skin. is anti-inflammatory. anti-oxidants. including dry skin. but not aqueous extracts of fermented juice. wrinkles become more abundant and pronounced. In contrast. The same pomegranate seed oil that stimulated keratinocyte proliferation was without effect on fibroblast function. has natural estrogenic properties. assist with wrinkles.63 - COSMECEUTICAL Pomegranate seed oil is commonly used in cosmetic products to revitalize dull or mature skin.

35% and 1. questionnaires were conducted regarding the condition of the skin before the start of the test food intake and at the termination of the test food intake. and at 2 and 3 wk (p<0. Each group received the respective test foods for 4 wk. as compared to the control group.5 MED (minimum erythema dose) of UV irradiation on an inside region of the right upper arm. respectively) and 4 wk (p<0.. Thirteen healthy volunteers per group were randomly assigned to three groups. 2 (p<0. 2. melanin and erythema values were measured before the start of the test food intake. decreasing rates of L values from the baseline in the low. in some items such as "brightness of the face" and "stains and freckles. using female subjects in their 20s to 40s. namely. low dose (100 mg/d ellagic acid) and control (0 mg/d ellagic acid: placebo).73% respectively. a stratified analysis using subjects with a slight sunburn revealed an inhibited decrease of L values compared with the control group at 1. based on the MED value measured on the previous day. Further.05) after the start of the test food intake in the low-dose group.05) in the high-dose group. and after 1. Luminance (L). namely aqueous extracts (especially of pomegranate peel) promoting regeneration of dermis." . Further. and pomegranate seed oil promoting regeneration of epidermis.64 - polar manner. Each subject received a 1. high dose (200 mg/d ellagic acid).01.and high-dose groups were inhibited by 1. As a result. 3 and 4 wk following the start of the test food intake. placebo-controlled trial was performed in Japan to clinically evaluate the protective and ameliorative effects of ellagic acid-rich pomegranate extract on pigmentation in the skin after ultraviolet ray (UV) irradiation. the results of questionnaires showed ameliorating tendencies due to the test food. A double-blind. Furthermore.

The goal of this study was to determine whether PFE affords protection against UVA-mediated activation of STAT3. it is suggested that ellagic acid-rich pomegranate extract. Immunoblot analysis demonstrated that 4 J/cm2 of UVA exposure to NHEK led to an increase in phosphorylation of STAT3 at Tyr705. . One studiy has shown that PFE treatment of normal human epidermal keratinocytes (NHEK) inhibits UVB-mediated activation of MAPK and NF-kappaB pathways. AKT and extracellular signalregulated kinase (ERK1/2). Pomegranate fruit extract (PFE) possesses strong antioxidant and anti-inflammatory properties.. Signal transducers and activators of transcription 3 (STAT3). modulate cell proliferation. Cosmeceutical/Skin Cancer UVA is the major portion (90-99%) of solar radiation reaching the surface of the earth and has been described to lead to formation of benign and malignant tumors. Protein Kinase B/AKT and Map Kinases (MAPKs). apoptosis and other biological activities. AKT at Ser473 and ERK1/2. photo-dermatoses.65 - Based on the above-mentioned results. has an inhibitory effect on a slight pigmentation in the human skin caused by UV irradiation. ingested orally. which are activated by a variety of factors. photo-aging and photo-carcinogenesis. UVAmediated cellular damage occurs primarily through the release of reactive oxygen species and is responsible for immuno-suppression. Pretreatment of NHEK with PFE (60-100 microg/mL) for 24 h before exposure to UVA resulted in a dose-dependent inhibition of UVA-mediated phosphorylation of STAT3 at Tyr705. AKT at Ser473 and ERK1/2. mTOR.

. PFE pre-treatment of NHEK was found to increase the cell-cycle arrest induced by UVA in the G1 phase of the cell cycle and the expression of Bax and Bad (proapoptotic proteins). is involved in the regulation of p70S6K. which in turn phosphorylates the S6 protein of the 40S ribosomal subunit. The data further demonstrate that PFE pre-treatment of NHEK resulted in significant inhibition of UVA exposure-mediated increases in Ki-67 and PCNA. The data suggest that PFE is an effective agent for ameliorating UVA-mediated damages by modulating cellular pathways and merits further evaluation as a photochemopreventive agent. with downregulation of Bcl-X(L) expression (antiapoptotic protein).66 - structurally related to PI3K. The study found out that UVA radiation of NHEK resulted in the phosphorylation of mTOR at Thr2448 and p70S6K at Thr421/Ser424. . PFE pre-treatment resulted in a dose-dependent inhibition in the phosphorylation of mTOR at Thr2448 and p70S6K at Thr421/Ser424.

Consequently. In general. new infections can occur in hospitals resulting in high mortality. This means: A stricter control in the use of antibiotics and increased research for a better understanding of the genetic mechanisms of microbial drug-resistance. but resistance to these drugs by micro-organisms has increased dramatically.. bacteria have the genetic ability to transmit and acquire resistance to drugs. the development of new drugs is very important… but not only synthetic. From 1980 to 1990. Of course. and due to new bacterial strains. The problem of microbial resistance is growing and the outlook for the use of antimicrobial drugs in the future is uncertain. The pharmacological industries have produced a number of new antibiotics in the last three decades. This causes of course concern. Therefore. which are multi-resistant. This fact has also been verified in other clinics around all over world. Montelli and Levy documented a high incidence of resistant micro-organisms in clinical microbiology in Brazil. but also natural drugs. which are utilized as therapeutic agents. actions must be taken to reduce this problem. . because of the number of patients in hospitals who have suppressed immunity.67 - ANTIMICROBIAL Introduction: The necessity of natural antimicrobials.

In Argentina. The use of plant extracts and phytochemicals.68 - For a long period of time. for example. especially in the last decade. which are due to compounds synthesized in the secondary metabolism of the plant. Many plants have been used because of their antimicrobial activity. which has compounds derived from medicinal plants. especially in Latin America. Therefore. In the last few years. Inhibition of bacterial growth was observed with the following bacteria: Staphylococus aureus Escherichia coli Aspergillus niger . According to the World Health Organization (WHO) medicinal plants would be the best source to obtain a variety of drugs. a research tested 122 known plant species used for therapeutic treatments. The antimicrobial properties of plants have been investigated by a number of researchers world wide. About 80% of individuals from developed countries use traditional medicine. These products are known by their active substances. plants have been a valuable source of natural products for maintaining human health. both with known antimicrobial properties. safety and efficiency.. The use of plant compounds for pharmaceutical purposes has gradually increased. can be of great significance in therapeutic treatments. the phenolic compounds. a number of studies have been conducted in different countries to prove such efficiency. such plants should be investigated to better understand their properties. with more intensive studies for natural therapies.

. which is interesting enough to focus more on antimicrobial research and the promicrobial use of pomegranate. coli O157 : H 7 illness have been . Although outbreaks of food-associated E. Its reported mortality rates was as high as 31% . In recent years there has been a dramatic increase in the number of reported cases of foodborne illness. Its transmission to human has recently been epidemiologically associated with the consumption of contaminated foods. This pathogen is an etiological agent of haemorrhagic colitis and haemolytic uremic syndrome in humans.69 - Candida albicans Klebsiella pneumoniae Pseudomonas aeruginosa Bacillus subtilis Styaphylococcus aureus Staphylococcus epidermidis Bacillus cereus Escherichia coli Salmonella typhi Salmonella paratyphi Some research suggest that the potential systemic biological effects of pomegranate juice ingestion should be attributed to the colonic microflora metabolites rather than to the polyphenols present in the juice. A very important theory. Escherichia coli O157 : H 7 is a highly virulent bacterium.

Darmstadt) at 37C and maintained on TSA at 4C. this bacterium has been isolated from drinking water. Thailand. tofu. TSA (Merck. the present study provides data on antibacterial activity of various spices and medicinal plants that are available in Thailand against Y. coli O157 : H 7 ATCC 43889 and Y. Yersinia enterocolitica is an important foodborne pathogen because of its ability to grow at refrigeration temperatures. attention is shifting towards alternatives that the consumers perceive as natural.70 - associated with the consumption of undercooked ground beef. Foodborne outbreaks due to Y. cultures were activated by two successive transfers in Tryptic Soy Broth (TSB) at 37C. chocolate–flavoured milk. Of very high interest is the In searching for natural antimicrobial products. Ministry of Public Health. mayonnaise and mayonnaise-based sauces. cheese and pork. Inoculum population was determined by serially diluting the suspension with . enterocolitica ATCC 23715 were btained from the National Institute of Health. enterolitica. Yersinia readily withstand freezing and can survive in frozen foods for extended periods even after repeated freezing and thawing. apple cider. Cultures of E. Cultures were grown in Tryptic Soy Agar.. Before preparing inoculum for the test media. spices and medicinal plants or herbs. pork. powdered milk. bean sprouts. an in particular. Due to negative consumer perceptions of artificial preservatives. Bangkok. enterocolitica have been related to raw milk.

The inoculated tubes were incubated for 24 hr at 37C and the lowest concentrations showing no growth in tubes were recorded as the MICs. Sensitivity testing was done by well assay technique. The aqueous extract was sterilized by millipore membrane.71 - phosphate buffer.1 ml from an 18 hr culture to give approximately 3. Spices and medicinal plants used in this experiment are shown in Table 1. sappan and betle leaf were inoculated with 0. They were dried and ground before extracting with 95% ethyl alcohol for 48 hr.0104 or 3. One ml of an overnight culture in TSB was inoculated into 100 ml of a TSA maintained at 45C. while pomegranate had the highest inhibitory effects on Yersinia enterocolitica. Briefly.0104 or 6. Clove showed the highest inhibitory effect for E.0106 cfu/ml Y. The volumes were adjusted to obtain the concentration of 100 mg/ml.0106 cfu/ml E. 9. All plates were incubated at 37C for 48 hr and the diameters of the inhibition zones were measured. coli O157 : H 7 and 6. Wells (4 mm in diameter) were cut into the agar and filled with 40 l of filtered aqueous extracted of spices and medicinal plants. Approximately 25 ml was poured into a petri dish and allowed to solidify. pour plating with TSA and incubating at 37C for 48 hr before counting colonies.9 ml volumes of TSB containing a series of dilutions of cloves. cinnamon. MICs were determined according to the method of Richards et al. . leadwort. coli. pomegranate. roselle. enterocolitica..

Synergic activity was detected between PGME and the 5 antibiotics tested. The bactericidal activity of PGME (0. In conclusion.5% in MSSA and MRSA populations.1 × MIC) in combination with ampicillin (0. cell viability was reduced by 99. tetracycline. Susceptibility testing of the isolates to PGME and antibiotics was performed by the broth dilution method. aureus resistant to PGME was low and they did not survive. In Brazil. pomegranate (Punica granatum L. respectively. PGME increased the post-antibiotic effect (PAE) of ampicillin from 3 to 7 h. Studies evaluated the effect of pomegranate extract on Staphylococcus aureus FRI 722 growth and subsequent enterotoxin production. and they confirmed the synergic activity. The detection of in vitro variant colonies of S. gentamicin. ampicillin.9% and 72.5 × MIC) was assessed using chosen isolates by time-kill assays. PGME dramatically enhanced the activity of all antibiotics tested. Bacterial susceptibility was determined by tube dilution method and production of . Using this combination. and thus.72 - One study evaluated the interaction between Punica granatum (pomegranate) methanolic extract (PGME) and antibiotics against 30 clinical isolates of methicillinresistant Staphylococcus aureus (MRSA) and methicillin-sensitive Staphylococcus aureus (MSSA). (Punicaceae)) is widely used as a phytotherapeutic agent. PGME did not interfere with the action of any of the antibiotics tested. In addition. chloramphenicol. offers an alternative for the extension of the useful lifetime of these antibiotics. ranging from 38% to 73%. For some isolates. and oxacillin.. PGME demonstrated the potential to either inhibit the efflux pump NorA or to enhance the influx of the drug.

XAD-BuOH. punicalins (3). Gallagic acid (2) and punicalagins (4) exhibited antiplasmodial activity against Plasmodium falciparum D6 and W2 clones with IC50 values of 10.2. At a low extract concentration (0.01% v/v) bacterial growth was delayed. These data further implicate pomegranate extracts as potential antibacterial therapeutics with the added ability to inhibit enterotoxin production. Aspergillus fumigatus and Mycobacterium . 3.1. XAD-H2O and XAD-PJ compounds 1 . Compounds 1 .05% (v/v) concentration of extract was found to inhibit Staphylococcal enterotoxin (SE) A production. respectively. respectively. The mixture of tannins (TPT). Candida albicans. and punicalagins (4).25 microg/mL against HL-60 cells. a 0. The Punica granatum L.6. while a higher concentration (1% v/v) eliminated bacterial growth.4 and the mixture of tannin fractions (XAD-16 eluates) were evaluated for antioxidant.5 and 8. 10. antiplasmodial.3 and 1. methicillin-resistant Staphylococcus aureus (MRSA). and antimicrobial activities in cell-based assays. exhibited IC50 values against reactive oxygen species (ROS) generation at 0. Cryptococcus neoformans.73 - enterotoxin was assessed using membrane-over-agar (MOA) plates.9.4 revealed antimicrobial activity when assayed against Escherichia coli. EtOAc and n-BuOH.8 .4 showed IC50 values of 1. The EtOAc and n-BuOH extracts were purified by XAD16 and Sephadex LH-20 column chromatography to afford ellagic acid (1). 2. XAD-H2O.. XAD-PJ and XAD-BuOH. Pseudomonas aeruginosa. Most interestingly. Compounds 1 . against ROS generation and no toxicity up to 31.4 microM.19 microg/mL. XADEtOAc. 7.8 microM. (pomegranate) by-product POMx was partitioned between water. Fractions XAD-EtOAc. gallagic acid (2).

Rifampicin combined with kaempferol or quercetin exhibited good beta-lactamase inhibitory effects (57. The in vitro activities of 10 antibiotics and 15 natural polyphenols against the isolates were evaluated by determining minimum inhibitory concentrations (MICs).4. Our results suggest a beneficial effect from the daily intake of POMx and pomegranate juice (PJ) as dietary supplements to augment the human immune system's antioxidant.7-tetrahydroxyflavone) and quercetin (3. The study results and ready availability and low toxicity of plant polyphenols warrant further investigations on the therapeutic potential of combination therapies for MRSA infections.8 % and 75. Compounds 2 and 4 showed activity against P. aeruginosa. C. In checkerboard assays.7-pentahydroxyflavone) showed the lowest MICs.5. including structure-activity relationships (SAR) of the free radical inhibition activity of compounds 1 .8 %. FIC:fractional inhibitory concentration MIC:minimum inhibitory concentration MRSA:methicillin-resistant . researcher determined the effects of combinations of antibiotics and plant polyphenols against 20 clinical isolates of MRSA. All isolates were susceptible to vancomycin and resistant to rifampicin. neoformans. antimalarial and antimicrobial capacities. and MRSA. kaempferol (3. combinations of rifampicin and either kaempferol or quercetin acted synergistically or partially synergistically against the clinical MRSA isolates.5.74 - intracellulare. antiplasmodial and antimicrobial activities of POMx isolates. respectively) against a representative isolate according to nitrocefin analysis.4'.3'.4'. Among the 15 natural polyphenols.. while susceptibilities to ciprofloxacin varied. In another study. This is the first report on the antioxidant.

A selected complex was assayed for inhibition of infection by primary HIV-1 isolates. Methods Fruit juices were screened for inhibitory activity against HIV-1 IIIB using CD4 and CXCR4 as cell receptors. The resulting complex blocks virus binding to CD4 and CXCR4/CCR5 and inhibits infection by primary virus clades A to G and group O. topical microbicides. may provide this option. with established safety records and adequate anti-HIV-1 activity. microbicides should be: acceptable. Antiretroviral chemotherapeutics have decreased AIDS mortality in industrialized countries. expected to block virus transmission. affordable. and accelerative in transition from development to marketing. accessible. To remove most colored juice components. . utilizing CCR5 as the cellular coreceptor. Results: HIV-1 entry inhibitors from pomegranate juice adsorb onto corn starch.75 - New therapies are needed to address the public health problem posed by methicillinresistant Staphylococcus aureus. The best juice was tested for inhibition of: (1) infection by HIV-1 BaL. and (2) binding of gp120 IIIB and gp120 BaL.. respectively. To prevent an analogous dichotomy. but only minimally in developing countries. Already marketed pharmaceutical excipients or foods. HIV In the absence of vaccines. to CXCR4 and CCR5. the adsorption of the effective ingredient(s) to dispersible excipients and other foods was investigated. offer hope for controlling the pandemic.

provided that its quality is adequately standardized and monitored.76 - Conclusion: These results suggest the possibility of producing an anti-HIV-1 microbicide from inexpensive.. widely available sources. whose safety has been established throughout centuries. .

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