Fats and degeneration.

Fats and degeneration. A R T I C L E

Fats and degeneration

50 years ago, in the first phase of marketing the polyunsaturated fatty acids (PUFA), linoleic acid was “heart protective,” and the saturated fats raised cholesterol and caused heart disease.

In the second phase, the other “essential fatty acid,” linolenic acid, was said to be even better than linoleic acid.

In the third phase, the longer chain omega -3 (omega minus three, or n minus three) fatty acids, DHA and EPA, are said to be even better than linolenic acid.

Along the way, the highly unsaturated arachidonic acid, which we and other animals make out of the linoleic acid in foods, was coming to be identified with the “harmful animal fats.” But we just didn’t hear much about how the amount of arachidonic acid in the tissues depended on the amount of linoleic acid in the diet.

U.S. marketing dominates the world economy, including of course the communication media, so we shouldn’t expect to hear much about the role of PUFA in causing cancer, diabetes, obesity, aging, thrombosis, arthritis and immunodeficiency, or to hear about the benefits of the saturated fats.

The saturated fats include the “tropical fats,” because they are synthesized in very warm organisms, and are very stable at those temperatures. Their stability offers some protection against the unstable PUFA.

Several of the degenerative conditions produced by the 'essential fatty acids' can be reversed by use of saturated fats, varying in length from the short chains of coconut oil to the very long chains of waxes.

When a person uses a drug, there is generally an awareness that the benefit has to be weighed against the side effects. But if something is treated as a “nutrient,” especially an “essential nutrient,” there is an implication that it won’t produce undesirable side effects.

Over the last thirty years I have asked several prominent oil researchers what the evidence is that there is such a thing as an “essential fatty acid.” One professor cited a single publication about a solitary sick person who recovered from some sickness after being given some unsaturated fat. (If he had known of any better evidence, wouldn’t he have mentioned it?) The others (if they answered at all) cited “Burr and Burr, 1929.” The surprising thing about that answer is that these people can consider any nutritional research from 1929 to be definitive. It’s very much like quoting a 1929 opinion of a physicist regarding the procedure for making a hydrogen bomb. What was known about nutrition in 1929? Most of the B vitamins weren’t even suspected, and it had been only two or three years since “vitamin B” had been subdivided into two factors, the “antineuritic factor,” B₁, and the “growth factor,” B₂. Burr had no way of really understanding what deficiencies or toxicities were present in his experimental diet.

A few years after the first experiments, Burr put one of his “essential fatty acid deficient” rats under a bell jar to measure its metabolic rate, and found that the deficient animals were metabolizing 50% faster than rats that were given linoleic and linolenic acids as part of their diet. That was an important observation, but Burr didn’t understand its implications. Later, many experiments showed that the polyunsaturated fats slowed metabolism by profoundly interfering with the function of the thyroid hormone and the cellular respiratory apparatus. Without the toxic fats, respiratory energy metabolism was very intense, and a diet that was nutritionally sufficient for a sluggish animal wouldn’t necessarily be adequate for the vigorous animals.

Several publications between 1936 and 1944 made it very clear that Burr’s basic animal diet was deficient in various nutrients, especially vitamin B₆. The disease that appeared in Burr’s animals could be cured by fat free B-vitamin preparations, or by purified vitamin B6 when it became available. A zinc deficiency produces similar symptoms, and at the time Burr did his experiments, there was no information on the effects of fats on mineral absorption. If a diet is barely adequate in the essential minerals, increasing the metabolic rate, or decreasing intestinal absorption of minerals, will produce mineral deficiencies and metabolic problems.

Although “Burr’s disease” clearly turned out to be a B-vitamin deficiency, probably combined with a mineral deficiency, it continues to be cited as the basis justifying the multibillion dollar industry that has grown up around the “essential” oils.

Two years before Burr’s experiment, German researchers found that a fat-free diet prevented almost all spontaneous cancers in rats. Later work showed that the polyunsaturated fats both initiate and promote cancer. With that knowledge, the people who kept claiming that “linoleic, linolenic, and maybe arachidonic acid are the essential fatty acids,” should have devoted some effort to finding out how much of that “essential nutrient” was enough, so that people could minimize their consumption of the carcinogenic stuff.

Between the first and second world wars, cod liver oil was recommended as a vitamin supplement, at first as a source of vitamin A, and later as a source of vitamins A and D. But in the late 1940s, experimenters used it as the main fat in dogs’ diet, and found that they all died from cancer, while the dogs on a standard diet had only a 5% cancer mortality. That sort of information, and the availability of synthetic vitamins, led to the decreased use of cod liver oil.

But around that time, the seed oil industry was in crisis because the use of those oils in paints and plastics was being displaced by new compounds made from petroleum. The industry needed new markets, and discovered ways to convince the public that seed oils were better than animal fats. They were called the “heart protective oils,” though human studies soon showed the same results that the animal studies had, namely, that they were toxic to the heart and increased the incidence of cancer.

The “lipid hypothesis” of heart disease argued that cholesterol in the blood caused atherosclerosis, and that the polyunsaturated oils lowered the amount of cholesterol in the blood. Leaving behind the concept of nutritional essentiality, this allowed the industry (and their academic supporters, such as Frederick Stare at Harvard) to begin promoting the oils as having drug-like therapeutic properties. Larger amounts of polyunsaturated fat were supposed to be more protective by lowering the cholesterol, and were to be substituted for the saturated fats, which supposedly raised cholesterol and increased heart disease, producing atherosclerotic plaques in the blood vessels and increasing the formation of blood clots.

Since all ordinary foods contain significant amounts of the polyunsaturated fats, there was no reason to think that, even if they were essential nutrients, people were likely to become deficient in them. So the idea of treating the seed oils as drug-like substances, to be taken in large amounts, appealed to the food oil industry.

Prostaglandins, which are produced in the body by oxidizing the polyunsaturated fatty acids, provided an opportunity for the drug industry to get involved in a new market, and the prostaglandins offered a new way of arguing for the nutritional essentiality of linoleic and related acids: A whole system of “hormones” is made from these molecules. Since some of the prostaglandins suppress immunity, cause inflammation and promote cancer growth, some people have divided them into the “good prostaglandins” and the “bad prostaglandins.”

PGI2, or prostacyclin, is considered to be a good prostaglandin, because it causes vasodilatation, and so drug companies have made their own synthetic equivalents: Epoprostenol, iloprost, taprostene, ciprostene, UT-15, beraprost, and cicaprost. Some of these are being investigated for possible use in killing cancer.

But many very useful drugs that already existed, including cortisol and aspirin, were found to achieve some of their most important effects by inhibiting the formation of the prostaglandins. It was the body’s load of polyunsaturated fats which made it very susceptible to inflammation, stress, trauma, infection, radiation, hormone imbalance, and other fundamental problems, and drugs like aspirin and cortisone, which limit the activation of the stored “essential fatty acids,” gain their remarkable range of beneficial effects partly by the restraint they impose on those stored toxins.

Increasingly, the liberation of arachidonic acid from tissues during stress is seen as a central factor in all forms of stress, both acute (as in burns or exercise) or chronic (as in diabetes or aging). And, as the fat stores become more toxic, it seems that they more readily liberate the free fatty acids. (For example, see Iritani, et al., 1984)

During this same period, a few experimenters were finding that animals which were fed a diet lacking the “essential” fatty acids had some remarkable properties: They consumed oxygen and calories at a very high rate, their mitochondria were unusually tough and stable, their tissues could be transplanted into other animals without provoking immunological rejection, and they were very hard to kill by trauma and a wide variety of toxins that easily provoke lethal shock in animals on the usual diet. As the Germans had seen in 1927, they had a low susceptibility to cancer, and new studies were showing that they weren’t susceptible to various fibrotic conditions, including alcoholic liver cirrhosis.

In 1967 a major nutrition publication, Present Knowledge in Nutrition, published Hartroft and Porta’s observation that the “age pigment,” lipofuscin, was formed in proportion to the amount of polyunsaturated fat and oxidants in the diet. The new interest in organ transplantation led to the discovery that the polyunsaturated fats prolonged graft survival, by suppressing the immune system. Immunosuppression was considered to have a role in the carcinogenicity of the “essential” fatty acids.

Around the same time, there were studies that showed that unsaturated fats retarded brain development and produced obesity.

Substances very much like the prostaglandins, called isoprostanes and neuroprostanes, are formed spontaneously from highly unsaturated fatty acids, and are useful as indicators of the rate of lipid peroxidation in the body. Most of the products of lipid peroxidation are toxic, as a result of their reactions with proteins, DNA, and the mitochondria. The age-related glycation products that are usually blamed on sugar, are largely the result of peroxidation of the polyunsaturated fatty acids.

Through the 1970s, this sort of information about the harmful effects of the PUFA was being slowly assimilated by the culture, though many dietitians still spoke of “the essential fatty acids, vitamin F.” By 1980, it looked as though responsible researchers would see the promotion of cancer, heart disease, mitochondrial damage, hypothyroidism and immunosuppression caused by the polyunsaturated fats as their most important feature, and would see that there had never been a basis for believing that they were essential nutrients.

But then, without acknowledging that there had been a problem with the doctrine of essentiality, fat researchers just started changing the subject, shifting the public discourse to safer, more profitable topics. The fats that had been called essential, but that had so many toxic effects, were no longer emphasized, and the failed idea of “essentiality” was shifted to different categories of polyunsaturated fats.

The addition of the long chain highly unsaturated fats to baby food formulas was recently approved, on the basis of their supposed “essentiality for brain development.” One of the newer arguments for the essentiality of the PUFA is that “they are needed for making cell membranes.” But human cells can grow and divide in artificial culture solutions which contain none of the polyunsaturated fats, and no one has claimed that they are growing “without membranes.”

The long chain fats found in fish and some algae don’t interfere with animal enzymes as strongly as the seed oils do, and so by comparison, they aren’t so harmful. They are also so unstable that relatively little of them is stored in the tissues. (And when they are used as food additives, it’s necessary to use antioxidants to keep them from becoming smelly and acutely toxic.)

When meat is grilled at a high temperature, the normally spaced double bonds in PUFA migrate towards each other, becoming more stable, so that linoleic acid is turned into “conjugated linoleic acid.” This analog of the “essential” linoleic acid competes against the linoleic acid in tissues, and protects against cancer, atherosclerosis, inflammation and other effects of the normal PUFA. Presumably, anything which interferes with the essential fatty acids is protective, when the organism contains dangerous amounts of PUFA. Even the trans-isomers of the unsaturated fatty acids (found in butterfat, and convertible into conjugated linoleic acid) can be protective against cancer.

In the 1980s the oil promoters were becoming more sophisticated, and were publishing many experiments in which the fish oils were compared with corn oil, or safflower, or soy oil, and in many of those experiments, the animals’ health was better when they didn’t eat the very toxic seed oils, that contained the “essential fatty acids,” linoleic and linoleic acids.

Besides comparing the fish oils to the stronger toxins, another trick is to take advantage of the same immunosuppressive property that had seemed troublesome, and to emphasize their ability to temporarily alleviate some autoimmune or allergic diseases. X-rays were once used that way, to treat arthritis and ringworm, for example.

And, knowing that cancer cells have the ability to consume large amounts of fatty acids, they would test these fats in tissue culture dishes, and demonstrate that they were poisonous, cytotoxic, to the fast growing cancer cells. Although they caused cancer in animals, if they could be shown to kill cancer cells in a dish, they could be sold as anticancer drugs/nutrients, with the special mystique of being “essential fatty acids.” Strangely, their ability to kill cancer cells under some circumstances and to suppress some immunological reactions is being promoted in close association with the doctrine that these fats are nutritionally essential.

Arachidonic acid is made from linoleic acid, and so those two oils were considered as roughly equivalent in their ability to meet our nutritional needs, but a large part of current research is devoted to showing the details of how fish oils protect against arachidonic acid. The “balance” between the omega -3 and the omega -6 fatty acids is increasingly being presented as a defense against the toxic omega -6 fats. But the accumulation of unsaturated fats with aging makes any defense increasingly difficult, and the extreme instability of the highly unsaturated omega -3 fats creates additional problems.

PUFA and x-rays have many biological effects in common. They are immunosuppressive, but they produce their own inflammatory reactions, starting with increased permeability of capillaries, disturbed coagulation and proteolysis, and producing fibrosis and tumefaction or tissue atrophy. This isn’t just a coincidence, since ionizing radiation attacks the highly unstable polyunsaturated molecules, simply accelerating processes that ordinarily happen more slowly as a result of stress and aging.

Prolonged stress eventually tends to be a self-sustaining process, impairing the efficient respiratory production of energy, converting muscle tissue to amino acids, suppressing the thyroid, and activating further mobilization of fatty acids. Fatty acids are mobilized from within the structure of cells by phospholipases, and from fat tissues by other lipases.

The highly unsaturated fatty acids, as well as the ordinary “essential fatty acids,” act directly to increase capillary permeability, even without conversion into prostaglandins, and they interefere in many ways with the clotting and clot removal systems. The effects of PUFA taken in a meal probably disturb the clotting system more than the same quantity of saturated fat, contrary to many of the older publications. The PUFA are widely believed to prevent clotting, but when cod liver oil is given to “EFA deficient” animals, it activates the formation of clots (Hornstra, et al., 1989). An opposite effect is seen when a long chain fatty acid synergizes with aspirin, to restrain clotting (Molina, et al., 2003).

Fibrosis is a generalized consequence of the abnormal capillary permeability produced by things that disrupt the clotting system. Estrogen, with its known contribution to the formation of blood clots and edema and fibrosis and tumors, achieves part of its effect by maintaining a chronically high level of free fatty acids, preferentially liberating arachidonic acid, rather than saturated fatty acids.

Butter, beef fat, and lamb fat are the only mostly saturated fats produced on a large scale in the U.S., and the cheapness/profitability of the seed oils made it easy to displace them. But, in the face of the immense amount of propagandistic “health” claims that have been made against the saturated fats, it’s instructive to look at some of their actual effects, especially on the clotting system, and the related fibrotic reactions.

The saturated fatty acids are very unreactive chemically. Coconut oil, despite containing about 1% of the unstable PUFA, can be left in a bucket at room temperature for a year or more without showing any evidence of deterioration, suggesting that the predominance of saturated fat acts as an antioxidant for the unsaturated molecules. In the body, the saturated fats seem to act the same way, preventing or even reversing many of the conditions caused by oxidation of fats.

The stress-induced liberation of arachidonic acid causes blood vessels to leak, and this allows fibrin to escape from the blood stream, into the basement membrane and beyond into the extracellular matrix, where it produces fibrosis. (Cancer, autoimmune diseases, and heart disease involve the same inflammatory, thrombotic, fibrotic processes as the nominal fibroses.) Scleroderma, liver cirrhosis, fibrosis of the lungs, heart, and other organs, and all the diseases in which fibrous tissue becomes dense and progressively contracts, involve similar processes, and the treatments which are successful are those that stop the inflammation produced by the oxidation of the polyunsaturated fatty acids.

Retroperitoneal fibrosis is now known to be produced by estrogen, and is treated by antiestrogenic and antiserotonergic drugs, but as early as 1940 Alejandro Lipschutz demonstrated that chronic exposure to very low doses of estrogen produced fibromas in essentially every part of the body. Earlier, Loeb had studied the action of large doses of estrogen, which produced fibrosis of the uterus, as if it had accelerated aging. Following Lipschutz’ work, in which he demonstrated the “antifibromatogenic” actions of pregnenolone and progesterone, several Argentine researchers showed that progesterone prevented and cured abdominal adhesions and other fibrotic conditions, including retroperitoneal fibrosis.

Since estrogen produces both leakiness of the capillaries and excessive formation of fibrin, its effects will be seen first in the organs where it concentrates, but eventually anywhere capillaries leak fibrin. Estrogen activates the phospholipase which liberates arachidonic acid, and progesterone inhibits that phospholipase.

As the fat tissues become more burdened with arachidonic acid, they release it more easily in response to moderately lipolytic stress signals. This could explain the increased levels of free fatty acids and lipid peroxidation that occur with aging. In animals that are “deficient” in the polyunsaturated fatty acids, adrenalin doesn’t have the lipolytic effect that it does in animals on the standard diet. With aging, there is not only a tendency to have chronically higher free fatty acids in the blood, but for those fatty acids to be more unsaturated. The phospholipids of mitochondria and microsomes become more unsaturated with aging (Laganiere and Yu, 1993, Lee, et al., 1999). In the human retina there is a similar accumulation of PUFA with aging (Nourooz-Zadeh and Pereira, 1999), which implies that the aged retina will be more easily damaged by light.

Several studies suggest that a high degree of unsaturation in the fats is fundamentally related to the aging process, since long lived species have a lower degree of unsaturation in their fats. Caloric restriction decreases the age-related accumulation of the fatty acids with 4 and 5 double bonds.

Although publicity has emphasized the antiinflammatory effects of fish oil, experiments show that it is extremely effective in producing alcohol-related liver cirrhosis. Breakdown products of polyunsaturated fats (isoprostanes and 4-HNE) are found in the blood of people with alcoholic liver disease (Aleynik, et al., 1998). In the absence of polyunsaturated fats, alcohol doesn’t produce cirrhosis. Saturated fats allow the fibrosis to regress:

“A diet enriched in saturated fatty acids effectively reverses alcohol-induced necrosis, inflammation, and fibrosis despite continued alcohol consumption. The therapeutic effects of saturated fatty acids may be explained, at least in part, by reduced endotoxemia and lipid peroxidation....” (Nanji, et al., 1995, 2001)

In these studies, the animals were switched from fish oil to either palm oil or medium chain triglycerides (a major fraction of coconut oil). In other studies, Knittel, et al. (1995), show that fibrinogen, in “a clotting-like process,” is involved in the development of liver fibrosis, and that this appears to provide a basis for the growth of additional extracellular matrix.

Brown, et al. (1989), discussed this developmental process (leaky capillaries, fibrosis) in relation to wound healing, lung disease, and tumor growth.

The relatively few studies of fish oil and linoleic acid that compare them with palmitic acid or coconut oil have produced some very important results. For example, pigs exposed to endotoxin developed severe lung problems (resembling “shock lung”) when they had been on a diet with either fish oil or Intralipid (which is mostly linoleic acid, used for intravenous feeding in hospitals), but not after palmitic acid (Wolfe, et al., 2002).

Eating low-fat seafood (sole, whitefish, turbot, scallops, oysters, lobster, shrimp, squid, etc.) once in a while can provide useful trace minerals, without much risk. However, fish from some parts of the ocean contain industrial contaminants in the fat, and large fish such as tuna, swordfish, Chilean sea bass and halibut contain toxic amounts of mercury in the muscles. Chilean sea bass (Patagonian toothfish) is very high in fat, too.

About ten years ago I met a young man with a degenerative brain disease, and was interested in the fact that he (working on a fishing boat) had been eating almost a pound of salmon per day for several years. There is now enough information regarding the neurotoxic effects of fish oil to justify avoidance of the fatty fish.

Some of the current advertising is promoting fish oil to prevent cancer, so it’s important to remember that there are many studies showing that it increases cancer.

The developmental and physiological significance of the type of fatty acid in the diet has been established for a long time, but cultural stereotypes and commercial interests are threatened by it, so it can’t be discussed publicly.

REFERENCES

Alcohol Clin Exp Res 1998 Feb;22(1):192-6. Increased circulating products of lipid peroxidation in patients with alcoholic liver disease. Aleynik SI, Leo MA, Aleynik MK, Lieber CS

Ann N Y Acad Sci. 1976;275:28-46. Metabolic influences in experimental thrombosis. Antoniades HN, Westmoreland N.

Nutr Cancer. 2001;41(1-2):91-7. Vaccenic acid feeding increases tissue levels of conjugated linoleic acid and suppresses development of premalignant lesions in rat mammary gland. Banni S, Angioni E, Murru E, Carta G, Melis MP, Bauman D, Dong Y, Ip C.

Obstet Gynecol. 1987 Sep;70(3 Pt 2):502-4. The treatment of retroperitoneal fibromatosis with medroxyprogesterone acetate. Barnhill D, Hoskins W, Burke T, Weiser E, Heller P, Park R. Wide excision is the recommended primary therapy for retroperitoneal fibromatosis. Radiation therapy and a variety of medications have been used to treat patients with recurrent tumors, but the response to these agents has not been uniform. The patient presented was successfully treated with medroxyprogesterone acetate for recurrent retroperitoneal fibromatosis that was refractory to multiple operative resections and radiation therapy.

Medicina (B Aires). 1978 Mar-Apr;38(2):215-6. [Fibromatosis, relaxin and progesterone] [in Spanish] Barousse AP. [Letter]

Medicina (B Aires). 1985;45(2):159-63. Progesterone as therapy for retroperitoneal fibrosis. Bilder CR, Barousse AP, Mazure PA.

Adv Exp Med Biol. 1976;75:497-503. Effect of ionizing radiation on liver microcirculation and oxygenation. Bicher HI, Dalrymple GV, Ashbrook D, Smith R, Harris D.

Lipids. 1981 May;16(5):323-7. Iodination of docosahexaenoic acid by lactoperoxidase and thyroid gland in vitro: formation of an lodolactone. Boeynaems JM, Watson JT, Oates JA, Hubbard WC. “In the presence of iodide, hydrogen peroxide and lactoperoxidase, docosahexaenoic acid (22:6 omega 3) was converted into iodinated compounds.”

Am Rev Respir Dis 1989 Oct;140(4):1104-7. Leaky vessels, fibrin deposition, and fibrosis: a sequence of events common to solid tumors and to many other types of disease. Brown LF, Dvorak AM, Dvorak HF

Medicina (B Aires). 1979 Sep-Oct;39(5):652-4. [Effect of progesterone in the treatment of a patient with idiopathic retroperitoneal fibrosis] [in Spanish] Casadei DH, Najun Zarazaga C, Leanza HJ, Schiappapietra JH.

Biochem Mol Biol Int 1993 Jan;29(1):175-83. Influence of antioxidant vitamins on fatty acid inhibition of lymphocyte proliferation. Calder PC, Newsholme EA. “Vitamin E (10 microM) increased human lymphocyte proliferation by 35%. However, vitamin E did not prevent the inhibitory effects of fatty acids upon lymphocyte proliferation. It is concluded that inhibition of lymphocyte proliferation by fatty acids is not caused by their conversion to peroxidised products.”

Clin Sci (Lond). 1992 Jun;82(6):695-700. Polyunsaturated fatty acids suppress human peripheral blood lymphocyte proliferation and interleukin-2 production. Calder PC, Newsholme EA.

J Neurochem 1980 Oct;35(4):1004-7. Transient formation of superoxide radicals in polyunsaturated fatty acid-induced brain swelling. Chan PH, Fishman RA

Int J Cancer 2001 Mar 15;91(6):894-9. Tumor invasiveness and liver metastasis of colon cancer cells correlated with cyclooxygenase-2 (COX-2) expression and inhibited by a COX-2-selective inhibitor, etodolac. Chen WS, Wei SJ, Liu JM, Hsiao M, Kou-Lin J, Yang WK.

Free Radic Biol Med. 1999 Jul;27(1-2):51-9. Arachidonic acid interaction with the mitochondrial electron transport chain promotes reactive oxygen species generation. Cocco T, Di Paola M, Papa S, Lorusso M.

Clin Exp Metastasis 1997 Jul;15(4):410-7. Influence of lipid diets on the number of metastases and ganglioside content of H59 variant tumors. Coulombe J, Pelletier G, Tremblay P, Mercier G, Oth D.

BJU Int. 2003 Jun;91(9):830-8. Fibrin as an inducer of fibrosis in the tunica albuginea of the rat: a new animal model of Peyronie's disease. Davila HH, Ferrini MG, Rajfer J, Gonzalez-Cadavid NF.

Carcinogenesis 1994 Jul;15(7):1399-404. Peroxidation of linoleic, arachidonic and oleic acid in relation to the induction of oxidative DNA damage and cytogenetic effects. de Kok TM, ten Vaarwerk F, Zwingman I, van Maanen JM, Kleinjans JC.

Biochem Biophys Res Commun. 2000 Oct 14;277(1):128-33. Arachidonic acid causes cytochrome c release from heart mitochondria. Di Paola M, Cocco T, Lorusso M.

J Physiol. 1998 Mar 1;507 ( Pt 2):541-7. Arachidonic acid increases cerebral microvascular permeability by free radicals in single pial microvessels of the anaesthetized rat. Easton AS, Fraser PA.

Am J Physiol. 1992 May;262(5 Pt 1):E637-43. ATP depletion stimulates calcium-dependent protein breakdown in chick skeletal Muscle. Fagan JM, Wajnberg EF, Culbert L, Waxman L.

Cancer Res 1998 Aug 1;58(15):3312-9. Dietary omega-3 polyunsaturated fatty acids promote colon carcinoma metastasis in rat liver. Griffini P, Fehres O, Klieverik L, Vogels IM, Tigchelaar W, Smorenburg SM, Van Noorden CJ.

J Indian Med Assoc 1997 Mar;95(3):67-9, 83. Association of dietary ghee intake with coronary heart disease and risk factor prevalence in rural males. Gupta R, Prakash H

Transplantation 1995 Sep 27;60(6):570-7. The effect of dietary polyunsaturated fatty acids (PUFA) on acute rejection and cardiac allograft blood flow in rats. Haw MP, Linnebjerg H, Chavali SR, Forse RA. “The immunosuppressive effect of dietary PUFA warrants further investigation, and their use as a possible adjunctive treatment in organ transplantation should be considered.”

Dtsch Med Wochenschr. 2003 Jun 20;128(25-26):1395-8. [Rare cause of chronic abdominal pain: retractile mesenteritis] [in German] Hermann F, Speich R, Schneemann M. “Retractile mesenteritis is a rare cause of chronic abdominal pain with variable symptoms. Its aetiology is unknown. In case of bowel ischemia a surgical approach is preferred, milder forms may be treated with immunosuppressive agents as well as oral progesterone. Progesterone has exhibited positive effects on fatty tissue with successful treatment in desmoid tumors and retroperitoneal fibrosis. Here in we could demonstrate its safe and efficient use in a patient with retractile mesenteritis.”

Mech Ageing Dev 2001 Apr 15;122(4):427-43. Effect of the degree of fatty acid unsaturation of rat heart mitochondria on their rates of H2O2 production and lipid and protein oxidative damage. Herrero A, Portero-Otin M, Bellmunt MJ, Pamplona R, Barja G. “Previous comparative studies have shown that long-lived animals have lower fatty acid double bond content in their mitochondrial membranes than short-lived ones. In order to ascertain whether this trait protects mitochondria by decreasing lipid and protein oxidation and oxygen radical generation, the double bond content of rat heart mitochondrial membranes was manipulated by chronic feeding with semi-purified AIN-93G diets rich in highly unsaturated (UNSAT) or saturated (SAT) oils. UNSAT rat heart mitochondria had significantly higher double bond content and lipid peroxidation than SAT mitochondria. They also showed increased levels of the markers of protein oxidative damage malondialdehyde-lysine, protein carbonyls, and N(e)-(carboxymethyl)lysine adducts.” “These results demonstrate that increasing the degree of fatty acid unsaturation of heart mitochondria increases oxidative damage to their lipids and proteins, and can also increase their rates of mitochondrial oxygen radical generation in situations in which the degree of reduction of Complex III is higher than normal. These observations strengthen the notion that the relatively low double bond content of the membranes of long-lived animals could have evolved to protect them from oxidative damage.”

Biochem J. 1994 May 15;300 ( Pt 1):251-5. Regulation of fibrinolysis by non-esterified fatty acids. Higazi AA, Aziza R, Samara AA, Mayer M. “Examination of the fatty acid specificity showed that a minimal chain length of 16 carbon atoms and the presence of at least one double bond, preferably in a cis configuration, were required for inhibition of the fibrinolytic activity of plasmin.”

Science. 1976 Feb 27;191(4229):861-2. Nicotinic acid reduction of plasma volume loss after thermal trauma. Hilton JG, Wells CH. Intravenous administration of nicotinic acid to the anesthetized dog prior to thermal trauma reduced plasma loss at 10 minutes after burn from 7 milliliters per kilogram to less than 2 millimeters per kilogram. During the next 50 minutes plasma loss was the same in treated and untreated animals. An additional dose of nicotinic acid 30 minutes after burn prevented this further loss.

Z Gesamte Inn Med. 1976 Oct 15;31(20):838-43. [Age-dependence of catecholamine effects in man. IV. Effects of specific inhibitors on the lipolytic action of alpha and beta adrenergics] [in German] Hoffmann H.

Thromb Res. 1989 Jan 1;53(1):45-53. Normalization by dietary cod-liver oil of reduced thrombogenesis in essential fatty acid deficient rats. Hornstra G, Haddeman E, Don JA.

Radiographics. 2003 Nov-Dec;23(6):1561-7. CT Findings in Sclerosing Mesenteritis (Panniculitis): Spectrum of Disease. Horton KM, Lawler LP, Fishman EK.

Nutr Cancer. 1985;7(4):199-209. Isomeric fatty acids and tumorigenesis: a commentary on recent work. Hunter JE, Ip C, Hollenbach EJ. “Neither epidemiological nor experimental studies published to date have demonstrated any valid association between trans fatty acid ingestion and tumorigenesis. A recent study showed that under controlled conditions, a fat with a high content of trans fatty acids did not promote the development of mammary tumors induced in rats by 7,12-dimethylbenz[a]anthracene to any greater extent than did a comparable fat with a high content of cis fatty acids. In addition, in this study a high trans fat was less tumor promoting than was a blend of fats that simulated the dietary fat composition of the United States and had a lower level of trans fatty acids.”

Medicina (B Aires). 1978 Mar-Apr;38(2):215. [Progesterone and retroperitoneal fibrosis] [in Spanish] Introzzi A.[Letter]

Cancer Res. 1985 May;45(5):1997-2001. Requirement of essential fatty acid for mammary tumorigenesis in the rat. Ip C, Carter CA, Ip MM. “Mammary tumorigenesis was very sensitive to linoleate intake and increased proportionately in the range of 0.5 to 4.4% of dietary linoleate.”

Clin Exp Metastasis 2000;18(5):371-7. Promotion of colon cancer metastases in rat liver by fish oil diet is not due to reduced stroma formation. Klieveri L, Fehres O, Griffini P, Van Noorden CJ, Frederiks WM. “Recently, it was demonstrated that dietary omega-3 polyunsaturated fatty acids (PUFAs) induce 10-fold more metastases in number and 1000-fold in volume in an animal model of colon cancer metastasis in rat liver.”

Folia Haematol Int Mag Klin Morphol Blutforsch. 1977;104(1):1-10. [Review: hemorrhagic diathesis resulting from acute exposure to ionizing Radiation] [Article in German] Krantz S, Lober M. The symptoms of the acute radiopathy are chiefly characterized by a severe blood coagulation disorder. The main results and problems of research work on this haemorrhagic diathesis are shortly reviewed.

Gerontology 1993;39(1):7-18. Modulation of membrane phospholipid fatty acid composition by age and food restriction. Laganiere S, Yu BP. H.M. “Phospholipids from liver mitochondrial and microsomal membrane preparations were analyzed to further assess the effects of age and lifelong calorie restriction on membrane lipid composition.” “The data revealed characteristic patterns of age-related changes in ad libitum (AL) fed rats: membrane levels of long-chain polyunsaturated fatty acids, 22:4 and 22:5, increased progressively, while membrane linoleic acid (18:2) decreased steadily with age. Levels of 18:2 fell by approximately 40%, and 22:5 content almost doubled making the peroxidizability index increase with age.” “We concluded that the membrane-stabilizing action of long-term calorie restriction relates to the selective modification of membrane long-chain polyunsaturated fatty acids during aging.”

Medicina (B Aires). 1978 Mar-Apr;38(2):123-32. [Effective treatment of several types of fibromatosis with progesterone. Fibrous mediastinitis, desmoid tumors, paraneoplastic fibrosis] [in Spanish] Lanari A, Molinas FC, Castro Rios M, Paz RA.

Medicina (B Aires). 1979 Nov-Dec;39(6):826-35. [Progesterone in fibromatosis and atherosclerosis] [in Spanish] Lanari A.

Free Radic Biol Med 1999 Feb;26(3-4):260-5. Modulation of cardiac mitochondrial membrane fluidity by age and calorie intake. Lee J, Yu BP, Herlihy JT. “The fatty acid composition of the mitochondrial membranes of the two ad lib fed groups differed: the long-chain polyunsaturated 22:4 fatty acid was higher in the older group, although linoleic acid (18:2) was lower. DR eliminated the differences.” “Considered together, these results suggest that DR maintains the integrity of the cardiac mitochondrial membrane fluidity by minimizing membrane damage through modulation of membrane fatty acid profile.”

Lipids 2001 Jun;36(6):589-93. Effect of dietary restriction on age-related increase of liver susceptibility to peroxidation in rats. Leon TI, Lim BO, Yu BP, Lim Y, Jeon EJ, Park DK.

Acta Chir Scand. 1976;142(1):20-5. Induction of endogenous fibrinolysis inhibition in the dog. Effect of intravascular coagulation and release of free fatty acids. Lindquist O, Bagge L, Saldeen T. “In all groups subjected to infusion of thrombin an increase in plasma free fatty acids (FFA) was observed. The role of this increase for the development of fibrinolysis inhibition was tested by infusion of norepinephrine alone and in combination with nicotinic acid. Norepinephrine caused an increase of FFA after 2 hours and in urokinase inhibitor activity after 24-48 hours. Both of these were diminished by high doses of nicotinic acid, indicating that the release of FFA rather than intravascular coagulation might be the principal mechanism underlying the occurrence of fibrinolysis inhibition following trauma.”

Proc Natl Acad Sci U S A 1990 Nov;87(22):8845-9. Incorporation of marine lipids into mitochondrial membranes increases susceptibility to damage by calcium and reactive oxygen species: evidence for enhanced activation of phospholipase A2 in mitochondria enriched with n-3 fatty acids. Malis CD, Weber PC, Leaf A, Bonventre JV.

Prostaglandins Leukot Essent Fatty Acids 1994 Jul;51(1):33-40. Suppression of human T-cell growth in vitro by cis-unsaturated fatty acids: relationship to free radicals and lipid peroxidation. Madhavi N, Das UN, Prabha PS, Kumar GS, Koratkar R, Sagar PS.

Clin Exp Metastasis 1998 Jul;16(5):407-14. Diminution of the development of experimental metastases produced by murine metastatic lines in essential fatty acid-deficient host mice. Mannini A, Calorini L, Mugnai G, Ruggieri S.

Biochem Pharmacol. 1990 Mar 1;39(5):879-89. Histamine release from rat mast cells induced by metabolic activation of polyunsaturated fatty acids into free radicals. Masini E, Palmerani B, Gambassi F, Pistelli A, Giannella E, Occupati B, Ciuffi M, Sacchi TB, Mannaioni PF.

Journal of Lipid Research, Vol. 44, 271-279, February 2003. Arachidonic acid and prostacyclin signaling promote adipose tissue development : a human health concern? F. Massiera, P. Saint-Marc, J. Seydoux , T. Murata , T. Kobayashi , S. Narumiya , P. Guesnet, Ez-Zoubir Amri, R. Negrel and G. Ailhaud1.

Infection. 1994 Mar-Apr;22(2):106-12. Influence of dietary (n-3)-polyunsaturated fatty acids on leukotriene B4 and prostaglandin E2 synthesis and course of experimental tuberculosis in guinea pigs. Mayatepek E, Paul K, Leichsenring M, Pfisterer M, Wagner D, Domann M, Sonntag HG, Bremer HJ.

Biochim Biophys Acta 1994 Sep 15;1214(2):209-20. Reinvestigation of lipid peroxidation of linolenic acid. Mlakar A, Spiteller G. “Thus, a great number of previously unknown lipid peroxidation products was detected. It is assumed that these compounds also occur--at least as intermediates--in lipid peroxidation processes in mammalian tissue.”

Prostaglandins Leukot Essent Fatty Acids. 2003 May;68(5):305-10. Synergistic effect of D-003 and aspirin on experimental thrombosis models. Molina V, Arruzazabala ML, Carbajal D, Mas R.

Chem Res Toxicol. 2001 Apr;14(4):431-7. Defining mechanisms of toxicity for linoleic acid monoepoxides and diols in Sf-21 cells. Moran JH, Mon T, Hendrickson TL, Mitchell LA, Grant DF.

J Biochem (Tokyo). 1977 Aug;82(2):529-33. Effects of free fatty acids on fibrinolytic activity. Muraoka T, Okuda H. A novel method for the estimation of fibrinolytic activity is proposed. In this method, a fibrin clot suspension is used as a substrate (fibrin is known to be a physiological substrate of plasmin). The fibrin clot suspension was prepared by homogenization of human fibrin clots. With this method, we found that free fatty acids inhibited the plasmin activity, and long-chain, unsaturated free fatty acids had a particularly strong inhibitory action on plasmin. As regards the mechanism of the inhibitory action, free fatty acids may not inhibit complex formation between plasmin and fibirin, but may make it impossible for plasmin to act on fibrin due to deformation of the surface of the fibrin clot.

Alcohol Clin Exp Res. 1986 Jun;10(3):271-3. Dietary factors and alcoholic cirrhosis. Nanji AA, French SW.

Gastroenterology. 1995 Aug;109(2):547-54. Dietary saturated fatty acids: a novel treatment for alcoholic liver disease. Nanji AA, Sadrzadeh SM, Yang EK, Fogt F, Meydani M, Dannenberg AJ.

J Pharmacol Exp Ther. 1996 Jun;277(3):1694-700. Medium chain triglycerides and vitamin E reduce the severity of established experimental alcoholic liver disease. Nanji AA, Yang EK, Fogt F, Sadrzadeh SM, Dannenberg AJ.

Hepatology. 1997 Dec;26(6):1538-45. Dietary saturated fatty acids down-regulate cyclooxygenase-2 and tumor necrosis factor alfa and reverse fibrosis in alcohol-induced liver disease in the rat. Nanji AA, Zakim D, Rahemtulla A, Daly T, Miao L, Zhao S, Khwaja S, Tahan SR, Dannenberg AJ.

J Pharmacol Exp Ther. 2001 Nov;299(2):638-44. Dietary saturated fatty acids reverse inflammatory and fibrotic changes in rat liver despite continued ethanol administration. Nanji AA, Jokelainen K, Tipoe GL, Rahemtulla A, Dannenberg AJ.

Gastroenterology 1995 Apr;108(4):1124-35. Accumulation and cellular localization of fibrinogen/fibrin during short-term and long-term rat liver injury. Neubauer K, Knittel T, Armbrust T, Ramadori G “Fibrinogen/fibrin deposition in damaged livers was studied by immunohistology.” “Immunohistology showed striking amounts of fibrinogen and fibrin deposits in pericentral necrotic areas (short-term damage) and within fibrotic septa (long-term damage).” “The results show fibrinogen/fibrin deposition during short-term liver injury and liver fibrogenesis, which may suggest the involvement of a "clotting-like process" in short-term liver damage and liver fibrosis. The data might indicate that fibrin/fibronectin constitute a "provisional matrix," which affects the attraction and proliferation of inflammatory and matrix-producing cells.”

Ophthalmic Res. 1999;31(4):273-9. Age-related accumulation of free polyunsaturated fatty acids in human retina. Nourooz-Zadeh J, Pereira P.

Chem Res Toxicol. 2002 Mar;15(3):367-72. Formation of cyclic deoxyguanosine adducts from omega-3 and omega-6 polyunsaturated fatty acids under oxidative conditions. Pan J, Chung FL.

Radiobiologiia. 1985 Nov-Dec;25(6):763-7. [Mechanism of circulatory disorders in animals irradiated at high doses] [in Russian] Pozharisskaia TD, Vasil'eva TP, Sokolova EN, Alekseeva II. Some data are reported on pathoanatomical changes, a status of the microcirculatory channel and the coagulogram of animals affected by high doses of ionizing radiation. The signs of disseminated intravascular blood coagulation have been revealed.

J Biol Chem. 1998 May 29;273(22):13605-12. Formation of isoprostane-like compounds (neuroprostanes) in vivo from docosahexaenoic acid. Roberts LJ 2nd, Montine TJ, Markesbery WR, Tapper AR, Hardy P, Chemtob S, Dettbarn WD, Morrow JD.

Nutr Cancer 1995;24(1):33-45. Effects of linoleic acid and gamma-linolenic acid on the growth and metastasis of a human breast cancer cell line in nude mice and on its growth and invasive capacity in vitro. Rose DP, Connolly JM, Liu XH

Nutr Cancer 1998;30(2):137-43. Effects of dietary n-3-to-n-6 polyunsaturated fatty acid ratio on mammary carcinogenesis in rats. Sasaki T, Kobayashi Y, Shimizu J, Wada M, In'nami S, Kanke Y, Takita T. “An increase in the n-3/n-6 ratio did not suppress the incidence or reduce the latency of mammary tumor development. The number and weight of mammary tumors per tumor-bearing rat tended to be large in the group with an n-3/n-6 ratio of 7.84 compared with those in the other groups. As the n-3/n-6 ratios were elevated, the total number and weight of tumors increased gradually.”

J. Biol. Chem. 1940 132: 539-551. Essential fatty acids, vitamin B₆, and other factors in the cure of rat acrodynia. H. Schneider, H. Steenbock, and Blanche R. Platz

Science. 1988 May 20;240(4855):1032-3. Essential fatty acid depletion of renal allografts and prevention of rejection. Schreiner GF, Flye W, Brunt E, Korber K, Lefkowith JB.

Physiol Bohemoslov. 1990;39(2):125-34. Proportion of individual fatty acids in the non-esterified (free) fatty acid (FFA) fraction in the serum of laboratory rats of different ages. Smidova L, Base J, Mourek J, Cechova I.

Acta Neurochir Suppl (Wien) 1994;60:20-3. Mechanisms of glial swelling by arachidonic acid. Staub F, Winkler A, Peters J, Kempski O, Baethmann A.

Arch Biochem Biophys. 1991 Aug 15;289(1):33-8. A possible mechanism of mitochondrial dysfunction during cerebral ischemia: inhibition of mitochondrial respiration activity by arachidonic acid. Takeuchi Y, Morii H, Tamura M, Hayaishi O, Watanabe Y.

J Drug Target. 2003 Jan;11(1):45-52. Modulation of tumor-selective vascular blood flow and extravasation by the stable prostaglandin 12 analogue beraprost sodium. Tanaka S, Akaike T, Wu J, Fang J, Sawa T, Ogawa M, Beppu T, Maeda H.

Biochem Soc Trans. 2003 Oct;31(Pt 5):1075-9. Regression of pre-established atherosclerosis in the apoE-/- mouse by conjugated linoleic acid. Toomey S, Roche H, Fitzgerald D, Belton O.

Int J Biochem Cell Biol. 2003 May;35(5):749-55. Increased muscle proteasome activities in rats fed a polyunsaturated fatty acid supplemented diet. Vigouroux S, Farout L, Clavel S, Briand Y, Briand M. “Changes in the proteasome system, a dominant actor in protein degradation in eukaryotic cells, have been documented in a large number of physiological and pathological conditions.” “With the polyunsaturated fatty acid enriched diet, the chymotrypsin-like and peptidylglutamylpeptide hydrolase activities increased by 45% in soleus and extensor digitorum longus (EDL), and by 90% in the gastrocnemius medialis (GM) muscle. Trypsin-like activity of the proteasome increased by 250% in soleus, EDL and GM.” “Proteasome activities and level were less stimulated with a monounsaturated fatty acid supplemented diet.” “Unsaturated fatty acids are particularly prone to free radical attack. Thus, we suggest that alterations in muscle proteasome may result from monounsaturated and polyunsaturated fatty acid-induced peroxidation, in order to eliminate damaged proteins.”

J Am Coll Nutr. 2000 Aug;19(4):478S-486S. Conjugated linoleic acid and bone biology. Watkins BA, Seifert MF. “Recent investigations with growing rats given butter fat and supplements of CLA demonstrated an increased rate of bone formation and reduced ex vivo bone PGE2 production, respectively.”

Ups J Med Sci. 1979;84(3):195-201. Effect of nicotinic acid on the posttraumatic increase in free fatty acids and fibrinolysis inhibition activity in the rat. Wegener T, Bagge L, Saldeen T. Nicotinic acid effectively inhibited the posttraumatic increase in both free fatty acids (FFA) and fibrinolysis inhibition activity (FIA) in the blood in rats, indicating that FFA might be involved in the posttraumatic increase of FIA. The FIA in the liver was greater than that in other organs studied and was increased in the posttraumatic phase. The possible role of the liver in the posttraumatic increase of FIA is discussed.

Am J Physiol Regul Integr Comp Physiol. 2001 Mar;280(3):R908-12. CLA reduces antigen-induced histamine and PGE(2) release from sensitized guinea pig tracheae. Whigham LD, Cook EB, Stahl JL, Saban R, Bjorling DE, Pariza MW, Cook ME.

Toxicol Appl Pharmacol 1993 May;120(1):72-9. Essential fatty acid deficiency in cultured human keratinocytes attenuates toxicity due to lipid peroxidation. Wey HE, Pyron L, Woolery M.

Nutrition. 2002 Jul-Aug;18(7-8):647-53. Dietary fat composition alters pulmonary function in pigs. Wolfe RR, Martini WZ, Irtun O, Hawkins HK, Barrow RE.

Thromb Res. 1997 Nov 1;88(3):283-90. Enhanced blood coagulation and enhanced fibrinolysis during hemodialysis with prostacyclin. Novacek G, Kapiotis S, Jilma B, Quehenberger P, Michitsch A, Traindl O, Speiser W. In the present study the effect of unfractionated heparin (UFH) (Liquemin, 750-1000 IU/h), low molecular weight heparin (LMWH) (Fragmin, 3000-7250 IU bolus), and prostacyclin (Flolan, 5 ng/kg body weight/min) on the activation of blood coagulation and fibrinolysis, induced by polysulfone membrane dialyzers during hemodialysis, was compared. Plasma levels of thrombin-antithrombin III complex (TAT), fibrin split product D-dimer, and plasmin-plasmin inhibitor-complex (PPI) were measured in the arterial and venous line of the dialyzer at the beginning and at 10, 60, 120, and 180 minutes of hemodialysis. Five patients on chronic hemodialysis treatment were investigated in a cross over study. Clinically all three anticoagulation regimen were sufficient for hemodialysis treatment. Using UFH or LMWH TAT, PPI, and D-dimer levels were similar in the venous and the arterial line of the dialyzer. However, during prostacyclin treatment the levels of these activation markers were significantly higher in the venous line. Based on these data the dialyzer membrane can be considered as a site of activation of blood coagulation and of fibrinolysis during anticoagulation with prostacyclin in hemodialysis.

Am J Clin Nutr. 2003 May;77(5):1125-32. Effect of individual dietary fatty acids on postprandial activation of blood coagulation factor VII and fibrinolysis in healthy young men. Tholstrup T, Miller GJ, Bysted A, Sandstrom B. “When the data were pooled, the saturated (S, P, and M) test fats resulted in a smaller postprandial increase in FVIIa (P = 0.036, diet effect), a smaller increase in FVII:c (P < 0.001, diet x time interaction), a greater rise in tissue plasminogen activator concentrations (P = 0.028, diet effect), and a tendency to a greater postprandial decline in PAI-1 (P = 0.06, diet effect) compared with the unsaturated test fats (O, T, and L).” “Our results indicate a lesser increase in FVIIa after the consumption of saturated fats, especially the S fat, than after unsaturated test fats.”

Randomized Controlled Trial

Prostaglandins. 1978 Apr;15(4):557-64. Prostaglandin I2 as a potentiator of acute inflammation in rats. Komoriya K, Ohmori H, Azuma A, Kurozumi S, Hashimoto Y, Nicolaou KC, Barnette WE, Magolda RL. Prostaglandin I2 potentiated the paw swelling induced by carrageenin in rats. Prostaglandin I2 (0.1 microgram) showed similar activity to PGE1 (0.01 microgram). This potentiating property disappeared in 60 minutes and was completely abolished by diphenhydramine (25 mg kg-1, i.p.). In vascular permeability tests, PGI2 itself (2.5 X 10(-10) mol, 88 ng) caused no dye leakage reaction, but PGE1 (2.5 X 10(-10) mol, 88.5 ng) caused a significant dye leakage. This effect of PGE1 was statistically significant compared with vehicle- or PGI2-treated groups (p less than 0.05). Prostaglandin I2 potentiated the increased vascular permeability induced by 5-hydroxytriptamine (2.5 X 10(-10) mol), bradykinin (5 X 10(-10) mol) and histamine (2 X 10(-10) to 2 X 10(-8) mol). The potentiation was the most evident in the case of histamine.

Journal of Lipid Research, Vol. 44, 271-279, February 2003. Arachidonic acid and prostacyclin signaling promote adipose tissue development : a human health concern? F.lorence Massiera*, Perla Saint-Marc*, Josiane Seydoux , Takahiko Murata , Takuya Kobayashi , Shuh Narumiya , Philippe Guesnet**, Ez-Zoubir Amri*, Raymond Negrel* and Gérard Ailhaud1,* Institut de Recherche Signalisation, Biologie du Développement et Cancer, Centre de Biochimie (UMR6543CNRS), UNSA, Faculté des Sciences, Parc Valrose, 06108 Nice cedex 2, France Centre Médical Universitaire, Département de Physiologie, 1 rue Michel Servet, 1211 Genève 4, Switzerland Department of Pharmacology, Kyoto University, Faculty of Medicine, Yoshida, Sakyo-ku, Kyoto 606-8315, Japan** Laboratoire de Nutrition et Sécurité Alimentaire, INRA, 78352 Jouy-en-Josas, France 1 To whom correspondence should be addressed. e-mail: ailhaud@unice.fr High fat intake is associated with fat mass gain through fatty acid activation of peroxisome proliferator-activated receptors and , which promote adipogenesis. We show herein that, compared to a combination of specific agonists to both receptors or to saturated, monounsaturated, and -3 polyunsaturated fatty acids, arachidonic acid (C20:4, -6) promoted substantially the differentiation of clonal preadipocytes. This effect was blocked by cyclooxygenase inhibitors and mimicked by carbacyclin, suggesting a role for the prostacyclin receptor and activation of the cyclic AMP-dependent pathways that regulate the expression of the CCAAT enhancer binding proteins ß and implicated in adipogenesis. During the pregnancy-lactation period, mother mice were fed either a high-fat diet rich in linoleic acid, a precursor of arachidonic acid (LO diet), or the same isocaloric diet enriched in linoleic acid and -linolenic acid (LO/LL diet). Body weight from weaning onwards, fat mass, epididymal fat pad weight, and adipocyte size at 8 weeks of age were higher with LO diet than with LO/LL diet. In contrast, prostacyclin receptor-deficient mice fed either diet were similar in this respect, indicating that the prostacyclin signaling contributes to adipose tissue development. These results raise the issue of the high content of linoleic acid of i) ingested lipids during pregnancy and lactation, and ii) formula milk and infant foods in relation to the epidemic of childhood obesity.

Arthritis Rheum 2000 Apr;43(4):894-900

Abnormalities of erythrocyte membrane fluidity, lipid composition, and lipid peroxidation in systemic sclerosis: evidence of free radical-mediated injury. Solans R, Motta C, Sola R, La Ville AE, Lima J, Simeon P, Montella N,

Armadans-Gil L, Fonollosa V, Vilardell M

Medicina Interna, Vall d'Hebron University General Hospital, Barcelona, Spain.

OBJECTIVE: To elucidate whether oxidative injury occurs in systemic sclerosis

(SSc) and whether it affects the erythrocyte membrane (EM) properties. METHODS:

EM fluidity and lipid composition (cholesterol:phospholipid molar ratio [C:PL],

fatty acid composition) were studied in 52 patients with SSc and in 53 subjects

without SSc (32 with primary Raynaud's phenomenon [RP] and 21 healthy subjects

[controls]). Fluidity was measured as the fluorescence anisotropy of the

hydrophobic fluorescent probe DPH (1,6-diphenyl-1,3,5-hexatriene). Lipid

peroxidation products were determined as thiobarbituric acid-reactive substances

(TBARS). RESULTS: EM fluidity was significantly lower in SSc patients than in

primary RP patients and controls (P < 0.001). The EM C:PL molar ratio was

significantly higher in SSc patients than in primary RP patients and controls (P

< 0.05). Levels of EM polyunsaturated n6 fatty acids (PUFA n6) were

significantly lower in SSc patients than in primary RP patients and controls (P

< 0.001). TBARS were significantly increased in SSc patients compared with

primary RP patients and controls (P < 0.001). Multiple regression analyses indicated that the reduced EM fluidity was partly due to its greater C:PL molar ratio, lower PUFA n6 content, and higher TBARS levels. EM fluidity was lower among patients with nailfold capillary loss (P < 0.001) and digital ischemic ulcers (P < 0.05). EM lipid peroxidation products were higher among patients with pulmonary involvement (bibasal pulmonary fibrosis [P < 0.05] and reduced levels of diffusing capacity for carbon monoxide [P < 0.001]) and among patients who were positive for anti-topoisomerase I antibodies (P < 0.05) or negative for anticentromere antibodies (P < 0.001). CONCLUSION: Our findings support the idea that oxidative injury occurs in SSc and that, through lipid peroxidation, it

induces structural and functional changes of the EM that may contribute to the

development of the microvascular abnormalities that are seen in the disease.

Alcohol Clin Exp Res 1998 Feb;22(1):192-6

Increased circulating products of lipid peroxidation in patients with alcoholic liver disease.

Aleynik SI, Leo MA, Aleynik MK, Lieber CS

Alcohol Research and Treatment Center, Bronx Veterans Affairs Medical Center and

Mount Sinai School of Medicine, New York 10468, USA. F2-isoprostanes (F2-IP) and 4-hydroxynonenal (4-HNE), peroxidation products of polyunsaturated fatty acids (PUFA), are considered the most reliable indicators of endogenous lipid peroxidation in vivo. To determine to what extent these are

also altered in patients with alcoholic liver disease, plasma free and esterified F2-IP as well as 4-HNE were measured by GC/MS in 49 fasting subjects who underwent diagnostic percutaneous needle biopsies of the liver. Compared to

patients with mild steatosis and no fibrosis, free F2-IP and 4-HNE were strikingly increased in individuals with alcoholic hepatitis. There was also a significant but lesser rise of 4-HNE in patients with perivenular fibrosis. An

increase of F2-IP was also found in subjects with transition to, or complete, alcoholic cirrhosis, with a comparable trend for 4-HNE. By contrast, in patients who were drinking heavily up to 48 hr before admission, F2-IP were not abnormal,

but they increased later (p < 0.005). Contrasting with plasma free F2-IP, esterified F2-IP were not significantly changed with fibrosis. Thus, whereas

circulating esterified F2-IP were unchanged in patients with alcoholic liver disease, there was an increase in free F2-IP as well as 4-HNE during recovery from intoxication. The increase was not a result of accompanying hepatitis C but a function of the stage of alcoholic liver injury, possibly reflecting enhanced lipid peroxidation as well as interference with biliary excretion and/or hepatic esterification.

Gastroenterology. 1995 Aug;109(2):547-54. Dietary saturated fatty acids: a novel treatment for alcoholic liver disease. Nanji AA, Sadrzadeh SM, Yang EK, Fogt F, Meydani M, Dannenberg AJ Department of Pathology, New England Deaconess Hospital, Boston, Massachusetts, USA. BACKGROUND & AIMS: Lipid peroxidation may be important in the pathogenesis of alcoholic liver injury. The purpose of this study was to determine whether a saturated fatty acid-based therapy (palm oil) could decrease lipid peroxidation and alcoholic liver injury during ethanol withdrawal. METHODS: Three groups of male Wistar rats (5 rats/group) were studied. Rats in group 1 were fed a fish oil-ethanol diet for 6 weeks; rats in groups 2 and 3 were fed a fish oil-ethanol diet for 6 weeks before treatment with fish oil-dextrose (group 2) or palm oil-dextrose (group 3) for 2 weeks. Liver samples were analyzed for histopathology, lipid peroxidation, fatty acid composition, cytochrome P450 2E1 activity, and tocopherol levels. RESULTS: By 6 weeks, all rats had developed fatty liver, inflammation, and necrosis. Group 2 showed minimal histological improvement, whereas group 3 showed near normalization of the histology. The improvement in group 3 was associated with decreased lipid peroxidation and P450 2E1 activity. Higher levels of omega-3 fatty acids were detected in group 2 than group 3. Tocopherol levels were similar among the groups. CONCLUSIONS: A diet enriched in saturated but not unsaturated fatty acids reversed alcoholic liver injury. This effect may be explained by down-regulation of lipid peroxidation.Comment in: Gastroenterology 1995 Aug;109(2):617-20

Nutrition. 2002 Jul-Aug;18(7-8):647-53. Dietary fat composition alters pulmonary function in pigs. Wolfe RR, Martini WZ, Irtun O, Hawkins HK, Barrow RE. Department of Surgery, The University of Texas Medical Branch, Galveston, Texas, USA. Rwolfe@utmb.edu OBJECTIVES: We investigated the effect of various dietary fats on pulmonary surfactant composition and lung function changes that occur before and after endotoxin infusion in pigs. METHODS:Eighteen pigs were assigned to three groups (n = 6 per group) to receive a diet of protein (20% of calories), carbohydrate (20% of calories), and fat (40% of calories). In one group the fat content consisted entirely of palmitic acid. In the second group, fat came from Intralipid, which provided predominantly linoleic acid. The third group was fed fish oil. Pigs were maintained on these diets for 21 d before the experiment. Cardiovascular and pulmonary functions were determined on day 22. Pigs then were infused with endotoxin (80 mg. kg(-1). min(-1)) until the pulmonary arterial pressure reached a pressure similar to that found in trauma victims (45 to 50 mmHg). Cardiovascular and pulmonary function tests were then repeated, the animals killed, and the lungs removed for study. RESULTS: Compliance was reduced in the linoleate and fish-oil groups compared with the palmitate group before and after endotoxin. Compliance changes in pigs fed the linoleate and fish-oil diets were consistent with significant increases in lung wet:dry weight ratios, increased CO(2) retention, histologic evidence of vascular congestion, intra-alveolar edema, and alveolar septa thickening. Changes in surfactant phosphatidylcholine composition between groups were consistent with the notion that increased unsaturated fatty acids could affect surfactant function. CONCLUSIONS: We concluded that the common practice of providing calories in the form of polyunsaturated fatty acids to critically ill patients carries the risk of being detrimental to lung function.

Recent Dev Alcohol. 1985;3:101-22.

Synthesis of prostaglandins and leukotrienes. Effects of ethanol. Murphy RC, Westcott JY. Prostaglandins, thromboxane, and leukotrienes are metabolites of arachidonic acid that have a variety of physiological effects at low concentrations. Effects include smooth muscle contraction, platelet aggregation, platelet disaggregation, bronchoconstriction, increased capillary permeability, cardiac dysfunction, and polymorphonuclear leukocyte accumulation. Although their formation does not appear to be essential for life, these eicosanoid activities are wide ranging and have important roles in normal physiology as well as pathophysiology. At the center of eicosanoid biosynthesis lies the plasma cell membrane which serves as the arachidonic acid reservoir. It has been widely appreciated that ethanol exerts effects on the lipid bilayer, and it is not surprising that a growing body of evidence supports the concept that important interactions between ethanol and eicosanoid biosynthesis can occur. Furthermore, at various steps leading to ultimate prostaglandin, thromboxane and leukotriene production, reactive intermediates such as radicals are involved whose lifetime in the biological milieu can be profoundly modulated by ethanol.

J Nutr Sci Vitaminol (Tokyo). 1984 Apr;30(2):179-85. Physiological impairment in linoleic acid deficiency of rats and the effect of n-3 polyunsaturated fatty acids. Iritani N, Ikeda Y, Fukuda H. Some impairments related to membrane function were found in linoleic acid-deficient rats and the effects of fish oil feeding were investigated. In linoleic acid-deficient rats, glucose transport into erythrocytes was decreased. The concentrations of plasma free fatty acids were significantly reduced in the animals. Further, epinephrine-stimulated lipase was remarkably less sensitive to epinephrine in the deficient rat than in the corn oil-fed control rat. However, these impairments were relieved by fish oil feeding. Therefore, the impairments may be ascribed to the decrease of arachidonic acid as a polyunsaturated fatty acid in membrane phospholipids, since n-3 polyunsaturated fatty acids appear to take the place of arachidonic acid.

Alcohol Clin Exp Res. 2002 May;26(5):731-6. Pathogenesis of alcoholic liver disease--recent advances. Nanji AA, Su GL, Laposata M, French SW. Department of Pathology and Center for the Study of Liver Diseases, The University of Hong Kong and Queen Mary Hospital. Ananji@pathology.hku.hk The article summarizes the proceedings of a symposium on recent advances in research on the pathogenesis of alcoholic liver disease at the 2001 RSA meeting in Montreal, Canada. The chairs were Amin A. Nanji and Samuel W. French. The presentations were (1) Role of inflammatory mediators in alcoholic liver injury by Amin A. Nanji, (2) Role of endotoxin, lipopolysaccharide binding protein,

CD14 and Toll receptors in alcoholic liver injury by Grace Su, (3) Fatty acid

ethyl esters: toxicity, metabolism and markers of ethanol intake by Michael

Laposata, and (4) Cyclic changes in gene expression when rats are fed alcohol at

a constant rate by Samuel W. French.

Department of Pathology and Center for the Study of Liver Diseases, The University of Hong Kong, Hong Kong, China. Ananji@pathology.hku.hk We investigated the potential of dietary saturated fatty acids to reverse alcoholic liver injury despite continued administration of alcohol. Five groups (six rats/group) of male Wistar rats were studied. Rats in groups 1 and 2 were fed a fish oil-ethanol diet for 8 and 6 weeks, respectively. Rats in groups 3

and 4 were fed fish oil and ethanol for 6 weeks before being switched to

isocaloric diets containing ethanol with palm oil (group 3) or medium-chain triglycerides (MCTs, group 4) for 2 weeks. Rats in group 5 were fed fish oil and dextrose for 8 weeks. Liver samples were analyzed for histopathology, lipid

peroxidation, nuclear factor-kappaB (NF-kappaB) activation, and mRNAs for cyclooxygenase-2 (Cox-2) and tumor necrosis factor-alpha (TNF-alpha). Endotoxin in plasma was determined. The most severe inflammation and fibrosis were detected in groups 1 and 2, as were the highest levels of endotoxin, lipid peroxidation, activation of NF-kappaB, and mRNAs for Cox-2 and TNF-alpha. After the rats were switched to palm oil or MCT, there was marked histological improvement with decreased levels of endotoxin and lipid peroxidation, absence of NF-kappaB activation, and reduced expression of TNF-alpha and Cox-2. A diet enriched in saturated fatty acids effectively reverses alcohol-induced necrosis, inflammation, and fibrosis despite continued alcohol consumption. The therapeutic effects of saturated fatty acids may be explained, at least in part, by reduced endotoxemia and lipid peroxidation, which in turn result in decreased activation of NF-kappaB and reduced levels of TNF-alpha and Cox-2.

Hepatology. 1997 Dec;26(6):1538-45.

Dietary saturated fatty acids down-regulate cyclooxygenase-2 and tumor necrosis

factor alfa and reverse fibrosis in alcohol-induced liver disease in the rat.

Nanji AA, Zakim D, Rahemtulla A, Daly T, Miao L, Zhao S, Khwaja S, Tahan SR,

Dannenberg AJ.

Department of Pathology, Beth Israel Deaconess Medical Center and Harvard

Medical School, Boston, MA 02215, USA.

We investigated the potential of dietary saturated fatty acids to decrease

endotoxemia and suppress expression of cyclooxygenase 2 (Cox-2) and tumor

necrosis factor alpha (TNF-alpha) in established alcohol-induced liver injury.

Six groups (five rats/group) of male Wistar rats were studied. Rats in group 1

were fed a fish oil-ethanol diet for 6 weeks. Rats in groups 2, 3, and 4 were

fed fish oil and ethanol for 6 weeks. Ethanol administration was stopped at this

time, and the rats were switched to isocaloric diets containing dextrose with

fish oil (group 2), palm oil (group 3), or medium-chain triglycerides (group 4)

as the source of fat for an additional 2 weeks. Rats in groups 5 and 6 were fed

fish oil-ethanol and fish oil-dextrose, respectively, for 8 weeks. Liver samples

were analyzed for histopathology, lipid peroxidation, and levels of messenger

RNA (mRNA) for Cox-2 and TNF-alpha. Concentrations of endotoxin were determined

in plasma. The most severe inflammation and fibrosis were detected in groups 1

and 5, as were the highest levels of endotoxin, lipid peroxidation, and mRNA for

Cox-2 and TNF-alpha. After ethanol was discontinued, there was minimal

histological improvement in group 2 but near normalization of the histology,

including regression of fibrosis, in groups 3 and 4. Histological improvement

was associated with decreased levels of endotoxin, lipid peroxidation, and

reduced expression of Cox-2 and TNF-alpha. The data indicate that a diet

enriched in saturated fatty acids (groups 3 and 4) effectively reverses

alcohol-induced liver injury, including fibrosis. The therapeutic effects of

saturated fatty acids may be explained, at least in part, by reduced endotoxemia

and lipid peroxidation, which in turn result in decreased levels of TNF-alpha

and Cox-2.

4: J Pharmacol Exp Ther. 1996 Jun;277(3):1694-700.

Medium chain triglycerides and vitamin E reduce the severity of established

experimental alcoholic liver disease.

Nanji AA, Yang EK, Fogt F, Sadrzadeh SM, Dannenberg AJ.

Department of Pathology, New England Deaconess Hospital, Boston, Massachusetts,

USA.

Lipid peroxidation may be important in the pathogenesis of alcoholic liver

injury. We investigated the potential of medium chain triglycerides and vitamin

E to decrease lipid peroxidation and reverse established alcoholic liver injury.

Four groups (five rats/group) of male Wistar rats were studied. Rats in group 1

were fed a fish oil-ethanol diet for 6 weeks. Rats in groups 2, 3 and 4 were fed

the fish oil-ethanol diet for 6 weeks before being switched to fish oil-dextrose

(group 2), fish oil-dextrose plus vitamin E (group 3) or medium chain

triglycerides-dextrose (group 4) diets for 2 weeks. Liver samples were analyzed

for histopathology, lipid peroxidation, fatty acid composition and cytochrome

P450 2E1 activity. By 6 weeks, all rats developed fatty liver, inflammation and

necrosis. After switching to the dextrose-containing diets, there was minimal

histologic improvement in group 2, moderate improvement in group 3 and near

normalization of the histology in group 4. Histologic improvement was associated

with decreased lipid peroxidation and cytochrome P450 2E1 activity. Higher

levels of polyunsaturated fatty acids were seen in groups 2 and 3 than in group

4. Our results indicate that a diet enriched in saturated (group 4) but not

polyunsaturated (group 2) fatty acids effectively reverses alcoholic liver

injury. Treatment with vitamin E also led to histologic improvement. These

effects may be explained, at least in part, by down-regulation of lipid

peroxidation. Other effects of medium chain triglycerides, such as their

propensity for oxidation rather than esterification, may also be important.

5: Gastroenterology. 1995 Aug;109(2):547-54. Comment in:

Gastroenterology. 1995 Aug;109(2):617-20.

Dietary saturated fatty acids: a novel treatment for alcoholic liver disease.

Nanji AA, Sadrzadeh SM, Yang EK, Fogt F, Meydani M, Dannenberg AJ.

Department of Pathology, New England Deaconess Hospital, Boston, Massachusetts,

USA.

BACKGROUND & AIMS: Lipid peroxidation may be important in the pathogenesis of

alcoholic liver injury. The purpose of this study was to determine whether a

saturated fatty acid-based therapy (palm oil) could decrease lipid peroxidation

and alcoholic liver injury during ethanol withdrawal. METHODS: Three groups of

male Wistar rats (5 rats/group) were studied. Rats in group 1 were fed a fish

oil-ethanol diet for 6 weeks; rats in groups 2 and 3 were fed a fish oil-ethanol

diet for 6 weeks before treatment with fish oil-dextrose (group 2) or palm

oil-dextrose (group 3) for 2 weeks. Liver samples were analyzed for

histopathology, lipid peroxidation, fatty acid composition, cytochrome P450 2E1

activity, and tocopherol levels. RESULTS: By 6 weeks, all rats had developed

fatty liver, inflammation, and necrosis. Group 2 showed minimal histological

improvement, whereas group 3 showed near normalization of the histology. The improvement in group 3 was associated with decreased lipid peroxidation and P450 2E1 activity. Higher levels of omega-3 fatty acids were detected in group 2 than group 3. Tocopherol levels were similar among the groups. CONCLUSIONS: A diet

enriched in saturated but not unsaturated fatty acids reversed alcoholic liver injury. This effect may be explained by down-regulation of lipid peroxidation.

Toxicol Appl Pharmacol 1993 May;120(1):72-9. Essential fatty acid deficiency in cultured human keratinocytes attenuates toxicity due to lipid peroxidation. Wey HE, Pyron L, Woolery M Centers for Disease Control, National Institute for Occupational Safety and Health, Division of Biomedical and Behavioral Science, Cellular Toxicology Section, Cincinnati, Ohio 45226. Human keratinocytes are commonly grown in culture with a serum-free medium. Under these conditions, keratinocytes become essential fatty acid deficient (EFAD), as determined by gas chromatographic analysis of cell phospholipid fatty acid composition. Exposure of EFAD keratinocytes for 2 hr to concentrations of t-butyl hydroperoxide (tBHP) up to 2 mM did not result in toxicity assessed by lactate dehydrogenase (LDH) release and only a small indication of lipid peroxidation assessed by the release of thiobarbituric acid-reactive substances (TBARS). Addition of 10 microM linoleic acid (LA) to serum-free medium alleviated the EFAD condition by increasing the phospholipid content of LA and its elongation and desaturation products, arachidonic acid and docosatetraenoic acid. Exposure of LA-supplemented keratinocytes to tBHP resulted in significant LDH (at 1 and 2 mM tBHP) and TBARS (tBHP concentration dependent) release. TBARS release was also significantly elevated in unexposed LA-supplemented keratinocytes (basal release). Co-supplementation with the antioxidant, alpha-tocopherol succinate (TS) prevented tBHP (1 mM)-induced LDH release in LA-supplemented cultures. TS supplementation also attenuated the effect of tBHP on TBARS release, but when compared to TS-supplemented EFAD cultures, LA supplementation still led to increased tBHP-induced TBARS release. Keratinocyte cultures are potentially useful as an alternative to animals in toxicology research and testing. It is important, however, that the cell model provide a response to toxic insult similar to that experienced in vivo. Our results suggest that fatty acid and antioxidant nutrition of cultured keratinocytes are important parameters in mediating the toxic effects of lipid peroxidation.

Thromb Res. 1989 Jan 1;53(1):45-53. Normalization by dietary cod-liver oil of reduced thrombogenesis in essential fatty acid deficient rats. Hornstra G, Haddeman E, Don JA. Department of Nutrition and Safety, Unilever Research, Vlaardingen, The Netherlands. Rats, deficient in essential fatty acids (EFA), were given diets containing 5 energy% sunflowerseed oil (SO, rich in linoleic acid), cod-liver oil (CLO, rich in timnodonic acid and cervonic acid), or hydrogenated coconut oil (HCO), containing no EFAs at all. SO and CLO feeding resulted in normalization of the reduced arterial thrombus formation in EFA-deficient animals. SO feeding was associated with the normalization of the arachidonic acid content of platelet phospholipids. CLO feeding did not have this effect but greatly increased the availability of timnodonic acid (EPA) and cervonic acid (DHA). Further research is required to investigate whether these changes in fatty acid composition can be hold responsible for the normalizing effect of dietary CLO on the disturbed arterial thrombosis tendency in EFA deficient rats, possibly via the formation of eicosanoids.

Acta Neurochir Suppl (Wien) 1994;60:20-3. Mechanisms of glial swelling by arachidonic acid. Staub F, Winkler A, Peters J, Kempski O, Baethmann A Institute for Surgical Research, Ludwig-Maximilians-University, Munchen, Federal Republic of Germany. The effect of arachidonic acid (AA, 20:4) was analyzed in vitro by employment of C6 glioma cells and astrocytes from primary culture. The cells were suspended in an incubation chamber under continuous control of pH, pO2, and temperature. Cell swelling was quantified by flow cytometry. After a control period, the suspension was added with AA at concentrations of 0.01 to 1.0 mM. Administration of AA induced an immediate, dose dependent swelling in C6 glioma cells or astrocytes. AA-concentrations of 0.01 mM led to an increase of the glial cell volume to 103.0 +/- 1.0% of control, 0.1 mM to 110.0 +/- 1.5%, and 1.0 mM to 118.8 +/- 1.5% within 10 min. The swelling response to linoleic acid (18:2) was only about half of what was found when AA was administered at a concentration of 0.1 mM, whereas stearic acid (18:0) did not induce any cell volume changes. Inhibition of the cyclo- and lipoxygenase pathway by BW 755C did not prevent glial swelling from AA, whereas it was reduced by SOD, or almost completely abolished by the aminosteroid U-74389F, an antagonist of lipid peroxidation. Replacement of Na(+)- and Cl- -ions in the suspension medium by choline chloride was also associated with complete abolishment of cell swelling from AA. The results demonstrate an impressive efficacy of arachidonic acid to induce glial swelling which might be attributable to activation of lipid peroxidation by the fatty acid, leading to an increased Na(+)-permeability and subsequent influx of water into the cells.

Prostaglandins Leukot Essent Fatty Acids 1994 Jul;51(1):33-40. Suppression of human T-cell growth in vitro by cis-unsaturated fatty acids: relationship to free radicals and lipid peroxidation. Madhavi N, Das UN, Prabha PS, Kumar GS, Koratkar R, Sagar PS Department of Medicine, Nizam's Institute of Medical Sciences, Hyderabad, India. Cis-unsaturated fatty acids such as dihomogamma-linolenic acid (DGLA), arachidonic acid (AA) and eicosapentaenoic acid (EPA),

which form precursors to 1, 2 and 3 series prostaglandins (PGs), have been shown to suppress human T-cell growth in vitro by a prostaglandin

E (PGE)-independent mechanism. In an earlier study, we showed that these fatty acids can induce free radical generation in human neutrophils and tumor cells. Here we show that cis-unsaturated fatty acids augment free radical generation and lipid peroxidation in human T-cells. The growth suppressive action of cis-unsaturated fatty acids on human T-cells could be blocked by anti-oxidant, vitamin E and the superoxide anion quencher superoxide dismutase. These results suggest that c-UFAs-induced cell growth suppression is a free radical dependent process.

Carcinogenesis 1994 Jul;15(7):1399-404.Peroxidation of linoleic, arachidonic and oleic acid in relation to the induction of oxidative DNA damage and cytogenetic effects.

de Kok TM, ten Vaarwerk F, Zwingman I, van Maanen JM, Kleinjans JC

Department of Natural Sciences, Open University, Heerlen, The

Netherlands.

In the present study, the possible role of the polyunsaturated fatty acids

linoleic and arachidonic acid in the chemical induction of carcinogenesis

has been investigated. Analysis of

7,8-dihydro-8-oxo-2'-deoxyguanosine (8-oxodG) levels in

2'-deoxyguanosine (dG) and isolated DNA has demonstrated that

linoleic and arachidonic acid are capable of inducing this specific

genotoxic damage. This effect appears to be related to the degree of

fatty acid unsaturation, since it was not induced by monounsaturated

oleic acid. Enzymatic peroxidation of linoleic and arachidonic acid

resulted in a significant increase in oxidative DNA damage. Studies on

the interference of radical scavengers with the induction of 8-oxodG in

combination with electron spin resonance spectroscopy demonstrated

that the superoxide anion was generated during peroxidation of these

fatty acids and that singlet oxygen is most likely involved in the formation

of oxidative DNA damage. The level of oxidative damage in dG and single-stranded DNA was higher as compared to that in native DNA after equimolar treatment. Exposure of human lymphocytes to linoleic or arachidonic acid did not result in a significant increase in levels of 8-oxodG. This may indicate that the rate of intracellular peroxidation is relatively low and/or that nuclear DNA in intact cells is effectively protected against genetic damage induced by reactive oxygen species. It is therefore concluded that relatively short periods of linoleic or arachidonic acid administration are not likely to impose a direct genotoxic risk. It can, however, not be excluded that chronic exposure to polyunsaturated fatty acids induces oxidative DNA damage or is

related to cancer risk by epigenetic mechanisms, as is also indicated by the observed cytotoxic effects of linoleic and arachidonic acid.

Biochim Biophys Acta 1986 Apr

11;881(2):292-6

Effect of long-chain fatty acids on the binding of

thyroxine and triiodothyronine to human

thyroxine-binding globulin. Tabachnick M, Korcek L The effect of long-chain fatty acids on the binding of thyroxine to highly purified human thyroxine-binding globulin has been studied by

equilibrium dialysis performed at pH 7.4 and 37 degrees C. At a fixed molar ratio of 2000:1 of fatty acid to thyroxine-binding globulin, the

degree of binding inhibition based on the percent change in nK value relative to the control as determined from Scatchard plots was: palmitic,

0%; stearic, 0%; oleic, 76%; linoleic, 69%; and linolenic, 61%. At a 500:1 molar ratio of oleic acid to thyroxine-binding globulin, equivalent

to 0.125 mM free fatty acid in serum, thyroxine binding was inhibited by

18%, increasing to 93% at a 4500:1 molar ratio. At molar ratios of oleic

acid to thyroxine-binding globulin of 1000:1, 2000:1 and 4000:1, the

degree of inhibition of triiodothyronine binding was 24%, 41% and 76%, respectively. The results indicate that the unsaturated long-chain

fatty acids are potent inhibitors of thyroxine binding to thyroxine-binding

globulin, whereas the saturated fatty acids have little or no effect on thyroxine binding.

Biochim Biophys Acta 1993 Feb 10;1166(1):99-104. Inhibition of fatty acid synthesis in rat hepatocytes by exogenous polyunsaturated fatty acids is caused by lipid peroxidation. Mikkelsen L, Hansen HS, Grunnet N, Dich J Department of Biological Sciences, Royal Danish School of Pharmacy, Copenhagen. Rat hepatocyte long-term cultures were utilized to investigate the impact of different polyunsaturated fatty acids (PUFA) on the insulin-induced de novo fatty acid synthesis in vitro. The addition of 0.5 mM albumin-complexed oleic, linoleic, columbinic, arachidonic, eicosapentaenoic or docosahexaenoic acid resulted in a marked suppression of fatty acid synthesis. By evaluation of cell viability (determined as the leakage of lactate dehydrogenase (LDH) it turned out, that the antioxidant used (50 microM alpha-tocopherol phosphate) had a low antioxidant activity, resulting in cytotoxic effects by the peroxidized PUFA. Arachidonic acid and eicosapentaenoic acid showed a dose- and time-dependent cytotoxicity. Two other antioxidants: 50 microM alpha-tocopherol acid succinate and 1 microM N,N'-diphenyl-1,4-phenylenediamine, both proved more efficient than alpha-tocopherol phosphate. There was a significant correlation between LDH-leakage and inhibition of fatty acid synthesis. Lipid peroxidation, measured as thiobarbituric acid-reactive substances, also showed a significant correlation with the degree of inhibition of fatty acid synthesis. Furthermore, PUFA had no inhibitory effect on fatty acid synthesis when peroxidation was minimized by the use of proper antioxidants. These data indicate that PUFA in vitro inhibit the insulin-induced de novo fatty acid synthesis in hepatocytes from starved rats, due to cytotoxic effects caused by lipid peroxidation.

J Neurochem 1980 Oct;35(4):1004-7. Transient formation of superoxide radicals in polyunsaturated fatty acid-induced brain swelling. Chan PH, Fishman RA The involvement of superoxide free radicals and lipid peroxidation in brain swelling induced by free fatty acids has been studied in brain slices and homogenates. The polyunsaturated fatty acids linoleic acid (18:2), linolenic acid (18:3), arachidonic acid (20:4), and docosahexaenoic acid (22:6) caused brain swelling concomitant with increases in superoxide and membrane lipid peroxidation. Palmitic acid (16:0) and oleic acid (18:1) had no such effect. Furthermore, superoxide formation was stimulated by NADPH and scavenged by the addition of exogenous superoxide dismutase in cortical slice homogenates. These in vitro data support the hypothesis that both superoxide radicals and lipid peroxidation are involved in the mechanism of polyunsaturated fatty acid-induced brain edema.

Cell 1977 Sep;12(1):295-300. Decrease in adhesion of cells cultured in polyunsaturated fatty acids. Hoover RL, Lynch RD, Karnovsky MJ The addition of long chain unsaturated fatty acids (linoleic, linolenic and arachidonic acids) to BHK cells reduces the cell to substrate adhesion,

causes morphological changes and alters the cellular growth properties. The new characteristics are similar to those of transformed cells. The

data indicate that the effects are probably due to actual changes in the surface membrane lipids and not due to prostaglandin synthesis.

Biochem Mol Biol Int 1993 Jan;29(1):175-83

Influence of antioxidant vitamins on fatty acid

inhibition of lymphocyte proliferation. Calder PC, Newsholme EA Department of Biochemistry, University of Oxford, United Kingdom. Fatty acids, especially polyunsaturated fatty acids (PUFAs), inhibit a number of lymphocyte functions, including proliferation, cytokine

production and cytotoxicity, but their mechanism of action is not known. This study investigated whether fatty acids inhibit lymphocyte

proliferation by leading to the production of lipid peroxides, which are known to inhibit the growth of cells. The so-called "thiobarbituric

acid-reactive substances" (TBARS) and lipid hydroperoxide contents of lymphocytes (0.75 +/- 0.04 and 1.30 +/- 0.39 nmol/mg protein in fresh

cells, respectively) were increased by 48 h culture to 0.96 +/- 0.14 and 3.23 +/- 0.47 nmol/mg protein, respectively. The TBARS content was

increased by culture in the presence of 100 microM PUFAs to between 1.46 +/- 0.11 (linoleic acid) and 2.39 +/- 0.31 (docosahexaenoic acid)

nmol/mg protein. The lipid hydroperoxide content was increased by culture in the presence of 100 microM PUFAs to between 11.65 +/-

1.12 (linoleic acid) and 22.24 +/- 1.26 (docosahexaenoic acid) nmol/mg protein. These increases were partially prevented by inclusion of 10 microM vitamin E in the culture medium. Vitamin E (1 or 10 microM) enhanced concanavalin A-stimulated rat lymphocyte proliferation by approximately 45%. Vitamin E (10 microM) increased human lymphocyte proliferation by 35%. However, vitamin E did not

prevent the inhibitory effects of fatty acids upon lymphocyte proliferation. It is concluded that inhibition of lymphocyte proliferation

by fatty acids is not caused by their conversion to peroxidised products.

Biochim Biophys Acta 1994 Sep 15;1214(2):209-20. Reinvestigation of lipid peroxidation of linolenic acid. Mlakar A, Spiteller G Lehrstuhl fur Organische Chemie I, Universitat Bayreuth, Germany. Recently, we deduced a mechanism for lipid peroxidation of linoleic acid [1]. This mechanism was now applied to predict the occurrence of previously unknown lipid peroxidation products of linolenic acid. The proposed structures of peroxidation products allowed to search for these predicted compounds in reaction mixtures with the aid of 'ion trace' by mass spectrometry. Thus, a great number of previously unknown lipid peroxidation products was detected. It is assumed that these compounds also occur--at least as intermediates--in lipid peroxidation processes in mammalian tissue.

BJU Int. 2003 Jun;91(9):830-8. Fibrin as an inducer of fibrosis in the tunica albuginea of the rat: a new animal model of Peyronie's disease. Davila HH, Ferrini MG, Rajfer J, Gonzalez-Cadavid NF. Department of Urology, UCLA School of Medicine, Los Angeles, CA, USA. OBJECTIVES: To investigate the role of fibrin in inducing fibrosis in the tunica albuginea (TA) of the rat penis, to develop a new animal model for Peyronie's disease (PD). MATERIALS AND METHODS: The TA of rats (five per group per period) were injected with either saline, fibrin, transforming growth factor-beta1 (TGF-beta1) or TGF-beta1 plus fibrin; the rats were killed at 1, 3, and 6 weeks after injection. Images were analysed quantitatively from tissue sections stained for collagen (Masson trichrome), fibrin (Verhoeff's stain) and elastin (Hart's stain), and immunostained for TGF-beta1, inducible nitric oxide synthase (iNOS), heme oxygenase 1 (HO1), alpha-smooth muscle actin (ASMA), apoptosis (TUNEL) and plasminogen activator inhibitor (PAI). C

Arch Esp Urol. 2003 Sep;56(7):814-9.

[In Process Citation] [Article in Spanish] Lemourt Oliva M, Rodriguez Barroso A, Bordonado Ramirez R, Gonzalez Oramas E, Molina Castillo F. Servicio de Urologia, Hospital Docente Quirurgico Freyre de Andrade, La Habana, Cuba. Malem@infomed.sld.cu OBJECTIVES: To determine the optimal dose for propoleum caps in Peyronie's disease. METHODS: We evaluated two groups of 17 patients each, analyzing pain and plaque size on physical and ultrasound examination, before and after treatment. Group A patients received caps with 300 mg of propoleum powder daily for six months, and Group B 900 mg. RESULTS: Predominant ages were between 41 and 70 years. More than half the patients in group A had pain; more than half the patients in group B did not have pain; considering both groups there were more patients without pain. Physical reduction of the plaque: Group A: 0.6 cm, Group B: 0.8 cm. Ultrasound reduction of the plaque: Group A: 1.3 cm, Group B: 0.8 cm. In the higher dose group (B) clinical and ultrasound improvement started earlier. 79.4% of the treated patients evaluated preferred improvement or slight improvement, being this result statistically significant. CONCLUSIONS: Pain is not always an accompanying symptom in Peyronie's disease. Both doses administered reduced the size of the plaque, but the higher one has an earlier improvement. We may consider propoleum administered in this way another treatment option for this disease, with good results and low cost.

Horm Metab Res. 1988 Feb;20(2):86-90. The inhibition of PB125I formation in calf thyroid caused by 14-iodo-15-hydroxy-eicosatrienoic acid is due to decreased H2O2 availability. Krawiec L, Chazenbalk GD, Puntarulo SA, Burton G, Boveris A, Valsecchi RM, Pisarev MA.

Depto. Aplicaciones Biologicas, Comision Nacional de Energia Atomica, Buenos

Aires, Argentina. Previous work from our laboratory has shown that 14-iodo-15-hydroxy-5,8,11-eicosatrienoic acid (I-HO-A) is a potent inhibitor of

iodine organification in calf thyroid slices. The present studies were performed in order to clarify the mechanism of this action. Incubation of thyroid slices with 10(-4)M I-HO-A caused a 47 and 53% decrease in PB125I formation after 30 and 60 min incubation, respectively. In a series of experiments an inverse relationship between the degree of inhibition caused by I-HO-A and total iodine content and basal iodoprotein formation was observed. Chromatographic analysis

of the labeled compounds showed a significant decrease in 125I incorporation into MIT, DIT, T3 and total iodolipid. The site of the inhibitory effect of I-HO-A was then sought. TPO was measured by three different methods. When TPO

was solubilized from I-HO-A treated slices, no change in enzymatic activity was observed. Moreover, the same lack of action was found when solubilized TPO was incubated with I-HO-A. The production and release of H2O2 into the incubation medium was measured by chemiluminiscence technique. In control slices the values increased during the first 10 min and reached a plateau. Pretreatment of the slices with 10(-4)M KI caused a 51% inhibition, while the same concentration of I-HO-A produced a 59% inhibition. The possibility that I-HO-A might exert its action through a putative protein inhibitor was also explored. Incubation of slices with 10(-5)M I-HO-A caused a 46% decrease in PB125I formation and addition of actinomycin D or puromycin failed to alter this effect.(ABSTRACT

TRUNCATED AT 250 WORDS)

Arch Esp Urol. 2003 Sep;56(7):805-13. [In Process Citation] [Article in Spanish] Lemourt Oliva M, Rodriguez Barroso A, Puente Guillen M, Vega Guerrero C, Navarro Cutino M, Perez Monzon A. Servicio de Urologia, Hospital Docente Quirurgico Freyre de Andrade, La Habana, Cuba. Malem@infomed.sld.cu OBJECTIVES: Peyronie's disease is a plastic penile induration which represents a well-known problem affecting men in their middle age, who suffer penile incurvation during erections, pain and poor consistency at the site of incurvation. There is no satisfactory treatment for this disease. A patient with Peyronie's disease who received propoleum treatment for a giardiasis infection referred spontaneous improvement of this penile incurvation as treatment cycles were repeated. So, we decided to treat a group of patients with this pathologic entity with propoleum to evaluate its efficacy. METHODS: Initially, 25 patients were studied, only 13 completed the study 6 months later. Variables: age, penile incurvation, presence or not of pain with erections, detection and measurement (cm) of the fibrous plaque on physical examination and ultrasound, and description of its radiologic characteristics. Propoleum powder (300 mg) was administered for 6 months. RESULTS: Patient age was predominantly between 51 and 60 years. Upward deviation was the most frequent presentation; it disappeared in 2 patients in the "Improved" group. Most patients had no pain. 3 patients out of 13 had no plaque on examination at the end of treatment. Regarding incurvation 10 patients improved, 2 improved slightly, and only one improved somewhat during the first three months but then after remained the same. Penile x-ray was not useful as a diagnostic test; penile ultrasound was greatly useful. CONCLUSIONS: Penile incurvation improved in 77% of the patients. Fibrous plaques were reduced more than 0.5 cm on ultrasound (average 0.64 cm).

Arch Esp Urol. 1998 Mar;51(2):171-6.

[Clinical evaluation of the use of propoleum in Peyronie's disease]

[Article in Spanish] Lemourt Oliva M, Filgueiras Lopez E, Rodriguez Barroso A, Gonzalez Oramas E, Bordonado R.

Servicio de Urologia, Radiologia, Hospital de Andrade, Centro Habana, Cuba. OBJECTIVE: To analyze the clinical efficacy of propoleum in Peyronie's disease. METHODS: A controlled, double-blind clinical trial based on a previous pilot study was conducted in 34 patients with Peyronie's disease treated with propoleum (Cuban Academy of Sciences patent no. 21875). The study was performed at the Freyre de Andrade Hospital in Habana. The clinical course and ultrasound features of the fibrous plaque were evaluated during a period of 6 months. RESULTS: A marked improvement was observed for penile curvature and pain in the treated group. Furthermore, plaque consistency and size was reduced five-fold after treatment and two-fold at ultrasound evaluation. No clinical or ultrasound changes were observed in the control group. CONCLUSIONS: Our results demonstrate the efficacy of propoleum in Peyronie's disease.

Prostaglandins Leukot Essent Fatty Acids. 2003 May;68(5):305-10. Synergistic effect of D-003 and aspirin on experimental thrombosis models. Molina V, Arruzazabala ML, Carbajal D, Mas R. Center of Natural Products, National Center for Scientific Research, Ave 25 and 158, Cubanacan, 6880, Havana, Cuba. D-003 is a mixture of higher primary aliphatic saturated acids purified from sugarcane wax, with antiplatelet and antithrombotic effects experimentally demonstrated. Octacosanoic acid is the main component of D-003, followed by triacontanoic, dotriacontanoic, and tetracontanoic acids, while other acids are minor components. This work investigates the effects of combination therapy D-003+aspirin (ASA) on arachidonic acid (AA)-induced sudden death in mice and bleeding time in rats. In addition, the effects of D-003 on serum levels of two metabolites of AA: thromboxane A(2) and prostacyclin, assessed through the measurement of their stable metabolites: thromboxane B(2) (TxB(2)) and 6 keto PgF1alpha by radioimmunoassay kits, were also investigated. Combination therapy of D-003 (50mg/kg) and ASA (3mg/kg) significantly increased bleeding time in rats in a synergistic manner compared with D-003 or ASA alone. Moreover, the combined treatment of D-003 (200mg/kg) and ASA (5mg/kg) in mice protected against AA-induced sudden death (83% survivors) in a synergistic manner which was compared with each treatment alone (33% survivors). These results indicate that antiplatelet effects of D-003 are not mediated by a cyclooxygenase inhibition. D-003 and ASA monotherapies reduced serum TxB(2) levels, whereas D-003, but not ASA, significantly increased 6 keto PgF1alpha levels.

Drugs Exp Clin Res. 2002;28(5):177-83. Effect of D-003, a mixture of high molecular weight primary acids from sugar cane wax, on CL4C-induced liver acute injury in rats. Noa M, Mendoza S, Mas R, Mendoza N. Center for Natural Products, National Center for Scientific Research, Ave. 25 and 158 Street, Post Box 6990, Havana City, Cuba. D-003 is a mixture of very high molecular weight aliphatic primary acids purified from sugar cane (Saccharum officinarum, L.) wax, in which octacosanoic acid is the most abundant component. Previous experimental studies have shown

that D-003 not only shows cholesterol-lowering and antiplatelet effects, but also offers strong protection against plasma lipoprotein oxidation. Acute hepatotoxicity induced by CCL4 in rats has been related to an increased rate of lipid peroxidation, and different antioxidant compounds have been revealed to be effective in this model. The aim of this study was to investigate the effects of

D-003 in acute hepatotoxicity induced by CCL4 in rats. Male Sprague Dawley rats

were randomly distributed in four experimental groups as follows: group 1:

negative control rats; group 2: positive control rats (CCL4-treated); groups 3

and 4 rats with liver damage induced by CCL4 and treated with D-003 at 25 and

100 mg/kg, respectively. Acute liver injury was induced by CCL4 suspended in

olive oil and intraperitoneally administered at 1 ml/kg. Eighteen hours after CCL4 dosing, the rats were anesthetized with ether and their livers were removed for histopathological studies. D-003 at 25 and 100 mg/kg significantly (p < 0.01) decreased the percentage of ballooned cells and hepatocytes with lipidic inclusions and increased the percentage of normal hepatocytes compared with that in positive controls in a dose-dependent manner. The percent inhibitions of the

occurrence of ballooned cells and hepatocytes with lipids were marked (75% and 50%, respectively) with the high dose (100 mg/kg). The percent of turgent

hepatocytes was also significantly reduced compared with that in positive

controls, but this effect was not dose-dependent. No histological alterations in the liver sections of negative controls were found. Necrotic areas and inflammatory infiltrate were observed in the liver of 7/8 (87.5%) of positive controls. In turn, D-003 dramatically reduced both necrotic areas and inflammatory infiltrate and was present in only 1/8 (12.5%) animals treated with D-003 25 mg/kg and in none (0%) of the animals treated with 100 mg/kg. D-003 protected against the histological changes characteristic of CCL4-induced hepatic injury in rats, in which the process of lipid peroxidation plays the main role. The relationship between this protective action of D-003 on this experimental model and its antioxidant effects needs to be further investigated before definitive conclusions are drawn.

J Biochem (Tokyo). 1977 Aug;82(2):529-33. Effects of free fatty acids on fibrinolytic activity. Muraoka T, Okuda H. A novel method for the estimation of fibrinolytic activity is proposed. In this method, a fibrin clot suspension is used as a substrate (fibrin is known to be a physiological substrate of plasmin). The fibrin clot suspension was prepared by homogenization of human fibrin clots. With this method, we found that free fatty acids inhibited the plasmin activity, and long-chain, unsaturated free fatty acids had a particularly strong inhibitory action on plasmin. As regards the mechanism of the inhibitory action, free fatty acids may not inhibit complex formation between plasmin and fibirin, but may make it impossible for plasmin to act on fibrin due to deformation of the surface of the fibrin clot.

Mol Cell Biochem. 2003 Nov;253(1-2):141-9. Arachidonic acid and colorectal carcinogenesis. Jones R, Adel-Alvarez LA, Alvarez OR, Broaddus R, Das S. Department of Biological Sciences, University of Texas at El Paso, El Paso, TX 79968-0519, USA. Colorectal carcinoma is a leading cause of cancer related death worldwide. This deadly disease advances through a series of clinical and histopathological stages, initiated by single crypt lesions to small benign tumors and finally to malignancy. Although some progress has been made in elucidating the formation of colorectal tumors at molecular/genetic levels, the possible mechanisms of dietary lipids in inducing and promoting colorectal tumorigenesis are poorly understood. Recent epidemiological studies, however, indicate that lipid-rich diet containing omega-6 fatty acids (i.e. linoleic acid, arachidonic acid, etc.) may somehow be related with the disease process. Rapid metabolism of arachidonic acid, increased activities of phospholipases (i.e. phospholipase-A2s), and the elevated levels of cyclooxygenase (COX) and lipoxygenase (LOX) in colonic cells were reported in various stages of the malignancy, suggesting a possible link between dietary lipids and the incidence of colorectal cancer. The major focus of this review is to delineate the recent findings on enhanced arachidonic acid metabolism and its conversion into eicosanoids during the initiation and progression of colorectal carcinogenesis. In addition, the identification and participation of various phospholipases are also discussed. It is speculated that many of these phospholipases can be used as targets for developing new drugs against colorectal as well as other adenocarcinomas.

Gerontology 1993;39(1):7-18. Modulation of membrane phospholipid fatty acid composition by age and food restriction. Laganiere S, Yu BP. H.M. “Phospholipids from liver mitochondrial and microsomal membrane preparations were analyzed to further assess the effects of age and lifelong calorie restriction on membrane lipid composition.” “The data revealed characteristic patterns of age-related changes in ad libitum (AL) fed rats: membrane levels of long-chain polyunsaturated fatty acids, 22:4 and 22:5, increased progressively, while membrane linoleic acid (18:2) decreased steadily with age. Levels of 18:2 fell by approximately 40%, and 22:5 content almost doubled making the peroxidizability index increase with age.” “We concluded that the membrane-stabilizing action of long-term calorie restriction relates to the selective modification of membrane long-chain polyunsaturated fatty acids during aging.”

Lipids 2001 Jun;36(6):589-93. Effect of dietary restriction on age-related increase of liver susceptibility to peroxidation in rats. Leon TI, Lim BO, Yu BP, Lim Y, Jeon EJ, Park DK.

Nutr Cancer. 2001;41(1-2):91-7. Vaccenic acid feeding increases tissue levels of conjugated linoleic acid and

suppresses development of premalignant lesions in rat mammary gland. Banni S, Angioni E, Murru E, Carta G, Melis MP, Bauman D, Dong Y, Ip C. Dipartimento di Biologia Sperimentale, Sezione di Patologia Sperimentale, Universita degli Studi di Cagliari, Cittadella Universitaria, 09042 Monserrato, Cagliari, Italy. The objective of this report was to determine whether vaccenic acid (t11-18:1) is converted efficiently to conjugated linoleic acid (c9,t11-18:2, CLA) in rats via the delta 9-desaturase reaction and, if so, whether vaccenic acid could substitute for CLA as an anticancer agent. In Study 1, rats were fed 1%, 2%, or 3% vaccenic acid in their diet, and tissue levels of CLA and CLA metabolites were determined in liver and mammary gland. In general, concentrations of CLA and CLA metabolites increased proportionately with an increase in vaccenic acid intake, at least up to the 2% dose level. Beyond this dose, there was clearly a plateauing effect. Thus vaccenic acid concentration increased from an undetectable level in the control to 78.5 nmol/mg lipid in the liver of rats fed a 2% vaccenic acid diet. This was accompanied by an increase in CLA from 2.3 to 33.6 nmol/mg lipid. These changes were also mirrored in the mammary gland, where increases in vaccenic acid (from 27.5 to 163.2 nmol/mg lipid) and CLA (from 17.8 to 108.9 nmol/mg lipid) were similarly observed. Vaccenic acid at 2% produced a CLA concentration in the mammary gland that was historically associated with a positive response in tumor inhibition based on our past experience. This provided the basis for selecting 2% vaccenic acid in Study 2, which was designed to evaluate its efficacy in blocking the development of premalignant lesions in the rat mammary gland. In this experiment, formation of histologically identifiable pathology due to intraductal proliferation of terminal end bud cells of mammary epithelium was used as the end point of analysis at 6 wk after carcinogen administration. Treatment with vaccenic acid reduced the total number of these premalignant lesions by approximately 50%. We hypothesize that the anticancer response to vaccenic acid is likely to be mediated by its endogenous conversion to CLA via delta 9-desaturase.

J Nutr Biochem. 2001 Sep;12(9):536-549.

Dietary polyunsaturated n-6 lipids effects on the growth and fatty acid composition of rat mammary tumors.

Escrich E, Solanas M, Soler M, Ruiz de Villa MC, Sanchez JA, Segura R.

Department of Cell Biology, Physiology and Immunology, Medical Physiology Unit,

Universitat Auto'noma de Barcelona, 08193, Bellaterra --Barcelona, Spain

The aim of this study was to analyze the effects of a polyunsaturated n-6

high-fat diet on rat DMBA-induced breast cancer at different stages of the

carcinogenesis and to investigate if changes in the tumor fatty acid composition are one of the mechanisms by which dietary lipids could exert their effects. 14 fatty acids were evaluated in 6 lipid fractions. The results firstly showed that this high-fat diet stimulated the malignant mammary tumor growth, mainly all in the promotion group. The tumor lipid analysis indicated: 1) that each lipid fraction presented distinct major fatty acids (>5%) which were not the most abundant in the diet, except in the case of the triacylglicerides, suggesting the different resistance to dietary fatty acid modification of the tumor lipid fractions; 2) a higher arachidonic acid content in the fractions with less linoleic acid, above all in phospholipids, particularly in the phosphatidylethanolamine, indicating a different efficiency of conversion; 3) the three most abundant fatty acids in the dietary lipid (18:2n-6, 18:1n-9 and 16:0) were those which essentially displayed the differences between groups; thus, the high-fat diet changed the tumor lipid profile, increasing the 18:2n-6 relative content and decreasing that of the 18:1n-9; differences were significant in phosphatidylcholine, free fatty acids and triacylglycerides. Any change was obtained in the phosphatidylinositol. The greatest number of differences was found in the promotion group. Taken as a whole, our results suggest the different roles of lipid fractions in breast cancer cells and an association between cancer malignancy and the content of linoleic and oleic acids.

J Natl Cancer Inst. 1977 Feb;58(2):445-7. Reduced growth rate of transplantable mammary adenocarcinoma in C3H mice fed eicosa-5,8,11,14-tetraynoic acid. Rao GA, Abraham S. Female 3-month-old C3H mice were given sc injections of 5-mg pieces of mammary

adenocarcinoma and fed a linoleate-containing (15% corn oil) diet in the presence or absence of eicosa-5,8,11,14-tetraynoic acid (TYA), an inhibitor of prostaglandin synthesis. After 6 weeks, the weights of tumors of mice fed the TYA-free linoleate diet were three to five georgia,palatino,times,serif,Times New Roman greater than those of mice fed the TYA-containing linoleate diet. Dietary TYA caused a reduction in the levels of arachidonate and an elevation in the levels of linoleate in mammary tumors and livers. Aspirin, another known inhibitor of prostaglandin synthesis, when added to the linoleate diet, did not affect the tumor size or the composition of

fatty acids in the tumors and livers. Thus we concluded that a) the growth of

mammary tumors was not related to prostaglandin synthesis but was related to the availability of arachidonate, and b) TYA was an effective inhibitor for the conversion of linoleate to arachidonate.

Int J Biochem Cell Biol. 2003 May;35(5):749-55. Increased muscle proteasome activities in rats fed a polyunsaturated fatty acid supplemented diet. Vigouroux S, Farout L, Clavel S, Briand Y, Briand M. Laboratoire de Biochimie Appliquee, Associe INRA, Universite Blaise Pascal, 63174, Aubiere, France. Changes in the proteasome system, a dominant actor in protein degradation in eukaryotic cells, have been documented in a large number of physiological and pathological conditions. We investigated the influence of monounsaturated or polyunsaturated fatty acids (PUFAs) supplemented diets on the proteasome system, in rat skeletal muscles. Thirty rats were randomly assigned to three groups. The control group received only a standard diet. The monounsaturated fatty acid (MUFA) enriched diet group was fed with 3% sunflower oil in addition to standard food, and the polyunsaturated fatty acid supplemented diet group received 9% Maxepa) in addition to the standard diet. We analyzed muscle proteasome activities and content. Monounsaturated or PUFAs supplemented diets given for 8 weeks induced a significant increase in proteasome activities. With the polyunsaturated fatty acid enriched diet, the chymotrypsin-like and peptidylglutamylpeptide hydrolase activities increased by 45% in soleus and extensor digitorum longus (EDL), and by 90% in the gastrocnemius medialis (GM) muscle. Trypsin-like activity of the proteasome increased by 250% in soleus, EDL and GM. This increase in proteasome activities was associated with a concomitant enhancement in the muscle content of proteasome. Proteasome activities and level were less stimulated with a monounsaturated fatty acid supplemented diet. This study provides evidence that a monounsaturated or polyunsaturated fatty acid supplemented diet may regulate muscle proteasomes. Unsaturated fatty acids are particularly prone to free radical attack. Thus, we suggest that alterations in muscle proteasome may result from monounsaturated and polyunsaturated fatty acid-induced peroxidation, in order to eliminate damaged proteins.

Radiobiologiia. 1985 Nov-Dec;25(6):763-7. [Mechanism of circulatory disorders in animals irradiated at high doses] [Article in Russian] Pozharisskaia TD, Vasil'eva TP, Sokolova EN, Alekseeva II. Some data are reported on pathoanatomical changes, a status of the microcirculatory channel and the coagulogram of animals affected by high doses of ionizing radiation. The signs of disseminated intravascular blood coagulation have been revealed. The authors discuss the relationship between clinical manifestations with coagulopathy development and circulatory disturbances during intestinal and cerebral forms of acute radiation sickness.

Adv Exp Med Biol. 1976;75:497-503. Effect of ionizing radiation on liver microcirculation and oxygenation. Bicher HI, Dalrymple GV, Ashbrook D, Smith R, Harris D. Platelet aggregation and adhesiveness, as well as TPO2 responses to hypoxia were measured as microcirculation parameters in beagle dogs subject to Co60 ionizing radiation to a dose of 4600 rads in 5 weeks. Simultaneously, changes in blood chemistry and coagulation were also determined. Marked changes in all studied parameters in the post radiation period lead to the conclusion that radiation liver damage is at least in part mediated through microcirculation disturbances.

Am J Clin Nutr. 2003 May;77(5):1125-32. Effect of individual dietary fatty acids on postprandial activation of blood coagulation factor VII and fibrinolysis in healthy young men. Tholstrup T, Miller GJ, Bysted A, Sandstrom B. Research Department of Human Nutrition and the Center of Advanced Food Research, the Royal Veterinary and Agricultural University, Frederiksberg, Denmark. R.mensink@hb.unimaas.nl BACKGROUND: Hypertriglyceridemia may represent a procoagulant state involving

disturbances to the hemostatic system. Plasminogen activator inhibitor type 1 (PAI-1) is increased in the presence of hypertriglyceridemia. Free fatty acids (FFAs) in plasma may promote factor VII (FVII) activation. OBJECTIVE: We tested the hypothesis that FVII activation would be less after consumption of saturated fatty acids than after other fatty acids. DESIGN: The effects of 6 matching dietary test fats, rich in stearic (S), palmitic (P), palmitic + myristic (M), oleic (O), trans 18:1 (T), and linoleic (L) acid, respectively, on the postprandial lipid and hemostatic profile (after 2, 4, 6, and 8 h) were investigated in 16 young men. High-fat meals (1 g fat/kg body wt; 43% from the test fatty acid) were served in the morning on 6 separate days. RESULTS: All fats increased FVII activation. The S fat resulted in a lower increase in activated FVII (FVIIa) than did the T fat and in a lower FVII coagulant activity (FVII:c) than did the O fat (P < 0.02, diet x time interaction). When the data were pooled, the saturated (S, P, and M) test fats resulted in a smaller

postprandial increase in FVIIa (P = 0.036, diet effect), a smaller increase in FVII:c (P < 0.001, diet x time interaction), a greater rise in tissue plasminogen activator concentrations (P = 0.028, diet effect), and a tendency to

a greater postprandial decline in PAI-1 (P = 0.06, diet effect) compared with the unsaturated test fats (O, T, and L). The increase in FVIIa was not significantly associated with the level of lipemia, plasma FFAs, or plasma lipoprotein lipase activity. CONCLUSION: Our results indicate a lesser increase in FVIIa after the consumption of saturated fats, especially the S fat, than after unsaturated test fats.

Alcohol Clin Exp Res 1998 Feb;22(1):192-6.

Increased circulating products of lipid peroxidation in patients with alcoholic liver disease.

Aleynik SI, Leo MA, Aleynik MK, Lieber CS. Alcohol Research and Treatment Center, Bronx Veterans Affairs Medical Center and Mount Sinai School of Medicine, New York 10468, USA. F2-isoprostanes (F2-IP) and 4-hydroxynonenal (4-HNE), peroxidation products of polyunsaturated fatty acids (PUFA), are considered the most reliable indicators of endogenous lipid peroxidation in vivo. To determine to what extent these are also altered in patients with alcoholic liver disease, plasma free and esterified F2-IP as well as 4-HNE were measured by GC/MS in 49 fasting subjects who underwent diagnostic percutaneous needle biopsies of the liver. Compared to patients with mild steatosis and no fibrosis, free F2-IP and 4-HNE were strikingly increased in individuals with alcoholic hepatitis. There was also a significant but lesser rise of 4-HNE in patients with perivenular fibrosis. An

but they increased later (p < 0.005). Contrasting with plasma free F2-IP, esterified F2-IP were not significantly changed with fibrosis. Thus, whereas

Journal of Lipid Research, Vol. 44, 271-279, February 2003

Arachidonic acid and prostacyclin signaling promote adipose tissue development : a human health concern?

Florence Massiera*, Perla Saint-Marc*, Josiane Seydoux , Takahiko Murata , Takuya Kobayashi , Shuh Narumiya , Philippe Guesnet**, Ez-Zoubir Amri*, Raymond Negrel* and Gérard Ailhaud1,*

How big a foundation is needed to erect a multibillion dollar industry? Animal feed, paint; cholesterol; used as a drug, to prevent heart disease; but they were heart toxic and carcinogenic. The cholesterol/oil scam was lost by the time the anticholesterol drugs were established in the market, and the failed argument for “linoleic acid the essential fatty acid, the heart protective fat,” was quietly set aside. The researchers who now knew the toxicity of the “EFA” began to use animals fed corn oil or soy oil as the controls, against which to compare animals fed new oils, which were now being offered as alternatives to the previously marketed oils.

FFA stress; fibrosis; amiodarone, iodinated antithyroid;’ Dugrillon;

Pufa fibrosis, leak/angiogenesis

Exp Mol Pathol. 2003 Jun;74(3):282-90.

The role of COX-2 in angiogenesis and rheumatoid arthritis.

Woods JM, Mogollon A, Amin MA, Martinez RJ, Koch AE.

How Essential Are Fatty Acids? Ralph T. Holman. J. Am. Med. Assoc. 178, No. 9, 930-933 (1961).

Horrobin’s primrose path

Running an electrode through a single muscle cell, I noticed slight voltage differences at each depth.

Several georgia,palatino,times,serif,Times New Roman, when I have read an article by a prestigious professor on the subject of the essentiality of polyunsaturated fatty acids, I have written them asking what the evidence is that they are essential. One professor put the whole burden on a publication about one patient in a hospital. Others site G.O. Burr, or Burr and Burr, implying that the issue was settled by 1929. What was the state of nutritional knowledge in 1929? Over the last twenty years I have asked several prominent oil researchers what the evidence is that there is such a thing as an “essential fatty acid.” One man cited a single publication about a solitary sick person who recovered from some sickness after being given some unsaturated fat.

W.S. Hartroft and E.A. Porta, "Present Knowledge of Ceroid Pigment," Chapter VIII, p. 28, in Present Knowledge in Nutrition, 3rd edition, Nutrition Foundation, 1967.

FEBS Lett. 1974 Sep 15;46(1):39-41. On a slow inhibitory effect of free fatty acids on the respiratory chain of non-phosphorylating submitochondrial particles from beef heart. Schewe T, Ludwig P, Rapoport S.

Biochim Biophys Acta. 2001 Feb 14;1535(2):145-52.

Acrolein inhibits respiration in isolated brain mitochondria.

Picklo MJ, Montine TJ. Department of Pathology, Vanderbilt University Medical Center, Nashville, TN

37232, USA. matthew.picklo@mcmail.vanderbilt.edu

Lipid peroxidation is elevated in diseased regions of brain in several neurodegenerative diseases. Acrolein (2-propenal) is a major cytotoxic product of lipid peroxidation and its adduction to neuronal proteins has been demonstrated in diseased brain regions from patients with Alzheimer's disease. Mitochondrial abnormalities are implicated in several neurodegenerative disorders, and mitochondria are targets of alkenal adduction in vivo. We

examined the effects of acrolein upon multiple endpoints associated with the

mitochondrial involvement in neurodegenerative disease. Acrolein inhibited state

3 respiration with an IC(50) of approx. 0.4 micromol/mg protein; however, there

was no reduction in activity of complexes I-V. This inhibition was prevented by

glutathione and N-acetylcysteine. Acrolein did not alter mitochondrial calcium

transporter activity or induce cytochrome c release. These studies indicate that

acrolein is a potent inhibitor of brain mitochondrial respiration.

Biochim Biophys Acta. 1984 Nov 6;802(1):17-23. Activation of bovine platelets induced by long-chain unsaturated fatty acids at just below their lytic concentrations, and its mechanism. Kitagawa S, Endo J, Kametani F. The effects of long-chain unsaturated fatty acids such as linoleic acid on bovine platelets were examined. Not only linoleic acid, but also oleic and linolenic acid, at just below the concentrations causing marked cell lysis, induced an absorbance decrease of the platelet suspension in the presence of Ca2+. Since this absorbance decrease was reversed by the addition of EDTA and

moreover aggregate formation was found by macroscopic and microscopic

observation, it was concluded that unsaturated fatty acids at just below their lytic concentrations caused platelet aggregation. Unsaturated fatty acids also caused release of adenine nucleotides, but there was a lag time between the release and the aggregation, just as with ADP-induced release, suggesting that the aggregation was independent of the release of ADP. It was revealed that this activation of platelets by unsaturated fatty acids was caused by marked Ca2+ uptake into the cytoplasm, resulting from significant membrane perturbation.

3: Cancer Res. 1989 Apr 15;49(8):1931-6. Effects of fish oil and corn oil diets on prostaglandin-dependent and myelopoiesis-associated immune suppressor mechanisms of mice bearing metastatic Lewis lung carcinoma tumors. Young MR, Young ME. Department of Research Services, Edward J. Hines, Jr., Veterans Administration Hospital, Hines, Illinois 60141. The effects of a fish oil diet on the myelopoietic and immunological parameters of normal mice and of mice bearing metastatic Lewis lung carcinoma (LLC-C3) tumors were compared to the effects of a corn oil or a mixed-fat rodent chow diet. This was studied soon after tumor appearance, on Day 17, when immune suppression was mediated by prostaglandin E2 (PGE2)-producing suppressor cells, and late in tumor development, on Day 28 when immune suppression was associated with myelopoiesis and the appearance of bone marrow-derived suppressor cells whose activity was not dependent on PGE2. Feeding a fish oil diet from Days 10 to 17 of tumor growth partially restored splenic T-cell blastogenesis, reduced spleen cell secretion of PGE2, and alleviated splenic suppressor activity. When fed from Days 21 to 28 of tumor growth, a fish oil diet neither restored T-cell blastogenesis nor alleviated suppressor cell activity. The fish oil diet increased the frequency of myeloid progenitor cells in normal mice and in mice bearing small or large tumors. Concurrently, the fish oil diet stimulated the appearance of bone marrow-derived suppressor cells. When administered after the establishment of palpable primary tumors, a fish oil diet also increased the formation of pulmonary lung nodules. In contrast to the fish oil stimulation of myelopoiesis and the associated suppressor cells, feeding a corn oil diet to tumor-bearing mice during Days 21 to 28 after tumor implantation reduced myelopoiesis and the presence of the associated bone marrow suppressor cells. These data show that a fish oil diet can minimize the immune suppression in tumor bearers when suppression is mediated by PGE2-producing suppressor cells, but can also induce myelopoietic stimulation leading to the appearance of bone marrow-derived suppressor cells and increased tumor metastasis.

Am J Clin Nutr. 1987 Jan;45(1 Suppl):218-24. Fat and essential fatty acid in mammary carcinogenesis. Ip C.

Cancer Res. 1986 Feb;46(2):757-62. Comparative effects of different animal and vegetable fats fed before and during carcinogen administration on mammary tumorigenesis, sexual maturation, and endocrine function in rats. Sylvester PW, Russell M, Ip MM, Ip C. The purpose of this investigation was to determine whether diets high in animal or vegetable fat affected mammary tumorigenesis when fed to rats only prior to and during the initiation phase of carcinogenesis. Weanling 21-day-old female Sprague-Dawley rats were divided into different dietary treatment groups and were allowed to feed and libitum on one of the following diets: 5% (normal fat) corn oil; 20% (high fat) corn oil; 20% palm oil; 20% beef tallow; or 20% lard. At 52 days of age, all rats were given p.o. 7.5 mg 7,12-dimethylbenz(a)anthracene (DMBA). One week following DMBA administration, all rats were switched to the 5% corn oil control diet and were maintained on this diet for the duration of the experiment. Rats fed a 20% lard diet during the treatment period showed a significant increase in mammary tumor incidence and number 19 weeks after DMBA administration, when compared to all other dietary treatment groups. Rats fed a 20% beef tallow diet during this same time period also demonstrated enhanced mammary tumor development, during the 10- to 19-week time period after DMBA. Mammary tumor development in rats fed 20% corn oil or palm oil diets during this treatment period was similar to that of normal fat controls. Estrogens are potent stimulators of mammary tumor growth and development in rats. Because mammary tumorigenesis was enhanced in rats fed high

animal, but not vegetable fat diets, it was possible that estrogens present in

animal fat might be responsible for this stimulation. Further studies demonstrated however, that increased mammary tumorigenesis in rats fed diets high in animal fat could not be explained on the basis of endocrine stimulation. Average day of vaginal opening for all groups fed 20% fat diets was similar and occurred earlier than in normal fat controls. In addition, 50- to 65-day-old

rats in the different dietary treatment groups showed no differences in basal or

surge levels of serum prolactin, luteinizing hormone, or estradiol. Rat diestrus

uterine weight also showed no significant differences among dietary treatment

groups. Thus diets containing high levels of animal fat caused little if any

increased estrogenic activity in rats. In conclusion, high dietary intake of lard and beef tallow, but not vegetable fat, fed from weaning until only 1 week after DMBA administration, significantly enhances mammary tumorigenesis in rats. The mechanism(s) by which animal fat induces this stimulation is not clear, but it does not appear to result from endogenous or exogenous endocrine stimulation.

Cancer Res. 1986 Feb;46(2):763-9. Effects of high dietary fat on the growth and development of ovarian-independent carcinogen-induced mammary tumors in rats. Sylvester PW, Ip C, Ip MM. This study examined the influence of high dietary fat intake on the development

of ovarian-independent mammary tumors in both vehicle-treated controls and rats

made deficient in estrogen and prolactin during tumor induction. The majority of

7,12-dimethylbenz(a) anthracene (DMBA)-induced mammary tumors in rats are

dependent on estrogen and prolactin for growth, and suppression of prolactin and

estrogen at the time of tumor initiation causes a reduction in tumor incidence and increase in tumor latency. However, the majority of mammary tumors which do develop in these animals exhibit ovarian-independent growth. Sprague-Dawley rats were given 7.5 mg DMBA p.o. at 57 days of age. Starting 1 day prior to and continuing for 7 days after DMBA administration, rats were given daily injection of vehicle or the combination of tamoxifen (20 micrograms/rat) plus bromocryptine (5 mg/kg). At the end of drug treatment, rats in each treatment group were equally divided and placed on normal fat (5% corn oil) or high fat (20% corn oil) diets for the duration of the experiment. Vehicle-treated rats were ovariectomized 27 wk and drug-treated rats 47 wk after DMBA administration to determine tumor ovarian dependency. Vehicle-treated rats fed high fat diets showed significant increases in mammary tumor incidence and number as compared to similarly treated rats fed a normal fat diet, with approximately 80% of the tumors in each group being ovarian dependent. Likewise, tamoxifen-bromocryptine-treated rats fed a high fat diet showed a significant enhancement in mammary tumor number, although not incidence, as compared to similarly treated rats fed a normal diet. Tumors in these drug-treated groups displayed essentially the same incidence of ovarian dependence (23%). Tamoxifen-bromocryptine-treated groups displayed a 2-fold increase in latency of tumor appearance as compared to vehicle-treated controls; however, this long latency was not reduced when these rats were fed a high fat diet. These results demonstrate that high dietary fat stimulates ovarian-dependent and -independent mammary tumorigenesis in rats but does not influence the hormonal responsiveness

of these tumors.

Prog Clin Biol Res. 1986;222:283-94. Relevance of trans fatty acids and fish oil in animal tumorigenesis studies. Ip C, Ip MM, Sylvester P. Cancer Res. 1985 May;45(5):1997-2001. Requirement of essential fatty acid for mammary tumorigenesis in the rat. Ip C, Carter CA, Ip MM. In an attempt to determine the requirement of essential fatty acid for dimethylbenz(a)anthracene-induced mammary tumorigenesis, rats were fed diets containing different levels of linoleate: 0.5, 1.1, 1.7, 2.2, 3.5, 4.4, 8.5, or 11.5%. Each diet contained 20% of fat by weight, with varying amounts of coconut oil and corn oil added to achieve the desired levels of linoleate. Mammary tumorigenesis was very sensitive to linoleate intake and increased proportionately in the range of 0.5 to 4.4% of dietary linoleate. Regression analysis indicated that a breakpoint occurred at 4.4%, beyond which there was a very poor linear relationship, suggesting the possibility of a plateau. From the intersection of the regression lines in both the upper and lower ranges, the level of linoleate required to elicit the maximal tumorigenic response was estimated to be around 4%. The differences in tumor yield could not be correlated with changes in prostaglandin E concentration in the mammary fat pads of normal animals maintained on similar diets, suggesting that linoleate may act by some other mechanism to stimulate mammary tumorigenesis.

Nutr Cancer. 1985;7(4):199-209. Isomeric fatty acids and tumorigenesis: a commentary on recent work. Hunter JE, Ip C, Hollenbach EJ. This article critically reviews the existing, although limited, literature concerning trans fatty acids and tumorigenesis. Neither epidemiological nor experimental studies published to date have demonstrated any valid association between trans fatty acid ingestion and tumorigenesis. A recent study showed that under controlled conditions, a fat with a high content of trans fatty acids did not promote the development of mammary tumors induced in rats by 7,12-dimethylbenz[a]anthracene to any greater extent than did a comparable fat with a high content of cis fatty acids. In addition, in this study a high trans fat was less tumor promoting than was a blend of fats that simulated the dietary fat composition of the United States and had a lower level of trans fatty acids. Another study using comparable cis and trans fats demonstrated that the high

trans fat did not affect the growth and metastasis of implanted mammary tumors in mice relative to the high cis fat. Also, two recent studies reported no significant difference in the development of induced colon tumors in rats fed diets high in cis or trans fatty acids. The results of these and other studies are consistent with the conclusion that trans fatty acids are not uniquely related to tumor development.

Vitamins: (hypothesis of disease deficiency) Sir F. G. Hopkins, Casimir Funk, England, 1912; (vitamin A) Elmer V. McCollum, M. Davis, U.S., 1912–1914; (vitamin B) McCollum, U.S., 1915–1916; (thiamin, B1) Casimir Funk, England, 1912; (riboflavin, B2) D. T. Smith, E. G. Hendrick, U.S., 1926; (niacin) Conrad Elvehjem, U.S., 1937; (B6) Paul Gyorgy, U.S., 1934; (vitamin C) C. A. Hoist, T. Froelich, Norway, 1912; (vitamin D) McCollum, U.S., 1922; (folic acid) Lucy Wills, England, 1933.

Arch Toxicol. 1997;71(9):563-74. Impaired cellular immune response in rats exposed perinatally to Baltic Sea herring oil or 2,3,7,8-TCDD. Ross PS, de Swart RL, van der Vliet H, Willemsen L, de Klerk A, van Amerongen G, Groen J, Brouwer A, Schipholt I, Morse DC, van Loveren H, Osterhaus AD, Vos JG. Seal Rehabilitation and Research Centre, Pieterburen, The Netherlands. While the immunotoxicity of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) has been well established, the effects of complex environmental mixtures of polyhalogenated aromatic hydrocarbons (PHAHs) are poorly understood. Many PHAHs, including the polychlorinated-biphenyls (PCBs), -dibenzofurans (PCDFs), and dibenzo-p-dioxins (PCDDs), possess 'dioxin-like' activities, and accumulate in the aquatic food chain. Organisms occupying high trophic levels may therefore be exposed to concentrations which may present an immunotoxic risk. In this study, pregnant PVG rats were administered a daily oral dose of 1 ml of the following during pregnancy and lactation: (1) oil extracted from herring caught in the relatively uncontaminated Atlantic Ocean; (2) oil extracted from herring caught in the contaminated Blastic Sea; or (3) the Atlantic herring oil extract spiked with 2,3,7,8-TCDD. The daily intakes of aryl hydrocarbon (Ah)-receptor dependent toxic equivalents (TEQ) for mothers were 0.3 in the Atlantic group, 2.1 in the Baltic group, and 134 ng/kg body wt. in the 2,3,7,8-TCDD positive control group. Immune function and host resistance to rat cytomegalovirus (RCMV) were assessed in offspring aged 11, 25, 46 or 59 days. Rat pups in the positive control

TCDD-spiked group exhibited immunosuppression characterized by reduced thymus weight and cellularity, reduced thymocyte and splenocyte proliferative responses to T-dependent mitogens in vitro, reduced virus-associated natural killer (NK) cell and specific antibody responses. While less pronounced, a similar pattern of effects was observed in the rat pups exposed only to the Baltic Sea herring oil. These immunotoxic effects were transient in both exposure groups, with a time-related recovery in immune function possibly due to the half-life of TCDD in rats and the waning exposure levels in the rapidly growing pups. We previously demonstrated that the same Baltic Sea herring led to impaired natural killer cell and T-lymphocyte function in harbour seals during the course of a long-term captive feeding study. The collective results of these studies in rats and seals indicate the immunotoxic potential of environmental mixtures at current levels in the aquatic environment, and suggest that the developing immune system of young mammals may be at particular risk.

Plasmin FA inhibit

J Clin Invest. 1985 Feb;75(2):456-61. Plasmin inhibition of platelet function and of arachidonic acid metabolism. Schafer AI, Adelman B. To study interactions between platelets and the fibrinolytic system, we examined the effects of human plasmin on human platelets washed by gel filtration. Plasmin concentrations that did not affect platelet shape change, release, or aggregation (less than 1.0 caseinolytic units [CU]/ml) caused a dose- and time-dependent inhibition of platelet aggregation in response to thrombin, ionophore A23187, and collagen. Complete loss of aggregation occurred at 0.1-0.5 CU/ml of plasmin. In a parallel dose-dependent manner, plasmin likewise inhibited thrombin, ionophore, and collagen-stimulated thromboxane B2 production. In contrast, neither aggregation nor thromboxane B2 formation induced by arachidonate was inhibited by plasmin pretreatment of the platelets. Plasmin blocked the thrombin-induced release of [3H]arachidonic acid from platelet membrane phospholipids and the thrombin-induced platelet oxygen burst. However, plasmin did not inhibit the arachidonate-induced oxygen burst. Inhibition of arachidonic acid release by plasmin was not mediated by increase in platelet cyclic AMP. These results suggest that plasmin inhibits platelet function, at least in part, by blocking the mobilization of arachidonic acid from membrane phospholipid pools. The effects of plasmin on platelets may contribute to the hemostatic abnormalities seen in pathologic and pharmacologic fibrinolysis.

J Biochem (Tokyo). 1977 Aug;82(2):529-33. Effects of free fatty acids on fibrinolytic activity. Muraoka T, Okuda H. A novel method for the estimation of fibrinolytic activity is proposed. In this method, a fibrin clot suspension is used as a substrate (fibrin is known to be a physiological substrate of plasmin). The fibrin clot suspension was prepared by homogenization of human fibrin clots. With this method, we found that free fatty acids inhibited the plasmin activity, and long-chain, unsaturated free fatty acids had a particularly strong inhibitory action on plasmin. As regards the mechanism of the inhibitory action, free fatty acids may not inhibit complex formation between plasmin and fibirin, but may make it impossible for plasmin to act on fibrin due to deformation of the surface of the fibrin clot.

Biochem J. 1994 May 15;300 ( Pt 1):251-5. Regulation of fibrinolysis by non-esterified fatty acids. Higazi AA, Aziza R, Samara AA, Mayer M. Department of Clinical Biochemistry, Hadassah Medical Center, Jerusalem, Israel. The ability of oleic acid to modulate fibrinolysis was measured by following the urokinase-mediated and plasminogen-dependent cleavage of 125I-labelled fibrin clots. Oleic acid levels within the physiological range exerted a concentration-dependent inhibition of urokinase-mediated fibrinolytic activity. SDS/PAGE revealed that oleic acid enhances urokinase activity but simultaneously increases the autolytic cleavage of the newly formed low-molecular-mass subunit of plasmin. Oleic acid-induced cleavage of this subunit containing the catalytic site of plasmin was suppressed by the plasmin substrate H-D-valyl-L-leucyl-L-lysine-p-nitroanilide (S-2251) and was prevented by alpha 2-antiplasmin. A concentration-dependent inhibition of the activity of purified plasmin on 125I-labelled fibrin clot was also observed; 93% and 50% inhibition was noted with 150 microM and 32 microM oleic acid respectively. Oleic acid at 200 microM also effectively displaced plasmin prebound to a polylysine-Sepharose column. Examination of the fatty acid specificity showed that a minimal chain length of 16 carbon atoms and the presence of at least one double bond, preferably in a cis configuration, were required for inhibition of the fibrinolytic activity of plasmin. Oleic acid at a concentration that produced only a minimal inhibition of plasmin activity induced a marked inhibition by

palmitic acid, while palmitic acid alone is ineffective. The findings suggest that oleic acid stimulates plasminogen activation and modulates the fibrinolytic and autolytic activities of plasmin.

J Burn Care Rehabil. 1990 Jan-Feb;11(1):1-6. Fibrinolysis inhibition in human burn blister fluid. Rockwell WB, Ehrlich HP. Shriners Burns Institute, Boston, MA 02114. Fibrin plate assays revealed that human burn blister fluid represses plasmin-induced fibrinolysis. While this repression is significant, it is not as potent as that of human serum. Ibuprofen, a nonsteroidal anti-inflammatory drug, antagonizes blister fluid inhibition of fibrinolysis. The activity of ibuprofen appears to be unrelated to the synthesis of prostaglandins because other nonsteroidal drugs that were tested, such as indomethacin, imidazole, and tolmetin had significantly less antagonistic activity. This plasmin inhibition, which is contained in burn blister fluid, may contribute to vascular occlusion after burn injury, which leads to secondary dermal ischemia and necrosis in patients traumatized by burns.

Neurosurgery. 1987 Oct;21(4):523-31. Pharmacodynamic evaluation of human cerebral arteries in the genesis of Vasospasm. White RP, Robertson JT. Department of Pharmacology, University of Tennessee, Memphis. Experiments were performed on isolated human cerebral arteries to evaluate the role desensitization and tachyphylaxis might play in preventing certain agonists from producing prolonged vasoconstriction after subarachnoid hemorrhage. In addition, the antiproteases leupeptin and pepstatin were studied to ascertain whether these peptides might inhibit contraction as does antithrombin III. The maximal contraction to KCl was used as a standard for comparing the responses elicited by the agonists, the decay of the responses to the agonists over 15 minutes was used as an index of desensitization, and the percentage of decrease in response to a second application of the agonist over the first was a measure of tachyphylaxis. The results showed that desensitization and tachyphylaxis greatly reduced or abolished the contractile responses to norepinephrine, serotonin, angiotensin II, arginine vasopressin, substance P, neuropeptide Y, neurotensin, thrombin, uridine triphosphate, linoleic acid, melittin, and cathepsin D. Moreover, some arteries failed to respond to some of these agonists, and no contractile response was elicited by acetylcholine or bradykinin. In contrast, prostaglandins E2, D2, and F2 alpha, as well as plasmin, produced sustained contractions, without tachyphylaxis, but only prostaglandin E2 and plasmin produced contractions at concentrations of 10(-7) M or less that were comparable to those of KCl. None of the antiprotease peptides inhibited the responses to KCl whereas small concentrations (6 X 10(-8) M) of antithrombin III did. The results support the hypotheses that the phenomenon of desensitization and tachyphylaxis would prevent many diverse agents from acting as spasmogens and that substances like antithrombin III present in the cerebrospinal fluid after hemorrhage could immediately protect patients from cerebral vasospasm.(ABSTRACT TRUNCATED AT 250 WORDS)

Neurochem Res. 2000 Feb;25(2):269-76. Cortical impact injury in rats promotes a rapid and sustained increase in polyunsaturated free fatty acids and diacylglycerols. Homayoun P, Parkins NE, Soblosky J, Carey ME, Rodriguez de Turco EB, Bazan NG. Louisiana State University Health Sciences Center, Neuroscience Center of Excellence, New Orleans, USA. Neurotrauma activates the release of membrane phospholipid-derived second messengers, such as free arachidonic acid (20:4n-6, AA) and diacylglycerols (DAGs). In the present study, we analyze the effect of cortical impact injury of low-grade severity applied to the rat frontal right sensory-motor cortex (FRC) on the accumulation of free fatty acids (FFAs) and DAGs in eight brain areas 30 min and 24 hours after the insult. At these georgia,palatino,times,serif,Times New Roman, accumulation of FFAs and DAGs occurred mainly in the damaged FRC. The cerebellum was the only other brain area that displayed a significant accumulation of DAGs by day one post-injury. By 30 min, accumulation of free AA in the FRC displayed the greatest relative increase (300% over sham value), followed by free docosahexaenoic acid (22:6n-3, DHA, 150%), while both 20:4-DAGs and 22:6-DAGs were increased 100% over sham values. At day one, free 22:6 and 22:6-DAGs showed the greatest increase (590% and 230%, respectively). These results suggest that TBI elicits the hydrolysis of phospholipids enriched in excitable membranes, targeting early on 20:4-phospholipids (by 30 min post- trauma) and followed 24 hours later by preferential hydrolysis of DHA-phospholipids. These lipid metabolic changes may contribute to the initiation and maturation of neuronal and fiber track degeneration observed following cortical impact injury.

Glia. 2003 Dec;44(3):275-82. Extracellular phospholipase A2 inhibitors suppress central nervous system inflammation. Pinto F, Brenner T, Dan P, Krimsky M, Yedgar S. Laboratory of Neuroimmunology, Department of Neurology, Hadassah University Hospital and Hebrew University Medical School, Jerusalem, Israel. Phospholipase A(2) (PLA(2)) plays a key role in the production of proinflammatory mediators, namely the arachidonic acid-derived eicosanoids, lysophospholipids, and platelet-activating factor, and indirectly influences the generation of cytokines, nitric oxide (NO), and free radicals. Accordingly, regulation of its activity is important in the treatment of inflammation. Since the main site of PLA(2) action in inflammatory processes is the cell membrane, we synthesized extracellular PLA(2) inhibitors (ExPLIs) composed of N-derivatized phosphatidyl-ethanolamine linked to polymeric carriers. These membrane-anchored lipid conjugates do not penetrate the cell and interfere with vital phospholipid metabolism or cell viability. The ExPLIs markedly inhibited central nervous system inflammation. This was reflected by the suppressed production and secretion of lipopolysaccharide-induced sPLA(2), prostaglandin E(2), and NO by glial cells and by the amelioration of experimental autoimmune encephalomyelitis in rats and mice. Copyright 2003 Wiley-Liss, Inc.

J Pediatr. 2003 Oct;143(4 Suppl):S26-34. In vivo approaches to quantifying and imaging brain arachidonic and docosahexaenoic acid metabolism. Rapoport SI. Brain Physiology and Metabolism Section, National Institute on Aging, National Institutes of Health, Bethesda, Maryland, USA. A novel in vivo fatty acid method has been developed to quantify and image brain metabolism of nutritionally essential polyunsaturated fatty acids (PUFAs). In unanesthetized rodents, a radiolabeled PUFA is injected intravenously, and its rate of incorporation into brain phospholipids is determined by chemical analysis or quantitative autoradiography. Results indicate that about 5% of brain arachidonic acid (20:4 n-6) and of docosahexaenoic acid (22:6 n-3) acid are lost daily by metabolism and are replaced from dietary sources through the plasma. Calculated turnover rates of PUFAs in brain phospholipids, due to deesterification by phospholipase A(2) (PLA(2)) followed by reesterification, are very rapid, consistent with active roles of PUFAs in signal transduction and other processes. Turnover rates of arachidonate and docosahexaenoate are independent of each other and probably are regulated by independent sets of enzymes. Brain incorporation of radiolabeled arachidonate can be imaged in response to drugs that bind to receptors coupled to PLA(2) through G proteins, thus measuring PLA(2)-initiated signal transduction. The in vivo fatty method is being extended for human studies using positron emission tomography.

J Cell Physiol. 2004 Jan;198(1):48-52. cPLA2 activator peptide, PLAP, increases arachidonic acid release and apoptosis of vascular smooth muscle cells. Pilane CM, LaBelle EF. Department of Pharmacology and Physiology, Drexel University College of Medicine, Philadelphia, Pennsylvania. Apoptosis of vascular smooth muscle cells (VSMCs) has recently drawn a lot of interest in various laboratories due to its importance in atherogenesis. We have shown previously that nitric-oxide (NO) can induce apoptosis of VSMCs and that the NO-induced apoptosis is accompanied by an increase in arachidonic acid release via cytoplasmic Ca(2+)-dependent phospholipase A(2) (cPLA(2)). We have demonstrated here that NO-induced activation of cPLA(2) leading to increased arachidonic acid release can be mimicked via direct activation of cPLA(2) with a cPLA(2) activator peptide, PLAP. The PLAP induced arachidonic acid release and apoptosis is inhibitable by a cPLA(2)-specific inhibitor, AACOCF(3), indicating the direct involvement of cPLA(2). In this study, activation of cPLA(2) appears to be preceded by activation and binding by PLAP indicating that the cPLA(2) functions are mediated via PLAP. J. Cell. Physiol. 198: 48-52, 2004. Copyright 2003 Wiley-Liss, Inc.

Placenta. 2003 Nov;24(10):965-73. Augmented PLA(2)Activity in Pre-eclamptic Decidual Tissue-A Key Player in the Pathophysiology of 'Acute Atherosis' in Pre-eclampsia? Staff AC, Ranheim T, Halvorsen B. Department of Obstetrics and Gynecology, Ulleval University Hospital, Kirkeveien 166, 0450, Oslo, Norway Decidual acute atherosis is associated with pre-eclampsia, but the underlying mechanism is still unclear. We have previously demonstrated elevated level of the oxidative stress marker 8-isoprostaglandin F(2alpha)(8-isoprostane) and lipids in pre-eclamptic decidual tissue. Arachidonic acid (AA) in tissue phospholipids is a source for 8-isoprostane generation, and 8-isoprostane is liberated from tissue phospholipids by phospholipase A(2)(PLA(2)). The aims of this study were to explore whether AA content or PLA(2)expression in pre-eclamptic decidual tissue differed from controls. Decidua basalis tissues were obtained by vacuum aspiration at Caesarean delivery in pre-eclamptic and control pregnancies. We demonstrated a statistically significantly higher total PLA(2)activity in pre-eclamptic decidua compared to control tissue. On the other hand, no differences in AA content of tissue phospholipids or protein expression of secretory and cytosolic PLA(2)between pre-eclamptic and control decidual tissue were found. In conclusion, the elevated level of free 8-isoprostane in pre-eclamptic decidual tissue could be caused by augmented PLA(2)activity. We speculate that an elevated PLA(2)enzyme activity in pre-eclamptic decidual tissue could be of importance in the pathogenesis of 'acute atherosis', comparable to the atherogenesis in cardiovascular diseases.

Antioxid Redox Signal. 2003 Oct;5(5):647-54. Phospholipase a(2), hydroxyl radicals, and lipid peroxidation in transient cerebral ischemia. Adibhatla RM, Hatcher JF, Dempsey RJ. Department of Neurological Surgery, Cardiovascular Research Center, University of Wisconsin, Madison, WI 53792. Phospholipid degradation is an important promoter of neuronal death after transient cerebral ischemia. Phospholipid hydrolysis by phospholipase A(2) (PLA(2)) after transient cerebral ischemia releases arachidonic acid. Arachidonic acid metabolism results in formation of reactive oxygen species, lipid peroxides, and toxic aldehydes (malondialdehyde, 4-hydroxynonenal, and acrolein). Citicoline (cytidine-5'-diphosphocholine), an intermediate in phosphatidylcholine synthesis, has undergone 13 phase III clinical trials for stroke, and is being evaluated for treatment of Alzheimer's and Parkinson's diseases. Here we examined the effect of citicoline on PLA(2) activity in relationship to attenuating hydroxyl radical (OH*) generation and lipid peroxidation after transient forebrain ischemia in gerbil. High Ca(2+) dependency (millimolar range) of PLA(2) activity suggests that secretory PLA(2) is the predominant isoform in membrane and mitochondria. Citicoline attenuated the increase in PLA(2) activity in both membrane and mitochondrial fractions. In vitro, citicoline and its components choline and cytidine had no effect on the PLA(2) activity. Thus, citicoline is not a "direct PLA(2) inhibitor." Citicoline also significantly attenuated loss of cardiolipin and arachidonic acid release from phosphatidylcholine and phosphatidylethanolamine. Transient cerebral ischemia resulted in significant formation of OH* and malondialdehyde, and citicoline significantly attenuated their formation. These results suggest that citicoline provides neuroprotection by attenuating the stimulation of PLA(2).

Am J Clin Nutr. 2003 May;77(5):1125-32. Effect of individual dietary fatty acids on postprandial activation of blood coagulation factor VII and fibrinolysis in healthy young men. Tholstrup T, Miller GJ, Bysted A, Sandstrom B. Research Department of Human Nutrition and the Center of Advanced Food Research, the Royal Veterinary and Agricultural University, Frederiksberg, Denmark. r.mensink@hb.unimaas.nl

BACKGROUND: Hypertriglyceridemia may represent a procoagulant state involving disturbances to the hemostatic system. Plasminogen activator inhibitor type 1 (PAI-1) is increased in the presence of hypertriglyceridemia. Free fatty acids (FFAs) in plasma may promote factor VII (FVII) activation. OBJECTIVE: We tested the hypothesis that FVII activation would be less after consumption of saturated fatty acids than after other fatty acids. DESIGN: The effects of 6 matching dietary test fats, rich in stearic (S), palmitic (P), palmitic + myristic (M), oleic (O), trans 18:1 (T), and linoleic (L) acid, respectively, on the postprandial lipid and hemostatic profile (after 2, 4, 6, and 8 h) were investigated in 16 young men. High-fat meals (1 g fat/kg body wt; 43% from the test fatty acid) were served in the morning on 6 separate days. RESULTS: All fats increased FVII activation. The S fat resulted in a lower increase in activated FVII (FVIIa) than did the T fat and in a lower FVII coagulant activity (FVII:c) than did the O fat (P < 0.02, diet x time interaction). When the data were pooled, the saturated (S, P, and M) test fats resulted in a smaller postprandial increase in FVIIa (P = 0.036, diet effect), a smaller increase in FVII:c (P < 0.001, diet x time interaction), a greater rise in tissue plasminogen activator concentrations (P = 0.028, diet effect), and a tendency to a greater postprandial decline in PAI-1 (P = 0.06, diet effect) compared with the unsaturated test fats (O, T, and L). The increase in FVIIa was not significantly associated with the level of lipemia, plasma FFAs, or plasma lipoprotein lipase activity. CONCLUSION: Our results indicate a lesser increase in FVIIa after the consumption of saturated fats, especially the S fat, than after unsaturated test fats.

Randomized Controlled Trial

Prostaglandins. 1978 Apr;15(4):557-64.

Prostaglandin I2 as a potentiator of acute inflammation in rats. Komoriya K, Ohmori H, Azuma A, Kurozumi S, Hashimoto Y, Nicolaou KC, Barnette WE, Magolda RL. Prostaglandin I2 potentiated the paw swelling induced by carrageenin in rats. Prostaglandin I2 (0.1 microgram) showed similar activity to PGE1 (0.01 microgram). This potentiating property disappeared in 60 minutes and was completely abolished by diphenhydramine (25 mg kg-1, i.p.). In vascular permeability tests, PGI2 itself (2.5 X 10(-10) mol, 88 ng) caused no dye leakage reaction, but PGE1 (2.5 X 10(-10) mol, 88.5 ng) caused a significant dye leakage. This effect of PGE1 was statistically significant compared with vehicle- or PGI2-treated groups (p less than 0.05). Prostaglandin I2 potentiated the increased vascular permeability induced by 5-hydroxytriptamine (2.5 X 10(-10) mol), bradykinin (5 X 10(-10) mol) and histamine (2 X 10(-10) to 2 X

10(-8) mol). The potentiation was the most evident in the case of histamine.

Journal of Lipid Research, Vol. 44, 271-279, February 2003. Arachidonic acid and prostacyclin signaling promote adipose tissue development : a human health concern? Florence Massiera*, Perla Saint-Marc*, Josiane Seydoux , Takahiko Murata , Takuya Kobayashi , Shuh Narumiya , Philippe Guesnet**, Ez-Zoubir Amri*, Raymond Negrel* and Gérard Ailhaud1,* Institut de Recherche Signalisation, Biologie du Développement et Cancer, Centre de Biochimie (UMR6543CNRS), UNSA, Faculté des Sciences, Parc Valrose, 06108 Nice cedex 2, France Centre Médical Universitaire, Département de Physiologie, 1 rue Michel Servet, 1211 Genève 4, Switzerland Department of Pharmacology, Kyoto University, Faculty of Medicine, Yoshida, Sakyo-ku, Kyoto 606-8315, Japan** Laboratoire de Nutrition et Sécurité Alimentaire, INRA, 78352 Jouy-en-Josas, France 1 To whom correspondence should be addressed. e-mail: ailhaud@unice.fr High fat intake is associated with fat mass gain through fatty acid activation of peroxisome proliferator-activated receptors and , which promote adipogenesis. We show herein that, compared to a combination of specific agonists to both receptors or to saturated, monounsaturated, and -3 polyunsaturated fatty acids, arachidonic acid (C20:4, -6) promoted substantially the differentiation of clonal preadipocytes. This effect was blocked by cyclooxygenase inhibitors and mimicked by carbacyclin, suggesting a role for the prostacyclin receptor and activation of the cyclic AMP-dependent pathways that regulate the expression of the CCAAT enhancer binding proteins ß and implicated in adipogenesis. During the pregnancy-lactation period, mother mice were fed either a high-fat diet rich in linoleic acid, a precursor of arachidonic acid (LO diet), or the same isocaloric diet enriched in linoleic acid and -linolenic acid (LO/LL diet). Body weight from weaning onwards, fat mass, epididymal fat pad weight, and adipocyte size at 8 weeks of age were higher with LO diet than with LO/LL diet. In contrast, prostacyclin receptor-deficient mice fed either diet were similar in this respect, indicating that the prostacyclin signaling contributes to adipose tissue development. These results raise the issue of the high content of linoleic acid of i) ingested lipids during pregnancy and lactation, and ii) formula milk and infant foods in relation to the epidemic of childhood obesity.

J Clin Invest. 1985 Feb;75(2):456-61.

Plasmin inhibition of platelet function and of arachidonic acid metabolism. Schafer AI, Adelman B.

To study interactions between platelets and the fibrinolytic system, we examined the effects of human plasmin on human platelets washed by gel filtration. Plasmin concentrations that did not affect platelet shape change, release, or aggregation (less than 1.0 caseinolytic units [CU]/ml) caused a dose- and time-dependent inhibition of platelet aggregation in response to thrombin, ionophore A23187, and collagen. Complete loss of aggregation occurred at 0.1-0.5 CU/ml of plasmin. In a parallel dose-dependent manner, plasmin likewise inhibited thrombin, ionophore, and collagen-stimulated thromboxane B2 production. In contrast, neither aggregation nor thromboxane B2 formation induced by arachidonate was inhibited by plasmin pretreatment of the platelets. Plasmin blocked the thrombin-induced release of [3H]arachidonic acid from

platelet membrane phospholipids and the thrombin-induced platelet oxygen burst. However, plasmin did not inhibit the arachidonate-induced oxygen burst. Inhibition of arachidonic acid release by plasmin was not mediated by increase in platelet cyclic AMP. These results suggest that plasmin inhibits platelet function, at least in part, by blocking the mobilization of arachidonic acid from membrane phospholipid pools. The effects of plasmin on platelets may contribute to the hemostatic abnormalities seen in pathologic and pharmacologic

fibrinolysis.

J Biochem (Tokyo). 1977 Aug;82(2):529-33. Effects of free fatty acids on fibrinolytic activity. Muraoka T, Okuda H.

A novel method for the estimation of fibrinolytic activity is proposed. In this method, a fibrin clot suspension is used as a substrate (fibrin is known to be a physiological substrate of plasmin). The fibrin clot suspension was prepared by homogenization of human fibrin clots. With this method, we found that free fatty acids inhibited the plasmin activity, and long-chain, unsaturated free fatty acids had a particularly strong inhibitory action on plasmin. As regards the

mechanism of the inhibitory action, free fatty acids may not inhibit complex formation between plasmin and fibirin, but may make it impossible for plasmin to act on fibrin due to deformation of the surface of the fibrin clot.

Biochem J. 1994 May 15;300 ( Pt 1):251-5. Regulation of fibrinolysis by non-esterified fatty acids.

Higazi AA, Aziza R, Samara AA, Mayer M. Department of Clinical Biochemistry, Hadassah Medical Center, Jerusalem, Israel. The ability of oleic acid to modulate fibrinolysis was measured by following the urokinase-mediated and plasminogen-dependent cleavage of 125I-labelled fibrin clots. Oleic acid levels within the physiological range exerted a concentration-dependent inhibition of urokinase-mediated fibrinolytic activity.

SDS/PAGE revealed that oleic acid enhances urokinase activity but simultaneously increases the autolytic cleavage of the newly formed low-molecular-mass subunit

of plasmin. Oleic acid-induced cleavage of this subunit containing the catalytic site of plasmin was suppressed by the plasmin substrate

H-D-valyl-L-leucyl-L-lysine-p-nitroanilide (S-2251) and was prevented by alpha 2-antiplasmin. A concentration-dependent inhibition of the activity of purified

plasmin on 125I-labelled fibrin clot was also observed; 93% and 50% inhibition was noted with 150 microM and 32 microM oleic acid respectively. Oleic acid at 200 microM also effectively displaced plasmin prebound to a polylysine-Sepharose column. Examination of the fatty acid specificity showed that a minimal chain length of 16 carbon atoms and the presence of at least one double bond, preferably in a cis configuration, were required for inhibition of the fibrinolytic activity of plasmin. Oleic acid at a concentration that produced

only a minimal inhibition of plasmin activity induced a marked inhibition by palmitic acid, while palmitic acid alone is ineffective. The findings suggest that oleic acid stimulates plasminogen activation and modulates the fibrinolytic

and autolytic activities of plasmin.

J Burn Care Rehabil. 1990 Jan-Feb;11(1):1-6. Fibrinolysis inhibition in human burn blister fluid. Rockwell WB, Ehrlich HP.

Shriners Burns Institute, Boston, MA 02114. Fibrin plate assays revealed that human burn blister fluid represses plasmin-induced fibrinolysis. While this repression is significant, it is not as

potent as that of human serum. Ibuprofen, a nonsteroidal anti-inflammatory drug,

antagonizes blister fluid inhibition of fibrinolysis. The activity of ibuprofen

appears to be unrelated to the synthesis of prostaglandins because other nonsteroidal drugs that were tested, such as indomethacin, imidazole, and tolmetin had significantly less antagonistic activity. This plasmin inhibition,

which is contained in burn blister fluid, may contribute to vascular occlusion after burn injury, which leads to secondary dermal ischemia and necrosis in patients traumatized by burns.

Neurosurgery. 1987 Oct;21(4):523-31. Pharmacodynamic evaluation of human cerebral arteries in the genesis of

Vasospasm. White RP, Robertson JT.

Department of Pharmacology, University of Tennessee, Memphis.

Experiments were performed on isolated human cerebral arteries to evaluate the role desensitization and tachyphylaxis might play in preventing certain agonists

from producing prolonged vasoconstriction after subarachnoid hemorrhage. In addition, the antiproteases leupeptin and pepstatin were studied to ascertain

whether these peptides might inhibit contraction as does antithrombin III. The maximal contraction to KCl was used as a standard for comparing the responses elicited by the agonists, the decay of the responses to the agonists over 15 minutes was used as an index of desensitization, and the percentage of decrease in response to a second application of the agonist over the first was a measure of tachyphylaxis. The results showed that desensitization and tachyphylaxis greatly reduced or abolished the contractile responses to norepinephrine, serotonin, angiotensin II, arginine vasopressin, substance P, neuropeptide Y, neurotensin, thrombin, uridine triphosphate, linoleic acid, melittin, and cathepsin D. Moreover, some arteries failed to respond to some of these

agonists, and no contractile response was elicited by acetylcholine or bradykinin. In contrast, prostaglandins E2, D2, and F2 alpha, as well as plasmin, produced sustained contractions, without tachyphylaxis, but only

prostaglandin E2 and plasmin produced contractions at concentrations of 10(-7) M

or less that were comparable to those of KCl. None of the antiprotease peptides inhibited the responses to KCl whereas small concentrations (6 X 10(-8) M) of

antithrombin III did. The results support the hypotheses that the phenomenon of desensitization and tachyphylaxis would prevent many diverse agents from acting as spasmogens and that substances like antithrombin III present in the

cerebrospinal fluid after hemorrhage could immediately protect patients from cerebral vasospasm.(ABSTRACT TRUNCATED AT 250 WORDS)

http://www.wesleyan.edu/synthesis/GROUP4/FINALVERSIONS/neutrino.html.

http://www.google.com/search?q=cache:uN3B0g4R6EMJ:www.wesleyan.edu/synthesis/GROUP4/FINALVERSIONS/neutrino.html+neutrino+beta+recoil+history&hl=en&ie=UTF-8

Neutrinos, born of decay, are given off as one particle deteriorates into a more stable state. Neutrinos are emitted in positron (another type of subnuclear particle) beta decay while the anti-neutrino is emitted from electron beta decay. As a pion decays into a muon, the muon neutrino emerges along side the muon. When a pion decays, a neutral particle must be emitted in the direction opposite that of the muon in order to conserve momentum. The original assumption was that this particle was the neutrino that conserves momentum in beta decay. In 1962, however, researchers proved that the neutrino accompanying pion decay is different. At this point, little is known about the tau neutrino.

Neutrinos come from a variety of sources, nuclear reactions like those caused by nuclear warheads and our own sun create neutrinos. Super-novas, that is, exploding distant stars, create neutrinos. Neutrinos also come from the earth's own atmosphere, as cosmic rays bombard atmospheric particles to create new particles, some of them the unstable pions that deteriorate into muons, that further deteriorate into electrons. At each of these deteriorations, neutrinos are given off.

Clever scientists first caught wind of the idea for the neutrino by a perplexing flaw in their formerly-solid physics equationing. Understanding neutrinos are radiated by nuclear reactions and the sun itself is a "giant atom-smashing machine" powered by nuclear reactivity, the sun then is a source for neutrinos. However, the math doesn't work out. The predicted number of neutrinos is off, scientists can only observe only 50% of the neutrinos that "should be" there. Why? This is the "solar neutrino problem" being explored by scientists at the Super-Kamiokande detector, but first one should look at the past before diving into the future.

A History of the Neutrino

Nobel Prize winner Wolfgang Pauli proposed in 1930 that the missing energy in nuclear beta decays was carried by a neutral particle. Three years later, Nobel-Prize-Winning nuclear physicist Enrico Fermi named the particle "neutrino," meaning "little neutral one," to distinguish it from the much larger neutron and developed a theory calculating that a neutrino and electron were both emitted in decay. However, both scientists were just blowing smoke at this point as no one had yet detected the theoretical neutrino. The problem was that little could stop a neutrino long enough to detect it and it plowed through light-years worth of matter making containment seemingly impossible.

In 1956, UC Irvine physicists Frederick Reines and Clyde Cowan stepped up to the challenge of detecting the elusive neutrino and succeeded.

Biochim Biophys Acta. 1975 Aug 26;397(2):318-30. Palmitic acid activation of peroxidase and its possible significance in mango Ripening. Mattoo AK, Modi VV. Palmitic acid stimulated the activity of mango peroxidase and reversed the inhibition due to the peroxidase inhibitor present in the preclimacteric fruit. The palmitic acid effect appeared to saturate in the range of 45 to 60 muM palmitic acid. Crude fatty acid extract of the mango exerted similar effect. The percentage stimulation was pH-dependent. Palmitic acid stimulated the enzyme by 18 percent at its optimum pH (5) but the stimulation was in excess of 63 percent at pH 2.5. At pH 2.5 the enzyme concentration versus velocity plot was non-linear and the activation by palmitic acid appeared to saturate between 32 and 48 muM concentration of the effector. The inhibition of the enzyme at and above 0.86 muM concentration of substrate (H202) was not found in the presence of palmitic acid. The effector also changed the heat inactivation kinetics of the enzyme and activated only two out of the four peroxidase isoenzymes present in the climacteric fruit extracts. The results presented indicate the regulatory nature of the enzyme and support its significance in fruit ripening.

fatty acid proteolysis

Whitehouse AS, Tisdale MJ.

Galban VD, Evangelista EA, Migliorini RH, do Carmo Kettelhut I.

Farooqui AA, Yi Ong W, Lu XR, Halliwell B, Horrocks LA.

Del Roso A, Vittorini S, Cavallini G, Donati A, Gori Z, Masini M, Pollera M, Bergamini E.

Kee aj

Voisin L, Breuille D, Combaret L, Pouyet C, Taillandier D, Aurousseau E, Obled C, Attaix D. 3 types Hasselgren PO, Wray C, Mammen J.

Pickering WP, Price SR, Bircher G, Marinovic AC, Mitch WE, Walls J.

Yamada S, Yamada J, Sato K, Tokumoto T, Yasutomi M, Ishikawa K.

Biochem Biophys Res Commun. 2001 Jul 20;285(3):598-602. Downregulation of ubiquitin-dependent proteolysis by eicosapentaenoic acid in acute starvation. Whitehouse AS, Tisdale MJ. Pharmaceutical Sciences Research Institute, Aston University, Birmingham B4 7ET, United Kingdom. A number of acute wasting conditions are associated with an upregulation of the ubiquitin-proteasome system in skeletal muscle. Eicosapentaenoic acid (EPA) is effective in attenuating the increased protein catabolism in muscle in cancer cachexia, possibly due to inhibition of 15-hydroxyeicosatetraenoic acid (15-HETE) formation. To determine if a similar pathway is involved in other catabolic conditions, the effect of EPA on muscle protein degradation and activation of the ubiquitin-proteasome pathway has been determined during acute fasting in mice. When compared with a vehicle control group (olive oil) there was a significant decrease in proteolysis of the soleus muscles of mice treated with EPA after starvation for 24 h, together with an attenuation of the proteasome "chymotryptic-like" enzyme activity and the induction of the expression of the 20S proteasome alpha-subunits, the 19S regulator and p42, an ATPase subunit of the 19S regulator in gastrocnemius muscle, and the ubiquitin-conjugating enzyme E2(14k). The effect was not shown with the related

(n-3) fatty acid docosahexaenoic acid (DHA) or with linoleic acid. However,

2,3,5-trimethyl-6-(3-pyridylmethyl)1,4-benzoquinone (CV-6504), an inhibitor of

5-, 12- and 15-lipoxygenases also attenuated muscle protein catabolism, proteasome "chymotryptic-like" enzyme activity and expression of proteasome 20Salpha-subunits in soleus muscles from acute fasted mice. These results suggest that protein catabolism in starvation and cancer cachexia is mediated through acommon pathway, which is inhibited by EPA and is likely to involve a lipoxygenase metabolite as a signal transducer. Copyright 2001 Academic Press.

Cancer Res. 2001 May 1;61(9):3604-9. Mechanism of attenuation of skeletal muscle protein catabolism in cancer cachexia by eicosapentaenoic acid. Whitehouse AS, Smith HJ, Drake JL, Tisdale MJ.Pharmaceutical Sciences Research Institute, Aston University, Birmingham B4 7ET, United Kingdom. Cancer cachexia is characterized by selective depletion of skeletal muscle protein reserves. Soleus muscles from mice bearing a cachexia-inducing tumor (MAC16) showed an increased protein degradation in vitro, as measured by tyrosine release, when compared with muscles from nontumor-bearing animals. After incubation under conditions that modify different proteolytic systems, lysosomal, calcium-dependent, and ATP-dependent proteolysis were found to contribute to the elevated protein catabolism. Treatment of mice bearing the MAC16 tumor with the polyunsaturated fatty acid, eicosapentaenoic acid (EPA), attenuated loss of body weight and significantly suppressed protein catabolism in soleus muscles through an inhibition of an ATP-dependent proteolytic pathway. The ATP-ubiquitin-dependent proteolytic pathway is considered to play a major role in muscle catabolism in cachexia, and functional proteasome activity, as determined by "chymotrypsin-like" enzyme activity, was significantly elevated in gastrocnemius muscle of mice bearing the MAC16 tumor as weight loss progressed. When animals bearing the MAC16 tumor were treated with EPA, functional proteasome activity was completely suppressed, together with attenuation of the expression of 20S proteasome alpha-subunits and the p42 regulator, whereas there was no effect on the expression of the ubiquitin-conjugating enzyme (E2(14k)). These results suggest that EPA induces an attenuation of the up-regulation of proteasome expression in cachectic mice, and this was correlated with an increase in myosin expression, confirming retention of contractile proteins. EPA also inhibited growth of the MAC16 tumor in a dose-dependent manner, and this correlated with suppression of the expression of the 20S proteasome alpha-subunits in tumor cells, suggesting that this may be the mechanism of tumor growth inhibition. Thus EPA antagonizes loss of skeletal muscle proteins in cancer cachexia by down-regulation of proteasome expression, and this may also be the mechanism for inhibition of tumor growth.

Biochem Biophys Res Commun. 2001 Jul 20;285(3):598-602. Downregulation of ubiquitin-dependent proteolysis by eicosapentaenoic acid in acute starvation. Whitehouse AS, Tisdale MJ. Pharmaceutical Sciences Research Institute, Aston University, Birmingham B4 7ET, United Kingdom. A number of acute wasting conditions are associated with an upregulation of the ubiquitin-proteasome system in skeletal muscle. Eicosapentaenoic acid (EPA) is effective in attenuating the increased protein catabolism in muscle in cancer cachexia, possibly due to inhibition of 15-hydroxyeicosatetraenoic acid (15-HETE) formation. To determine if a similar pathway is involved in other catabolic conditions, the effect of EPA on muscle protein degradation and activation of the ubiquitin-proteasome pathway has been determined during acute fasting in mice. When compared with a vehicle control group (olive oil) there was a significant decrease in proteolysis of the soleus muscles of mice treated with EPA after starvation for 24 h, together with an attenuation of the proteasome "chymotryptic-like" enzyme activity and the induction of the expression of the 20S proteasome alpha-subunits, the 19S regulator and p42, an ATPase subunit of the 19S regulator in gastrocnemius muscle, and the ubiquitin-conjugating enzyme E2(14k). The effect was not shown with the related (n-3) fatty acid docosahexaenoic acid (DHA) or with linoleic acid. However, 2,3,5-trimethyl-6-(3-pyridylmethyl)1,4-benzoquinone (CV-6504), an inhibitor of 5-, 12- and 15-lipoxygenases also attenuated muscle protein catabolism, proteasome "chymotryptic-like" enzyme activity and expression of proteasome 20S alpha-subunits in soleus muscles from acute fasted mice. These results suggest that protein catabolism in starvation and cancer cachexia is mediated through a common pathway, which is inhibited by EPA and is likely to involve a lipoxygenase metabolite as a signal transducer. Copyright 2001 Academic Press.

protein reserves. Soleus muscles from mice bearing a cachexia-inducing tumor (MAC16) showed an increased protein degradation in vitro, as measured by tyrosine release, when compared with muscles from nontumor-bearing animals.

After incubation under conditions that modify different proteolytic systems,

lysosomal, calcium-dependent, and ATP-dependent proteolysis were found to

contribute to the elevated protein catabolism. Treatment of mice bearing the MAC16 tumor with the polyunsaturated fatty acid, eicosapentaenoic acid (EPA), attenuated loss of body weight and significantly suppressed protein catabolism

in soleus muscles through an inhibition of an ATP-dependent proteolytic pathway.

The ATP-ubiquitin-dependent proteolytic pathway is considered to play a major

role in muscle catabolism in cachexia, and functional proteasome activity, as determined by "chymotrypsin-like" enzyme activity, was significantly elevated in gastrocnemius muscle of mice bearing the MAC16 tumor as weight loss progressed. When animals bearing the MAC16 tumor were treated with EPA, functional proteasome activity was completely suppressed, together with attenuation of the expression of 20S proteasome alpha-subunits and the p42 regulator, whereas there was no effect on the expression of the ubiquitin-conjugating enzyme (E2(14k)). These results suggest that EPA induces an attenuation of the up-regulation of proteasome expression in cachectic mice, and this was correlated with an increase in myosin expression, confirming retention of contractile proteins. EPA also inhibited growth of the MAC16 tumor in a dose-dependent manner, and this correlated with suppression of the expression of the 20S proteasome alpha-subunits in tumor cells, suggesting that this may be the mechanism of tumor growth inhibition. Thus EPA antagonizes loss of skeletal muscle proteins in cancer cachexia by down-regulation of proteasome expression, and this may also be the mechanism for inhibition of tumor growth.

More MS news articles for June 2003

Impaired fibrinolysis in multiple sclerosis: a role for tissue plasminogen activator inhibitors

http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=12805124&dopt=Abstract

Brain. 2003 Jun 4

Gveric D, Herrera B, Petzold A, Lawrence DA, Cuzner ML.

Tissue plasminogen activator (tPA), a neuronal as well as the key fibrinolytic enzyme, is found concentrated on demyelinated axons in multiple sclerosis lesions together with fibrin(ogen) deposits.

The decreased tPA activity in normal-appearing white and grey matter and lesions of multiple sclerosis is reflected in diminished fibrinolysis as measured by a clot lysis assay.

Nonetheless, peptide products of fibrin, including D-dimer, accumulate on demyelinated axons--the result of fibrinogen entry through a compromised blood-brain barrier (BBB).

As total tPA protein remains unchanged in acute lesions and the concentration of PAI-1 rises several fold, complex formation is a probable cause of the impaired fibrinolysis.

This is G o o g l e's cache of http://saturn.med.nyu.edu/groups/ChaoLab/personnel/katerina/Akassoglou.html.

G o o g l e's cache is the snapshot that we took of the page as we crawled the web.

The page may have changed since that time. Click here for the current page without highlighting.

To link to or bookmark this page, use the following url: http://www.google.com/search?q=cache:zGSqF0X_aigJ:saturn.med.nyu.edu/groups/ChaoLab/personnel/katerina/Akassoglou.html+multiple+sclerosis+fibrinogen&hl=en&ie=UTF-8

Google is not affiliated with the authors of this page nor responsible for its content.

These search terms have been highlighted: multiple sclerosis fibrinogen

Katerina Akassoglou, Ph.D.

Ph.D., University of Athens, Athens, Greece

Postdoc, The Rockefeller University & SUNY at Stony Brook

Keywords: Multiple Sclerosis, extracellular matrix, fibrin, regeneration

picture

RESEARCH INTERESTS:

Regeneration failure in the adult mammalian CNS is not due to an inability of central neurons to elongate and remyelinate, but rather to the non-permissive nature of the CNS environment. Our studies identified fibrin as a novel inhibitory protein that delays nerve regeneration after sciatic nerve injury and showed that fibrin degradation correlates with nerve regeneration, while decreased proteolytic activity in the nervous tissue exacerbates damage. Fibrin, derived from the blood protein fibrinogen, is deposited in the nervous tissue after injury or disease associated with blood-brain barrier leakage. For example, in Multiple Sclerosis fibrin deposition correlates with demyelination and persists in plaques that do not show signs of repair. My research interests focus on the cellular mechanisms and molecular pathways that fibrin uses to affect remyelination and inflammation in the nervous system in an attempt to develop novel therapeutic strategies for neuroinflammatory diseases.

SELECTED PUBLICATIONS:

Akassoglou K., Yu W-M, Akpinar P, Strickland S. 2002. Fibrin inhibits peripheral nerve regeneration by arresting Schwann cell differentiation. Neuron, 33:861-875. pdf

Akassoglou K, Strickland S. 2002. Nervous system pathologies: The fibrin perspective. Biol Chem, 383:37-45. pdf

Probert L, Akassoglou K. 2001. Glial expression of cytokines in transgenic animals- how do these models reflect the “normal situation”. Glia, 36:212-219. pdf

Akassoglou K, Kombrink KW, Degen JL, Strickland S. 2000. Tissue plasminogen activator-mediated fibrinolysis protects from axonal demyelination after sciatic nerve injury, J Cell Biol, 149:1157-1166. pdf

Akassoglou K, Bauer J, Kassiotis G, Pasparakis M, Lassmann H, Kollias G and Probert L. 1998. Oligodendrocyte apoptosis and primary demyelination induced by local TNF/p55TNF receptor signaling in the CNS of transgenic mice: models for multiple sclerosis with primary oligodendrogliopathy. Am J Pathol, 153:801-813. pdf

Akassoglou K, Kassiotis G, Kollias G, Probert L. Role for TNF in CNS inflammation, demyelination and neurodegeneration studied in transgenic mice, in Neuroimmunodegeneration Edited by P. Wong, B. Lynn, LANDES Bioscience, 1998, pp. 133-149. (book chapter).

Akassoglou K, Probert L, Kontogeorgos G and Kollias G. 1997. Astrocyte, but not neuron-specific, transmembrane TNF triggers inflammation and degeneration in the CNS of transgenic mice. J Immunol 158:438-445. pdf

Probert L, Akassoglou K, Pasparakis M, Kontogeorgos G and Kollias G. 1995. Spontaneous inflammatory and demyelinating disease in transgenic mice showing central nervous system-specific tumor necrosis factor ? expression. Proc Natl Acad Sci USA 92:11294-11298. pdf

AWARDS

2002-2005 Research Grant, National Multiple Sclerosis Society

2002-2005 Young Investigator Award, The Wadsworth Foundation

2002 Young Investigator Award, International Society for Neurochemistry .

2000 Young Investigator Award, International Society for Fibrinolysis and Proteolysis.

1999-2001 Human Frontier Science Program (HFSP), long-term postdoctoral fellowship

1998 Women in Neuroimmunology Award, International Society of Neuroimmunology .

1998 Young Investigator Award, European League Against Rheumatism (EULAR).

1997 Best Poster Award, UK Multiple Sclerosis Society.

1996 European Molecular Biology Organization (EMBO) short-term fellowship

1996 Young Investigator Award, University of Ioannina, Greece.

4: Med Hypotheses 1994 Dec;43(6):415-7

Can linoleic acid and gamma-linolenic acid be important in cancer treatment?

Van Aswegen CH, Du Plessis DJ.

Department of Urology, HF Verwoerd Hospital, University of Pretoria, South

Africa.

This hypothesis proposes that the essential fatty acids (EFAs), linoleic acid

(LA) and gamma-linolenic acid (GLA), play important roles in cancer treatment.

Oxidation of LA by lipoxidase especially increases tumour cell death, whilst GLA

inhibits urokinase-type plasminogen activator (uPA) activity. Increased uPA

activity is: firstly, responsible for cancer invasion and metastasis and

secondly, responsible for proteolysis of lipoxidase which favours a decrease in

cancer cell death. Addition of LA and GLA to available therapeutic regimens may

be worth considering in cancer treatment.

CONTACT

Molecular Neurobiology

Skirball Institute of Biomolecular Medicine

NYU Medical Center

540 First Avenue

New York, NY 10016

Tel. 212 263 0722 Fax. 212 263 8214

e-mail: akass@saturn.med.nyu.edu

Return to the Chao Lab HomePage

"Pregnant animals remain healthy if inoculated with tetanus spore. . . Most beautiful experiment."

For several decades, most professors dogmatically asserted that neutrinos have zero rest mass. Then it was learned otherwise. Holding a belief for which there is no evidence is a major feature of culture as we know it.

Lipids. 1981 May;16(5):323-7. Iodination of docosahexaenoic acid by lactoperoxidase and thyroid gland in vitro: formation of an lodolactone. Boeynaems JM, Watson JT, Oates JA, Hubbard WC.

In the presence of iodide, hydrogen peroxide and lactoperoxidase,

docosahexaenoic acid (22:6 omega 3) was converted into iodinated compounds. The

major product was identified as 5-iodo-4-hydroxy-7, 10, 13, 16,

19-docosapentaenoic acid, gamma-lactone, on the basis of 125 I incorporation,

mass spectrometry, chemical modifications and proton nuclear magnetic resonance

spectroscopy. Iodolactonization of docosahexaenoic acid occurred in the rat

thyroid in vitro and was inhibited by the peroxidase inhibit or methimazole.

These data indicate that formation of an idolactone constitutes one pathway of

docosahexaenoic acid metabolism which could be expressed in tissues containing

an iodide peroxidase.

11: Science. 1988 May 20;240(4855):1032-3. Essential fatty acid depletion of renal allografts and prevention of rejection. Schreiner GF, Flye W, Brunt E, Korber K, Lefkowith JB.

Department of Medicine, Washington University School of Medicine, St. Louis, MO

63110.

A central hypothesis in transplantation biology is that resident leukocytes

expressing class II histocompatibility antigens may determine the immunogenicity

of an organ. By means of a novel method to deplete the kidney of resident

leukocytes, essential fatty acid deficiency (EFAD), this hypothesis was tested

in an intact, vascular organ. Kidneys subjected to EFAD and thus depleted of

resident Ia-positive macrophages survived and functioned when transplanted

across a major histocompatibility antigen barrier in the absence of

immunosuppression of the recipient. Control allografts were rejected promptly.

Allografts from donors subjected to EFAD normalized their lipid composition and

were repopulated with host macrophages by 5 days. Administration of Ia-positive

cells at the time of transplantation established that the resident leukocyte

depletion induced by EFAD was responsible for the protective effect. These

observations may provide insights into the mechanisms underlying tissue

immunogenicity and the population of normal tissues with resident leukocytes.

Proc Soc Exp Biol Med. 2000 Jan;223(1):88-95.

Omega-3 fatty acids enhance ligament fibroblast collagen formation in

association with changes in interleukin-6 production.

Hankenson KD, Watkins BA, Schoenlein IA, Allen KG, Turek JJ.

Department of Basic Medical Sciences, Lipid Chemistry Laboratory, Purdue

University, West Lafayette, Indiana 47907, USA.

Altering dietary ratios of n-3 and n-6 polyunsaturated fatty acids (PUFA)

represents an effective nonpharmaceutical means to improve systemic inflammatory

conditions. An effect of PUFA on cartilage and bone formation has been

demonstrated, and the purpose of this study was to determine the potential of

PUFA modulation to improve ligament healing. The effects of n-3 and n-6 PUFA on

the in vitro healing response of medial collateral ligament (MCL) fibroblasts

were investigated by studying the cellular coverage of an in vitro wound and the

production of collagen, PGE2, IL-1, IL-6, and TNF. Cells were exposed to a

bovine serum albumin (BSA) control or either eicosapentaenoic acid (EPA,

20:5n-3) or arachidonic acid (AA, 20:4n-6) in the form of soaps loaded onto BSA

for 4 days and wounded on Day 5. AA and EPA improved the healing of an in vitro

wound over 72 hr. EPA increased collagen synthesis and the overall percentage of

collagen produced, but AA reduced collagen production and total protein. PGE2

production was increased in the AA-treated group and decreased in the

EPA-treated group, but was not affected by wounding. IL-1 was not produced at

the time point evaluated, but TNF and IL-6 were both produced, and their levels

varied relative to the PUFA or wounding treatment. There was a significant

linear correlation (r2 = 0.57, P = 0.0045) between IL-6 level and collagen

production. These results demonstrate that n-3 PUFA (represented by EPA in this

study) positively affect the healing characteristics of MCL cells and therefore

may represent a possible noninvasive treatment to improve ligament healing.

Additionally, these results show that MCL fibroblasts produce PGE2, IL-6, and

TNF and that IL-6 production is related to MCL collagen synthesis.

16: Trans Am Ophthalmol Soc. 2001;99:319-43.

Enhancement of scleral macromolecular permeability with prostaglandins.

Weinreb RN.

Glaucoma Center, University of California, San Diego, School of Medicine, La

Jolla, USA.

PURPOSE: It is proposed that the sclera is a metabolically active and

pharmacologically responsive tissue. These studies were undertaken to determine

whether prostaglandin exposure can enhance scleral permeability to

high-molecular-weight substances. METHODS: Topical prostaglandin F2 alpha (PGF2

alpha) was administered to monkeys to determine if this altered the amount of

scleral matrix metalloproteinases (MMPs). Experiments also were performed to

determine whether the prostaglandin F (FP) receptor and gene transcripts are

expressed in normal human sclera. Permeability of organ-cultured human sclera

following prostaglandin exposure then was studied and the amount of MMP released

into the medium measured. Finally, the permeability of human sclera to basic

fibroblast growth factor (FGF-2) was determined following prostaglandin

exposure. RESULTS: Topical prostaglandin administration that reduced scleral

collagen also increased scleral MMP-1, MMP-2, and MMP-3 by 63 +/- 35%, 267 +/-

210%, and 729 +/- 500%, respectively. FP receptor protein was localized in

scleral fibroblasts, and FP receptor gene transcript was identified in sclera.

Exposure to prostaglandin F2 alpha, 17-phenyltrinor, PGF2 alpha, or latanoprost

acid increased scleral permeability by up to 124%, 183%, or 213%, respectively.

In these cultures, MMP-1, MMP-2, and MMP-3 were increased by up to 37%, 267%,

and 96%, respectively. Finally, transscleral absorption of FGF-2 was increased

by up to 126% with scleral exposure to latanoprost. CONCLUSIONS: These studies

demonstrate that the sclera is metabolically active and pharmacologically

responsive to prostaglandins. Further, they demonstrate the feasibility of

cotreatment with prostaglandin to enhance transscleral delivery of peptides,

such as growth factors and high-molecular-weight substances, to the posterior

segment of the eye.

2: Biochem J. 1998 Feb 1;329 ( Pt 3):469-75.

Identification of cardiac oxidoreductase(s) involved in the metabolism of the

lipid peroxidation-derived aldehyde-4-hydroxynonenal.

Srivastava S, Chandra A, Ansari NH, Srivastava SK, Bhatnagar A.

Department of Human Biological Chemistry and Genetics, University of Texas

Medical Branch, Galveston 77555-1067, USA.

The aim of this study was to identify the cardiac oxidoreductases involved in

the metabolism of 4-hydroxy-2-trans-nonenal (HNE), an alpha,beta unsaturated

aldehyde generated during the peroxidation of omega-6 polyunsaturated fatty

acids. In homogenates of bovine, human and rat ventricles the primary pyridine

coenzyme-linked metabolism of HNE was associated with NADPH oxidation. The

NADPH-dependent enzyme catalysing HNE reduction was purified to homogeneity from

bovine heart. The purified enzyme displayed kinetic and immunological properties

identical with the polyol pathway enzyme aldose reductase (AR), and catalysed

the reduction of HNE to its alcohol 1,4-dihydroxynonene (DHN), with a Km of

7+/-2 microM. In the presence of NADP the enzyme did not catalyse the oxidation

of DHN. During catalysis, HNE did not cause inactivation of AR. Nevertheless

when the apoenzyme was incubated with HNE a dissociable complex was formed

between the enzyme and HNE, followed by irreversible loss of activity.

Inactivation of the enzyme by HNE was prevented by NADP. Partial modification of the enzyme with HNE led to a 17-fold increase in the KHNEm and Kglyceraldehydem,

and the HNE-modified enzyme had a 500-fold higher IC50 for sorbinil than for the reduced enzyme, whereas the IC50 for tolrestat increased 25-fold. Incubation of the enzyme with radiolabelled HNE resulted in the incorporation of 2 mol of the aldehyde per mol of the enzyme. Sequence analysis of the radiolabelled peptides revealed modification of Cys-298 and Cys-187. The amino acid sequence of the

HNE-modified peptides confirmed that the HNE-reducing cardiac enzyme is AR and

not a related protein such as the fibroblast-growth-factor-regulated protein FR-1 or the mouse vas deferens protein MVDP. These results indicate that AR represents the only major oxidoreductase in the heart capable of utilizing HNE. The high affinity of the enzyme for HNE, the lack of inactivation during catalysis, and the lack of significant alcohol dehydrogenase activity of the

protein suggests that AR-mediated catalysis of HNE is unlikely to be limited by substrate/product inhibition. Thus AR might constitute an antioxidative enzyme

involved in myocardial protection against endogenous and exogenous cytotoxic

aldehydes and against oxidative stress.

PMID: 9445372 [PubMed - indexed for MEDLINE]

3: Exp Clin Endocrinol Diabetes. 1996;104 Suppl 4:41-5.

Iodolactones and iodoaldehydes--mediators of iodine in thyroid autoregulation.

Dugrillon A.

Central Clinical Laboratory, University of Heidelberg, Germany.

Within the last decades multiple iodolipid-classes have been identified in

thyroid tissue. For a long time they have been supposed to be involved in

thyroid autoregulation, but for the time being no specific compounds could be

isolated. A new approach was stimulated by the finding that thyroid cells were

able to iodinate polyunsaturated fatty acids to form iodolactones and by the

identification of alpha-iodohexadecanal (alpha-IHDA) as the major compound of an

iodolipid fraction. alpha-IHDA exerts multiple inhibitory effects on adenylate

cyclase, NADPH-oxidase and thyroid peroxidase. Therefore, it is speculated as a

mediator of the Wolff-Chaikoff-effekt and to be involved in the autoregulation

of specific thyroid functions mediated by the cyclic

adenosine-3',5'-monophosphate (cAMP)-pathway. Meanwhile

6-iodo-5-hydroxy-8,11,14-eicosatrienoic acid delta-lactone (delta-iodolactone)

has been identified in human thyroid tissue and it could be demonstrated that

this iodoeicosanoid specifically inhibits signal transduction pathways induced

by local growth factors such as epidermal growth factor (EGF) and basic

fibroblast growth factor (bFGF). Therefore, delta-iodol-actones seem to act as

mediators of iodine, especially in the autoregulation of cAMP-independent

thyroid cell proliferation. We will summarize these important new findings and

discuss the role of these iodolipids on thyroid cell growth regulation.

28: Alcohol Clin Exp Res. 1986 Jun;10(3):271-3. Dietary factors and alcoholic cirrhosis. Nanji AA, French SW. Mortality from cirrhosis in many countries deviates markedly from that expected for a given per capita alcohol intake. We investigated the possibility that dietary factors might explain the deviation expected and actual mortality rates

in different countries. Deviations from expected cirrhosis mortality was

calculated as a percentage for 17 different countries, all of whom had carrier

rates for hepatitis B virus of less than 2%. The percentage of deviation was correlated with dietary intake of saturated fat, polyunsaturated fat, cholesterol, and also with mortality from ischemic heart disease. The percentage of deviation correlated inversely with dietary cholesterol (r = -0.86, p 0.001) and saturated fat (r = -0.80, p 0.001) and positively with polyunsaturated fats (r = -0.55 p 0.05). This suggests that both saturated fat and cholesterol protect against alcoholic cirrhosis while polyunsaturated fats promote cirrhosis. The correlation between percentage of deviation and ischemic heart disease (r = -0.78, p 0.002) suggests that those factors that promote ischemic

heart disease protect against alcoholic cirrhosis.

6: J Pharmacol Exp Ther. 2001 Nov;299(2):638-44.

Dietary saturated fatty acids reverse inflammatory and fibrotic changes in rat

liver despite continued ethanol administration.

Nanji AA, Jokelainen K, Tipoe GL, Rahemtulla A, Dannenberg AJ.

Department of Pathology and Center for the Study of Liver Diseases, The

University of Hong Kong, Hong Kong, China. ananji@pathology.hku.hk

We investigated the potential of dietary saturated fatty acids to reverse

alcoholic liver injury despite continued administration of alcohol. Five groups

(six rats/group) of male Wistar rats were studied. Rats in groups 1 and 2 were

fed a fish oil-ethanol diet for 8 and 6 weeks, respectively. Rats in groups 3

and 4 were fed fish oil and ethanol for 6 weeks before being switched to

isocaloric diets containing ethanol with palm oil (group 3) or medium-chain

triglycerides (MCTs, group 4) for 2 weeks. Rats in group 5 were fed fish oil and

dextrose for 8 weeks. Liver samples were analyzed for histopathology, lipid

peroxidation, nuclear factor-kappaB (NF-kappaB) activation, and mRNAs for

cyclooxygenase-2 (Cox-2) and tumor necrosis factor-alpha (TNF-alpha). Endotoxin

in plasma was determined. The most severe inflammation and fibrosis were

detected in groups 1 and 2, as were the highest levels of endotoxin, lipid

peroxidation, activation of NF-kappaB, and mRNAs for Cox-2 and TNF-alpha. After

the rats were switched to palm oil or MCT, there was marked histological

improvement with decreased levels of endotoxin and lipid peroxidation, absence

of NF-kappaB activation, and reduced expression of TNF-alpha and Cox-2. A diet

enriched in saturated fatty acids effectively reverses alcohol-induced necrosis,

inflammation, and fibrosis despite continued alcohol consumption. The

therapeutic effects of saturated fatty acids may be explained, at least in part,

by reduced endotoxemia and lipid peroxidation, which in turn result in decreased

activation of NF-kappaB and reduced levels of TNF-alpha and Cox-2.

29: J Stud Alcohol. 1986 May;47(3):253-5.

Correlations between deviations from expected cirrhosis mortality and serum uric acid and dietary protein intake.

Nanji AA, French SW.

Mortality from cirrhosis in many countries deviates markedly from that expected

for a given per capita alcohol intake. Since lipid peroxidation is thought to be

one of the mechanisms involved in the pathogenesis of liver disease and since

uric acid is a natural antioxidant, we investigated the possibility that

deviation in mortality rates from cirrhosis in different countries might be

related to serum uric acid levels in the populations studied. Deviations from

expected cirrhosis mortality were calculated as a percentage and correlated with

the serum uric acid levels in nine countries. Since animal protein is a major

source of dietary purines, the percentage deviation was also correlated with

animal protein intake (g/capita/day) for 15 countries. Significant Pearson r

correlations were obtained between the percentage deviation in cirrhosis

mortality and serum uric acid (-.70, p less than .05) and animal protein

ingestion (-.67, p less than .02). This suggests that higher levels of serum

uric acid or higher intake of protein may protect against the development of

cirrhosis.

9: Alcohol Clin Exp Res. 1998 Oct;22(7):1493-500.

Fatty acid omega- and (omega-1)-hydroxylation in experimental alcoholic liver

disease: relationship to different dietary fatty acids.

Amet Y, Adas F, Nanji AA.

Laboratoire de Biochimie-Nutrition, Faculte de Medecine, Brest, France.

Arachidonic acid concentrations in liver are decreased in response to ethanol

administration. In addition, the oxygenated products of arachidonic acid

metabolites could affect the severity of alcoholic liver injury. Selective

utilization of arachidonic acid by the cytochrome P-450 system could, in part,

account for the decrease in arachidonic acid. To evaluate this pathway further,

male Wistar rats were fed different dietary fats: medium chain triglycerides,

palm oil, and corn oil or fish oil with either ethanol or isocaloric amounts of

dextrose. Histopathology, cytochrome P-4502E1 (CYP2E1) and cytochrome P-4504A

(CYP4A), and omega- and (omega-1)-hydroxylation products of lauric and

arachidonic acids were evaluated. Ethanol induction of CYP2E1 was related to the

concentration of polyunsaturated fatty acids in the diet; induction of CYP4A by

ethanol was seen in all groups. The highest levels of 11-hydroxy-lauric acid and

19-hydroxyarachidonic acid (omega-1) were seen in rats fed ethanol with palm oil

and corn oil. Highly significant correlations were seen between the

(omega-1)-hydroxylation products and CYP2E1 activity. No correlation was seen

between the omega-hydroxylation products and CYP2E1 activity. In contrast, the

levels of omega-hydroxylation products correlated with CYP4A. The overall

results showed a significant increase in (omega-1)-hydroxylation products in

rats fed diets containing significant amounts of linoleic acid (i.e., palm oil

and corn oil).

Hepatology. 1998 May;27(5):1317-23. Increased lipid peroxidation and impaired antioxidant enzyme function is

associated with pathological liver injury in experimental alcoholic liver

disease in rats fed diets high in corn oil and fish oil. Polavarapu R, Spitz DR, Sim JE, Follansbee MH, Oberley LW, Rahemtulla A, Nanji AA. Department of Pharmacology, Pennsylvania State University, Hershey, USA. Increased hepatic oxidative stress with ethanol administration is hypothesized

to be caused either by enhanced pro-oxidant production or decreased levels of antioxidants or both. We used the intragastric feeding rat model to assess the relationship between hepatic antioxidant enzymes and pathological liver injury in animals fed different dietary fats. Male Wistar rats (5 per group) were fed ethanol with either medium-chain triglycerides (MCTE), palm oil (PE), corn oil (CE), or fish oil (FE). Control animals were fed isocaloric amounts of dextrose

instead of ethanol with the same diets. The following were evaluated in each

group: liver pathology, lipid peroxidation, manganese superoxide dismutase (MnSOD) levels, copper-zinc SOD (CuZnSOD) levels, glutathione peroxidase (GPX) levels, and catalase (CAT) levels. All enzymes were evaluated using activity assays and immunoblots. Rats fed FE showed the most severe pathology (fatty liver, necrosis, and inflammation), those fed CE showed moderate changes, those fed PE showed fatty liver only, and those fed MCTE were normal. Parameters indicative of lipid peroxidation (conjugated dienes and thiobarbituric acid-reactive substances) were also greater in rat livers from animals fed the diets high in polyunsaturated fatty acids (CE and FE). CuZnSOD, GPX, and CAT activities showed an inverse correlation (r=-.92, P < .01) with severity of pathological injury, with the lowest levels for both enzymes found in FE-fed rats. Decreased enzyme activity in CE- and FE-fed rats was accompanied by similar decreases in immunoreactive protein. Ethanol administration did not cause significant decreases in enzyme activity in groups that showed no necroinflammatory changes (MCTE and PE). MnSOD activity showed no significant change in any ethanol-fed group. Our results show that decreases in CuZnSOD, GPX, and CAT occur in rats showing pathological liver injury and also having the highest levels of lipid peroxidation. These results suggest that feeding dietary substrates that enhance lipid peroxidation can exacerbate both ethanol-induced oxidative damage as well as necroinflammatory changes. The decrease in activity of antioxidant enzymes observed in animals fed diets high in polyunsaturated fatty acids and ethanol could possibly increase the susceptibility to oxidative

damage and further contribute to ethanol-induced liver injury.

Department of Pathology, Beth Israel Deaconess Medical Center and Harvard

Medical School, Boston, MA 02215, USA.

We investigated the potential of dietary saturated fatty acids to decrease

endotoxemia and suppress expression of cyclooxygenase 2 (Cox-2) and tumor

necrosis factor alpha (TNF-alpha) in established alcohol-induced liver injury. Six groups (five rats/group) of male Wistar rats were studied. Rats in group 1 were fed a fish oil-ethanol diet for 6 weeks. Rats in groups 2, 3, and 4 were fed fish oil and ethanol for 6 weeks. Ethanol administration was stopped at this time, and the rats were switched to isocaloric diets containing dextrose with fish oil (group 2), palm oil (group 3), or medium-chain triglycerides (group 4) as the source of fat for an additional 2 weeks. Rats in groups 5 and 6 were fed fish oil-ethanol and fish oil-dextrose, respectively, for 8 weeks. Liver samples were analyzed for histopathology, lipid peroxidation, and levels of messenger RNA (mRNA) for Cox-2 and TNF-alpha. Concentrations of endotoxin were determined in plasma. The most severe inflammation and fibrosis were detected in groups 1 and 5, as were the highest levels of endotoxin, lipid peroxidation, and mRNA for Cox-2 and TNF-alpha. After ethanol was discontinued, there was minimal