Anti-Inflammatory Diet

All health care starts with diet. My recommendations for a healthy diet are here:
Anti-Inflammatory Diet and Lifestyle.
There are over 190 articles on diet, inflammation and disease on this blog
(find topics using search [upper left] or index [lower right]), and
more articles by Prof. Ayers on Suite101 .

Wednesday, December 24, 2008

Synuclein and Amyloid Diseases

NSAIDs, such as ibuprofen and aspirin are possible treatments to inhibit the aggregation of proteins (synuclein, beta amyloid) on charged polymers in amyloid diseases, such as Parkinson’s disease, Alzheimer’s disease, etc. Contradictory studies show that intracellular aggregate formation may be protective, since dimers are more toxic than aggregates.

The list of amyloid diseases is long and there are few effective treatments. In each case a protein starts to accumulate in fibers that form amyloid plaques inside or outside the cells. The large aggregates outside are toxic. Inside it appears that the large aggregates are not as toxic as small clumps, oligomers, of the protein.

The amyloid proteins are stacked up in the fibers in a very organized way, so that the same portions of the protein are lined up on each side of the fibers. Outside the cell, the regions with basic amino acids interact with heparin, and in Alzheimer’s disease, for example, the beta amyloid plaque is half heparin. In test tube experiments, fiber formation from protein solutions is accelerated by adding heparin.

Amyloid fibers also form inside cells in the case of the tau fibers of Alzheimer’s disease or the synuclein aggregates in Parkinson’s disease. In theses cases, there should not be any intracellular heparin, and it is not known what polyanion (RNA?) serves to accelerate fiber formation in these cases.

Non-steroidal anti-inflammatory drugs (NSAIDs), such as ibuprofen and aspirin, reduce the incidence of Parkinson’s and Alzheimer’s diseases. It has recently been shown that in test tube experiments, NSAIDs also decrease the formation of amyloid fibers from synuclein.

Amyloid fibers can be stained by Congo Red and thioflavin. Curcumin is the active component of tumeric and it has a structure related to Congo Red. Curcumin has been shown in recent studies to block synuclein amyloid formation.

In addition, the heparin in the fiber complexes can be stained with berberine. Berberine is a traditional herbal treatment for arthritis. It would not be surprising if it was also effective against Alzheimer’s amyloid plaque.

The large extracellular plaque aggregates appear to be toxic, but the small, oligomeric aggregate of protein appear to be the toxic form in cells. Recent experiments show that facilitating the formation of large intracellular aggregates minimizes the toxicity in animal models of Huntington’s and Parkinson’s diseases. It appears that the large visible aggregates are not the form that kills the cell.

For the time being, the only safe treatments that focus on amyloid fiber formation are the NSAIDs, curcumin and perhaps berberine.

Hirohata M, Ono K, Morinaga A, Yamada M. 2008. Non-steroidal anti-inflammatory drugs have potent anti-fibrillogenic and fibril-destabilizing effects for alpha-synuclein fibrils in vitro. Neuropharmacology 54(3):620-7.

Pandey N, Strider J, Nolan WC, Yan SX, Galvin JE. 2008. Curcumin inhibits aggregation of alpha-synuclein. Acta Neuropathol. 115(4):479-89.

Bodner RA, Outeiro TF, Altmann S, Maxwell MM, Cho SH, Hyman BT, McLean PJ, Young AB, Housman DE, Kazantsev AG. 2006. Pharmacological promotion of inclusion formation: a therapeutic approach for Huntington's and Parkinson's diseases. Proc Natl Acad Sci U S A. 103(11):4246-51.

Outeiro TF, Kontopoulos E, Altmann SM, Kufareva I, Strathearn KE, Amore AM, Volk CB, Maxwell MM, Rochet JC, McLean PJ, Young AB, Abagyan R, Feany MB, Hyman BT, Kazantsev AG. 2007. Sirtuin 2 inhibitors rescue alpha-synuclein-mediated toxicity in models of Parkinson's disease. Science. 317(5837):516-9.

Monday, December 22, 2008

Is a Belly Bad?

A protruding midsection can mean many different things. It can be obesity, starvation/kwashiorkor or sarcopenia. It can mean there is a layer of fat outside, sagging organs poorly supported by weak abdominal muscles or fat surrounding the organs and stretching the muscles outward.

Alternatively having the same “lean” profile at 60 as at 16 could be a sign of a decline of muscle and an increase in fat surrounding organs. You can always have a porthole put in to see what is going on, or you can check the calendar and your C-reactive protein level.

If you are over fifty and your doctor has told you that your CRP, a measure of inflammation, is starting to creep up, then you are starting to suffer from age-related loss of muscle, sarcopenia. That is, you have begun to replace your muscle mass with fat, and the fat is producing inflammatory signal molecules, cytokines, that are the same as those produced by your immune system cells in response to an infection.

Most of the symptoms we associate with aging are just poorly managed chronic inflammation, as a result of replacing muscle with fat. The fat is metabolically lethargic, so you actually need less food, as your muscle mass declines. The result is that it is progressively easier to put pounds on. That is the bad news. The good news is that building muscle requires at least one of the inflammatory cytokines produced by fat, IL-6, so gaining muscle should be easier until you get back into shape. Losing weight is also anti-inflammatory, if the loss is fast enough to produce a fasting physiology.

All of the diseases associated with increasing age are just the accumulation of problems that result from increasing levels of chronic inflammation. Most of the increase is the result of inflammatory diet, but the increasing inflammation also decreases the desire to be physically active, with the result being a loss of muscle mass. Adjustment to an anti-inflammatory diet and lifestyle, can reverse the aging process.

It is also possible to be physically active and have a youthful muscle mass, but still have an inflammatory diet. The result will still be chronic inflammation, and cryptic inflammatory diseases that accumulate with time and produce sudden failures of joints, coronary arteries or cancer. Estrogens are also naturally anti-inflammatory, so women will find that menopause reveals any inflammatory diet/lifestyle that has been hormonally camouflaged. Thus, menopause may produce any of the typical signs of inflammation, e.g. acne, depression, back problems, arthritis, etc.

The bottom line is that a weak gut and/or extra body fat will cause problems at any age. And at any age, the diseases that are associated with inflammation can be minimized or avoided by an anti-inflammatory diet and lifestyle/exercise.

Friday, December 19, 2008

HIV TAT and Methamphetamine -- TNF

HIV infection and methamphetamine both cause inflammation of the brain and together they are paralyzing.

What’s worse than a TNF typhoon resulting from methamphetamine use? The answer is a TNF typhoon resulting in dementia from HIV infection of the brain.
Combine methamphetamine with HIV in the brain and the result is a Parkinson’s type of paralysis.

This sounds like very morbid subject matter to pursue out of curiosity, but if you put heparin into the equation, as I always do, it all becomes very interesting.

Here are pieces to the big picture:

HIV, the AIDS virus, infects cells of the immune system and causes chronic inflammation. The inflammation causes a disruption of heparin metabolism and since heparin is a major part of the matrix that holds together the endothelial cells that line the capillaries that feed the brain, the capillaries leak, i.e. there is a leak in the blood brain barrier. HIV-infected cells can pass out of the capillaries and into the brain. Here comes the insidious part, HIV produces a protein called TAT.

I drew a graphic of TAT with the basic amino acids in blue. The sequence of this nasty little protein shows how it gets around. It is secreted from cells, attached to heparan sulfate proteoglycans. It sticks tightly to heparin (yellow and red stick figures, sticking to ribbon of TAT), because of the patches with three and four adjacent basic amino acids. Frequently, the TAT will just be secreted and then sweep over the surface of the infected cell and be brought back into the cell on the circulating HSPG.


A protein with three adjacent basic amino acids will get swept into a cell. All allergens that I have examined have this internalization triplet of basic amino acids. TAT is so powerful, that if it is chemically linked to the larger fluorescent protein from jelly fish (left with green, fluorescent amino acid derivative down the center), the whole fluorescent protein is dragged into cells.

The TAT can move from the HSPGs of an HIV-infected cell to neighboring cells with HSPGs. The TAT then gets taken into the cytoplasm of the next cell. Four adjacent basic amino acids are the signal that transports a protein to the nucleus and into the nucleus. It is in the nucleus that TAT really causes trouble. The TAT can move from an infected immune cell in the brain to neurons. TAT can kill neurons and stimulate other cells to produce TNF.

Methamphetamine also causes a TNF storm in the brain. This is a quick way to start the wasting symptoms that TNF in known for -- it is also call cachexin, after the wasting process of cachexia. If methamphetamine is given to someone with neurological symptoms of HIV, then the neuropathology is further exaggerated into a Parkinson’s type of paralysis. The TNF production of both is additive.

TNF production by methamphetamine brings up the consequences of the very closely related compound amphetamine (Adderall, Dexedrine) used by children and young adults (college age) as a treatment for ADHD.

Theodore S, Cass WA, Nath A, Maragos WF. 2007. Progress in understanding basal ganglia dysfunction as a common target for methamphetamine abuse and HIV-1 neurodegeneration. Curr HIV Res. May;5(3):301-13.

Tuesday, December 16, 2008

What If Medicine Was Scientific?

A view of the future of medicine, if the health of each patient was the highest priority.

2020 Somebody read the biomedical literature and decided to save a few bucks on the national healthcare system.

It has been a couple of years since everyone began favoring their right butt cheek. The “BioMed” subcutaneous, WiFi, microchip implants were controversial, but the economic argument was compelling. In the first year after the BioMed became mandatory and implantation accompanied every office visit, the expenditure on medical treatment in the US dropped by 10%. There was a lot of complaining about what a literal “pain in the butt” the prevention program was, but the prodding of the BioMed got results.

Once subQed, the BioMed enforced a simple dietary and exercise regimen. Violation of the regimen produced progressively more severe itching of the cheek, until the real “pain in the ass” set in. Surcease from this sorrow was Poetic in its justice. Sufferers could pay for temporary relief in conveniently marked toll stalls, called “Craigs.” There was a certain amount of public opprobrium associated with use of these facilities, but they brought relief. Those compelled to use the stalls complained of being “Idahoed.”

The prescribed diet was high in omega-3 oils and complex carbohydrates, but proscribed large amounts of starch, sugar and other hyperglycemic carbohydrates. Ten minutes of daily solar exposure (vitamin D) on the cheeks was monitored by the BioMed and “mooning” was replaced in the vernacular by “BMing.” Twenty minutes of daily exercise was enforced based on the BioMed’s analysis of physiological (pulse, blood pressure, blood sugar) and physical (GPS, accelerometer) measurements. Sustained, sedentary activity was not possible.

The BioMed provided identity verification for each patient and every datum on the patient was accessible by any medical official. Absolute privacy was provided by a variant of Apple’s DRM. Prescriptions for each patient were verified at purchase and were confirmed by access to a wireless expert system. Correct timing of medication was enforced by BioMed reminders. Compliance soared.

In the second year, 2 BM, office visits began to plummet. Doctors in private practice saw their incomes drop by 20% and there was already talk of a surplus of medical staff. Prices of the stocks of the major food processors began to drop as people shifted away from fast food, common vegetable oils, corn and soy products, and began to purchase more local meat and produce. Breastfeeding gained new popularity spurred by the introduction of the BioMedNip, a nipple implant to enhance the health of newborns.

The third year, 3 BM, brought appreciation for the BioMed and “kiss my cheek” was first used as an invocation of good luck. The decline in chronic inflammation decimated cases of depression, obesity, arthritis and allergies. Detection rates for cancer and degenerative diseases of the elderly plateaued. Cancer and geriatric specialists took credit for the efficacy of their latest therapies. Infant mortality in America began to approach that observed in developed countries. Pharmaceutical stocks tanked. Lactation fashions reach the catwalks. Hospital closures begin. Advertising for cold medications, drugs and baby formula is banned. American worker productivity soars.

It is amazing that at any given time, the US is just a couple of years away from medical utopia. The cure for chronic inflammation, diet and exercise, is available to all.

Medicine Can Treat. Why Can’t It Cure or Prevent?

Prescription for all diseases: Anti-inflammatory Shock Treatment

Cancer death rates may be finally slowing. Why has it taken so long? Medicine has some cobbled-together treatments for allergies, asthma, Alzheimer’s and atherosclerosis, but all of these are still increasing. Cures and prevention are always ten years away. Something is fundamentally wrong with medicine and the fatal flaw is obvious in the biomedical literature. Causes are not mentioned. Treatments are tested and evaluated. Pharmaceuticals are developed for therapy, not for cures.

Examination of medical websites reveals a public interested in why they get sick, but there is no explanation. There is a recent tendency connected to the concept of personal medical solutions, to attribute sickness to the unique genetics of the individual. Certainly there are some molecular genetic diseases, but this obscures the issue. In most cases the genetics only gives a predisposition. Biomedical research does not pursue why some people get the disease and others with the same genes do not.

The research literature shows abundant evidence that various diets and lifestyles dominate the outcome in health, but the medical industry fails to apply these factors. Why should someone spend a lifetime on drugs, if two weeks of intensive diet/lifestyle therapy provides a cure? Is a surgical intervention preferable over a gut flora exchange?

There is compelling evidence that chronic inflammation due to diet and lifestyle is the foundation for most degenerative and autoimmune diseases, and yet simple changes to avoid or eliminate inflammation are not emphasized by doctors to prevent or cure most diseases. It is quite possible to design a one-size-fits-all primary treatment for all diseases, an “Anti-inflammatory Shock Treatment.“ It would be a two week controlled diet and exercise program that provided the anti-inflammatory benefits of controlled carbohydrates, adequate protein, balanced omega-3/6 fatty acids, anti-oxidants, vitamins, exercise and meditation. Careful control of probiotic gut flora would be a priority. This system could be designed based on current research and would be generally applicable. The prescribed system for each patient would match severity of the disease with intensity of the anti-inflammatory intervention.

Monday, December 15, 2008

Lectins - Heat’em and Eat’em

Lectins are proteins common in seeds. They bind to sugars attached in chains to proteins, i.e. glycoproteins, and are displayed on the surfaces of cells that line the gut. Lectins could inhibit digestion of raw beans, but cooking makes them digestible.

Fear of lectins is puzzling. Lectins are proteins that have binding sites on their surfaces for specific single or small sequences of sugars. They are present in seeds to protect the seeds from herbivores.

A seed is mostly food (starch, protein, fat) for the plant embryo that will grow from it. This is also true of a chicken egg and just like the egg, the seed contains defensive proteins to inhibit the growth of bacteria, fungi and egg/seed eaters.

The egg has enzymes to degrade bacterial walls and proteins that bind iron, vitamins, etc. needed by bacteria and humans. Eating many raw eggs can lead to vitamin deficiencies. Boiling the eggs, unravels the defensive proteins and makes them digestible and nutritious.

Seeds block being digested by containing proteins that foul the digestion system of would be devourers. For example, soybeans have trypsin inhibitor that binds to our digestive enzyme and makes eating raw soybeans nonproductive and uncomfortable. Boiling soybean meal to produce a curd, i.e. tofu, agglutinates the denatured soy proteins, including the lectins and washes away the soy trypsin inhibitor. Tofu is free of digestion inhibitors and lectin activity.

It is not an accident that lectins bind to human red blood cells. The sugars displayed on the surface of red blood cells are the blood group antigens. Different sugars on the end of the sugar chains decorating RBCs characterize the A, B and O antigens. These same sugars are present on the surfaces of various bacteria. Immune systems don’t produce antibodies to self antigens, so a person with type A blood produces antibodies only to B antigen sugars it encounters on bacteria. A person who is type AB doesn’t produce antibodies to A or B sugar antigens. There aren’t antibodies to O, because that sugar structure is the basis upon which both A and B are made, and some of the
structure is present on all RBCs. Lectins are specific for A or B or other common bacterial sugar antigens.

I did some modeling to show a lectin with lactose (red and gray) bound to sticky tryptophans (yellow) in two places on the surface. In one case a lysine (blue) is draped on the other side. That shows that sugars bind both to aromatic amino acids and to the hydrophobic arms of basic amino acids.

Some people think that humans and other mammals must be protected from lectins and that this protection is shown in human and cow’s milk in the form of antibodies against lectins. This seems to be a misunderstanding. For example, human antibodies secreted in breast milk are secretory IgAs. These antibodies are glycoproteins, i.e. they are proteins with attached sugar chains. Some lectins will bind to these antibodies, because of the attached sugars. These are not lectin-specific antibodies, but rather glyco-specific lectins. The lectins are binding to the glycoprotein antibodies, not the other way around.

It is possible for people to be allergic to lectins, but this is unlikely. For example, peanut allergies involve proteins other than the peanut lectins.

There are some dangerous lectins. For example, ricin is a very nasty, but effective, toxin produced by the castor plant. Ricin is a lectin, in that it binds very specifically to sugars found on the surface of gut cells of insects and humans. After the ricin binds to surface proteins, it is brought into the cells where it chops up the protein synthesizing machinery. That is a dangerous lectin. It takes very little ricin to kill each cell and only a tiny amount to kill a human. Ricin is a terrorist toxin. Yet oil extracted from castor beans contains so little contaminating ricin that it is safe to eat. [Castor oil is wonderful to apply to aching feet overnight for painfree, luxuriously soft feet in the morning.]

The bottom line is that seed lectins add to the nutrition of cooked beans and grains that have been the foundations for several thriving civilizations. The longest living members of the bean and grain cultures are typically older and more fit than comparable individuals with a modern, inflammatory diet based on omega-6 oils.

Thursday, December 11, 2008

A Paleolithic Perspective on Biomedical Literature

Homo sapiens seems to be inflammation prone, based on its assortment of biochemical deficiencies. All of the following lead to inflammation: hyperglycemia, vitamin C deficiency, fish deficiency, vitamin D deficiency, grilling meat. Is this an adaptation to agriculture and high population/communicable disease risk?

I was just visiting Diet Rosso and his article on the paleolithic diet flashed me back to some thoughts that I had on the evolutionary benefits of inflammation. So, Rosso made me think about this.

Inflammation is a big health problem in the US. All of the major diseases are inflammatory and all are exacerbated by the inflammatory US diet. But why is the fast food diet so inflammatory? Why is our corn/soy agricultural economy so hazardous to our health?

Corn and soybeans provide a good balance between carbohydrates, fats and protein. The amino acid composition of the combo is also fairly good, and corn and soy oils are high is unsaturated fats. So why does a corn/soy diet lead to degenerative and autoimmune diseases?

I think that the answer is that inflammation is getting a bad rap; as long as our immune system produces effective local responses to pathogens, we are pleased, but when the immune system cranks it up in response to a deluge of disease, we complain. I argue that our current inflammatory response to fast food is just a slight embellishment of the first dietary-based increase in inflammation that provided adaptive protection against the dangers of agriculture.

As I see it, agriculture had a series of dramatic impacts on the evolution of plants, animals and humans, in particular. Taming of plants and animals altered the human diet. Agriculture also institutionalized grilling and grouling, which meant bringing together carbs and protein at high temperatures. The result was an increase in dietary starch, seed/grain oils and advanced glycation endproducts (AGEs). There was also a decrease in fish, leafy vegetables and complex carbohydrates/fiber. Agriculture also led to a dramatic increase in population density.

I imagine that the first villages or very large family groups that resulted from sustained planting of harvestable crops resulted in plagues. Lots of people in close proximity with minimal hygiene is a prescription for infectious disease. Agricultural development required an immunological adaptation to higher loads of communicable diseases. That adaptation was inflammation triggered by agriculturally-associated diets high in starch and low in browsed veggies.

Hunting/gathering, especially along coasts, provided dietary vitamin C, as well as a high ratio of omega-3 to omega-6 fatty acids. Early humans defective in the ability to synthesize vitamin C or omega-3 fatty acids, would not suffer if they ate plenty of leaf veggies. Wild fish and game, as well as leafy veggies, have a high ratio omega-3 to omega-6 fatty acids, so these people would be safe from inflammation-based disease and infertility.

Agriculture focused on seed harvest results in a dramatic shift in diet and disease. Communicable disease was not a problem for hunter/gatherers, because of the necessarily widely distributed small population groups. Agriculture concentrates populations around the crop lands and increases the benefits of physiological energy expenditures on heightened immune activity to provide consistent protection against pathogens. Agriculture required chronic inflammation for disease protection.

Inflammation triggered by cues in the agricultural diet would have a high selective advantage. Individuals who increased their chronic level of inflammation in response to high blood sugar, compounds produced during cooking, i.e. AGE, vitamin C deficiency, vitamin D deficiency (low exposure to sun) and/or omega-3 oil deficiency, would have survival advantage in high population densities associated with agriculture.

The fast food diet is nothing more than an exaggeration of the agricultural diet and it produces and an exaggeration in the human adaptation to agriculture, high chronic inflammation and a suite of inflammatory diseases. Metabolic syndrome is another name for high chronic inflammation. Obesity is inflammatory. A sedentary lifestyle is inflammatory and aging is a suite of symptoms associated with inflammation. Hunter/gatherers didn’t show the same signs of aging as modern humans, and probably had comparable longevity (although there were many other risks.)

Lastly, I want to ponder the modern decline in fertility. Fertilization and implantation requires suppression of inflammation in the female reproductive system. Semen is uniquely rich in omega-3 oils and women who have a high frequency of unprotected coitus with a man with a high level of omega-3 in his semen, are much more likely to become pregnant and carry a pregnancy full term. The fetus requires high levels of omega-3 fatty acids for brain development and can rapidly deplete the omega-3 fatty acid a mother with a deficient diet. Omega-3 deficiency is associated with preeclampsia.

Early males and females with an inflammatory agricultural diet would tend to be infertile, because of omega-3 oil deficiencies and chronic elevated inflammation. Periodic exposure to an abundance of omega-3 fatty acids, such as feasting on migrating salmon, would synchronize fertility and subsequent births. It is humbling that a current research program in land-locked regions of South America uses cans of anchovies to remedy the same pregnancy problems that plague North America and its inflammatory fast food diets.

Saturday, December 6, 2008

Niacin Flush

Niacin is a B vitamin that is cheap and highly effective at raising HDL and lowering LDL. HDL and LDL were previously called good and bad cholesterol, resp., but since the data from numerous studies show that they don’t have a big impact on health, it is probably easier to just call them heavy and light reflecting less and more lipid content. If you still want to adjust your blood lipids, then niacin is more effective than the costly statins. Unfortunately, niacin also causes an uncomfortable (itchy and hot) flush.

The niacin flush is part of the inflammatory process that includes the classic tetrad of symptoms: rubor (redness), calor (increased heat), tumor (swelling), dolor (pain). Flushing in response to niacin shows that the immune cells in the skin respond to ingested niacin that is flowing through the capillaries. Mast cells in the skin have receptors that bind niacin and the cells secrete inflammatory prostaglandins. The prostaglandins act on the capillaries to cause dilation and flushing. Mast cells have secretory granules that fuse to the cytoplasmic membrane and release their contents outside. The granules contain histamine, heparin and tryptase. The histamine stimulates histamine receptors on pain/itch nerves and the tryptase stimulates receptors on a second set of pain/itch nerves.

Prostaglandins are produced by membrane bound enzymes on the surface of mast cells. When the mast cells are stimulated, additional enzymes are added to the surface through fusion of the secretory granules. The combined enzyme complex produces prostaglandins by releasing arachidonic acid (ARA) from phospholipids of the membrane (phospholipase A2, PLA2), converting the ARA to an epoxide prostaglandin (cyclooxygenase, COX-1) and stepwise producing additional prostaglandins. These prostaglandins cause the dilation of capillaries that is seen as flushing.

Niacin also binds to receptors on fat cells, adipocytes, and blocks release of fatty acids from the triglycerides stored in these cells. It is this action that is responsible for the increase in HDL and the lowering of LDL in blood serum.

An extension of the niacin skin flushing reaction is the use of this response to demonstrate the presence of arachidonic acid and a functional immune system in the skin. A recent study used topical application of niacin and skin reddening to test the idea that schizophrenia exhausts ARA as a result of inflammatory processes in the brain. Tests showed a tendency for schizophrenic episodes to be accompanied by a diminished flushing response to niacin. This result also suggests that a lowered system-wide ARA level should show up in a predisposition to gut problems.

It would be very interesting to test the interplay between inflammatory provocations, e.g. infection, serum omega-6/omega-3 fatty acids, and measures of inflammation, e.g. C-reactive protein on niacin flushing. Inflammatory depletion of ARA may be important in the decline in the integrity of tissues that is observed in inflammatory diseases of the gut (Helicobacter-based ulcers, IBD, Crohn’s disease, celiac), autoimmune diseases (arthritis, atherosclerosis), skin diseases (vitiligo), etc. It would also be interesting to test the impact of helminth infections to reverse ARA depletion.

Benyó Z, Gille A, Kero J, Csiky M, Suchánková MC, Nüsing RM, Moers A, Pfeffer K, Offermanns S. 2005. GPR109A (PUMA-G/HM74A) mediates nicotinic acid-induced flushing. J Clin Invest. 2005 Dec;115(12):3634-40.

Friday, December 5, 2008

Antioxidants or Fermenting Gut Flora?

Long life is the result of a good diet and exercise. Plant antioxidants are eaten to sop up the reactive oxygen species (ROS) and omega-3 fatty acid-rich fish oils are consumed to reduce the production of inflammatory prostaglandins. Avoiding inflammation that has been linked to essentially all degenerative and autoimmune diseases should make us live better and longer. Unfortunately it is more complex than that, and studies in the simple nematode, Caenorhabditis elegans, show that your gut flora may have something to say about your longevity.

C. elegans nematodes live in the laboratory fed on Escherichia coli, the common colon bacterium. Since the bacteria are the sole diet of the worms, to change the worm’s diet, mutant bacteria must be used. To study the impact of various vitamins on longevity, mutant bacteria unable to synthesize particular vitamins were fed to worms and their average length of life was measured.

Deleting coenzyme Q (10) resulted in a surprising increase in longevity. It was assumed that since coQ10 was needed for effective bacterial electron transport, the disruption would result in an increase in inflammatory ROS. What actually happened was that the bacteria shut down their use of aerobic metabolism and turned on fermentation.

Fermenting bacteria are probiotic in human guts and it appears that the same is true of worms. These results suggest that probiotic, fermenting gut flora may be profoundly important in determining longevity. What we eat may determine how long we keep eating.

Saiki R, Lunceford AL, Bixler T, Dang P, Lee W, Furukawa S, Larsen PL, Clarke CF. 2008. Altered bacterial metabolism, not coenzyme Q content, is responsible for the lifespan extension in Caenorhabditis elegans fed an Escherichia coli diet lacking coenzyme Q. Aging Cell. 7(3):291-304.

Wednesday, December 3, 2008

Menstrual Pain is Inflammatory

Inflammation is essential to the menstrual cycle. At key points inflammatory prostaglandins are made from omega-6 arachidonic acid to trigger ovulation and menses, the discharge of the blood-engorged lining of the uterus. Chronic diet-based inflammation can result in disrupted ovulation, infertility due to an inability to suppress an inflammatory response to egg implantation, menstrual pain/cramps and premature birth.

Several studies have shown that reducing diet-based inflammation by eating supplements containing long chain omega-3 oils, e.g. fish oil, decreased menstrual pain and cramps. The reduction in chronic inflammation was associated with decreased production of inflammatory prostaglandins that are the cause of the pain and intense uterine contractions. Normally, the diet would provide a balance of omega-3 and -6 fatty acids, which would yield a mixture of anti-inflammatory and inflammatory prostaglandins, and produce an effective discharge through more moderate uterine contractions.

A more recent evaluation of numerous studies on the impact of omega-3 oils on pain associated with menstruation, arthritis, inflammatory bowel disease, etc., showed a uniform decrease in inflammation and pain. The simple summary is that an inflammatory diet rich in omega-6 vegetable oils leads to pain, suffering and premature aging. A more normal diet with a balance of omega-3 and omega-6 fatty acids leads to health and reduced aging.

Typical symptoms of an inflammatory diet are: menstrual cramps, infertility (gestational problems: preeclampsia, prematurity), joint pain, back pain/sciatica, acne, allergies, asthma, autoimmune diseases. There is increasing evidence that obesity not only produces inflammation, but that an inflammatory diet can lead to obesity. An inflammatory diet, especially if augmented with antibiotics, disrupts the normal gut flora and leads to an inflammatory replacement flora that supports chronic inflammation throughout the body.

Chronic inflammation and much of the damage caused by chronic inflammation is reversible by a shift to an anti-inflammatory diet and lifestyle (described elsewhere on this blog.)

Deutch B. 1995. Menstrual pain in Danish women correlated with low n-3 polyunsaturated fatty acid intake. Eur J Clin Nutr. 49(7):508-16.

Goldberg RJ, Katz J. 2007. A meta-analysis of the analgesic effects of omega-3 polyunsaturated fatty acid supplementation for inflammatory joint pain. Pain 129(1-2):210-23.

Bell RF. 2007. Food and pain: should we be more interested in what our patients eat? Pain. 129(1-2):5-7.

Tuesday, December 2, 2008

Leptin and Diabetes

Fat cells produce the hormone leptin. Early experiments showed that leptin could ameliorate some of the effects of mouse models of type I diabetes, but it was always thought that the leptin just improved the activity of the small residual amounts of insulin produced by the rats. A recent experiment with genetically insulin deficient NOD mice, shows that leptin expressed by a virus infection reverses the symptoms of diabetes.

The NOD mouse strain is defective in the hormone, IL-2, that preserves tolerance to self proteins. As a result, a high percentage of the NOD mice develop symptoms of type I diabetes melitis, high blood glucose and ketosis. These animals will die without injected insulin.

In recent experiments, the NOD mice showing symptoms of diabetes were injected with a strain of adenovirus that had been engineered to express leptin. The leptin-expressing NOD mice displayed a rapid reversal of diabetes symptoms and began to gain weight normally. The high blood sugar and ketosis in NOD mice was stimulated by glucagon release in response to lack of insulin. Leptin stopped the glucagon release and reduced additional production of glucose by the liver and lipid oxidation that yielded the ketosis.

Muscle access to glucose in the NOD mice was restored by leptin by the induction of insulin-like growth factor 1 and its receptor. The NOD + leptin mice had normal blood sugar, but ate half the normal amount and had no body fat.

These are very promising results for type I diabetics, because it indicates that insulin injections are not the only approach to treatment. Stimulation of leptin production may be very useful and this new approach is being intensely investigated.

Yu X, Park BH, Wang MY, Wang ZV, Unger RH. 2008. Making insulin-deficient type 1 diabetic rodents thrive without insulin. Proc Natl Acad Sci U S A. 105(37):14070-5.

Monday, December 1, 2008

Anti-inflammatory Octopus

I always envied the neurophysiologists who studied the giant axons of lobster, because they could always eat their failed experiments. I was working on a fungal disease of soybeans. It had no gourmet prospects. My revenge came decades later, when I took a “research” trip to Tuscany to study the impact of Mediterranean cuisine on inflammation. Fish, cephalopods, red wine and tiramisu were my test materials and I was the test organism. For a month, I felt no stress, no inflammation and no pain. Recent research articles support my subjective conclusion that seafood and red wine from Siena to Venice are anti-inflammatory.

Recent tests of the fatty acid composition of Mediterranean fish and octopus show that the ratio of omega-3 to omega-6 fatty acids in the fish and octopus is roughly 2:1. (In contrast, the US ratio is 1:20) The omega-7 fatty acid, palmitoleic acid, which appears to act as an anti-inflammatory hormone, was also present. The prominent place of seafood in the Mediterranean diet and the high omega-3-rich fatty acid composition of that seafood, combined with the absence of the inflammatory, omega-6-rich vegetable oils, corn, soy, safflower, of the inflammatory US dietand the use of olive oil, go a long way to explain the relatively low incidence of inflammatory diseases in people who eat a Mediterranean diet. Just add some sunshine and exercise, and you will live better and longer.

Oh, by the way, the tiramisu was my daughters’ project to evaluate local variation of that dessert throughout Italy. So we ate tiramisu at a different restaurant each day for a month. The differences were amazing and the best tiramisu in Italy is ...

But the bottom line is the same. The Mediterranean diet is nothing more than another variant of the anti-inflammatory diet and lifestyle that I have been discussing throughout this blog. Biomedical research says that the US diet is killing us. You can go to Tuscany or you can just shop, cook and live wisely wherever you are.

Ozogul Y, Ozogul F, Cicek E, Polat A, Kuley E. 2008. Fat content and fatty acid compositions of 34 marine water fish species from the Mediterranean Sea. Int J Food Sci Nutr. Oct 29:1-12.

Ben-Youssef S, Selmi S, Ezzeddine-Najai S, Sadok S. 2008. Total lipids and fatty acids composition of the coastal and the deep-sea common octopus (Octopus vulgaris) populations: a comparative study. Nutr Health. 19(3):195-201.

Friday, November 28, 2008

Melamine Contaminated Formula

Baby formula is called, in the medical profession, an artificial milk substitute. It is made of highly processed food additives, and all of these additives are prone to contamination. The recent melamine scare is just the most recent example of dangerous contamination. With breast milk, the mother’s body provides another detoxification barrier to protect babies from food contaminants. Mother’s who eat melamine-contaminated food don’t pass the toxin on to their babies, but melamine-contaminated formula can kill babies.

When you look at the structure of melamine, you see a lot of nitrogen atoms, and since protein is frequently measured in food products by measuring nitrogen content, melamine mimics protein. Since protein is expensive and melamine is cheap, people can make money by diluting protein solutions, such as milk, and adding melamine to make it look as if it has the normal protein composition. In China, that little bit of subterfuge resulted in the hospitalization of thousands of children. Melamine may act like protein in simple nitrogen assays, but the body doesn’t have enzymes to react with it or transport proteins to excrete it. Melamine forms crystals, blocks kidneys and kills small people.

The newest threat is based on the report of a trace amount of melamine in formula in the US. It is hard to determine how the melamine got into the formula, because formula is made of dozens of highly processed and synthetic ingredients. Production of this artificial milk substitute from so many different sources has resulted in dozens, if not hundreds of contamination incidents.

Formula is a gut flora disrupter. In contrast to the trend toward prebiotics to support healthy anti-inflammatory probiotic bacteria in the gut, research shows that even a single feeding of formula will shift the gut flora of a newborn to an inflammatory composition. Gut inflammation, along with delayed intestinal development and absence of breast milk’s immunity enhancers, are considered to be the reasons why formula lowers intelligence (5-10 IQ pts.) and increases the incidence of allergic, autoimmune and infectious disease. Formula can be tolerated by babies in developed countries, because of the high hygiene. Formula use in developing countries is unsafe. In fact even though there is a small risk of transfer of HIV from an HIV positive mother to baby by nursing, it is safer for HIV positive mothers to breastfeed in some countries than to use formula.

The presence of melamine in formula would result in the introduction of a toxic compound into a immune compromised baby. It is very important to test formula very carefully for even seemingly innocuous contaminants. Breastfed babies would be protected from melamine even if the mother ingests melamine-contaminated food, because toxins, including most pharmaceuticals, are not passed into breast milk. Breast milk is the anti-inflammatory diet for babies and should be encouraged at least until solid food is started after the emergence of two teeth, top and bottom. The American Academy of Family Practice Physicians recommends breastfeeding for at least two years with exclusive breastfeeding (no formula or solid food) for the first six months. The worldwide average weaning age is 4 years.

Thursday, November 27, 2008

Turkey, Tryptophan & Transmitters

Does eating turkey make you stuffed and drowsy? It must the be tryptophan... or not. Sure there is tryptophan in turkey and tryptophan is the starting point for making some neurotransmitters and hormones, but turkey meat is simply protein and fat. Tryptophan (left) is just one of the twenty amino acids found in most proteins, so the drowsiness after the big Turkeyday dinner is more about the “big” and less about the turkey.

Turkey meat is muscle and muscle, as the diagram shows is made of protein molecules that use energy in the form of ATP to move past each other and contract. We chew up the turkey muscle and our stomach juices contain enzymes (these “proteases” are also proteins) that reverse the process of protein synthesis and produce protein fragments called peptides. The specificity of the proteases in the stomach, e.g. pepsin, results in peptides containing intact heparin-binding domains that are also antimicrobial. In the intestines, a new group of proteases are added by the pancreas and the peptides are further reduced in size and heparin-binding domains are degraded. [Pathogens need heparin-binding domains to bind to the intestines.] The remaining peptides bind to the microvilli of the endocytes lining the small intestines, surface bound peptidases release individual amino acids and transport proteins bring amino acids into the endocytes and on to the blood stream.

In the brain, tryptophan is converted by a series of enzymes into serotonin and the serotonin is stored in secretory vesicles adjacent to the synapse that controls signals between nerves. A nerve action potential moves down the axon from the cell body to the synapse. The change in electrical potential reaching the synapse causes the secretory vesicles to fuse with the cytoplasmic membrane and release the serotonin into the synapse. The serotonin binds to the receptors of the adjacent nerve and starts a new action potential that travels to the next nerve body to repeat the process. The synapse is reset by reuptake or degradation of the serotonin. The degradation product, 5-HIAA, is removed into the blood and excreted in urine.

Turkey tryptophan does get converted into serotonin and high serotonin could make you mellow, but turkey is just like any other meat source of tryptophan. The big meal just makes you groggy, because there is less blood to your brain when the mysenteric blood flow is enhanced for digestion. There may also be a rise in blood sugar as the starch off your plate is rapidly converted into glucose in your blood. The potentially damaging high blood sugar is controlled by a rise in insulin that lowers glucose in the blood by stimulating transport into fat cells for immediate conversion into fat. The starch from the meal is rapidly depleted, blood sugar rises and then sudden falls. The low blood sugar also leaves you groggy.

So it was the size of the meal (decreased brain blood flow) and the sweet potatoes and rolls (starch-induced hypoglycemia) that induced you to kickback on the sofa and pass out with the big game lulling you to sleep. Tryptophan from the big bird is in the background waiting for you to awaken. Before you take the first mouthful, check to make sure that your meal follows the anti-inflammation guidelines. Planning ahead can help you to enjoy a meal that won't be a pain later.

Wednesday, November 26, 2008

CIDP, Inflammation and Autoantigens

Chronic Inflammatory Demyelinating Polyneuropathy (CIDP) is chronic, relapsing autoimmune disease in which the immune system attacks the insulating myelin sheaths of nerves in the limbs. The result is numbness and pain. An acute version of this myelin compromising disease is Guillain-Barre syndrome (GBS) typically initiated by Campylobacter jejuni infection. A related disease of brain tissue is multiple sclerosis (MS). In each of these cases antibodies bind to protein or glycolipid antigens unique to myelin and target the attack by lymphocytes.

Autoimmune diseases represent a disruption of the normal function of the immune system that arises from a breakdown of the self/non-self tolerance process. Immune responses by antibodies or lymphocytes is normally reserved for molecules that don’t exist in the body, i.e. self. Differentiation of lymphocytes in the thymus or bone marrow involves systematic elimination of all lymphocytes that interact with self antigens. There are, however, self antigens that are not normally encountered by the immune system, e.g. nuclear antigens, brain antigens, since these antigens are only excluded from the cytoplasm or result from specialized differentiation. Thus, you can find in mystery novels the use of letters coated with powdered brain antigen to slowly kill targets by induction of a lethal autoimmune disease.

The self antigens that are the typical targets of the CIDP, GB and MS are gangliosides, myelin glycolipid also produced by C. jejuni, and basic proteins such as peripheral myelin protein 22. The glycolipid makes sense, because the initial exposure is during a bacterial infection that also produces inflammation. The basic proteins are a different case.

I have examined numerous self or autoantigens that are the targets for antibodies produced in autoimmune diseases and allergies. In each case the same amino acid sequence or domain is present, three basic (positively charged, arginine or lysine) amino acids. This is true for the myelin autoantigens; allergens from ragweed, mites, peanut, etc.; diabetes, lupus and arthritis.

Here is the example of the CIDP autoantigen, peripheral myelin protein 22:

The three basic amino acids, RKR, are at the end of the protein amino acid sequence.

A dramatic instance of tissue damage caused by physical injury or infection, is superimposed on chronic dietary inflammation to provide the environment for an inappropriate response of the immune system to a self or identical foreign antigen. The only proteins that seem to be presented in this way to the immune system are those with the strong heparin-binding domain, BBB (B = basic amino acid). This indicates that perhaps the reduction in heparin production that accompanies inflammation, may be responsible for the aberrant immune response. It is also noteworthy that heparin and anti-inflammatory agents can reduce the symptoms of many autoimmune diseases.

There are many complex therapies to deal with the symptoms of these autoimmune diseases. In all of these cases, one of the most effective approaches is an anti-inflammatory diet and lifestyle. A novel additional approach is helminth therapy, infection with parasites to suppress the T-lymphocyte autoimmune attack. This approach is effective with MS, and I think would be an excellent, relatively safe approach to be examined for the other myelin autoimmune diseases.

Tuesday, November 25, 2008

Hazards of Air Travel: DVT

Deep Vein Thrombosis (DVT) -- clots in your veins

Air travel during the holidays means sitting quietly for hours while the blood pools in the major veins of your legs. This is a test. How have you been eating lately? If you stuck to an anti-inflammatory diet and got your exercise, just fidgeting a little and flexing your legs ever once in a while should avoid clots. If you are the typical sedentary American with an inflammatory diet, then worry. One tenth of you will typically have clots in your leg veins after a long flight.

Rolling stones gather no moss, and the same is true for rapidly moving red blood cells (RBCs). Keep them moving and they don’t stick together. Slow down RBCs traveling along sticky vessel walls and you have problems. RBCs have no nuclei and since the intracellular secretory system originates from the outer membrane of the nucleus, red blood cells don’t secrete anything. RBCs just age until they are removed by the spleen. So RBCs just move passively with the rest of the blood.

Another player in clot formation is the platelet. Platelets are cell fragments. They are formed by extrusion and shearing. The process is like bubbles forming as you blow air through a child’s bubble wand. Cells in the bone marrow are squeezed through a grid and the extruded fingers of the cells are blown away in the blood flow as platelets. The electron micrograph shows a platelet between and RBC and a white blood cell. Platelets don’t have any active cell machinery, so they are just little bags containing secretory vesicles that can be released by triggering of receptors on their surface. Platelets are only good for one shot of release.

Platelet release of secretory contents is triggered by norepinephrin, ADP and PGI2, an inflammatory prostaglandin produced from the omega-6 arachidonic acid. Norepinephrin is one of the fight-or-flight hormones that prepares the vascular system for damage control. ADP is released from other activated platelets and insures that isolated platelets are not randomly activated.

One of the proteins released is platelet factor 4. I have illustrated PF4 and the strip of basic amino acids (blue) that girdles the protein are readily apparent. PF4 binds strongly to heparin. Since the clotting process is normally under heparin inhibition, PF4 release from platelets removes the heparin inhibition and promotes clotting. ADP is also released and promotes further activation of other platelets.

Clot formation occurs in response to stress (norepinephrin), damage (vascular inflammation) and a consensus of platelets (ADP). Chronic inflammation can mimic this combination of signals through its impact on heparin metabolism. My research suggests that inflammation lowers heparin synthesis. An example of this effect is kidney damage caused by diabetes. High blood sugar causes inflammation of the kidney blood vessels, this reduces heparin production and since heparin lining the vessels is needed to retain proteins as blood is filtered in the kidney, protein is lost into the urine, i.e. proteinuria. Similarly, chronic inflammation can disrupt the blood brain barrier that is also made up of heparin.

A major source of chronic inflammation is an inflammatory diet. A recent research study indicated that a typical inflammatory American diet leads to elevated risk for deep vein thrombosis. Alternatively, an anti-inflammatory diet rich in B vitamins and omega-3 oils minimized DVT. Saturated fats had no impact, consistent with the lack of evidence supporting the shift from saturated fats to toxic omega-6-rich polyunsaturated vegetable oils.

So, the best thing that you can do to protect yourself from clots when you travel over the holidays, is to eat right and get your exercise, before you travel. Avoid starch (in large amounts) and polyunsaturated vegetable oils (except olive oil.) Corn oil, soy oil, cottonseed oil and safflower oil are particularly inflammatory. Eat plenty of veggies and fruits and enjoy the turkey and cranberries. Make sure that the only sweeteners used are sugar and honey (avoid high fructose corn syrup.) Light corn syrup is the stealth form of HFCS -- it may be lower in calories, since fructose is sweeter than sugar, but it is highly inflammatory! (Research also indicates that fructose causes premature wrinkling and skin aging, by enhancing the crosslinking of collagen. HFCS also causes type II diabetes in lab animals.)

Monday, November 24, 2008

Crohn’s Disease and Cryptidins

The intestines produce enzymes to digest food, antimicrobial peptides to kill pathogens and have lots of surface area to absorb nutrient molecules released from the food macromolecules (protein, polysaccharides, fats). The epithelial cells that line the intestines, enterocytes, must communicate with bacteria in the gut, the gut flora, to maintain bacteria helpful in food digestion, i.e. probiotic bacteria, and trigger an immune response to eliminate pathogens. Probiotic bacteria are tolerated and pathogens are identified and attacked.

Enterocytes are produced by division of stem cells at the bottom of the crypts that are in the valleys between the villi that project into the lumen where the digesting food is. New enterocytes are added at the base of the villi and old enterocytes are sloughed off at the top of the villi. As the new enterocytes move up the villi, they differentiate to produce the dramatic surface of microvilli, the furry brush border that further expands their surface area. The mature enterocytes produce transport proteins on their microvilli to take up sugars, amino acids and fats. The small nutrient molecules pass through the base of the enterocytes and bath the cells below, the lamina propia. The nutrients enter the capillaries of the villi and travel to the liver. Fats are transported through the lymphatic system.

Bacteria that slip through the enterocyte layer encounter macrophages and other types of white blood cells of the lamina propia. Among these cells are the Paneth cells. Fragments of the cell walls of bacteria bind to the NOD proteins of the Paneth cells and trigger the secretion of antimicrobial peptides, the cryptidins. Cryptidins are antimicrobial because of their array of basic amino acids surrounded by hydrophobic amino acids. These short proteins are able to disrupt the membrane function of most bacteria. I think they work on bacteria the same way that amyloid proteins, e.g. amyloid plaque proteins of Alzheimer’s disease, kill human cells. In fact, amyloid fibers bind to heparin and so do antibiotic peptides.

Here is an example of an antibiotic peptide, cryptidin 4,


Note the pairs of basic amino acids (blue). These amino acids are necessary for toxicity to bacteria. Heparin binding domains from proteins are produced naturally as proteins are digested to peptides in the stomach by pepsin. Pepsin hydrolyzes proteins next to the basic amino acids and leaves antimicrobial peptides that sterilize incoming food. I have illustrated the cryptidin protein to show how the basic amino acids (blue) are displayed on its surface.

With each meal, the fat content normally stimulates the production of a hormone, cholecystokinin, that binds to a receptor and causes an anti-inflammatory release of cytokines from the vagus nerves that reach the villi. Thus, food normally makes the intestines more tolerant of food antigens.

If the intestines become chronically inflamed, then exposure to normal probiotic bacteria can lead to cycles of inflammation that damage the integrity of the intestines. The intestines lose the ability to discriminate between probiotic and pathogen.

Crohn’s disease is an inflammatory, autoimmune disease of the bowel. The chronic inflammation of the lamina propia eliminate the ability of the Paneth cells to produce cryptidins and bacteria set up residence in the crypts and cause continual inflammation. This disease is typically treated by suppressing inflammation and treating with antibiotics.

Other treatment approaches that have been found effective are omega-3 oils to stimulate production of anti-inflammatory prostaglandins, pre- and probiotics, heparin and helminth eggs, e.g. wireworm.

Crohn’s disease would seem to benefit from the standard recommendation of an anti-inflammatory diet and lifestyle.

Friday, November 21, 2008

Fecal Transplants

Grey’s Anatomy, “In the Midnight Hour,” episode 8 of season 5 includes a fecal transplant from a recalcitrant husband to his anxious, antibiotic-compromised wife. Drama aside, fecal transplants, the introduction of gut bacteria from a healthy donor into a patient for treatment, represent a powerful potential therapy. Your gut flora may be your health destiny.

Here is a smattering of the recent biomedical research literature that is an indication of the influence of gut flora on health:

  • The appendix has been identified as a bacterial reservoir to reestablish gut flora after diarrhea clears the gut.
  • Fecal transplants between obese and lean patients reverses weight gain patterns.
  • Hospital-acquired Clostridium infections can be treated with fecal transplants.
  • Success of total gut transplants is improved by retaining the gut flora of the donor.
  • Formula disrupts the gut flora of newborns leading to lower intelligence and increased morbidity.
  • Prebiotics change gut flora and enhance beneficial bacteria, probiotics, that reduce inflammation and promote health.
  • The National Institutes of Health have launched a major initiative to identify the bacteria that live in and on the human body.

Most of the migratory cells of the immune system are in residence in intestinal tissues that communicate with gut flora and ingested contents. What we eat determines our gut flora, but our gut bacteria also communicate with the intestines and alter our inflammatory/immune status. This gut/bacteria communication controls the shift of the gut from tolerant suppression of immunological responses to mundane food molecules, to alarmed defense against pathogens. Mistakes in communication can lead to susceptibility to pathogens or autoimmune diseases of the gut, joints, brain, etc.

The dawning recognition of the importance of the symbiotic relationship between gut and bacteria on health and disease, is also spawning numerous interventions that involve injecting donor bacteria into patient intestines. This type of procedure may be the next fad. Imagine going to a health spa for a gut flora replacement. It may soon be possible to pick your donor from a catalogue. The possibilities for celebrity gut flora are mind boggling. There may even be an upsurge in nerd transplants in preparation for college finals. You are what you eat is still true, but the potty is proving to be a measure of more than passing performance.

Thursday, November 20, 2008

Brain Arachidonic Acid: Alzheimer’s, Bipolar, Parkinson’s

A recent review article on brain lipid metabolism discussed the results obtained by looking at how the major omega-6 fatty acid, arachidonic acid is imported and used in brain tissue. Arachidonic acid conversion to inflammatory prostaglandins was monitored by extracting lipids from rat brains after a variety of treatments. Similarly, isotopes (13C) of fatty acids were imaged by PET scans in patients treated for Alzheimer’s, bipolar disorder and Parkinson’s disease.

The major findings on brain arachidonic acid (AA, omega-6) and docosahexaenoic acid (DHA, omega-3) are:

  • Ca. 5% of daily dietary AA and DHA are converted to make prostaglandins in the brain. Converted AA and DHA are rapidly replaced by serum AA and DHA.
  • Brain DHA and AA metabolisms are independent.
  • AA and DHA are rapidly circulated into phospholipids (R2 on the diagram) on the endoplasmic reticulum, move to the cytoplasmic membrane (see diagram, gray and white strands) removed by phospholipase A2 in synapses, converted to prostaglandins, leukotrienes, etc., or recycled to phospholipids. Enzymes that catalyze these reactions are usually different for DHA and for AA.
  • Drugs used to treat bipolar disorder (lithium, carbamazepine, valproic acid, lamotrigine) lower AA conversion in rats, but do not affect DHA conversion.
  • Experimentally induced brain inflammation or neurotoxicity increases AA conversion, but not DHA conversion to prostaglandins.
  • An omega-3 fatty acid deficient diet also increases AA, but not DHA conversion.
  • More AA is converted in Alzheimer’s patients. This is consistent with increased inflammation and neurotoxicity in postmortem examinations.
  • Mice that have been genetically manipulated to eliminate alpha-synuclein, a protein implicated in Parkinson’s disease, also show an increase in AA conversion and a decrease in DHA conversion.

Interpretation: Inflammation in the brain is separate from the rest of the body, but is the foundation of many brain disorders, including Alzheimer’s disease, bipolar disorder and Parkinson’s disease. In these disorders, arachidonic acid is rapidly converted into inflammatory prostaglandins and leukotrienes. Drugs that reduce symptoms, reduce AA conversion.

A diet rich in omega-3 DHA and reduced omega-6 arachidonic acid reduces the symptoms of these diseases -- an anti-inflammatory diet and lifestyle should be the first line of defense against brain/mental disorders.

Rapoport SI. 2008. Brain arachidonic and docosahexaenoic acid cascades are selectively altered by drugs, diet and disease. Prostaglandins Leukot Essent Fatty Acids. Oct 28. [Epub ahead of print]

Wednesday, November 19, 2008

Inflammation Causes Disease

Human diets have changed dramatically over the last few hundred years, and as a consequence so have our diseases. The most recent shift in diet over the last hundred years has resulted in a shift from infectious diseases to degenerative diseases. This trend is summarized in the following Wikipedia entry.

Lifestyle diseases, from Wikipedia:

"Lifestyle diseases (also called diseases of longevity or diseases of civilization) are diseases that appear to increase in frequency as countries become more industrialized and people live longer. They include Alzheimer's disease, atherosclerosis, asthma, cancer, chronic liver disease or cirrhosis, Chronic Obstructive Pulmonary Disease, Type 2 diabetes, heart disease, nephritis or chronic renal failure, osteoporosis, acne, stroke, depression and obesity.

Death statistics in the United States
In 1900, the top three causes of death in the United States were pneumonia/influenza, tuberculosis, and diarrhea/enteritis. Communicable diseases accounted for about 60 percent of all deaths. In 1900, heart disease and cancer were ranked number four and eight respectively. Since the 1940s, the majority of deaths in the United States have resulted from heart disease, cancer, and other degenerative diseases. And, by the late 1990s, degenerative diseases accounted for more than 60 percent of all deaths.
National Center for Health Statistics, National Office of Vital Statistics, 1947 for the year 1900 (page 67), for the year 1938 (page 55)."

My point here is that all of the so-called lifestyle diseases are also based on inflammation. I checked the research literature for studies of the response of each of these diseases to diets supplemented with omega-3 fish oils. Studies had been performed in each case. Reduction of inflammation by fish oil treatment was uniformly effective in reducing symptoms of all of the degenerative diseases. Other diseases that can be added to the inflammatory list are spinal disc problems and hypertension. It is interesting that disc dislocations are associated with coeliac, an inflammatory/autoimmune disease. It is also interesting that acne and depression are listed. Acne is indirectly associated with diet, but if sufferers shift to an anti-inflammatory diet, acne symptoms disappear. Depression associated with childbirth is particularly responsive to anti-inflammatory drugs, diet and exercise. Most of the symptoms associated with aging are just due to inflammation and are similarly responsive to anti-inflammatory lifestyle changes

To summarize:
  • Modern degenerative diseases are caused by modern inflammatory diets (and insufficient exercise.)
  • Anti-inflammatory diet and lifestyle reduce degenerative diseases.
  • Aging is predominantly mismanaged inflammation.

Gulf War Syndrome

Gulf war syndrome (GWS)is now officially a disease. It shares many of the symptoms of chronic fatigue syndrome, fibromyalgia, multiple chemical sensitivity and post traumatic shock syndrome. In other words, GWS is the body’s response to neurotoxins (acetylcholinesterase inhibitors) under inflammatory conditions.

The nervous system has to detect external signals, transmit the sensory information to the brain, process the information, transmit signals to peripheral organs/muscles, and stimulate responses in peripheral organs/muscles. At the molecular level, all of this detection and transmission of signals is accomplished by small molecules binding to and changing the shape/function of proteins. Disrupting the interaction between the small signal molecules (neurotransmitters) and their protein receptors is used throughout nature as an attack on the most vulnerable animal characteristic, the nervous system. Examples of nerve system attackers include mushroom toxins, plant alkaloids (atropine), spider/insect/frog/snake venoms, organophosphate/chloride pesticides, and mustard/nerve gas.

During the Gulf War, several chemicals that mimic neurotransmitters were used with the intent to protect soldiers. The neurotransmitter mimicked was acetylcholine, which is the signal used between peripheral nerves and muscles. Nerve gas targets acetylcholine signaling by blocking the enzyme, acetylcholinesterase, that sits in the space between the nerve and muscle and destroys the acetylcholine in preparation for the next signal. I have illustrated a model of the acetylcholine enzyme and have an inhibitor (grey and red) bound in the cavity of the active site of the enzyme. The active site is lined with hydrophobic tryptophan (yellow) and tyrosine (orange) amino acids.

The pesticides used by the soldiers were designed to kill or repel insects, but they also act as acetylcholine mimetics and bind to acetylcholinesterase. These include Lindane (left), Permethrin (right) and DEET (lower left).

The American soldiers were also forced to take a chemical to protect them from nerve gas. This compound, pyridostigmine (lower right) bind reversibly to acetylcholinesterase and since nerve gas is very reactive, presumably the pyridostigmine will protect some of the acetylcholinesterase until the nerve gas is depleted.

Unfortunately it appears that both the insecticides and the pyridostigmine had a permanent impact on the nervous systems of many of the Gulf War soldiers. Soldiers from other countries did not use the same chemicals on their soldiers, so they were not similarly affected.

It is now fairly well established that pesticides and pyridostigmine were involved in causing Gulf War Syndrome, but very little research has been performed to explain how these chemicals cause the damage or what therapy can reverse the damage.

It appears to me that GWS occurs in similar manner to related illnesses, e.g. chronic fatigue syndrome, multiple chemical sensitivity, fibromyalgia, which can also be induced by organophosphate pesticide exposure. Two insults to the body appear to be needed: acetylcholine signal disruption and inflammation. The acetylcholine mimetics (pesticides, pyridostigmine) disrupted the nervous system and numerous immunological, infectious, chemical and emotional stresses generated a high level of chronic inflammation. The vaccine against anthrax and exposure to burning oil wells may have contributed to inflammation.

Sunday, November 16, 2008

Topoisomerase Inhibitors

Inhibiting enzymes involved in DNA synthesis should stop cancer cells, because cancer is uncontrolled cell division. Topoisomerases are enzymes that help to relieve the twists on double helical DNA as it unwinds preparatory to replication. It appears logical that topoisomerase inhibitors should be cancer inhibitors. Unfortunately targeting DNA-binding proteins also targets most of the signal receptors that are the targets for the evolution of plant alkaloids.

Drugs are designed to be specific in their interactions with a particular target protein, but they are too small to be specific and end up binding to many other related proteins. Hence, drugs have side reactions that are to some extent unpredictable, because the interacting proteins are not known.

Aspirin, for example, is supposed to bind to and inhibit COX-2, the enzyme that converts omega-3 and omega-6, long-chain fatty acids into corresponding anti-inflammatory and inflammatory prostaglandins, resp. Aspirin also binds to proteins that inhibit NFkB, the transcription factor that controls expression of inflammatory genes. Aspirin binds to dozens of other proteins. Aspirin does lots of other things than just blunt inflammation, but those side reactions are usually not significant enough to get our attention.

Heparin is one of the most commonly used drugs. It binds to and activates an inhibitor of thrombin, an enzyme that activates fibrin and mediates clotting. Heparin also binds to other components of the clotting system, as well as a dozen components of the complement system, and most of the cytokines that control communications throughout the body. When patients are given heparin injections, heparin binds continually to all of these components and must be constantly supplemented and monitored. Inflammation depletes the heparin components throughout the body, so it is not known prior to injection, how much heparin will be needed to saturate other serum proteins before the desired level of clotting inhibition is achieved. This illustrates rather dramatically that most drugs have only limited specificity.

One of my students provided another example of the minimal specificity of small molecules, especially the alkaloids and phenolics produced by plants. He brought to me a research article espousing the use of phenolics from yerba mate, which serves as a coffee-like stimulant in Argentina, as a topoisomerase inhibitor and potential anti-tumor treatment. Sure enough, phenolics extracted from this plant inhibit topoisomerase, and they may well be able to inhibit the growth of tumors, but it is doubtful that the binding of the phenolics to topoisomerase in the tumor nuclei has anything to do with inhibition of tumor growth.

Topoisomerase binds to nuclear DNA as the DNA unwinds during replication to produce two new double helical DNA molecules. Topoisomerase is a DNA-binding protein, i.e. a protein that binds to a negatively charged polymer of small deoxyribose sugars and flat purine and pyrimidine bases. Proteins bind to DNA in two ways. Amino acids of the protein either bind along the edges of the hydrophobic stack of base pairs, e.g. sequence-specific transcription factors, or they provide hydrophobic, flat surfaces that bind to the hydrophobic faces of the separated bases. Topoisomerase does both, because it deals with single-stranded regions of DNA and therefore binds to both the phosphates, as well as the bases. The important point here is that both aromatic amino acids, with flat hydrophobic rings, and the hydrophobic tails of basic amino acids, i.e. lysine and arginine, bind to the hydrophobic faces of nucleic acid bases.

I have illustrated the binding of a “topoisomerase inhibitor” to show the arginine (blue) in the active site cleft of the topoisomerase that binds across the hydrophobic face of the inhibitor (grey and red). Many plant phenolics and alkaloids would be expected to similarly bind and act as inhibitors of topoisomerase. This observation and the ease by which alkaloids enter cells (attached to circulating heparan sulfate?) suggests that a major function of the nuclear envelope may be to minimize access of alkaloid and related molecules to the nucleic acid binding proteins of the nucleus.

The binding promiscuity of secondary plant products is further exemplified by berberine. Berberine is an alkaloid found in goldenseal and is an herbal remedy used to treat a variety of inflammatory diseases. It also binds to heparin (and nucleic acids) to produce a fluorescent complex. Thus, mast cells that store and secrete histamine and heparin to produce the symptoms of allergy, can be vividly stained with berberine.

I could not resist the temptation to check to see if berberine also binds to topoisomerase. A quick search of the research literature showed that berberine is in fact a topoisomerase inhibitor.

The numerous cross reactions of drugs are further illustrated by metformin, the common drug used in the treatment of type II diabetes. Metformin is approximately planar and provides a surface that cannot hydrogen bond, i.e. it is hydrophobic. I expected that metformin would bind to tryptophans that I observed as common substrate-binding amino acids in the active sites of proteins that bound to polysaccharides, e.g. lectins, glycosidases and glycanases. To test this, I had students in one of my courses examine the inhibitory activity of metformin on E. coli beta-galactosidase. They found measurable inhibition and support for competitive binding to the active site that contains a pair of the predicted tryptophans.

My protein modeling and structural studies show the basis for numerous interactions between plant secondary compounds, drugs, nucleic acids, polysaccharides (glycosaminoglycans, e.g. heparin) and proteins. Unpredicted cross reactions abound and every drug can be expected to interact with multiple proteins. This provides a note of caution to the use of any drug and encourages minimal exposure, since many unobserved and unanticipated side effects are occurring. These observations also question routine ingestion of herbal remedies, after all, plants use their secondary products as potent defenses against being eaten. Alkaloids disrupt nervous systems and cellular signaling. Plants are not naturally safe.

Friday, November 14, 2008

Fiber Phytate

Some polysaccharides are only digested by the action of colon bacteria. Some of these fiberous plant polysaccharides (and associated macromolecules, such as lignan) are insoluble, whereas the soluble fiber is made of beta-glucans. Insoluble fiber also contains the active ingredient, phytic acid (inositol hexasulfate).

Fiber stimulates the passage of ingested material through the colon, i.e. it has a laxative effect. Growth of beneficial (probiotic) bacteria are enhanced by fiber. Beta-glucans also enhance immune function and have been reported to inhibit tumor expansion and enhance resistance to some bacterial infections, e.g. anthrax.

My personal experience with fiber components began with my graduate research. I ended up determining the structure of the beta-glucans from fungi that trigger the disease resistance of plants. Other researchers soon found that the beta-glucans also triggered the immune system of crayfish and it wasn’t long before beta-glucans were recognized as the active ingredients of a number of foods associated with reduction in diseases.

More recently, I began using yeast beta-glucan to stimulate inflammation in my study of rat chondrocyte cultures. It appears that these glucans are recognized by a cell surface receptor and trigger NFkB-mediated expression of typical inflammatory genes, COX-2, iNOS, IL-1, IL-6, TNF, etc. This form of inflammation also reduces the expression of heparan sulfate proteoglycans.

Beta-glucan-stimulated inflammation may also enhance infiltration of tissue with neutrophils and other leukocytes that provide cellular immunity against some tumors.

Immunity may also be altered by the impact of soluble and insoluble fiber on the bacteria of the gut. Fructose oligosaccharide, such as those in milk, enhance the growth of lactobacilli that are anti-inflammatory. Soluble and insoluble beta-glucans are also prebiotic. The prebiotic effects on the gut flora may be partially responsible for the enhanced resistance to infectious bacteria resulting from ingestion of soluble beta-glucans.

I encountered another activity of fiber in my work on heparin and inflammation. Heparin is derived from a polysaccharide made from glucose and glucosamine. Several enzymes add sulfates and the heavily sulfated, negatively charged fragments, oligosaccharides are secreted from mast cells as heparin.

Other negatively charged polysaccharides, e.g. nucleic acids and sugars also have some of the properties of heparin. One of these molecules that mimics heparin is inositol hexaphospate, phytate. As I pursued the binding of tryptophan and related structures to proteins, I remembered a similar molecule, auxin or indole-acetic acid, from my plant work and examined an auxin receptor. I was pleasantly shocked when I looked at the auxin receptor sequence. I expected to find that a heparin-binding domain was used to bind auxin to this protein receptor. That proved to be the case and substantiated the generalized observation that tryptophan and similar molecules bind to basic amino acids, and that sugars also bind to both aromatic and basic amino acids. What was amazing about the auxin receptor was that the purified and crystalized protein had a phytate bound to an assortment of basic amino acids in the center of its structure.

I have illustrated the structure of the auxin receptor here to show how the blue arginines extend as inward pointing spokes from a wheel of alpha-helices, and the orange/red sulfates of the phytate attach to the arginines. Only the lower right quarter of the image is shown in the closeup, with the whole molecule shown in the smaller picture.

Phytate is important, because it holds much of the phosphate content of the fiber, and the binding of divalent cations, e.g. calcium, magnesium, to the phytate may prevent these minerals from being absorbed. In ruminants, e.g. cattle, the gut bacteria produce and enzyme, phytase, that removes the phosphates and solves the absorption problem. This is frequently not the case in humans.

Inositol with two or more phosphates is attached to the lipids of membranes to make phospholipids. Signal cascades may activate enzymes that release the inositol phosphates (IP3) into the cytoplasm where they can cause the elevation of cytoplasmic calcium and the release of secretory granule contents, e.g. neurotransmitters.

Phytate is considered to be responsible for the reduction in colon cancer attributed to fiber from grains and legumes. The heparin-mimicking actions are probably the basis for its activity, since phytate can partially replace or inhibit surface receptor interactions that normally involve heparan sulfate proteoglycans. Phytate can also be internalized along with heparin and it should be very aggressive if released into the cytoplasm.

Phytate has been found to be very safe, but may be more effective if ingested in the soluble form, since it is frequently not released from fiber. Extensive cooking is required to release phytate from insoluble fiber. Soybeans are one of the best sources of phytate, so maybe it would be wise to cook some for hours just to release the phytate.