Lactoferrin: Natural Anti-Microbial Milk Protein

Nosocomial infections happen when the immune system is compromised through a medical procedure and common bacteria, such as Staphlococcus aureus, get introduced. From my perspective, that means that the walking bacterial reservoirs, i.e. gut flora of healthcare practitioners, provide an inoculum directly to the damaged tissue, or indirectly by contaminating the patients gut flora and then spreading the pathogens from the patient’s digestive tract to the damage site. This is what happens with ventilator-associated pneumonia and sepsis in critically ill patients.

The trick is to keep the patient’s gut flora healthy -- healthy as a breastfed baby’s. The typical medical approach is to kill off lurking pathogens with a dose of antibiotics. The problem with this approach is that it is both indiscriminant and selective, i.e. it kills both pathogens and beneficial bacteria, but it also provides a selective advantage for the antibiotic resistant hospital strains of opportunistic pathogens.

Humor break: Why do babies spit up half-digested breastmilk and then smile? Answer: Pepsin produces antimicrobial peptides from milk proteins. The baby smugly acknowledges that she knows that she has just protected her upper respiratory and digestive tracts against bacterial pathogens.

Pepsin hydrolyzes proteins next to aromatic amino acids and away from the basic amino acids, arginine and lysine. That means that heparin-binding domains, which consist of groups of basic amino acids in a hydrophobic environment, are clipped out intact from proteins by pepsin. Thus, babies sucking down milk make their own isolated peptides with heparin-binding domains.

Lactoferricin with basic amino acids in blue.

Many organisms, from fruit flies to frogs to humans, produce anti-microbial peptides. They also produce proteins with nucleic acid-binding domains and nuclear localization signals and heparin-binding domains and IP3-binding domains. If all of those binding domains are clipped out by pepsin and the peptides are compared to the anti-microbial defensive peptides, amazingly they are all the same. All have groups of basic amino acids among hydrophobic neighbors, and all are toxic to bacteria.

Lactoferrin is a major component of milk whey. It binds iron and heparin. It can be digested by pepsin into an an anti-microbial peptide, lactoferricin. Baby’s smile and spit-up on your shoes when you say lactoferrin.

Transgenic mice that produce porcine lactoferrin in their milk, transfer extra lactoferrin their little suckling mouse pups and that extra lactoferrin gives extra protection against bacterial and yeast pathogens. That is the experimental justification to suggest that treating patients at risk of nosocomial infections (I guess that would mean every patient in contact with a nurse or doctor) with oral lactoferrin should selectively eliminate the pathogens. Lactoferrin is prebiotic and supports the growth of probiotic gut flora.

ref:
Yen CC, Lin CY, Chong KY, Tsai TC, Shen CJ, Lin MF, Su CY, Chen HL, Chen CM. Lactoferrin as a natural regimen for selective decontamination of the digestive tract: recombinant porcine lactoferrin expressed in the milk of transgenic mice protects neonates from pathogenic challenge in the gastrointestinal tract .J Infect Dis. 2009 Feb 15;199(4):590-8.

Cure Acne, Back Pain, Tendonitis, Depression

Remedies Include Vicks Vaporub, Castor Oil and Anti-Inflammatory Diet

Simple anti-inflammatory treatments cure some of the most common health complaints. The big question is why people tolerate the problems rather than applying the readily available remedies.

It seems to me that one reason people don’t simply live anti-inflammatory lives and avoid health problems is that attacking the underlying inflammation by approaches that would have prevented the health problem in the first place, is inadequate for fixing the problem after it becomes established.

Health problems based on inflammation may have many different sources of inflammation. Many dietary deficiencies, for example, contribute to inflammation, so what we eat or don’t eat is a major health risk. Other common contributors to inflammation are dental cavities/infections and inadequate exercise.

Common Symptoms of Chronic Inflammation: Acne, Back Pain, Tendonitis, Depression

I have started to ask casual acquaintances if they have any aches or pains, because eliminating dietary sources of inflammation will be evident in relief of these problems. Common complaints are sore joints and tendons related to repeated use. An example is my barber who complained of pain in all of the tendons used to raise his arms to cut hair. Another friend just had her second child and suffered from shooting pains in the tendons of the arm she used to cradle the youngest when she used the other arm on some task.

Simple Anti-Inflammatory Diet Adjustments Get Quick Results

In many cases, a simple change in diet can lower chronic inflammation enough to provide relief from symptoms. Vitamin D deficiency is probably an underlying source of inflammation of most people in the US. So a simple supplement of 2000-5000 IU per day will have noticeable, anti-inflammatory impact on most people.

I recommended vitamin D and fish oil supplements to a friend suffering from chronic back pain. The back pain persisted, but his acne resolved. He stopped taking the supplements, but after physical therapy relieved the back pain, he returned to the supplements as an acne treatment. Now he has long term relief from all of his pains.

Elimination of Dietary Inflammation May Not Resolve Inflammation Based Health Problems

Health problems that start from aggrevated inflammation, may not be eliminated with resolution of the initial cause. My friend’s back ache, for example, didn’t respond to just elimination of deficiencies in his diet. It seemed that the back problems were self-sustaining. After he did exercises to remove the physical aggravation of his back, lack of dietary inflammation prevented the return of the back ache.

Complex Inflammatory Webs

A student of mine suffers from celiac. This is a complex autoimmune disorder of the intestines that is triggered by wheat gluten and is self-perpetuating. Of interest in this context is that celiacs frequently also have back problems. This indicates that the inflammation of the disease is systemic and impacts other tissues. Clearly, reducing dietary inflammation can go only so far in relieving this complex web of reinforcing sources of inflammation.

Simple Anti-inflammatory Interventions

My friend with tendonitis from holding her child got immediate relief from topical application of castor oil and dietary supplements eliminated the problem. Castor oil and capsaicin react with skin heat-sensing neurons to initiate an anti-inflammatory response in adjacent tissue. In a similar way, menthol acts on cold-sensing neurons and relieves pain by reducing inflammation. Vicks Vaporub is a common commercial source of menthol (other sources are blue Listerine mouthwash and Noxema lotion), which give faster relief than longer lasting castor oil for many connective tissue/joint aches. Exercise is another source of relief for inflammation-based aches and pains.

Health: Combinations of Interventions and an Anti-Inflammatory Diet

Anti-inflammatory drugs, such as aspirin, disrupt the molecular signals that produce inflammation and result in relief from inflammation and pain. The common ailments discussed here respond to anti-inflammatory drugs. Depression was mentioned to point out the psychological dimensions of inflammation. Reproduction/birth is controlled at many points by the processes that we call inflammation and the most inflammatory stage is birth. It is not surprising that disruption of the normally rapid resolution of inflammation following birth leads to postpartum depression. It is surprising that postpartum depression can be relieved by anti-inflammatory drugs.

Fighting Inflammation-Based Diseases

Complex diseases such as allergies, asthma, arthritis, vascular/heart diseases, inflammatory bowel diseases, cancers, etc. are all based on chronic inflammation, but they are also self-reinforcing inflammatory diseases. Cures will require elimination of sources of chronic inflammation, e.g. diet, plus disruption of the disease-supporting inflammation, e.g. food/gut flora-stimulation of inflammation of the bowel.

Fundamental to the cure of all diseases is a supporting anti-inflammatory diet and lifestyle.

Extreme Flu Remedies

Experimental Therapies for ARDS, Cytokine Storms

Do not do this at home. There are doctors and hospitals. Use them.

....But, if a doctor emailed me pleading for any ideas that I had to save a bunch of patients suffering from acute respiratory distress syndrome (ARDS) from Tamiflu-resistant H1N1, my first response would be to suggest therapies designed for ARDS from other origins, e.g. burns, septicemia, etc.

Cytokine Storms Are Out of Control
When too much tissue is injured, the local, molecular communication that normally occurs just between cells, spills into the blood stream and becomes potentially lethal. That is what happens in anaphylactic shock. It is also what happens in cytokine storms, where inflammatory cytokines that are normally short-lived and processed locally to progress into recovery, erupt into the blood stream and impact distant organs.

Major disruption of body function by aggressive blood infections or burns over most of the body, will be lethal without heroic medical interventions. These are injuries beyond the evolved adaptations of mammals.  Until recently there were no survivors.

Influenza has been around for a long time. Humans, other mammals and birds get the flu and get over it. Many body cells become infected, antibodies specific to the virus are produced within about a week, the infected cells are killed, the virus is digested and life goes on.

People die from the flu, because an opportunistic pathogen causes a lethal secondary infection, or the body over-reacts and damages itself in attempts to attack its own infected cells. This is a cytokine storm.

Silence the Storms
Cytokine storms can be weathered by blocking the signaling system. Cytokines are just small proteins that are complementary in shape to corresponding protein receptors that penetrate through the surface membranes of cells throughout the body. Binding of cytokine to receptor changes the shape of the receptor and transmits a signal into the cytoplasm of the receptive cell. This turns on aggressive behavior of immune cells and triggers more inflammatory signaling in other cells. This causes fever, malaise, etc.

...But, I was the one the doctor is pleading with to save the people. And I know that there is more to cytokine signaling than just cytokines and receptors. There are also heparan sulfate proteoglycans (HSPGs). Cytokines are not supposed to be broadcast throughout the body. Cytokines function in the space between cells, the extracellular matrix. Polysaccharides attached to membrane proteins, HSPGs, are secreted at one end of the cells, sweep across the surface and are taken back up at the other end. Cytokines have heparan-binding domains and so they stick to the heparan and are swept along. Cytokines can move from one cell to another as the sweeping HSPGs of adjacent cells come in contact.

HSPGs Mediate Cytokine Signaling
The critical point here is that cytokines bind to their receptors with the heparan between -- the cytokine and receptor are like two halves of a bun and the hot dog is the heparan. In fact the heparan bridges two cytokine/receptor complexes to make an active, signaling pentamer.

Heparin Can Block Cytokine Signaling
Heparin is a fragment of heparan sulfate produced by enzymatic degradation of HSPG. Commercial heparin, used to block blood clotting, is obtained from the mast cells of lungs and intestines of hogs and cattle. The mast cells release heparin and histamine in response to parasites or pollen. Since heparin is a short version of heparan sulfate, it can block the formation of active cytokine/receptor complexes.

Heparin is used in a mist to treat the lungs of burn patients. It is also injected into some infertility patients to suppress inflammation that is inhibiting implantation and gestation. It is also effective in treatment of autoimmune inflammation in Crohn’s disease. I think it should be tested as a therapy for H1N1 cytokine storms. It may be useful in nebulizing mists and oral treatment of intestines.

Berberine Binds to HSPG
Berberine is a phytochemical from Barberry traditionally used in the treatment of intestinal infections and arthritis. It also binds to heparan sulfate to form fluorescent complexes visible in microscopy. Berberine-treated mast cells glow brightly. Heparan sulfate can also be detected in Alzheimer’s plaque, atherosclerotic plaque and prion complexes. Because berberine binds to heparan sulfate, it should also disrupt cytokine signaling. It has been used successfully in treatment of septicemic ARDS.

Curcumin Blocks NFkB
One of the most potent chemicals that blocks inflammatory signaling via the inflammatory transcription factor, NFkB, is curcumin. Curcumin is a major component of the spice turmeric. Oral curcumin can be enhanced by co-administration of black pepper, because the piperine in pepper inhibits intestinal inactivation.

Anti-Inflammatory Diet
Of course, I would also recommend vigorous implementation of an anti-inflammatory diet and lifestyle to support any medical treatment.

Inflammatory Protection from Swine Flu

Inflammation causes allergies, autoimmune and degenerative diseases, and is the foundation for cancer, but it does provide protection against infection.

One of the reasons that older people are less prone to infectious diseases, is that their immune systems have already been exposed to earlier versions of pathogens. Another reason for the relative good health of the parents of baby boomers, is that chronic inflammation increases with age and protection against pathogens is the adaptive advantage of inflammation.

As of now, the new swine flu pre-epidemic is very scary in Mexico City, but doesn’t pack much of a punch north of the border. Maybe it is too early to be optimistic, but it is tantalizing to speculate that the diet-based chronic inflammation that stuffs the coffins in the US, may be good for something besides stuffing the coffers of agribusiness.

One of the symptoms of high chronic inflammation is many years without a day of sick leave. It is hard for a pathogen to get a foothold when the immune system is already provoked by inflammation. Even food and harmless pollen is attacked with vigor and lots of sneezing.

Swine flu is attacking the young and healthy in Mexico. King Corn has already wiped out those targets in the US. Obesity fed by starch, vegetable oils and corn-fed meat, produces chronic inflammation and a potential resistance to swine flu. The US diet provides the inflammatory benefits of old age for the young.

This explains why I live in Idaho. Here it is easy to maintain an island of health in a sea of inflammation. Few visitors with infectious diseases can travel from healthy countries fast enough to transmit the diseases before they show symptoms. Everyone else who reads my advice, lowers their inflammation and lives longer, healthier lives free of chronic diseases, had better start washing their hands and wearing face masks.

Stem Cells Using HSPG Uptake of Recombinant Transcription Factors

Stem Cells from Adult Cells using Transcription Factor Genes
Stem cells have been produced from adult cells using transformation with genes for transcription factors. The problem with this approach was that the embryonic transcription factors had a tendency to promote cancer-like proliferation. What was needed was a temporary push toward embryonic gene expression by temporarily introducing a dose of embryonic transcription factors to dominate gene expression long enough to convert adult, differentiated cells into pluripotent stem cells.

Transcription Factors Synthesized by Recombinant Bacteria
The technical solution was tested and successful results were announced in a prior to publication paper in the journal Cell Stem Cell. Four transcription factors successfully used in prior experiments to induce stem cell transformation were synthesized using recombinant bacteria. The problem was getting the proteins into skin cells that were already growing in cell culture.

Protein Uptake via Triplets of Basic Amino Acids (Heparin-Binding Domains, NLS)
Transcription factors bind to DNA via basic amino acids and many of those basic amino acids are parts of the nuclear localization signals (NLS, quartet or two neighboring pairs of basic amino acids) that bind to importin and transport transcription factors from the cytoplasm to the nucleus.

HSPG Circulation Should Take in Transcription Factors
By inspection, I have demonstrated that proteins observed to be taken up by cells, without specific receptors, e.g. HIV-TAT, lactoferrin, heparanase, allergens, autoantigens, have triplets (or neighboring pairs) or basic amino acids, and this uptake is inhibited by heparin. One would expect that transcription factors would be naturally taken into cells by HSPG circulation. Just adding recombinant transcription factors to cultured skin cells should transform them into stem cells. I don’t believe that this was tested. Instead, more powerful heparin-binding domains were added.

Poly Arginine was used for Uptake of Transcription Factors
The investigators ensured a high efficiency of uptake by adding potent poly arginine sequences to the ends of the transcription factors and synthesized them in recombinant bacteria. The recombinant, arg-tailed transcription factors were taken up by the cultured skin cells and changed the pattern of gene expression in the skin cells. The cultured cells reverted to embryonic patterns of gene expression of pluripotent stem cells. The recombinant proteins were eventually metabolized, but the stem cells had been stably transformed.

reference:
Zhou, H. et al., Generation of Induced Pluripotent Stem Cells Using Recombinant Proteins, Cell Stem Cell (2009), ahead of publication 04.005

Allergy, Asthma, Autoimmunity Start the Same Way

Inflammation is the current medical buzzword. Name the disease and inflammation is there.

Reproduction Requires Controlled Inflammation
Aspirin blocks many of the steps in triggering inflammation and thus, aspirin administration can be used to reveal a role of inflammation in many unexpected places. Aspirin is effective in blocking some forms of infertility, inhibiting miscarriages and ameliorating postpartum depression. So inflammation is a critical part of reproduction. But, also notice that depression is a symptom of chronic inflammation.

Cancer Requires Inflammation
High dose (IV) aspirin has been successfully used to treat cancer. Inflammation is required for cancer growth, because both use the same transcription factor, NFkB. The aberrant signaling of cancer cells would normally lead to programed cell death, apoptosis, but inflammation blocks apoptosis. Aspirin can in turn block NFkB and in the absence of inflammation, cancer cells die by apoptosis.

Inflammation is Self-Limiting
Aspirin also transforms the COX/lipoxidase system to produce anti-inflammatory prostaglandins/eicosinoids. Inflammation normally progresses into anti-inflammation. Blocking this progression leads to chronic inflammation and a shift from local to systemic inflammation with the rise of inflammatory interleukins in the blood stream.

Immune Response Requires Inflammation
The signal molecules (IL-1, IL-6, TNF) and transcription factor, NFkB, associated with inflammation were all initially identified in the development of lymphocytes. Hence, IL stands for interleukin, a hormone that triggers leukocyte (literally white blood cells or cells associated with the lymphatic immune system, i.e. lymphocytes) development. The nuclear factor, i.e. transcription factor, involved in expression of the large chain, kappa, of immunoglobulins in B cells, was called NFkB.

Genes Expressed by NFkB Cause Symptoms of Inflammation
About five dozen genes are under control of NFkB. Among these are COX-2, the enzyme that converts omega-6 arachidonic acid to inflammatory prostaglandins; iNOS, the enzyme that produces nitric oxide that dilates blood vessels to produce hot, red skin; and the inflammatory interleukins, IL-1, IL-6 and TNF, associated with autoimmune disease, fatigue and cachexia (wasting).

Autoimmunity and Allergy Start with Inflammation
Medical treatments focus on symptom abatement and ignore cause. What causes obesity, allergy or autoimmune disease? The answer appears to be chronic systemic inflammation plus exposure to unusual proteins. The unusual proteins are immunogenic, i.e. interact with the immune system to produce antibodies or reactive T-cell receptors, and are subsequently recognized as autoantigens or allergens, that are the targets for immune attack. Inspection of these autoantigens and allergens shows that they all have one thing in common, they bind to heparin via a strong heparin-binding protein domain that is typically a triplet of adjacent basic amino acids.

Heparin is a Short, Highly Sulfated Fragment of Heparan Sulfate
Commercial heparin is purified from the intestines of hogs and cattle. Heparin is released from mast cells (made fluorescent for microscopy using berberine) along with histamine and is released into the intestines to block pathogens from binding to the heparan sulfate that is part of the intestine surface. The heparin is anti-inflammatory and it contributes to minimizing the inflammatory response of the intestines to food.

Inflammation Reduces Heparan Sulfate Production
Pathogen-generated inflammation of the intestines reduces heparan sulfate production and increases immune response to food antigens. NFkB activation by inflammation turns off the production of some genes needed for heparan sulfate proteoglycan (HSPG) synthesis. Since HSPG is a major component of the basement membrane that holds tissues together, the reduction of HSPG results in protein loss (proteinuria) from kidneys, leaking of intestines, and disruption of the blood/brain barrier.

Reduction of HSPG Results in Immunological Presentation of Autoantigens/Allergens
Proteins are brought into cells by specific binding to protein receptors. In many cases, particularly involving signaling or growth factors, both the signal molecules and the receptors bind to heparin. In addition, there is a robust circulation of HSPG, which is secreted and internalized with a half-life of approximately six hours. The sweep of the HSPGs take heparin-binding proteins with them for internalization, e.g. HIV-TAT, heparanase, tissue transglutaminase. I think that this HSPG sweep under inflammatory conditions also internalizes basic autoantigens and allergens with strong heparin-binding domains. This internalization is the first step toward immunological presentation and the immune response to autoantigens and allergens.

Autoantigen/autoantibody/HSPG Complexes Kill Cells
Antibodies against self-antigens, autoantigens form antigen/antibody complexes that also bind to and cross-link HSPGs, because of the heparin-binding domains of the autoantigens. The large complexes may disrupt HSPG circulation and trigger apoptosis or abnormal physiology. There are many other examples of heparin-based complexes that are toxic, e.g. Alzheimer’s amyloid plaque, diabetic beta cell antibody complexes, celiac gluten/tRG antibody complexes, multiple sclerosis myelin antibody complexes, atherosclerotic plaque.

Anti-Inflammatory Diet and Lifestyle Protects
Dietary and lifestyle adjustments that minimize inflammation, e.g. low starch, no HFCS, low vegetable oil (except olive) and supplements of vitamins D & C, fish oil (omega-3) and glucosamine, reduce the risk of allergies/asthma, degenerative diseases and cancers. Simple, high level supplements with fish oil reduce numerous mental disorders, e.g. depression, ADHD; infertility, pre-eclampsia and postpartum depression; allergies, asthma; arthritis, atherosclerosis; burn recovery, septicemia and head injury.

Reducing Inflammation is a Panacea for Modern Diseases
Most modern diseases have an inflammatory component, because modern diets are rich in inflammatory components, e.g. starch/sugar, corn/soy oil, HFCS, trans fats, and exercise is minimal. The medical industry has not successfully promoted healthy eating and exercise; and in fact has promoted the devastating replacement of saturated fats with inflammatory polyunsaturated vegetable oils. Meat production has moved away from grazing on omega-3-rich plant vegetation to omega-6-rich corn and soy. Replacement of the corn/soy based agricultural economy would have predictably immense beneficial impact in reducing inflammation-based degenerative autoimmune diseases and cancers.

Cure for Cancer, Autoimmunity, Allergies, etc.

The immune system is powerful enough to provide protection from disease. Unfortunately, to act decisively the cells of the immune system have to be able to discriminate between self and non-self. Poor discrimination can lead to autoimmunity, cancer or infection. New approaches promise the precise use of interleukins, to reset self-recognition, eliminate a wide range of diseases and liberalize organ transplantation.

IL-2 is the Cytokine Responsible for Suppression of Autoimmunity -- Tolerance

Self/non-self discrimination is dependent on cellular communication and much of that communication takes place via small proteins called interleukins. First and foremost among the interleukins is interleukin-2 (IL-2). IL-2 is made by cells of the immune system, lymphocytes. Mice that are either defective in producing IL-2 or the lymphocyte receptor for IL-2, IL2R alpha, also called CD25, rapidly develop autoimmune diseases, such as type I diabetes or inflammatory bowel disease. Thus IL-2 is necessary for both effective immunological defenses against pathogens and suppression of immune attacks on self tissues, i.e. autoimmunity.

IL-2 Balance Achieved with Complex of IL-2 and Anti-IL-2 Antibodies

Direct injection of IL-2 has some impact on cancers, but is very difficult to control. This should be expected, because local environments should determine if the IL-2 will stimulate aggressive immunological attacks or development of regulatory T cells, Tregs, that produce tolerance.

More subtle control is achieved by using antibodies that bind to particular regions of the IL-2. The resulting IL-2/anti-IL-2 complexes can be used to stimulate immunological reactions to an antigen, which is useful for vaccines, or can stimulate tolerance for use in organ transplantation.

Future applications may be in the cure of a wide variety of autoimmune diseases, e.g. type I diabetes, inflammatory bowel diseases, allergies, asthma; degenerative diseases, such as arthritis or athersclerosis, and cancers.

reference:
Webster KE, Walters S, Kohler RE, Mrkvan T, Boyman O, Surh CD, Grey ST, Sprent J. 2009. In vivo expansion of T reg cells with IL-2-mAb complexes: induction of resistance to EAE and long-term acceptance of islet allografts without immunosuppression. J Exp Med. Mar 30. [Epub ahead of print]

Enteroviruses, Autoimmunity, Diabetes

Insulin-producing cells of the pancreases of diabetics have been found to harbor viruses common to the gut. Antibodies to the virus coat proteins also bind to pancreas proteins.

A recent paper, referenced below, shows that in a collection of samples from the pancreases of individuals that had been diagnosed with type I diabetes less than one year prior to the sampling, insulin producing cells are also infected with enterovirus. The same association between enterovirus infection was found to a lesser extent in type II diabetics, but not in non-diabetic controls.

Enteroviruses have been repeated associated with diabetes over the last decade and antigenic determinants of the enterovirus protein coat also bind, i.e. cross react, with antigenic determinants of human cellular proteins.

I examined the enterovirus coat protein, VP1, and found the same three amino acid sequence (three basic amino acids, lysine [K] or arginine [R], highlighted) that I also found in all allergens (peanut, ragweed, dust mite, bee venom) and autoantigens of autoimmune diseases (lupus, MS), and is associated with heparan sulfate-based internalization and presentation of protein immunogens. This observation is consistent with my hypothesis that inflammation plus the presence of one of these proteins, results in production of B and T lymphocytes specific for antigenic determinants on the surface of the immunogen protein. Note that the antigenic determinants usually do not include the three basic amino acid sequence, e.g. RRK, that is involved in uptake and presentation of the protein.

VP1 [Human enterovirus B]
HVINYHTRSESSVENFMGRAACVYIAQYATEKVNDELDR
YTNWEITTRQVAQLRRKLEMFTYMRFDLEVTFVITSSQR
TSTTYASDSPPLTHQVM

reference:
Richardson SJ, Willcox A, Bone AJ, Foulis AK, Morgan NG. 2009. The prevalence of enteroviral capsid protein vp1 immunostaining in pancreatic islets in human type 1 diabetes. Diabetologia. Mar 6. [Epub ahead of print]

Omega-3 Fatty Acids, Antioxidants and Cancer

It is hard to sort out the inflammatory effects of short/long-chain omega-6 and omega-3 fatty acids. Vegetable antioxidants make the picture even worse. The absolute, as well as relative amounts, of the various types of fatty acids make a difference. It also now appears that oxidation prior and during digestion may be important to the impact of polyunsaturated fatty acids (PUFAs). The source (perhaps even the meal composition) of the PUFAs was as important as omega-3 versus omega-6, for common, short chain PUFAs.

In some studies, omega-3 PUFAs, such as the short-chain alpha linolenic acid (ALA) common in flax seed or the long-chain fish oil FUFAs, eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), reduced cancer in human and mice. Earlier work in cell cultures showed that all of PUFAs suppressed the growth of cancer cells.

A large French study (reference below) began in 1993. Approximately 100,000 women between the ages of 40 and 65 volunteered to provide dietary and breast malignancy information and ca. 75,000 qualified for the study (the French component of EPIC, European Investigation of Cancer and Nutrition) . The dietary data provided information on the fatty acid composition of meals and revealed who was eating vegetable antioxidants and vitamins.

Major findings:

• Neither omega-6 nor omega-3 fatty acids were related directly to breast cancer risk.
• Long chain omega-3 fatty acids (EPA and DHA) reduced breast cancer in the group of women with the highest consumption of omega-6 PUFAs.
• High LA or ALA consumption in the form of vegetable oils or vegetables reduced cancer incidence.
• High LA or ALA consumption in the form of processed foods or nuts was associated with a higher incidence of breast cancer.
• Longer chain PUFAs were not associated with increased risk, regardless of source.

What does this mean?

• The source of the PUFA is of paramount importance. This study may apply more specifically to cancers and less to other inflammation-based degenerative diseases. The general anti-inflammatory diet may need refinement. I would suggest the following additions:
• Retain the preference for the more omega-3 friendly olive oil or perhaps flax oil versus the omega-6 rich vegetable oils (corn, soy, safflower), but focus on freshness and do not heat these oils.
• The data seem to be in favor of saturated fats for cooking. That means a shift to coconut oil.
• Vegetable antioxidants may be most important in the gut during digestion. Do these antioxidants even enter the blood stream? Certainly some alkaloids get to the brain, but much of the impact of the less mobile, large molecules may be restricted to the gut.
• An extension of this discussion may be to encourage eating more leafy vegetables with meat. That may be the paleo-diet connection.

reference:
Thiébaut AC, Chajès V, Gerber M, Boutron-Ruault MC, Joulin V, Lenoir G, Berrino F, Riboli E, Bénichou J, Clavel-Chapelon F. 2009. Dietary intakes of omega-6 and omega-3 polyunsaturated fatty acids and the risk of breast cancer. Int J Cancer. Feb 15;124(4):924-31.

Healthcare Practitioners: Modern Flint-Knappers?

Deadly Unintended Consequences

With the resurgence of the paleolithic diet, perhaps we should also consider paleolithic hygiene. The shaping of flint points was accomplished by skillful removal of ultrasharp (in excess of razor-sharp) flakes of glassy material. The artisan flint-knappers were community treasures, but they were banned from living near others, because they were also habitual child killers. Ingestion of flint flakes is lethal and the inherent flakiness of flint-knappers made them personae non gratae. Children living near flint-knappers died at an alarming rate and flint-knappers never sired offspring who didn’t whither and die.

Wise communities cherished flint-knappers, but they got the point and kept the knappers at a distance.

Constant Selection for New Pathogens

Where do bacterial pathogens come from and how are they spread? Some pathogenic bacteria are harbored by other animal species that humans encounter. We can pick up bacteria from pets, barnyard animals and game. We routinely place large numbers of humans in contact with every ecological niche on the planet and transport any bacteria that can grow humans rapidly to population centers. The human gut provides a mixing bowl where newly acquired bacteria are systematically extracted for their genes and recombined with resident bacterial genomes. Within days new bacteria are ready for testing in the new environment.

The most potent selection agents are antibiotics. After antibiotic treatment, only bacteria, both old residents, new arrivals, with resistance, will survive. New arrivals that incorporate the antibiotic resistance of residents will be immediately successful. New arrivals that enter with previously acquired resistance will be immediate successes and spread to fill niches vacated by antibiotic-sensitive residents.

Should Nurses and Doctors Be Quarantined?

Nurses and doctors who routinely touch patients and inhale the atomized bacterial mist around patients, receive a continuous inoculum of problematic bacteria. Healthcare practitioners are, however, immune to most of these potential pathogens by virtue of their highly educated immune systems. Unfortunately they do not get sick, but they are still potentially infectious. Their gut flora and the bacteria on their other surfaces are potential sources of the pathogens that they have acquired in their duties. Every contact with healthcare practitioners or by chronic exposure, to members of their immediate families, is potentially compromising to anyone with a compromised immune system.

A Modest Proposal: Routine Screening, Purging and Healthful Fecal Transplants

The public deserves to be healthier after treatment with a healthcare professional. It is therefore mandatory that the bacteria received from a nurse or doctor through professional (or informal) contact be safe and healthy. It should be an expectation that healthcare professionals have guts that are clean-running and sanitary. To that end, it seems reasonable to routinely screen their resident bacteria and if necessary replace it with a health-promoting alternative.

We expect professional athletes to be free of performance enhancing drugs. It makes sense that doctors and nurses be free of pathogens.

Medical Advice Is Just Wrong

Medical advise says to avoid sun, fats and red meat, but to drink lots of water, eat polyunsaturated vegetable oils and focus on the grain-rich bottom of the food pyramid. The medical advice is simply wrong and is not supported by the biomedical literature. A recent article in a major medical journal claims that about 90% of medical advice is not based on clinical research studies, but rather represents the opinions of experts who are supported by the health industry. Most research is conducted to support products. Unfortunately the advice that comes from medical societies is not healthy.

Here I will provide a few examples to illustrate that medical advice is frequently, if not usually, wrong about diet, nutrition, cause of disease, appropriate drug use and whether to spend a few unprotected moments basking in sunshine.

The Sun Is Not the Enemy, but Sun Blockers Can Increase Skin Cancer

Medicine is supposed to provide instructions on how to handle dangerous chemicals and procedures safely and to enhance health. Solar radiation is dangerous and will cause skin cancer if used inappropriately, but solar radiation is also needed to produce vitamin D in skin. The public response to the medical mandate to limit solar exposure to reduce radiation-based skin cancer resulted in increased use of solar-blocking lotions. Unfortunately, the result was that some people spent more time in the sun, assuming that avoiding sun burns meant that they were avoiding skin cancer. Unknowingly they had shifted their skin exposure down from doses sufficient to kill cells and cause inflammation, to levels sufficient to just cause solar mutagenesis -- the lower exposures were optimal for skin cancer production.

Spare the Sun and Spoil the Child

Babies and children are the most sensitive to solar radiation induced skin cancer and need protection from over exposure, but the public response to medical advice has been to avoid prudent exposure to the sun. Now kids in the U.S. are showing symptoms of rickets, a vitamin D deficiency disease common during early industrialization, in which air pollution, urban poverty and factory work limited solar exposure. Babies in strollers are completely covered. One frightening consequence of this over-reaction could be a resurgence of poor bone growth that in the 1920’s resulted in the development of the now-trendy Cesarean section procedure to accommodate women with malformed pelvises due to rickets.

Rickets Is Rampant

Ubiquitous vitamin D deficiencies due to inadequate sun exposure is compounded by inadequate sources of dietary vitamin D and inappropriate medical interventions. Most vitamin D deficiencies go unnoticed, because the typical symptoms of deficiency mimic other forms of inflammation. When serum levels of vitamin D are actually measured and found to be inadequate, supplements of 600-1000 iu/day of vitamin D3 are prescribed. Unfortunately, there is seldom followup testing and a recent study indicates that most treatment for vitamin D deficiencies is inadequate -- much higher doses, ca. 2-5000 iu/day are required to reach optimum levels. Most people are and remain vitamin D deficient.

Scourge of Scurvy

Vitamin C deficiencies are also a problem. Most people get enough vitamin C to avoid losing their teeth (vitamin C is needed for collagen production), but subclinical deficiencies still produce chronic inflammation. The major cellular anti-oxidant is glutathione, but vitamin C is another major defense against reactive oxygen species (ROS). An increase in ROS triggers oxygen stress and inflammation. Deficiency of vitamin C indicates that more vitamin C is being used up than is being replenished in the diet. Numerous metabolic disturbances associated with other deficiencies or infections can result in vitamin C depletion and chronic inflammation. Most people are vitamin C deficient.

Vegetable Oils Are the Problem, Not the Cure

Medical advice to avoid saturated fats in meats and shift to omega-6-rich vegetable oils is a major contributor to chronic inflammation and modern degenerative diseases. The original claimed association between saturated fat consumption and cardiovascular disease was tenuous, but produced a glacial shift in diet toward consumption of omega-6 fatty acids, e.g. corn and soy oils. The medical dependence on measurements and treatments of LDL, has outweighed the actual data in the biomedical literature -- LDL levels are not important in cardiovascular disease. Drugs that lower LDL, serum cholesterol, are only effective in reducing heart disease, if they lower LDL by lowering inflammation. The risk factor is the inflammation, not the LDL level. Agricultural practices that use grain over grass further reduce the omega-3 fatty acid content of meat and increase the inflammatory omega-6 fatty acid level.

Statins Are a Problem, Not the Cure

Statins are broad spectrum disrupters of the function of many different enzymes and proteins. They were originally isolated from fungi based on their ability to poison bacteria, i.e. they are antibiotics. They disrupt fat metabolism and thereby lower LDL levels, but they also cause many undesirable and potentially dangerous side-effects. One of these actions is to block inflammation triggered by activation of the inflammation transcription factor, NFkB. By blocking NFkB activation, some statins lower inflammation and thereby decrease cardiovascular disease. This activity is similar to aspirin, which acts on COX-2 as well as directly on NFkB. Both statins and aspirin (NSAIDs) have multiple activities on numerous areas of cellular metabolism. The activities of both include reduction in inflammation, but they also produce other undesirable side effects. Chronic inflammation is better treated by diet, exercise and traditional herbs and spices, rather than more dangerous statins.

Water Is Miraculous, but just Satisfy Your Thirst

If you are thirsty drink tap water. There is no improvement in health by drinking some extra amount of water each day. Drinking water in plastic bottles from magical sources provides no improvement in health. Much of the “spring water” with designer labels is only locally bottled tap water. The plastic bottles are an ecological disaster and the “purified” water in the bottles is contaminated with compounds leaching from the bottles. If you want a constant source of water, bottle your own tap water. If you want to avoid the minor contaminants added to avoid bacterial contamination of municipal water supplies, use a simple point-of-use filter.

Starch Is the Problem

Starch is rapidly converted into blood glucose and that spike in blood sugar causes major problems. The foundation of the old food pyramid, grains, is no different than table sugar in being hyperglycemic, i.e. rapidly raising blood sugar. A large muscle mass and high physical activity can minimize the rise in blood sugar, by using up the sugar for muscle energy as it enters the blood. Unfortunately, most people do not have enough muscle and are not physically active enough to be protected from the starch and sugars in their diets. The result is chronic inflammation in the form of metabolic syndrome and degenerative diseases, e.g. diabetes, allergies, depression, acne, infertility, cardiovascular disease, autoimmune diseases and cancers.

One slice of white bread with a meal may be too much starch for some people. The maximum for most people is: one half of a ripe banana or one half cup of a starchy entree such as pasta, potato, rice, or one of the two buns on a burger. The starch needs to be spread over several meals. Eating too much starch with a meal produces intense hunger, as the blood sugar rapidly rises, triggers insulin release and a subsequent crash in blood sugar. Don’t believe any of the diets that recommend starches to replace fats. Many “lite” diet foods are more unhealthy than the higher fat originals that they replace. Replacing saturated fats with saturated starch is dangerous. The temporary high blood sugar level produces the increased health risks routinely associated with diabetes.

Insufficient Food Is the Problem -- Insufficient Minerals

It takes only 2-3000 Calories per day to energize most people. That means that most people can eat their day’s worth of calories with the sandwich plate at a fast food restaurant. That meal will provide an overdose of starch and sugar, but will be deficient in vitamins and minerals. A major dilemma is that it takes so little food to provide adequate energy for a low activity lifestyle, that the choice must be made between obesity and vitamin/mineral deficiencies. Eating just enough to satisfy energy needs results in deficiencies, but eating more to avoid vitamin/mineral deficiencies, results in obesity. The only solutions are to eat supplements to supply needed vitamins, minerals, antioxidants, etc. or increase physical activity and body muscle mass, so that more can be eaten without producing obesity. For most people the solution is a combination of increased physical activity and supplements. That combination is also found to reduce inflammation and the associated risk of degenerative diseases.

It’s the Stupid Diet

The obsession of medicine with drugs and invasive procedures provides additional health risks for patients. Many researchers complain in the biomedical literature that there is insufficient focus on the cause of disease and too much emphasis on the study of the impact of specific drugs on disease symptoms. The result is that in most cases the symptoms are treated and the disease becomes chronic. Of course this also means that the patient is a permanent consumer of health care.

The foundation of all healthcare should be to improve the lifestyle of the patient. Diseases don’t just happen. The biggest contributions of immediate family to disease of an individual are not defective genes, but rather defective diet and lifestyle habits. Our healthcare system is too no fault. People are sick because there is something wrong with how they live. They eat too much or they eat the wrong foods. They don’t get enough exercise to develop a healthy muscle system to support their joints. Most importantly, bad diet and lifestyle choices produce chronic inflammation. Drugs can reduce chronic inflammation, but will also produce additional side effects that will also require interventions. It makes more sense to attack the original causes of inflammation.

Every treatment program should address the pervasive contribution of chronic inflammation by including a diet and lifestyle inventory and an assessment of the cause of the disease that is being treated. An appropriate anti-inflammatory diet and a path toward a more active lifestyle should be the foundation of every treatment plan.

Aricept: dementia treatment

Aromatic Binding to Enzymes -- 

Aricept, an acetylcholine esterase inhibitor used to treat Alzheimer’s disease and other conditions that benefit from enhanced accumulation of acetylcholine, is an example of a molecule with multiple hydrophobic rings that binds to an enzyme.

I want to discuss aricept as an arbitrary example that I just looked up to illustrate the lack of specificity of statins that I will characterize in another article as little more than molecular skeleton keys that work on many different enzymes.

I have presented two diagrams of the structure of Aricept. It has two isolated rings on the left and then a fused pair of rings on the right. The major chemical feature here is the inability of the rings to hydrogen bond with water. The result is that water next to the faces of the rings is highly structured in a high energy configuration. Two rings will be at a much lower energy if they are stacked together, because two of the surfaces will no longer be exposed to water.

Typical low energy, noncovalent bonds in water, such as ionic bonds are readily broken by the thermal, kinetic energy of water -- they get knocked apart. The energy of these bonds is only 1-2 kcal/mol. In contrast, the stacked hydrophobic rings are quite stable, because it takes ten times the energy to separate them, 20 kcal/mol.

Aricept binds to acetylcholine esterase, the enzyme that degrades the neurotransmitter acetylcholine by at least three stacked rings. These ring structures are shown in the close up of the tunnel leading to the enzymes active site near the yellow tryptophan on the left. Part of the enzyme shown by the white, ribbon-like twists of the amino acid backbone have been removed over the tope of the grey-red and blue aricept molecule, to make it easier to see.

I also showed the aricept in the tunnel with the surface of the protein shown to indicate how the aricept slips and sticks in the enzyme and blocks its activity.

The aricept is bound to yellow tryptophans at both ends and the middle ring is bound to the hydrophobic ring of orange tyrosine. The geometry of the interaction is important, but many other molecules with fewer rings would also bind to the same hydrophobic, aromatic ring amino acids. Acetylcholine, which can form hydrogen bonds with the paired electons of the acetyl oxygens, will just slip across the surface of the hydrophobic rings on its way into the enzymatic tunnel.

Statins were found by testing fungal extracts for molecules that would inhibit an enzyme (HMG-CoA reductase) in lipid metabolism. The normal lipid substrates for that enzyme would also be expected to bind to the surface of rings in the acetylcholine esterase enzyme. In fact, I would expect to find molecules from fungal extracts that would inhibit acetylcholine esterase.

I demonstrated the nonspecificity of all of these binding events with the aromatic rings in the active sites of enzymes by having one of my students check for the binding of a flat hydrophobic molecule, metformin, one of the common drugs for treating type II diabetes, to a common bacterial enzyme, beta galactosidase. Kinetic studies demonstrated competitive inhibition of typical beta galactosidase substrates, which indicates that the metformin binds the aromatic amino acids that are known to be involved in binding of the sugar substrates, e.g. lactose, of the enzyme. I would not be surprised if the statins are transported into cells by the same organic cationic transporter that transports metformin.

I am setting the stage for a discussion in a future article of what kind of activities would be expected from fungal molecules that were identified by the statin screening. It is not surprising that the statins have many activities other than reducing LDL. The only statins that are effective in treating cardiovascular disease are those that also lower inflammation. It is also not surprising that statins have many side-effects.

American Heart Association OKs Linoleic Acid and Arachidonic Acid

Can the AHA be correct in promoting omega-6 PUFAs? Doesn't this conflict with the broad therapeutic action of omega-3 PUFAs, EPA/DHA, against inflammatory diseases?

The dietary shift from saturated animal fats to polyunsaturated fatty acids (PUFAs) from vegetable oils paralleled the shift from infectious diseases to inflammatory/degenerative diseases as predominant killers in the Western world. Treatments for degenerative diseases associated with aging have improved, but these diseases have become more prevalent and the age of onset has decreased. And medical costs have skyrocketed. Omega-6 vegetable oils seem to be the problem, but the American Heart Association (AHA) has recently given these PUFAs a clean bill of health.

Why the AHA Conclusions Seem Just Wrong

The rise of inflammatory/degenerative diseases follows the shift to processed foods rich in omega-6 PUFAs (corn, soy, cottonseed, safflower oils) and simple carbohydrates (grain starch, sugar, high fructose corn syrup), but the AHA presents scientific data to exonerate omega-6 PUFAs. The central problem is that the AHA’s conclusions are not based on a conceptual framework to explain cardiovascular disease. Instead, conclusions are derived from experiments in which various diets are fed to people and consequences are analyzed. With some diseases, in which there is a simpler cause and effect relationship, this approach might lead to useful answers, unfortunately, the inflammatory component central to cardiovascular disease can have multiple, alternative origins and simple experiments yield misleading conclusions.

Experimental Basis for AHA Support for Omega-6 PUFAs

• Conversion of short PUFAs found in the diet, e.g. LA, to the long PUFAs that serve as the precursors of cellular hormones. But conversion is thought to be inefficient, so that less than 1% conversion occurs and short PUFAs have little impact on cellular long PUFA concentrations. Moreover, the brain does not perform the conversion and the high brain content of DHA is supplied on demand from DHA circulating in the blood.
• There don’t appear to be any direct, inflammatory derivatives of LA (C-18), but after it is converted to AA (C-20), the arachidonic acid is the starting point for the conversion to most of the inflammatory and anti-inflammatory cellular hormones, e.g. prostaglandins, leukotrienes and lipoxins. Thus inflammation is initiated by AA-derived products, but resolution and return to normal physiology is also based on other AA-derived products.
• Increases in blood plasma AA are associated with anti-inflammation, not inflammation.
• Increases in dietary AA and/or LA result in a decrease in cardiovascular disease. Replacing dietary saturated fat with PUFA leads to a reduction of disease by 25-50%. Higher serum LA translates into less disease.
• Increases in dietary LA result in lower serum cholesterol and LDL, but paradoxically they also lead to a narrowing of arteries.
• The relative amounts of dietary PUFAs (USA) are LA 15grams/day, AA 0.15g/d, ALA (omega-3, C-18, linolenic acid) 1g/d, EPA/DHA <>

Statins Lower Cardiovascular Disease by Lowering Inflammation (LDL Not Important)

The JUPITER study showed that the statin Crestor was effective in lowering heart disease, because it lowered inflammation. Individuals with chronic inflammation responded to Crestor by lowering inflammation. Lowering of LDL levels, however, was not related to decreasing disease. Elevated LDL levels may reflect inflammation.

Relating the JUPITER results to the AHA conclusions suggests that LA and AA may reduce inflammation and as a consequence also reduce serum LDL.

Inflammation Is the Cellular and Tissue Response to Many Stresses

The list of pathogens that trigger inflammation is long and includes specific signals from viruses, bacteria, fungi and protozoa. Pathogen-caused damage, as well as physical trauma, cause inflammation. Disruption of cellular metabolism and energy flow by vitamin, mineral, amino acid, or fatty acid deficiencies or excesses all produce inflammation. One of the difficulties of diagnosis is the overlapping of symptoms originating from numerous sources of underlying inflammation. Herniation of vertebral disks, for example, can be triggered by physical trauma, but it also may be initiated by the intestinal inflammation of gluten-based celiac. Acne and depression are common symptoms of chronic inflammation that may result from dietary deficiencies, gum disease, gluten sensitivity, etc. All of these examples respond to anti-inflammatory diets.

It is difficult to identify the sources of inflammation in experimental studies. In cardiovascular disease, the sources of inflammation are commonly not known in individual cases and the cardiac symptoms are treated. In reality, these are actually many different diseases, all with different sources of inflammation, pigeon-holed under the same symptom, a cardiac event. The most effective long term treatment for the dispart group is general suppression of inflammation. Any specific treatment of a root cause only works on a small subset of the group and would be considered ineffective. Thus, statins are considered effective against heart disease, because they reduce inflammation that is common to the whole group. Reduction of LDL is inadvertently used as a measure of control of inflammation and has become inappropriately designated as a risk factor. Directly lowering LDL has no impact on heart disease, but it is easy to measure. Inflammation is hard to measure and finding the source of inflammation is harder still.

Omega-6 Vs. Omega-3 Is Another False Dichotomy

Just as there is no opposite to inflammation, omega-6 and omega-3 fatty acids are not in opposition. The action of aspirin is the big clue. Aspirin changes the structures of the enzymes involved in converting AA into inflammatory prostaglandins and leukotrienes, with the result that anti-inflammatory lipoxins are produced instead. Aspirin is a biochemical switch that mimics the natural transition of the cellular machinery from producing enzymes that accentuate inflammation, to enzymes and signals that are the next step in the cycle, repair and restoration of normalcy.

Omega-6 PUFAs are needed for both inflammation and restoration of normal cellular functions. Some of the enzymes produced during inflammation are needed for the reset to normalcy. The difficulty comes when inflammation is sustained, components are depleted and the cycle cannot be completed. The result then is chronic inflammation, the symptoms of metabolic syndrome and degenerative diseases.

Why Did Demonizing LA and AA Seem Right?

It seems wise not to trust medicine, dietitians and the food industry, because they have made so many lamentable mistakes making dietary suggestions that have shortened so many lives. Professional societies like the AHA also frequently give silly advice, because the advice doesn’t reflect the best information from the biomedical literature. So it makes sense to be skeptical.

In this case the AHA appears to be right, only because established views were supported by straightforward experiments. What determines if an excess of dietary LA and AA is going to be a problem with inflammation is the absolute amount of AA and EPA available on the surface of immune cells. PUFAs are attached as part of the phospholipids of the lipid rafts on the membrane surface of immune cells that have received a inflammatory signal, e.g. bacterial lipopolysaccharide. There is usually adequate AA to be converted by enzymes on the cell surface to produce further inflammatory signals. The problem comes if there is so much AA that the EPA never made it to the lipid rafts. The result would be inadequate EPA conversion to anti-inflammatory prostaglandins and failure to return to normalcy. This would be a particular weakness in the presence of a large depletion of the EPA pools during sustained inflammation and chronic inflammation would result.

Thus, the AHA promotion of omega-6 PUFAs is half right. They should have said that omega-6 fatty acids are not a problem, if there is adequate EPA/DHA and no sustained inflammation. Unfortunately, the Western diet provides inadequate EPA/DHA and deficiencies that constantly produce inflammation. Of course, those enjoying an anti-inflammatory diet and lifestyle have biochemical tolerance for the AHA’s suggestions. Others eat vegetable oils at their peril.

reference:

Harris WS, Mozaffarian D, Rimm E, Kris-Etherton P, Rudel LL, Appel LJ, Engler MM, Engler MB, Sacks F. 2009. Omega-6 Fatty Acids and Risk for Cardiovascular Disease. A Science Advisory From the American Heart Association Nutrition Subcommittee of the Council on Nutrition, Physical Activity, and Metabolism; Council on Cardiovascular Nursing; and Council on Epidemiology and Prevention. Circulation. 2009 Jan 26. [Epub ahead of print]

Where’s the Aspirin?

Aspirin is the traditional anti-inflammatory agent. Many of us grew up with the quintessential doctoring phrase, “Take two aspirin and call me in the morning.” Aspirin stops inflammation in several ways. Like all drugs, it interacts with many different proteins/enzymes. In fact it interacts so intimately with the inflammatory system that it suggests that the process of inflammation may require an aspirin-like molecule to function normally.

Aspirin Binds to Multiple Enzymes of Inflammation

Aspirin is observed to reduce inflammation. That means that ingested aspirin tablet dissolve in the stomach and pass through the intestines into the blood stream and subsequently bath cells of the blood and the endothelium that lines the blood vessels. In order to reach the blood stream, the aspirin must pass through the intestinal cells. That passage requires binding to a protein transport molecule.

Cells responding to an inflammatory signal (NFkB, transcription factor is activated) synthesize enzymes that release unsaturated fatty acids (ARA, EPA, DHA) from membrane phospholipids (PLA2, phospholipase A2), form a cyclic epoxide from the fatty acid (prostaglandin H2 synthase 2, also called cyclooxygenase 2, COX-2).

Aspirin binds to the inhibitor that normally inactivates NFkB and prevents NFkB activation that is required for inflammation. Aspirin also binds to PLA2 and prevents fatty acid release and thereby blocks activation of inflammation. Aspirin also binds to COX-2 and blocks the production of inflammatory prostaglandins from ARA. But that is not all that aspirin does.

Inflammation Resolution Uses Aspirin-COX-2 Interaction

The strange interaction that makes aspirin suspicious is that aspirin doesn’t just interfere with the action of enzymes, it subtly changes their specificity. Thus aspirin chemically transfers its acetyl group (CH3-COOH-) to an amino acid in the active site of COX-2 to produce a new group of anti-inflammatory lipoxins from ARA, EPA and DHA.

This raises the question of whether aspirin is a natural dietary modulator of inflammation. Recall that aspirin was initially obtained from willow (Salix) bark. Unfortunately, the data are conflicting. Initial research indicated that grains (naturally inflammatory) lacked aspirin, but many herbs, spices and leafy vegetables (naturally anti-inflammatory) contained aspirin. Subsequent tests refuted this work. It would be consistent with observations that some dietary components are anti-inflammatory, but candidate acetyl donors have not been identified.

Speculative acetyl candidates may include the menthol relatives, such as menthyl acetate (figure). Peppermint oil, which contains mostly menthol with some menthyl acetate, is more effective in the treatment of inflammatory bowel disease than most pharmaceuticals prescribed to treat the condition. This anti-inflammatory activity may be due in part to the aspirin-like chemical structure and function of the menthyl acetate. Also note that acetic acid/vinegar is sometimes suggested as a cure-all. This activity may be a consequence of its formation of ester linkages with alcohols that have structures similar to menthol.

Large Dose Aspirin as Cancer Treatment

The potent anti-inflammatory effects of aspirin have been compromised, because inflammation is an essential developmental activity. Thus, the integrity of the gut, for example, requires modest production of inflammatory prostaglandins and a pill of aspirin can disrupt gut tissue. Large doses of aspirin cannot be given orally. Intravenous administration of large doses of aspirin, however, is possible and the impact on process that require inflammation is dramatic.

Anecdotal evidence indicates that large dose aspirin is able to disrupt cancers, because proliferation of cancer cells requires NFkB activation and other inflammatory responses. High doses of aspirin also cause other potentially dangerous complications, such as short-circuiting oxidative phosphorylation of mitochondria and increasing nitric oxide free radical production. Still, the impact of high dose aspirin on some diseases is so amazing that it is being actively and carefully pursued.

What’s the Opposite of Inflammation?

I want to commemorate the writing of my 100th article on Blogspot by discussing a new insight into inflammation.

I have been searching the last several years for an anti-inflammatory system to balance inflammation. Now I realize that there is no opposite to inflammation. There is only completion of inflammation to return to the original state. Inflammation is a process that includes resolution or recovery from the defensive, destructive state of immunological activity.

Inflammation is the martialling of resources for battle by offloading lymphocytes from the blood stream, engaging the enemy by triggering the release of toxic secretory vesicles from leukocytes, and cleaning up the carnage by macrophages engulfing cellular fragments. Each step in the inflammatory process induces the next step until there is a return to the origin. Inflammation is not balanced by anti-inflammatory processes.

Inflammation is triggered by molecules characteristic of viruses, bacteria or fungi binding to membrane receptors (TLRs). The result is activation of the inflammatory transcription factor, NFkB (illustrated holding DNA), that turns on the expression of dozens of genes that code for cytokines (IL-1, IL-6, TNF) and enzymes (COX-2) that produce signal compounds. Among the signal compounds are the inflammatory eicosanoids (PGE2) produced from the omega-6 fatty acid arachidonic acid (ARA).

The complex signaling pathways that lead to PGE2 synthesis subsequently initiate transcription of genes that code for the enzymes that make lipoxins (resolvins and protectins) from eicosapentanaenoic acid (EPA) and docosahexaenoic acid (DHA). EPA and DHA are the two omega-3 fatty acid components of fish oil and a shortage of these dietary components blocks the next step, resolution of inflammation.

The lipoxins reduce the permeability of blood vessels, stop the offloading of lymphocytes, reduce responsiveness to inflammatory cytokines, recruit phagocytic macrophages to clean up debris and orchestrate a return to quiescence of the inflammatory system. Without adequate lipoxins, inflammation continues.

An interesting footnote to this discussion is the impact of aspirin on inflammation. Aspirin binds to the enzyme (COX-2) that converts ARA to inflammatory prostaglandins and leukotrienes. Acetylation of COX-2 by aspirin stops inflammatory eicosanoid synthesis and shifts the synthesis to anti-inflammatory lipoxins. Even ARA is used to make anti-inflammatory lipoxins in the presence of aspirin. This shift to anti-inflammatory signaling may occur naturally in the small intestines in response to aspirin-like compounds in vegetables. This would be a transitory response similar to taking aspirin with a meal. More constant use of aspirin would disrupt the normal and necessary actions of the inflammatory signaling to maintain the integrity of the gut.

Excess of dietary omega-6 oils and deficiency in omega-3 fatty acids corrupts inflammatory signaling by eliminating recovery and produces chronic inflammation. Another name for chronic inflammation is obesity/metabolic syndrome. Chronic inflammation is the foundation for the degenerative, autoimmune and cancer diseases that are so prevalent today.

Fortunately a shift to an anti-inflammatory diet and lifestyle provides a simple solution to chronic inflammation.

[Note added: Perhaps the opposite of anti-inflammatory is immunosuppressed, as in high use of omega-3 oils can increase the risk of tuberculosis of influenza.]

There Is More Than Antioxidants

Every time a plant product has an impact on a disease it seems to be attributed to its antioxidant activity. Plant products are active, because they bind to proteins. They bind to lots of different proteins.

Krill oil is a good example. The anti-inflammatory activity of krill oil is due to its omega-3 oil (DHA and EPA) content, but krill oil is more potent than expected. Krill oil also contains a terpene, astaxanthene, that is probably derived from its algae diet. Astaxanthene is labeled as an anti-oxidant, but that is much too easy.

Astaxanthene consists of two flat, hydrophobic paddles, connected by a flexible, hydrophobic chain. Those paddles are important, because of their inability to hydrogen bond with water, i.e. hydrophobicity, and therefore their propensity to get stuck in contact with other hydrophobic surfaces. The list of candidate hydrophobic surfaces includes the obvious smaller aromatic rings (e.g. phenylalanine), indole double rings (e.g. tryptophan), and the less obvious sugars (e.g. galactose), unsaturated lipid/prostaglandins and basic amino acids (lysine and arginine). These are dominant cellular interactions.

The interchangeability of the hydrophobic paddle-binders means that astaxanthene can get its paddles stuck in enzyme or receptor protein active sites that normally bind a wide range of ligands (target small molecules, e.g. enzyme substrates). It is likely, therefore, that astaxanthene has anti-inflammatory activity, because it blocks an inflammatory interaction.

The ubiquity of interactions of terpenoids, based on their general structural properties, also gives these molecules access to cellular cytoplasm. These molecules are too large to diffuse through membranes and if they got half way through, they would be permanently stuck in the membrane. Terpenoids will tend to stick to carrier proteins that have hydrophobic patches or slots. These carriers will transport and internalize terpenoids and other similarly shaped molecules, e.g. steroid hormones.

Metformin, the diabetes drug, is another example of a molecule with a flat, hydrophobic side. It is a stretch to call this an antioxidant, but it is useful for this discussion, since one of my students tested to see it it would stick to a tryptophan in the active site of a classic enzyme, beta-galactosidase. Galactose, in the typical substrate for this enzyme, lactose, will bind to the active site, because of a prominent tryptophan. The shocker is that my student showed that metformin also binds to that same site and competes with lactose. Astaxanthene would also be expected to bind in the same way.

Curcumin is one of the most potent anti-inflammatory compounds and the main ingredient in turmuric, binds to proteins that inhibit the inflammatory transcription factor, NFkB. I would expect astaxanthene to also inhibit NFkB.

Capsaicin is a related molecule that binds to the heat/pain sensor in skin and blocks pain sensation. That is how capsaicin is used as a topical analgesic. Castor oil, ricinoleate, binds to the same sensor and competes with capsaicin and also is an effective pain reliever. Note that ricinoleate is a modified fatty acid that could curl up on the same hydrophobic paddle surface as capsaicin.

The bottom line of this discussion is that if someone tries to convince you that resveratrol, the anti-aging ingredient in wine, is an anti-oxidant, be skeptical. Expect that resveratrol will have numerous interactions with proteins and many of those will not be known.

I Don’t Believe It ...

There are too many myths in biomedical science. I think that our fears are misplaced and if we just address the real threats, e.g. inflammatory vegetable oils, we can enter the New Year with realistic hopes for good health.

Don’t Worry about Genetic Predispositions to Disease

Mendel was wrong in asserting that there is one gene for each phenotype. Modern clearinghouses of genes reveal the relationships between gene sequences and numerous protein functions. In the Human Protein Reference Database (HPRD), for example, each human gene is listed with all of its synonyms. The synonyms frequently reveal that genes lead multiple, unrelated lives and have sundry tangled relationships with dozens of other proteins. So the idea that there is one gene for each physical trait, phenotype, is a misleading simplification.

The good news is that the complexity would only be a problem, if our individual genetic composition was important to our health. In most cases, I think that our individual genetic predispositions to disease would only be important, if we stress our bodies (usually with an inflammatory diet) to the point of failure. With a healthy diet and lifestyle, I don’t think that most genetic predispositions to disease would ever affect health. To be specific, most people should be able to live until 80 with few physical compromises. This is very hopeful.

Don’t Worry about Drug Side-Effects (avoid the need for drugs)

Drugs have many side effects, simply because we don’t know enough about the interactions of small drug molecules with all of the possible protein targets. Take heparin, for example. Heparin is one of the most commonly used drugs to inhibit blood clotting. There is a clotting assay to measure how well heparin blocks clotting and injecting more heparin makes clotting slower. It appears that heparin binds to a protein involved in clotting. Heparin does bind to anti-thrombin, which in turn inhibits thrombin that is needed for clotting. Heparin also binds to several other proteins required for clotting and also binds to a dozen proteins involved in the complement system needed to kill pathogens or infected cells. Heparin binds to hormones, growth factors and their cell surface receptors. There are hundreds of proteins that use heparin to facilitate attachment to other proteins, to change shape and activity or to enter or leave cells. The point here is that heparin is a vivid example of a drug used to treat blood clotting, while at the same time dozens of other major interactions are ignored.

All drugs have multiple interactions with myriad proteins, but during screening, a drug is identified as having a significant desired affect. All drugs have side effects, because they are not completely specific. The side effects are often exploited as off-label uses for the drugs. Statins, for example, can impact lipid metabolism and change LDL blood levels. Unfortunately, research shows that lowering LDL does not affect heart disease. Fortunately, statins also lower inflammation and inflammation causes heart disease, so statins can be used as very expensive anti-inflammatory drugs.

The hopeful side is that most drugs are not necessary and there are other approaches that are more effective and cheaper to prevent and treat disease.

Don’t Worry about Your Gut Flora

There is a major NIH initiative to identify all of the bacterial species that live in or on the human body. That sounds potentially very useful in the context of all of the hype about pre- and probiotics. Your gut flora are important and people are doing very dumb things by killing off the bacteria that protect their skin from pathogens. The problem is that the concept of species of bacteria is wrong. In your gut, there is so much transfer of DNA between all of the different types of bacteria, that the idea of bacterial species doesn’t work.

Consider the pathogenic, toxin-producing E. coli strain. E. coli gets a bum rap, because we have created a new bacterium by treating cattle with antibiotics. The antibiotics increase fat accumulating in the beef, by altering the gut flora. The antibiotics eliminate some bacteria that normally live by adhering to the rectal areas of the cattle. E. coli can colonize the vacancies, but only if the gut lining releases nutrients and the E. coli is resistant to the antibiotics. The solution comes in the form of a plasmid from another bacterium. The plasmid is a small segment of DNA that carries genes for resistance to several different antibiotics and a toxin that causes intestinal cells to leak. The cattle don’t care much about the new Frankenstein E. coli, but if that plasmid-toting E. coli goes from cowpie to hamburger, and replaces your natural E. coli, it can be deadly.

The point is that the bacteria needed to fill all of the niches in your gut environment are created by approximation from the myriad genes of your whole gut flora community. You are constantly creating new “species” of bacteria. The problem is that you produce your own gut flora based on what you eat and by interactions with your gut. Those interactions can make you healthy or keep you sick. A single bottle of formula, for example, can permanently compromise the gut flora of a breastfed baby. We are learning how to control the development of healthy gut flora, probiotics, by supplements containing particular polysaccharides and oligosaccharides, i.e. prebiotics.

The hopeful side is that diets that have been shown to be healthy also produce healthy gut flora. That isn’t much of a surprise.

Don’t Worry about Toxins

The world is a dangerous place, but people have been there and done that. Plants have been standing around synthesizing a witch’s brew of natural toxins for millions of years. They are so good at it that even the most highly evolved species, bacteria, can’t eat them and survive. You can find a plant toxin that will interact with every human protein. That is why most human drugs are derived from plant toxins.

We know not to eat green potatoes, but pregnant women who make the mistake of eating rotten potatoes will regret the birth defects in their babies. There are so many toxins in plants that we have to be careful to watch what toddlers put in their mouths and every poison control center has to be able to identify plants over the phone to recommend emergency treatment. Pregnant women have a built in system to avoid plant toxins during the embryo’s most vulnerable first trimester. It is called morning sickness. A healthy diet during this time is to avoid vegetables and stick to meat and starch. Women build up baby fat to get through this sensitive time without exposure to plant toxins that their bodies may not be able to detoxify.

The happy thought here is that environmental toxins of human origin are not what make us sick, any more than eating plants generally is not a problem. With a healthy diet, even one modestly contaminated, we will not get sick. With a diet that compromises our health, even natural toxins become a threat.

Don’t Worry about Environmental Estrogens (except to save the rest of the animals)

There is an abundance of synthetic molecules that mimic human sex hormones, but once again, plants have been producing estrogen mimetics for millions of years. The second most abundant biological polymer (macromolecule), after cellulose, is lignin. Lignin is what makes up a large part of plant tissue and is what permits plants to be stiff. Lignin is also the “organic” material in fertile soil. This polymer is black in soil, because it is so complex that it has molecular structural components that can absorb all wavelengths of light. Lignin has no defined structure, but rather it is essentially a combination of thousands of structures.

Now imagine the mucky, black, lignin-laden banks of a forest stream. Tons of complex organic molecules leach into the stream and many of those molecules can mimic the shape of estrogens and bind to human molecules, receptors, that would normally bind to estrogen. It is possible to have excess estrogen leach from a sewage treatment plant, because it is not specifically targeted for removal, and the estrogens could temporarily reach a level to affect sensitive aquatic species, but there are already so many natural sources of estrogenic compounds, that the impact on humans would be expected to be unmeasurably small.

It is certainly possible for humans to pollute streams with other molecules, antimicrobial toothpaste ingredients, for example, that have unusually high environmental stability and bind with unnatural strength to estrogen receptors. Those would be problems like DDT. It would make sense to avoid exotic compounds and use natural plant products where possible, just because biological systems can digest them more readily.

The hopeful perspective here is that we already have ways of avoiding most of the problems with manmade estrogens and although these may be environmental hazards, they probably won’t affect human fertility.

An Anti-Inflammatory Diet and Lifestyle Is the Simple Solution

Prevention is the easiest way to stay healthy and since chronic inflammation is the foundation of degenerative, autoimmune and cancer diseases, an anti-inflammatory diet and lifestyle avoids most of the diseases. These same diseases also provide a modern description of aging, i.e. the consequences of mismanaged chronic inflammation. A healthy body is also the best protection from environmental threats, genetic weaknesses and the dangers of modern medicine.

The hopeful perspective for the New Year is eating and living well can keep you healthy and physically fit into your eighth decade.