tag:blogger.com,1999:blog-10999742052222645382024-03-08T00:09:15.865-08:00Stephanie's PageStephaniehttp://www.blogger.com/profile/00273178904990742948noreply@blogger.comBlogger111125tag:blogger.com,1999:blog-1099974205222264538.post-62279709761382784272011-03-26T07:07:00.000-07:002011-03-26T07:08:15.288-07:00How Statins Really Work Explains Why They Don't Really Work<b>Introduction</b><br /><br />The statin industry has enjoyed a thirty year run of steadily increasing profits, as they find ever more ways to justify expanding the definition of the segment of the population that qualify for statin therapy. Large, placebo-controlled studies have provided evidence that statins can substantially reduce the incidence of heart attack. High serum cholesterol is indeed correlated with heart disease, and statins, by interfering with the body's ability to synthesize cholesterol, are extremely effective in lowering the numbers. Heart disease is the number one cause of death in the U.S. and, increasingly, worldwide. What's not to like about statin drugs? <br /><br />I predict that the statin drug run is about to end, and it will be a hard landing. The thalidomide disaster of the 1950's and the hormone replacement therapy fiasco of the 1990's will pale by comparison to the dramatic rise and fall of the statin industry. I can see the tide slowly turning, and I believe it will eventually crescendo into a tidal wave, but misinformation is remarkably persistent, so it may take years.<br /><br />I have spent much of my time in the last few years combing the research literature on metabolism, diabetes, heart disease, Alzheimer's, and statin drugs. Thus far, in addition to posting essays on the web, I have, together with collaborators, published two journal articles related to metabolism, diabetes, and heart disease (Seneff1 et al., 2011), and Alzheimer's disease (Seneff2 et al., 2011). Two more articles, concerning a crucial role for cholesterol sulfate in metabolism, are currently under review (Seneff3 et al., Seneff4 et al.). I have been driven by the need to understand how a drug that interferes with the synthesis of cholesterol, a nutrient that is essential to human life, could possibly have a positive impact on health. I have finally been rewarded with an explanation for an apparent positive benefit of statins that I can believe, but one that soundly refutes the idea that statins are protective. I will, in fact, make the bold claim that nobody qualifies for statin therapy, and that statin drugs can best be described as toxins.Stephaniehttp://www.blogger.com/profile/00273178904990742948noreply@blogger.com73tag:blogger.com,1999:blog-1099974205222264538.post-41450230529114250622011-03-26T07:06:00.000-07:002011-03-26T07:07:08.454-07:002. Cholesterol and StatinsI would like to start by reexamining the claim that statins cut heart attack incidence by a third. What exactly does this mean? A meta study reviewing seven drug trials, involving in total 42,848 patients, ranging over a three to five year period, showed a 29% decreased risk of a major cardiac event (Thavendiranathan et al., 2006). But because heart attacks were rare among this group, what this translates to in absolute terms is that 60 patients would need to be treated for an average of 4.3 years to protect one of them from a single heart attack. However, essentially all of them will experience increased frailty and mental decline, a subject to which I will return in depth later on in this essay. <br /><br />The impact of the damage due to the statin anti-cholesterol mythology extends far beyond those who actually consume the statin pills. Cholesterol has been demonized by the statin industry, and as a consequence Americans have become conditioned to avoid all foods containing cholesterol. This is a grave mistake, as it places a much bigger burden on the body to synthesize sufficient cholesterol to support the body's needs, and it deprives us of several essential nutrients. I am pained to watch someone crack open an egg and toss out the yolk because it contains "too much" cholesterol. Eggs are a very healthy food, but the yolk contains all the important nutrients. After all, the yolk is what allows the chick embryo to mature into a chicken. Americans are currently experiencing widespread deficiencies in several crucial nutrients that are abundant in foods that contain cholesterol, such as choline, zinc, niacin, vitamin A and vitamin D.<br /><br />Cholesterol is a remarkable substance, without which all of us would die. There are three distinguishing factors which give animals an advantage over plants: a nervous system, mobility, and cholesterol. Cholesterol, absent from plants, is the key molecule that allows animals to have mobility and a nervous system. Cholesterol has unique chemical properties that are exploited in the lipid bilayers that surround all animal cells: as cholesterol concentrations are increased, membrane fluidity is decreased, up to a certain critical concentration, after which cholesterol starts to <i>increase</i> fluidity (Haines, 2001). Animal cells exploit this property to great advantage in orchestrating ion transport, which is essential for both mobility and nerve signal transport. Animal cell membranes are populated with a large number of specialized island regions appropriately called lipid rafts. Cholesterol gathers in high concentrations in lipid rafts, allowing ions to flow freely through these confined regions. Cholesterol serves a crucial role in the non-lipid raft regions as well, by preventing small charged ions, predominantly sodium (Na+) and potassium (K+), from leaking across cell membranes. In the absence of cholesterol, cells would have to expend a great deal more energy pulling these leaked ions back across the membrane against a concentration gradient. <br /><br />In addition to this essential role in ion transport, cholesterol is the precursor to vitamin D3, the sex hormones, estrogen, progesterone, and testosterone, and the steroid hormones such as cortisone. Cholesterol is absolutely essential to the cell membranes of all of our cells, where it protects the cell not only from ion leaks but also from oxidation damage to membrane fats. While the brain contains only 2% of the body's weight, it houses 25% of the body's cholesterol. Cholesterol is vital to the brain for nerve signal transport at synapses and through the long axons that communicate from one side of the brain to the other. Cholesterol sulfate plays an important role in the metabolism of fats via bile acids, as well as in immune defenses against invasion by pathogenic organisms. <br /><br />Statin drugs inhibit the action of an enzyme, HMG coenzyme A reductase, that catalyses an early step in the 25-step process that produces cholesterol. This step is also an early step in the synthesis of a number of other powerful biological substances that are involved in cellular regulation processes and antioxidant effects. One of these is coenzyme Q10, present in the greatest concentration in the heart, which plays an important role in mitochondrial energy production and acts as a potent antioxidant (Gottlieb et al., 2000). Statins also interfere with cell-signaling mechanisms mediated by so-called G-proteins, which orchestrate complex metabolic responses to stressed conditions. Another crucial substance whose synthesis is blocked is dolichol, which plays a crucial role in the endoplasmic reticulum. We can't begin to imagine what diverse effects all of this disruption, due to interference with HMG coenzyme A reductase, might have on the cell's ability to function.Stephaniehttp://www.blogger.com/profile/00273178904990742948noreply@blogger.com26tag:blogger.com,1999:blog-1099974205222264538.post-40547263120289845222011-03-26T07:05:00.000-07:002011-03-26T07:06:37.536-07:003. LDL, HDL, and FructoseWe have been trained by our physicians to worry about elevated serum levels of low density lipoprotein (LDL), with respect to heart disease. LDL is not a type of cholesterol, but rather can be viewed as a container that transports fats, cholesterol, vitamin D, and fat-soluble anti-oxidants to all the tissues of the body. Because they are not water-soluble, these nutrients must be packaged up and transported inside LDL particles in the blood stream. If you interfere with the production of LDL, you will reduce the bioavailability of all these nutrients to your body's cells. <br /><br />The outer shell of an LDL particle is made up mainly of lipoproteins and cholesterol. The lipoproteins contain proteins on the outside of the shell and lipids (fats) in the interior layer. If the outer shell is deficient in cholesterol, the fats in the lipoproteins become more vulnerable to attack by oxygen, ever-present in the blood stream. LDL particles also contain a special protein called "apoB" which enables LDL to deliver its goods to cells in need. ApoB is vulnerable to attack by glucose and other blood sugars, especially fructose. Diabetes results in an increased concentration of sugar in the blood, which further compromises the LDL particles, by gumming up apoB. Oxidized and glycated LDL particles become less efficient in delivering their contents to the cells. Thus, they stick around longer in the bloodstream, and the measured serum LDL level goes up.<br /><br />Worse than that, once LDL particles have finally delivered their contents, they become "small dense LDL particles," remnants that would ordinarily be returned to the liver to be broken down and recycled. But the attached sugars interfere with this process as well, so the task of breaking them down is assumed instead by macrophages in the artery wall and elsewhere in the body, through a unique scavenger operation. The macrophages are especially skilled to extract cholesterol from damaged LDL particles and insert it into HDL particles. Small dense LDL particles become trapped in the artery wall so that the macrophages can salvage and recycle their contents, and this is the basic source of atherosclerosis. HDL particles are the so-called "good cholesterol," and the amount of cholesterol in HDL particles is the lipid metric with the strongest correlation with heart disease, where <i>less</i> cholesterol is associated with increased risk. So the macrophages in the plaque are actually performing a very useful role in increasing the amount of HDL cholesterol and reducing the amount of small dense LDL.<br /><br />The LDL particles are produced by the liver, which synthesizes cholesterol to insert into their shells, as well as into their contents. The liver is also responsible for breaking down fructose and converting it into fat (Collison et al., 2009). Fructose is ten times more active than glucose at glycating proteins, and is therefore very dangerous in the blood serum (Seneff1 et al., 2011). When you eat a lot of fructose (such as the high fructose corn syrup present in lots of processed foods and carbonated beverages), the liver is burdened with getting the fructose out of the blood and converting it to fat, and it therefore can not keep up with cholesterol supply. As I said before, the fats can not be safely transported if there is not enough cholesterol. The liver has to ship out all that fat produced from the fructose, so it produces low quality LDL particles, containing insufficient protective cholesterol. So you end up with a really bad situation where the LDL particles are especially vulnerable to attack, and attacking sugars are readily available to do their damage.Stephaniehttp://www.blogger.com/profile/00273178904990742948noreply@blogger.com20tag:blogger.com,1999:blog-1099974205222264538.post-22105390259023698452011-03-26T07:04:00.000-07:002011-03-26T07:05:46.068-07:004. How Statins Destroy MusclesEurope, especially the U.K., has become much enamored of statins in recent years. The U.K. now has the dubious distinction of being the only country where statins can be purchased over-the-counter, and the amount of statin consumption there has increased more than 120% in recent years (Walley et al, 2005). Increasingly, orthopedic clinics are seeing patients whose problems turn out to be solvable by simply terminating statin therapy, as evidenced by a recent report of three cases within a single year in one clinic, all of whom had normal creatine kinase levels, the usual indicator of muscle damage monitored with statin usage, and all of whom were "cured" by simply stopping statin therapy (Shyam Kumar et al., 2008). In fact, creatine kinase monitoring is not sufficient to assure that statins are not damaging your muscles (Phillips et al., 2002).<br /><br />Since the liver synthesizes much of the cholesterol supply to the cells, statin therapy greatly impacts the liver, resulting in a sharp reduction in the amount of cholesterol it can synthesize. A direct consequence is that the liver is severely impaired in its ability to convert fructose to fat, because it has no way to safely package up the fat for transport without cholesterol (Vila et al., 2011). Fructose builds up in the blood stream, causing lots of damage to serum proteins. <br /><br />The skeletal muscle cells are severely affected by statin therapy. Four complications they now face are: (1) their mitochondria are inefficient due to insufficient coenzyme Q10, (2) their cell walls are more vulnerable to oxidation and glycation damage due to increased fructose concentrations in the blood, reduced choleserol in their membranes, and reduced antioxidant supply, (3) there's a reduced supply of fats as fuel because of the reduction in LDL particles, and (4) crucial ions like sodium and potassium are leaking across their membranes, reducing their charge gradient. Furthermore, glucose entry, mediated by insulin, is constrained to take place at those lipid rafts that are concentrated in cholesterol. Because of the depleted cholesterol supply, there are fewer lipid rafts, and this interferes with glucose uptake. Glucose and fats are the main sources of energy for muscles, and both are compromised.<br /><br />As I mentioned earlier, statins interfere with the synthesis of coenzyme Q10 (Langsjoen and Langsjoen, 2003), which is highly concentrated in the heart as well as the skeletal muscles, and, in fact, in all cells that have a high metabolic rate. It plays an essential role in the citric acid cycle in mitochondria, responsible for the supply of much of the cell's energy needs. Carbohydrates and fats are broken down in the presence of oxygen to produce water and carbon dioxide as by-products. The energy currency produced is adenosine triphosphate (ATP), and it becomes severely depleted in the muscle cells as a consequence of the reduced supply of coenzyme Q10. <br /><br />The muscle cells have a potential way out, using an alternative fuel source, which doesn't involve the mitochondria, doesn't require oxygen, and doesn't require insulin. What it requires is an abundance of fructose in the blood, and fortunately (or unfortunately, depending on your point of view) the liver's statin-induced impairment results in an abundance of serum fructose. Through an anaerobic process taking place in the cytoplasm, specialized muscle fibers skim off just a bit of the energy available from fructose, and produce lactate as a product, releasing it back into the blood stream. They have to process a huge amount of fructose to produce enough energy for their own use. Indeed, statin therapy has been shown to increase the production of lactate by skeletal muscles (Pinieux et al, 1996).<br /><br />Converting one fructose molecule to lactate yields only two ATP's, whereas processing a sugar molecule all the way to carbon dioxide and water in the mitochondria yields 38 ATP's. In other words, you need 19 times as much substrate to obtain an equivalent amount of energy. The lactate that builds up in the blood stream is a boon to both the heart and the liver, because they can use it as a substitute fuel source, a much safer option than glucose or fructose. Lactate is actually an extremely healthy fuel, water-soluble like a sugar but not a glycating agent.<br /><br />So the burden of processing excess fructose is shifted from the liver to the muscle cells, and the heart is supplied with plenty of lactate, a high-quality fuel that does not lead to destructive glycation damage. LDL levels fall, because the liver can't keep up with fructose removal, but the supply of lactate, a fuel that can travel freely in the blood (does not have to be packaged up inside LDL particles) saves the day for the heart, which would otherwise feast off of the fats provided by the LDL particles. I think this is the crucial effect of statin therapy that leads to a reduction in heart attack risk: the heart is well supplied with a healthy alternative fuel.<br /><br />This is all well and good, except that the muscle cells get wrecked in the process. Their cell walls are depleted in cholesterol because cholesterol is in such short supply, and their delicate fats are therefore vulnerable to oxidation damage. This problem is further compounded by the reduction in coenzyme Q10, a potent antioxidant. The muscle cells are energy starved, due to dysfunctional mitochondria, and they try to compensate by processing an excessive amount of both fructose and glucose anaerobically, which causes extensive glycation damage to their crucial proteins. Their membranes are leaking ions, which interferes with their ability to contract, hindering movement. They are essentially heroic sacrificial lambs, willing to die in order to safeguard the heart. <br /><br />Muscle pain and weakness are widely acknowledged, even by the statin industry, as potential side effects of statin drugs. Together with a couple of MIT students, I have been conducting a study which shows just how devastating statins can be to muscles and the nerves that supply them (Liu et al, 2011). We gathered over 8400 on-line drug reviews prepared by patients on statin therapy, and compared them to an equivalent number of reviews for a broad spectrum of other drugs. The reviews for comparison were selected such that the age distribution of the reviewers was matched against that for the statin reviews. We used a measure which computes how likely it would be for the words/phrases that show up in the two sets of reviews to be distributed in the way they are observed to be distributed, if both sets came from the same probability model. For example, if a given side effect showed up a hundred times in one data set and only once in the other, this would be compelling evidence that this side effect was representative of that data set. <i>Table 1 shows several conditions associated with muscle problems that were highly skewed towards the statin reviews.</i> <br /><br />I believe that the real reason why statins protect the heart from a heart attack is that muscle cells are willing to make an incredible sacrifice for the sake of the larger good. It is well acknowledged that exercise is good for the heart, although people with a heart condition have to watch out for overdoing it, walking a careful line between working out the muscles and overtaxing their weakened heart. I believe, in fact, that the reason exercise is good is exactly the same as the reason statins are good: it supplies the heart with lactate, a very healthy fuel that does not glycate cell proteins.Stephaniehttp://www.blogger.com/profile/00273178904990742948noreply@blogger.com24tag:blogger.com,1999:blog-1099974205222264538.post-32542652370312328542011-03-26T06:52:00.000-07:002011-03-26T07:35:07.889-07:00Table 1: Statins and Muscle Damage<table><br /><caption><br />Counts of the number of reviews where phrases associated with various symptoms related to muscles appeared, for 8400 statin and 8400 non-statin drug reviews, along with the associated p-value, indicating the likelihood that this distribution could have occurred by chance.<br /></caption><br /><tr><br /><td><b>Side Effect</b></td><td align="right"><b>Statin Reviews</b></td><td align="right"><b>Non-Statin Reviews</b></td><td align="right"> <b>P-value</b> </td><br /></tr><br /><tr><br /><td>Muscle Cramps</td> <td align="right">678</td> <td align="right"> 193</td> <td align="right">0.00005</td></tr><br /><tr><br /><td>General Weakness</td> <td align="right">687</td> <td align="right">210</td> <td align="right">0.00006</td><br /></tr><br /><tr><br /><td>Muscle Weakness</td> <td align="right">302</td> <td align="right">45</td> <td align="right">0.00023</td><br /></tr><br /><tr><br /><td>Difficulty Walking</td> <td align="right">419</td> <td align="right">128</td> <td align="right">0.00044</td><br /></tr><br /><tr><br /><td>Loss of Muscle Mass</td> <td align="right">54</td> <td align="right">5</td> <td align="right">0.01323</td><br /></tr><br /><tr><br /><td>Numbness</td> <td align="right">293</td> <td align="right">166</td> <td align="right">0.01552</td><br /></tr><br /><tr><br /><td>Muscle Spasms</td> <td align="right">136</td> <td align="right">57</td> <td align="right">0.01849</td><br /></tr><br /></table>Stephaniehttp://www.blogger.com/profile/00273178904990742948noreply@blogger.com9tag:blogger.com,1999:blog-1099974205222264538.post-80201782883594432192011-03-26T06:49:00.000-07:002011-03-26T06:51:41.455-07:005. Membrane Cholesterol Depletion and Ion TransportAs I alluded to earlier, statin drugs interfere with the ability of muscles to contract through the depletion of membrane cholesterol. (Haines, 2001) has argued that the most important role of cholesterol in cell membranes is the inhibition of leaks of small ions, most notably sodium (Na+) and potassium (K+). These two ions are essential for movements, and indeed, cholesterol, which is absent in plants, is the key molecule that permits mobility in animals, through its strong control over ion leakage of these molecules across cell walls. By protecting the cell from ion leaks, cholesterol greatly reduces the amount of energy the cell needs to invest in keeping the ions on the right side of the membrane.<br /><br />There is a widespread misconception that "lactic acidosis," a condition that can arise when muscles are worked to exahustion, is <i>due to </i> lactic acid synthesis. The actual story is the exact opposite: the acid build-up is due to excess breakdown of ATP to ADP to produce energy to support muscle contraction. When the mitochondria can't keep up with energy consumption by renewing the ATP, the production of lactate becomes absolutely necessary to <i>prevent </i> acidosis (Robergs et al., 2004). In the case of statin therapy, excessive leaks due to insufficient membrane cholesterol require <i> more</i> energy to correct, and all the while the mitochondria are producing <i> less</i> energy.<br /><br />In in vitro studies of phospholipid membranes, it has been shown that the removal of cholesterol from the membrane leads to a nineteen fold increase in the rate of potassium leaks through the membrane (Haines, 2001). Sodium is affected to a lesser degree, but still by a factor of three. Through ATP-gated potassium and sodium channels, cells maintain a strong disequilibrium across their cell wall for these two ions, with sodium being kept out and potassium being held inside. This ion gradient is what energizes muscle movement. When the membrane is depleted in cholesterol, the cell has to burn up substantially more ATP to fight against the steady leakage of both ions. With cholesterol depletion due to statins, this is energy it doesn't have, because the mitochondria are impaired in energy generation due to coenzyme-Q10 depletion. <br /><br />Muscle contraction itself causes potassium loss, which further compounds the leak problem introduced by the statins, and the potassium loss due to contraction contributes significantly to muscle fatigue. Of course, muscles with insufficient cholesterol in their membranes lose potassium even faster. Statins make the muscles much more vulnerable to acidosis, both because their mitochondria are dysfunctional and because of an increase in ion leaks across their membranes. This is likely why athletes are more susceptible to muscle damage from statins (Meador and Huey, 2010, Sinzinger and O'Grady, 2004): their muscles are doubly challenged by both the statin drug and the exercise.<br /><br />An experiment with rat soleus muscles in vitro showed that lactate added to the medium was able to almost fully recover the force lost due to potassium loss (Nielsen et al, 2001). Thus, production and release of lactate becomes essential when potassium is lost to the medium. The loss of strength in muscles supporting joints can lead to sudden uncoordinated movements, overstressing the joints and causing arthritis (Brandt et al., 2009). <i>In fact, our studies on statin side effects revealed a very strong correlation with arthritis, as shown in the table</i>.<br /><br />While I am unaware of a study involving <i>muscle</i> cell ion leaks and statins, a study on <i>red blood cells</i> and <i>platelets</i> has shown that there is a substantial increase in the Na+-K+-pump activity after just a month on a modest 10 mg/dl statin dosage, with a concurrent decrease in the amount of cholesterol in the membranes of these cells (Lijnen et al., 1994). This increased pump activity (necessitated by membrane leaks) would require additional ATP and thus consume extra energy.<br /><br />Muscle fibers are characterized along a spectrum by the degree to which they utilize aerobic vs anaerobic metabolism. The muscle fibers that are most strongly damaged by statins are the ones that specialize in anaerobic metabolism (Westwood et al., 2005). These fibers (Type IIb) have very few mitochondria, as contrasted with the abundant supply of mitochondria in the fully aerobic Type 1A fibers. I suspect their vulnerability is due to the fact that they carry a much larger burden of generating ATP to fuel the muscle contraction and to produce an abundance of lactate, a product of anaerobic metabolism. They are tasked with both energizing not only themselves but also the defective aerobic fibers (due to mitochondrial dysfunction) and producing enough lactate to offset the acidosis developing as a consequence of widespread ATP shortages.Stephaniehttp://www.blogger.com/profile/00273178904990742948noreply@blogger.com6tag:blogger.com,1999:blog-1099974205222264538.post-13620756853111478132011-03-26T06:48:00.000-07:002011-03-26T07:25:20.988-07:006. Long-term Statin Therapy Leads to Damage EverywhereStatins, then, slowly erode the muscle cells over time. After several years have passed, the muscles reach a point where they can no longer keep up with essentially running a marathon day in and day out. The muscles start literally falling apart, and the debris ends up in the kidney, where it can lead to the rare disorder, rhabdomyolysis, which is often fatal. <i>In fact, 31 of our statin reviews contained references to "rhabdomyolysis" as opposed to none in the comparison set. Kidney failure, a frequent consequence of rhabdomyolysis, showed up 26 times among the statin reviews, as opposed to only four times in the control set.</i> <br /><br />The dying muscles ultimately expose the nerves that innervate them to toxic substances, which then leads to nerve damage such as neuropathy, and, ultimately Amyloid Lateral Sclerosis (ALS), also known as Lou Gehrig's disease, a very rare, debilitating, and ultimately fatal disease which is now on the rise due (I believe) to statin drugs. People diagnosed with ALS rarely live beyond five years. <i>Seventy-seven of our statin reviews contained references to ALS, as against only 7 in the comparison set.</i> <br /><br />As ion leaks become untenable, cells will begin to replace the potassium/sodium system with a calcium/magnesium based system. These two ions are in the same rows of the periodic table as sodium/potassium, but advanced by one column, which means that they are substantially larger, and therefore it's much harder for them to accidentally leak out. But this results in extensive calcification of artery walls, heart valves, and the heart muscle itself. Calcified heart valves can no longer function properly to prevent backflow, and diastolic heart failure results from increased left ventricular stiffness. Research has shown that statin therapy leads to increased risk to diastolic heart failure (Silver et al., 2004, Weant and Smith, 2005). <i>Heart failure shows up 36 times in our statin drug data as against only 8 times in the comparison group.</i><br /><br />Once the muscles can no longer keep up with lactate supply, the liver and heart will be further imperilled. They're now worse off than they were before statins, because the lactate is no longer available, and the LDL, which would have provided fats as a fuel source, is greatly reduced. So they're stuck processing sugar as fuel, something that is now much more perilous than it used to be, because they are depleted in membrane cholesterol. Glucose entry into muscle cells, including the heart muscle, mediated by insulin, is orchestrated to occur at lipid rafts, where cholesterol is highly concentrated. Less membrane cholesterol results in fewer lipid rafts, and this leads to impaired glucose uptake. Indeed, it has been proposed that statins increase the risk to diabetes (Goldstein and Mascitelli, 2010, Hagedorn and Arora, 2010). <i>Our data bear out this notion, with the probability of the observed distributions of diabetes references happening by chance being only 0.006.</i>Stephaniehttp://www.blogger.com/profile/00273178904990742948noreply@blogger.com32tag:blogger.com,1999:blog-1099974205222264538.post-37591682723795775782011-03-26T06:33:00.000-07:002011-03-26T07:03:43.235-07:00Table 2: Statistics on Statins and Other Major Health IssuesCounts of the number of reviews where phrases associated with<br />various symptoms related to major health issues appeared, besides<br />muscle problems, for 8400 statin and 8400 non-statin drug reviews,<br />along with the associated p-value, indicating the likelihood that this<br />distribution could have occurred by chance.<br /> <br /><table><br /><tr><br /><td><b>Side Effect</b></td><td align="right"><b>Statin Reviews</b></td><td align="right"><b>Non-Statin Reviews</b></td><td align="right"> <b>P-value</b> <br /></td></tr> <br /><tr><br /><td>Rhabdomyolysis</td> <td align="right">31</td> <td align="right"> 0</td> <td align="right">0.02177</td></tr><br /><tr><br /><td>Liver Damage</td> <td align="right">326</td> <td align="right">133</td> <td align="right">0.00285</td><br /><br /></tr><br /><tr><br /><td>Diabetes</td> <td align="right">185</td> <td align="right">62</td> <td align="right">0.00565</td><br /></tr><br /><tr><br /><td>ALS</td> <td align="right">71</td> <td align="right">7</td> <td align="right">0.00819</td><br /></tr><br /><tr><br /><td>Heart Failure</td> <td align="right">36</td> <td align="right">8</td> <td align="right">0.04473</td><br /></tr><br /><tr><br /><td>Kidney Failure</td> <td align="right">26</td> <td align="right">4</td> <td align="right">0.05145</td><br /></tr><br /><br /><tr><br /><td>Arthritis</td> <td align="right">245</td> <td align="right">120</td> <td align="right">0.01117</td><br /></tr><br /><tr><br /><td>Memory Problems</td> <td align="right">545</td> <td align="right">353</td> <td align="right">0.01118</td><br /></tr><br /><br /><tr><br /><td>Parkinson's Disease</td> <td align="right">53</td> <td align="right">3</td> <td align="right">0.01135</td><br /></tr><br /><tr><br /><td>Neuropathy</td> <td align="right">133</td> <td align="right">73</td> <td align="right">0.04333</td><br /></tr><br /><tr><br /><td>Dementia</td> <td align="right">41</td> <td align="right">13</td> <td align="right">0.05598</td><br /></tr><br /></table>Stephaniehttp://www.blogger.com/profile/00273178904990742948noreply@blogger.com4tag:blogger.com,1999:blog-1099974205222264538.post-16737975347107003202011-03-26T06:31:00.002-07:002011-03-26T06:32:36.013-07:007. Statins, Caveolin, and Muscular DystrophyLipid rafts are crucial centers for transport of substances (both nutrients and ions) across cell membranes and as a cell signaling domain in essentially all mammalian cells. Caveolae ("little caves") are microdomains within lipid rafts, which are enriched in a substance called caveolin (Gratton et al., 2004). Caveolin has received increasing attention of late due to the widespread role it plays in cell signaling mechanisms and the transport of materials between the cell and the environment (Smart et al., 1999).<br /><br />Statins are known to interfere with caveolin production, both in endothelial cells (Feron et al., 2001) and in heart muscle cells, where they've been shown to reduce the density of caveolae by 30% (Calaghan, 2010). People who have a defective form of caveolin-3, the version of caveolin that is present in heart and skeletal muscle cells, develop muscular dystrophy as a consequence (Minetti et al., 1998). Mice engineered to have defective caveolin-3 that stayed in the cytoplasm instead of binding to the cell wall at lipid rafts exhibited stunted growth and paralysis of their legs (Sunada et al., 2001). Caveolin is crucial to cardiac ion channel function, which, in turn, is essential in regulating the heart beat and protecting the heart from arrhythmias and cardiac arrest (Maguy et al, 2006). In arterial smooth muscle cells, caveolin is essential to the generation of calcium sparks and waves, which, in turn, are essential for arterial contraction and expansion, to pump blood through the body (Taggart et al, 2010). <br /><br />In experiments involving constricting the arterial blood supply to rats' hearts, researchers demonstrated a 34% increase in the amount of caveolin-3 produced by the rat's hearts, along with a 27% increase in the weight of the left ventricle, indicating ventricular hypertrophy. What this implies is that the heart needs additional caveolin to cope with blocked vessels, whereas statins interfere with the ability to produce extra caveolin (Kikuchi et al., 2005).Stephaniehttp://www.blogger.com/profile/00273178904990742948noreply@blogger.com4tag:blogger.com,1999:blog-1099974205222264538.post-65617451014502658622011-03-26T06:31:00.001-07:002011-03-26T06:31:24.269-07:008. Statins and the BrainWhile the brain is not the focus of this essay, I cannot resist mentioning the importance of cholesterol to the brain and the evidence of mental impairment available from our data sets. Statins would be expected to have a negative impact on the brain, because, while the brain makes up only 2% of the body's weight, it houses 25% of the body's cholesterol. Cholesterol is highly concentrated in the myelin sheath, which encloses axons which transport messages long distances (Saher et al., 2005). Cholesterol also plays a crucial role in the transmission of neurotransmitters across the synapse (Tong et al, 2009). <i> We found highly skewed distribution of word frequencies for dementia, Parkinson's disease, and short term memory loss, with all of these occurring much more frequently in the statin reviews than in the comparison reviews.</i> <br /><br />A recent evidence-based article (Cable, 2009) found that statin drug users had a high incidence of neurological disorders, especially neuropathy, parasthesia and neuralgia, and appeared to be at higher risk to the debilitating neurological diseases, ALS and Parkinson's disease. The evidence was based on careful manual labeling of a set of self-reported accounts from 351 patients. A mechanism for such damage could involve interference with the ability of oligodendrocytes, specialized glial cells in the nervous system, to supply sufficient cholesterol to the myelin sheath surrounding nerve axons. Genetically-engineered mice with defective oligodendrocytes exhibit visible pathologies in the myelin sheath which manifest as muscle twitches and tremors (Saher et al, 2005). Cognitive impairment, memory loss, mental confusion, and depression were also significantly present in Cable’s patient population. Thus, his analysis of 351 adverse drug reports was largely consistent with our analysis of 8400 reports.Stephaniehttp://www.blogger.com/profile/00273178904990742948noreply@blogger.com4tag:blogger.com,1999:blog-1099974205222264538.post-2618629996581757982011-03-26T06:30:00.001-07:002011-03-26T06:30:58.020-07:009. Cholesterol's Benefits to LongevityThe broad spectrum of severe disabilities with increased prevalence in statin side effect reviews all point toward a general trend of increased frailty and mental decline with long-term statin therapy, things that are usually associated with old age. I would in fact best characterize statin therapy as a mechanism to allow you to <i>grow old faster</i>. A highly enlightening study involved a population of elderly people who were monitored over a 17 year period, beginning in 1990 (Tilvis et al., 2011). The investigators looked at an association between three different measures of cholesterol and manifestations of decline. They measured indicators associated with physical frailty and mental decline, and also looked at overall longevity. In addition to serum cholesterol, a biometric associated with the ability to synthesize cholesterol (lathosterol) and a biometric associated with the ability to absorb cholesterol through the gut (sitosterol) were measured. <br /><br /><i>Low</i> values of all three measures of cholesterol were associated with a <i>poorer</i> prognosis for frailty, mental decline and early death. A reduced ability to <i>synthesize</i> cholesterol showed the strongest correlation with poor outcome. Individuals with high measures of all three biometrics enjoyed a 4.3 year extension in life span, compared to those for whom all measures were low. Since statins specifically interfere with the ability to synthesize cholesterol, it is logical that they would also lead to increased frailty, accelerated mental decline, and early death. <br /><br />For both ALS and heart failure, survival benefit is associated with elevated cholesterol levels. A statistically significant inverse correlation was found in a study on mortality in heart failure. For 181 patients with heart disease and heart failure, half of those whose serum cholesterol was below 200 mg/dlk were dead three years after diagnosis, whereas only 28% of the patients whose serum cholesterol was above 200 mg/dl had died. In another study on a group of 488 patients diagnosed with ALS, serum levels of triglycerides and fasting cholesterol were measured at the time of diagnosis (Dorstand et al., 2010). High values for both lipids were associated with improved survival, with a p-value < 0.05.Stephaniehttp://www.blogger.com/profile/00273178904990742948noreply@blogger.com5tag:blogger.com,1999:blog-1099974205222264538.post-74114486060748277852011-03-26T06:28:00.000-07:002011-03-26T06:29:54.119-07:0010. What to do Instead to Avoid Heart DiseaseIf statins don't work in the long run, then what can you do to protect your heart from atherosclerosis? My personal opinion is that you need to focus on natural ways to reduce the number of small dense LDL particles, which feed the plaque, and alternative ways to supply the product that the plaque produces (more about that in a moment). Obviously, you need to cut way back on fructose intake, and this means mainly eating whole foods instead of processed foods. With less fructose, the liver won't have to produce as many LDL particles from the supply side. From the demand side, you can reduce your body's dependency on both glucose and fat as fuel by simply eating foods that are good sources of lactate. Sour cream and yogurt contain lots of lactate, and milk products in general contain the precursor lactose, which gut bacteria will convert to lactate, assuming you don't have lactose intolerance. Strenuous physical exercise, such as a tread machine workout, will help to get rid of any excess fructose and glucose in the blood, with the skeletal muscles converting them to the much coveted lactate. <br /><br />Finally, I have a set of perhaps surprising recommendations that are based on research I have done leading to the two papers that are currently under review (Seneff3 et al, Seneff4 et al.). My research has uncovered compelling evidence that the nutrient that is most crucially needed to protect the heart from atherosclerosis is cholesterol sulfate. The extensive literature review my colleagues and I have conducted to produce these two papers shows compellingly that the fatty deposits that build-up in the artery walls leading to the heart exist mainly for the purpose of extracting cholesterol from glycated small dense LDL particles and synthesizing cholesterol sulfate from it, providing the cholesterol sulfate directly to the heart muscle. The reason the plaque build-up occurs preferentially in the arteries leading to the heart is so that the heart muscle can be assured an adequate supply of cholesterol sulfate. In our papers, we develop the argument that the cholesterol sulfate plays an essential role in the caveolae in the lipid rafts, in mediating oxygen and glucose transport.<br /><br />The skin produces cholesterol sulfate in large quantities when it is exposed to sunlight. Our theory suggests that the skin actually <i>synthesizes</i> sulfate from sulfide, capturing energy from sunlight in the form of the sulfate molecule, thus acting as a solar-powered battery. The sulfate is then shipped to all the cells of the body, carried on the back of the cholesterol molecule. <br /><br />Evidence of the benefits of sun exposure to the heart is compelling, as evidenced by a study conducted to investigate the relationship between geography and cardiovascular disease (Grimes et al., 1996). Through population statistics, the study showed a consistent and striking inverse linear relationship between cardiovascular deaths and estimated sunlight exposure, taking into account percentage of sunny days as well as latitude and altitude effects. For instance, the cardiovascular-related death rate for men between the ages of 55 and 64 was 761 in Belfast, Ireland but only 175 in Toulouse, France. <br /><br />Cholesterol sulfate is very versatile. It is water soluble so it can travel freely in the blood stream, and it enters cell membranes ten times as readily as cholesterol, so it can easily resupply cholesterol to cells. The skeletal and heart muscle cells make good use of the sulfate as well, converting it back to sulfide, and synthesizing ATP in the process, thus recovering the energy from sunlight. This decreases the burden on the mitochondria to produce energy. The oxygen released from the sulfate molecule is a safe source of oxygen for the citric oxide cycle in the mitochondria. <br /><br />So, in my view, the best way to avoid heart disease is to assure an abundance of an alternative supply of cholesterol sulfate. First of all, this means eating foods that are rich in both cholesterol and sulfur. Eggs are an optimal food, as they are well supplied with both of these nutrients. But secondly, this means making sure you get plenty of sun exposure to the skin. This idea flies in the face of the advice from medical experts in the United States to avoid the sun for fear of skin cancer. I believe that the excessive use of sunscreen has contributed significantly, along with excess fructose consumption, to the current epidemic in heart disease. And the natural tan that develops upon sun exposure offers far better protection from skin cancer than the chemicals in sunscreens.Stephaniehttp://www.blogger.com/profile/00273178904990742948noreply@blogger.com20tag:blogger.com,1999:blog-1099974205222264538.post-59959211461020424812011-03-26T06:25:00.000-07:002011-03-26T06:28:19.645-07:0011. Concluding RemarksEvery individual gets at most only one chance to grow old. When you experience your body falling apart, it is easy to imagine that this is just due to the fact that you are advancing in age. I think the best way to characterize statin therapy is that it makes you grow older faster. Mobility is a great miracle that cholesterol has enabled in all animals. By suppressing cholesterol synthesis, statin drugs can destroy that mobility. No study has shown that statins improve all-cause mortality statistics. But there can be no doubt that statins will make your remaining days on earth a lot less pleasant than they would otherwise be.<br /><br />To optimize the quality of your life, increase your life expectancy, and avoid heart disease, my advice is simple: spend significant time outdoors; eat healthy, cholesterol-enriched, animal-based foods like eggs, liver, and oysters; eat fermented foods like yogurt and sour cream; eat foods rich in sulfur like onions and garlic. And finally, say "no, thank-you" to your doctor when he recommends statin therapy.Stephaniehttp://www.blogger.com/profile/00273178904990742948noreply@blogger.com5tag:blogger.com,1999:blog-1099974205222264538.post-25669023263467201292011-03-26T06:17:00.000-07:002011-03-26T06:23:52.659-07:00References for "Why Statins Don't Really Work"[1] K.D. Brandt, P. Dieppe, E. Radin, "Etiopathogenesis of osteoarthritis". <i>Med. Clin. North Am.</i> 93 (1): 1–24, 2009.<br /><br />[2] J. Cable, "Adverse Events of Statins - An Informal Internet-based Study," JOIMR, 7(1), 2009.<br /><br /> [3] S. Calaghan, "Caveolae as key regulators of cardiac myocyte beta2 adrenoceptor signalling: a novel target for statins" <i>Research Symposium on Caveolae: Essential Signalosomes for the Cardiovascular System, Proc Physiol Soc</i> 19, SA21, University of Manchester, 2010.<br /><br />[4] K.S. Collison, S.M. Saleh, R.H. Bakheet, R.K. Al-Rabiah, A.L. Inglis, N.J. Makhoul, Z.M. Maqbool, M. Zia Zaidi, M.A. Al-Johi and F.A. Al-Mohanna, "Diabetes of the Liver: The Link Between Nonalcoholic Fatty Liver Disease and HFCS-55" <i>Obesity</i>, 17(11), 2003-2013, Nov. 2009.<br /><br />[5] J. Dorstand, P. Ku ̈hnlein, C. Hendrich, J. Kassubek, A.D. Sperfeld, and A.C. Ludolph. "Patients with elevated triglyceride and cholesterol serum levels have a prolonged survival in amyotrophic lateral sclerosis," <i>J Neurol.</i> in Press:Published online Dec. 3 2010.<br /><br />[6] O. Feron, C. Dessy, J.-P. Desager, andJ.-L. Balligand, "Hydroxy-Metholglutaryl-Coenzyme A Reductase Inhibition Promotes Endothelial Nitric Oxide Synthase Activation Through a Decrease in Caveolin Abundance," <i>Circulation</i> 103, 113-118, 2001.<br /><br />[7] M.R. Goldstein and L. Mascitelli, "Statin-induced diabetes: perhaps, its the tip of the iceberg," <i>QJM</i>, Published online, Nov 30, 2010.<br /><br />[8] S.S. Gottlieb, M. Khatta, and M.L. Fisher. "Coenzyme Q10 and congestive heart failure." <i>Ann Intern Med</i>, 133(9):745–6, 2000.<br /><br />[9] J.-P. Gratton, P. Bernatchez, and W.C. Sessa, "Caveolae and Caveolins in the Cardiovascular System," <i>Circulation Research</i>, 94:1408-1417, June 11, 2004.<br /><br />[10] D.S. Grimes, E. Hindle and T. Dyer, "Sunlight, Cholesterol and Coronary Heart Disease," <i>Q. J. Med</i> 89, 579-589, 1996; http://www.ncbi.nlm.nih.gov/pubmed/8935479<br /><br />[11] J. Hagedorn and R. Arora, "Association of Statins and Diabetes Mellitus," <i>American Journal of Therapeutics</i>, 17(2):e52, 2010.<br /><br />[12] T.H. Haines, "Do Sterols Reduce Proton and Sodium Leaks through Lipid Bilayers?" <i>Progress in Lipid Research</i>, 40, 299-324., 2001; http://www.ncbi.nlm.nih.gov/pubmed/11412894<br /><br />[13] T. Kikuchi, N. Oka, A. Koga, H. Miyazaki, H. Ohmura, and T. Imaizumi, "Behavior of Caveolae and Caveolin-3 During the Development of Myocyte Hypertrophy," <i>J Cardiovasc Pharmacol.</i> 45:3, 204-210, March 2005. <br /><br />[14] P.H. Langsjoen and A.M. Langsjoen, "The clinical use of HMG CoA-reductase inhibitors and the associated depletion of coenzyme Q10. A review of animal and human publications." <i>Biofactors</i>, 18(1):101–111, 2003.<br /><br />[15] P. Lijnen, H. Celis, R. Fagard, J. Staessen, and A. Amery, "Influence of cholesterol lowering on plasma membrane lipids and cationic transport systems," <i>J. Hypertension</i>, 12:59-64, 1994.<br /><br />[16] J. Liu, A. Li and S. Seneff, "Automatic Drug Side Effect Discovery from Online Patient-Submitted Reviews: Focus on Statin Drugs." <i>Submitted to First International Conference on Advances in Information Mining and Management (IMMM)</i> Jul 17-22, 2011, Bournemouth, UK.<br /><br />[17] A. Maguy, T.E. Hebert, and S. Nattel, "Involvement of Lipid rafts and Caveolae in cardiac ion channel function," <i>Cardiovascular Research</i>, 69, 798-807, 2006.<br /><br />[18] B.M. Meador and K.A. Huey, "Statin-Associated Myopathy and its Exacerbation with Exercise," <i>Muscle and Nerve</i>, 469-79, Oct. 2010.<br /><br />[19] C. Minetti, F. Sotgia, C. Bruno, et al., "Mutations in the caveolin-3 gene cause autosomal dominant limb-girdle muscular dystrophy," <i>Nat. Genet.</i>, 18, 365-368, 1998.<br /><br />[20] O.B. Nielsen, F. de Paoli, and K. Overgaard, "Protective effects of lactic acid on force production in rat skeletal muscles." <i>J. Phhsiology</i> 536(1), 161-166, 2001.<br /><br />[21] P.S. Phillips, R.H. Haas, S. Bannykh, S. Hathaway, N.L. Gray, B.J. Kimura, G. D. Vladutiu, and J.D.F. England. "Statin-associated myopathy with normal creatine kinase levels," <i>Ann Intern Med</i>, October 1, 2002;137:581–5. <br /><br />[22] G. de Pinieux, P. Chariot, M. Ammi-Said, F. Louarn, J.L. LeJonc, A. Astier, B. Jacotot, and R. Gherardi, "Lipid-lowering drugs and mitochondrial function: effects of HMG-CoA reducase inhibitors on serum ubiquinone and blood lactate/pyruvate ratios." <i>Br. J. Clin. Pharmacol.</i> 42: 333-337, 1996.<br /><br />[23] R.A. Robergs, F. Ghiasvand, and D. Parker, "Biochemistry of exercise-induced metabolic acidosis." <i>Am J Physiol Regul Integr Comp Physiol</i> 287: R502–R516, 2004. <br><br /><br />[24] G. Saher, B. Brügger, C. Lappe-Siefke, et al. "High cholesterol level is essential for myelin membrane growth." <i>Nat Neurosci</i> 8:468-75, 2005.<br /><br />[25] S. Seneff, G. Wainwright, and L. Mascitelli, "Is the Metabolic Syndrome Caused by a High Fructose, and Relatively Low Fat, Low Cholesterol Diet?" <i>Archives of Medical Science</i>, 7(1), 8-20, 2011; DOI: 10.5114/aoms.2011.20598<br /><br />[26] S. Seneff, G. Wainwright, and L. Mascitelli, "Nutrition and Alzheimer's Disease: the Detrimental Role of a High Carbohydrate Diet," <i>In Press, European Journal of Internal Medicine</i>, 2011. <br /><br />[27] S. Seneff, G. Wainwright and B. Hammarskjold, "Cholesterol Sulfate Supports Glucose and Oxygen Transport into Erythrocytes and Myocytes: a Novel Evidence Based Theory," <i>submitted to Hypotheses in the Life Sciences.</i><br /><br />[28] S. Seneff, G. Wainwright and B. Hammarskjold, "Atherosclerosis may Play a Pivotal Role in Protecting the Myocardium in a Vulnerable Situation," <i>submitted to Hypotheses in the Life Sciences.</i><br /><br />[29] H. Sinzinger and J. O’Grady, "Professional athletes suffering from familial hypercholesterolaemia rarely tolerate statin treatment because of muscle problems." <i>Br J Clin Pharmacol</i> 57,525-528, 2004.<br /><br />[30] E.J. Smart, G.A. Graf, M.A. McNiven, W.C. Sessa, J.A. Engelman, P.E. Scherer, T. Okamoto, and M.P. Lisanti, "Caveolins, Liquid-Ordered Domains, and Signal Transduction," <i>Molecular and Cellular Biology</i>, 19, 7289–7304, Nov. 1999.<br /><br />[31] A.J. Shyam Kumar, S.K. Wong, and G. Andrew, "Statin-induced muscular symptoms : A report of 3 cases." <i>Acta Orthop. Belg.</i> 74, 569-572, 2008. <br /><br />[32] M.A. Silver, P.H. Langsjoen, S. Szabo, H. Patil, and A. Zelinger, "Effect of atorvastatin on left ventricular diastolic function and ability of coenzyme Q10 to reverse that dysfunction." <i>The American Journal of Cardiology</i>, 94(10):1306–1310, 2004.<br /><br />[33] Y. Sunada, H. Ohi, A. Hase, H. Ohi, T. Hosono, S. Arata, S. Higuchi, K. Matsumura, and T. Shimizu, "Transgenic mice expressing mutant caveolin-3 show severe myopathy associated with increased nNOS activity," <i>Human Molecular Genetics</i> 10(3) 173-178, 2001. http://hmg.oxfordjournals.org/content/10/3/173.abstract<br /><br />[34] M. J. Taggart, "The complexity of caveolae: a critical appraisal of their role in vascular function," <i>Research Symposium on Caveolae: Essential Signalosomes for the Cardiovascular System, Proc Physiol Soc</i> 19, SA21, University of Manchester, 2010.<br /><br />[35] P. Thavendiranathan, A.Bagai, M.A. Brookhart, and N.K. Choudhry, "Primary prevention of cardiovascular diseases with statin therapy: a meta-analysis of randomized controlled trials," <i>Arch Intern Med.</i> 166(21), 2307-13., Nov 27, 2006.<br /><br />[36] R.S. Tilvis, J.N. Valvanne, T.E. Strandberg and T.A. Miettinen "Prognostic significance of serum cholesterol, lathosterol, and sitosterol in old age; a 17-year population study," <i>Annals of Medicine</i>, Early Online, 1–10, 2011. <br /><br />[37] J. Tong, P.P. Borbat, J.H. Freed, and Y. Shin, "A scissors mechanism for stimulation of SNARE-mediated lipid mixing by cholesterol." <i>Proc Natl Acad Sci U S A</i> 2009;106:5141-6.<br /><br />[38] L. Vila, A. Rebollo, G.S. Ađalsteisson, M. Alegret, M. Merlos, N. Roglans, and J.C. Laguna, "Reduction of liver fructokinase expression and improved hepatic inflammation and metabolism in liquid fructose-fed rats after atorvastatin treatment," <i>Toxicology and Applied Pharmacology</i> 251, 32–40, 2011.<br /><br />[39] Walley T., Folino-Gallo P., Stephens P et al, "Trends in prescribing and utilisation of statins and other lipid lowering drugs across Europe 1997-2003," <i>Br J Clin Pharmacol</i> 60, 543-551, 2005.<br /><br />[40] K.A. Weant and K.M. Smith, "The Role of Coenzyme Q10 in Heart Failure," <i>Ann Pharmacother</i>, 39(9), 1522-6, Sep. 2005.<br /><br />[41] F. R. Westwood, A. Bigley, K. Randall, A.M. Marsden, and R.C. Scott, "Statin-induced muscle necrosis in the rat: distribution, development, and fibre selectivity," <i>Toxicologic Pathology</i>, 33:246-257, 2005.Stephaniehttp://www.blogger.com/profile/00273178904990742948noreply@blogger.com5tag:blogger.com,1999:blog-1099974205222264538.post-10865068616967207172010-09-27T16:28:00.002-07:002010-09-27T16:33:29.097-07:00Could Sulfur Deficiency be a Contributing Factor in Obesity, Heart Disease, Alzheimer's and Chronic Fatigue Syndrome?<b> 1. Introduction </b><br /><br />Obesity is quickly becoming the number one health issue confronting America today, and has also risen to epidemic proportions worldwide. Its spread has been associated with the adoption of a Western-style diet. However, I believe that the widespread consumption of food imports produced by U.S. companies plays a crucial role in the rise in obesity worldwide. Specifically, these "fast foods" typically include heavily processed derivatives of corn, soybeans, and grains, grown on highly efficient mega-farms. Furthermore, I will argue in this essay that one of the core underlying causes of obesity may be sulfur deficiency. <br /><br />Sulfur is the eighth most common element by mass in the <a href="http://en.wikipedia.org/wiki/Composition_of_the_human_body"> <font color="red"> human body</font></a>, behind oxygen, carbon, hydrogen, nitrogen, calcium, phosphorus, and potassium. The two sulfur-containing amino acids, <a href="http://www.biology.arizona.edu/biochemistry/problem_sets/aa/sulfur.html"><font color="red"> methionine and cysteine</font></a>, play essential physiological roles throughout the body. However, sulfur has been consistently overlooked in addressing the issues of nutritional deficiencies. In fact, the American Food and Drug Administration has not even assigned a minimum daily requirement (MDR) for sulfur. One consequence of sulfur's limbo nutritional status is that it is omitted from the long list of supplements that are commonly artificially added to popular foods like cereal. <br /><br />Sulfur is found in a large number of foods, and, as a consequence, it is assumed that almost any diet would meet the minimum daily requirements. Excellent sources are eggs, onions, garlic, and leafy dark green vegetables like kale and broccoli. Meats, nuts, and seafood also contain sulfur. Methionine, an essential amino acid, in that we are unable to synthesize it ourselves, is found mainly in egg whites and fish. A diet high in grains like bread and cereal is likely to be deficient in sulfur. Increasingly, whole foods such as corn and soybeans are disassembled into component parts with chemical names, and then reassembled into heavily processed foods. Sulfur is lost along the way, and there is a lack of awareness that this matters.<br /><br />Experts have recently become aware that sulfur depletion in the soil creates a serious deficiency for plants [Jez2008], brought about in part by improved efficiency in farming and in part, ironically, by successful attempts to clean up air pollution. Over the last two decades, the U.S. farming industry has steadily consolidated into highly technologized mega farms. The high yield per acre associated with these farms results in greater depletion of sulfur each year by the tall, densely planted crops. Plants require sulfur in the form of the sulfate radical (SO<sub>4</sub><sup>-2</sup>). Bacteria in well aerated soil, similar to nitrogen fixing bacteria, can convert elemental sulfur into sulfate through an oxidation process. Coal contains a significant amount of sulfur, and factories that burn coal for energy release sulfur dioxide into the air. Over time, sun exposure converts the sulfur dioxide to sulfate, a significant contributor to acid rain. Acid rain is a serious pollutant, in that hydrogen sulfate, a potent acid, penetrates lakes, making them too acidic for lifeforms to thrive. The Clean Air Act, enacted by congress in 1980, has led to substantial decreases in the amount of acid rain released into the atmosphere. Factories have introduced highly effective scrubbing technologies to comply with the law, and, as a consequence, less sulfate makes its way back into the soil.<br /><br />Modern farmers apply highly concentrated fertilizer to their soil, but this fertilizer is typically enriched in phosphates and often contains no sulfur. Excess phosphates interfere with sulfur absorption. In the past, organic matter and plant residues remained after the fruit and grain were harvested. Such accumulating organic matter used to be a major source of recyclable sulfur. However, many modern machinery-based methods remove a great deal more of the organic matter in addition to the edible portions of the plant. So the sulfur in the decaying organic matter is also lost.<br /><br />It is estimated that humans obtain about 10% of their sulfur supply from drinking water. Remarkably, people who drink soft water have an increased risk to heart disease compared to people who drink hard water [Crawford1967]. Many possible reasons have been suggested for why this might be true ( <a href="http://www.mgwater.com/exhibitb.shtml"> <font color = "red" >Proposed theories for soft water/hard water differences in heart disease</font></a>), and just about every trace metal has been considered as a possibility [Biorck1965]. However, I believe that the real reason may simply be that hard water is more likely to contain sulfur. The sulfate ion is the most useful form of sulfur for humans to ingest. Water softeners provide a convenient environment for sulfur-reducing bacteria, which convert sulfate (SO<sub>4</sub><sup>-2</sup>) into sulfide (S<sup>-2</sup>), emitting hydrogen sulfide gas. Hydrogen sulfide gas is a poison that has been known to cause nausea, illness and, in extreme cases, death. When the bacteria are thriving, the gas will diffuse into the air and give off a foul odor. Obviously, it is rare that the concentration is sufficiently high to cause severe problems. But the sulfate ion is lost through the process. Water that is naturally soft, such as water collected from rain run-off, also contains little or no sulfur, because it has gone through an evaporation-condensation cycle, which leaves behind all the heavier molecules, including sulfur.Stephaniehttp://www.blogger.com/profile/00273178904990742948noreply@blogger.com5tag:blogger.com,1999:blog-1099974205222264538.post-63378985695628244602010-09-27T16:28:00.001-07:002010-09-27T16:28:29.384-07:00 2. Sulfur Availability and Obesity Rates The ultimate source of sulfur is volcanic rock, mainly basalt, spewed up from the earth's core during volcanic eruptions. It is generally believed that humans first evolved from a common ape ancestor in the African rift zone, a region that would have enjoyed an abundance of sulfur due to the heavy volcanic activity there. The three principle suppliers of sulfur to the Western nations are Greece, Italy and Japan. These three countries also enjoy low rates of heart disease and obesity and increased longevity. In South America, a line of volcanoes tracks the backbone of Argentina. Argentinians have a much lower obesity rate than their neighbors to the east in Brazil. In the United States, Oregon and Hawaii, two states with significant volcanic activity, have among the lowest obesity rates in the country. By contrast, the highest obesity rates are found in the midwest and southern farm country: the epicenter of the modern agricultural practices (mega farms) that lead to sulfur depletion in the soil. Among all fifty states, <a href="http://www.cbsnews.com/stories/2010/05/03/health/main6456992.shtml"> <font color = "red" >Oregon</font></a> has the lowest childhood obesity rates. Significantly, <a href= "http://www.bizjournals.com/pacific/stories/2009/06/29/daily36.html"> <font color = "red" >Hawaii</font></a>'s youth are faring less well than their parents: while Hawaii ranks as the fifth from the bottom in obesity rates, its children aged 10-17 weigh in at number 13. As Hawaii has recently become increasingly dependent on food imports from the mainland to supply their needs, they have suffered accordingly with increased obesity problems.Stephaniehttp://www.blogger.com/profile/00273178904990742948noreply@blogger.com10tag:blogger.com,1999:blog-1099974205222264538.post-26279583564997728862010-09-27T16:27:00.004-07:002010-09-27T16:28:10.428-07:00 3. Why Does Sulfur Deficiency Lead to Obesity? To summarize what has been said thus far, (1) foods are becoming depleted in sulfur, and (2) locations with naturally high sulfur deposits enjoy protection against obesity. Now comes the difficult question: why does sulfur deficiency lead to obesity? The answer, like much of biology, is complicated, and part of what I theorize is conjecture.<br /><br />Sulfur is known as a healing mineral, and a sulfur deficiency often leads to pain and inflammation associated with various muscle and skeletal disorders. Sulfur plays a role in many biological processes, one of which is metabolism. Sulfur is present in insulin, the essential hormone that promotes the utilization of sugar derived from carbohydrates for fuel in muscle and fat cells. However, my extensive literature search has led me to two mysterious molecules found in the blood stream and in many other parts of the body: vitamin D3 <i>sulfate</i> and cholesterol <i>sulfate</i> [Strott2003]. Upon exposure to the sun, the skin synthesizes vitamin D3 sulfate, a form of vitamin D that, unlike unsulfated vitamin D3, is water soluble. As a consequence, it can travel freely in the blood stream rather than being packaged up inside LDL (the so-called "bad" cholesterol) for transport [Axelsona1985]. The form of vitamin D that is present in both human milk [Lakdawala1977] and raw cow's milk [Baulch1982] is vitamin D3 sulfate (pasteurization destroys it in cow's milk, and the milk is then artificially enriched with vitamin D2, an unsulfated plant-derived form of the vitamin). <br /><br /><i>Cholesterol</i> sulfate is also synthesized in the skin, where it forms a crucial part of the barrier that keeps out harmful bacteria and other microorganisms such as fungi [Strott2003]. Cholesterol sulfate regulates the gene for a protein called profilaggrin, by interacting like a hormone with the nuclear receptor ROR-alpha. Profilaggrin is the precursor to filaggrin, which protects the skin from invasive organisms [Sandilands2009, McGrath2008]. A deficiency in filaggrin is associated with asthma and arthritis. Therefore, cholesterol sulfate plays an important role in protection from asthma and arthritis. This explains why sulfur is a healing agent. <br /><br />Like vitamin D3 sulfate, cholesterol sulfate is also water-soluble, and it too, unlike cholesterol, does not have to be packaged up inside LDL for delivery to the tissues. By the way, vitamin D3 is synthesized through a couple of simple steps from cholesterol, and its chemical structure is, as a consequence, nearly identical to that of cholesterol. <br /><br />Here I pose the interesting question: where do vitamin D3 sulfate and cholesterol sulfate go once they are in the blood stream, and what role do they play in the cells? Surprisingly, as far as I can tell, nobody knows. It has been determined that the sulfated form of vitamin D3 is strikingly <i>ineffective</i> for calcium transport, the well-known "primary" role of vitamin D3 [Reeve1981]. However, vitamin D3 clearly has many other positive effects (it seems that more and more are being discovered every day), and these include a role in cancer protection, increased immunity against infectious disease, and protection against heart disease (<a href= "http://health.usnews.com/health-news/diet-fitness/diabetes/articles/2010/08/24/vitamin-d-may-influence-genes-for-cancer-autoimmune-disease.html"> <font color = "red" >Vitamin D Protects against Cancer and Autoimmune Diseases</font></a>). Researchers don't yet understand how it achieves these benefits, which have been observed empirically but remain unexplained physiologically. However, I strongly suspect it is the sulfated form of the vitamin that instantiates these benefits, and my reasons for this belief will become clearer in a moment.<br /><br />One very special feature of cholesterol sulfate, as opposed to cholesterol itself, is that it is very agile: due to its polarity it can freely pass through cell membranes almost like a ghost [Rodriguez1995]. This means that cholesterol sulfate can easily enter a fat or muscle cell. I am developing a theory which at its core proposes an essential role for cholesterol sulfate in the metabolism of glucose for fuel by these cells. Below, I will show how cholesterol sulfate may be able to protect fat and muscle cells from damage due to exposure to glucose, a dangerous reducing agent, and to oxygen, a dangerous oxidizing agent. I will further argue that, with insufficient cholesterol sulfate, muscle and fat cells become damaged, and as a consequence become glucose intolerant: unable to process glucose as a fuel. This happens first to muscle cells but eventually to fat cells, as well. Fat cells become storage bins for fats to supply fuel to the muscles, because the muscles are unable to utilize glucose as fuel. Eventually, fat cells also become too disabled to release their stored fats. Fatty tissue then accumulates on the body.Stephaniehttp://www.blogger.com/profile/00273178904990742948noreply@blogger.com4tag:blogger.com,1999:blog-1099974205222264538.post-65833009899248000292010-09-27T16:27:00.003-07:002010-09-27T16:27:46.237-07:00 4. Sulfur and Glucose Metabolism In order to understand my theory, you will need to know more about glucose metabolism. Skeletal muscle cells and fat cells break down glucose in the presence of oxygen in their mitochondria, and in the process they produce ATP, the basic energy currency of all cells. A glucose transporter called GLUT4 is present in the cytoplasm of muscle cells, and it migrates to the cell membrane upon stimulation by insulin. GLUT4 essentially acts as a key that unlocks the door, letting glucose into the cell, but, like a key, it only works when it's inserted in the membrane. Both glucose and oxygen, unless they are carefully managed, can cause harm to the cell's proteins and fats. The glucose enters the cell within special cholesterol rich sites in the cell wall called lipid rafts [Inoue2006]. This is likely orchestrated to protect the cell wall from damage, because extra cholesterol allows the vulnerable lipoproteins in the cell wall to pack more tightly and reduce their risk of exposure. In muscle cells, myoglobin is able to store additional oxygen, bound to an iron molecule safely sequestered in an interior cavity within the myoglobin protein. <br /><br />Sulfur is a very versatile molecule, because it can exist in several distinct oxidative states, ranging from +6 (in the sulfate radical) to -2 (in hydrogen sulf<i>ide</i>). Glucose, as a powerful reducing agent, can cause significant glycation damage to exposed proteins, leading to the formation of Advanced Glycation End Products (AGE's) that are extremely destructive to health: they are believed to be a major contributor to heart disease risk [Brownlee1988]. So, I hypothesize that, if sulfur (+6) is made available to glucose as a decoy, the glucose will be diverted into reducing the sulfur rather than glycating some vulnerable protein such as myoglobin.<br /><br />In searching the Web, I came across an article written in the 1930's about the striking ability of <i>iron sulfate</i>, in the presence of the oxidizing agent hydrogen peroxide, to break down starch into simple molecules, even in the absence of any enzymes to catalyze the reaction [Brown1936]. The article pointedly mentioned that <i>iron</i> works much better than other metals, and <i>sulfate</i> works much better than other anions. In the human body, starch is first converted to glucose in the digestive system. The muscle and fat cells only need to break down glucose. Thus, their task is easier, because the iron sulfate is now starting from an intermediate breakdown product of starch rather than from starch itself.<br /><br />Where would the iron sulfate come from? It seems to me that the cholesterol sulfate, having hopped across the cell membrane, could transfer its sulfate radical to the myoglobin, whose iron molecule could provide the other half of the formula. In the process, the sulfur molecule's charge would be driven down from +6 to -2, releasing energy and absorbing the impact of the reducing effects of glucose, and therefore serving as a decoy to protect the proteins in the cell from glycation damage. <br /><br />When the cell is exposed to insulin, its mitochondria are triggered to start pumping both hydrogen peroxide and hydrogen ions into the cytoplasm, essentially gearing up for the assault by glucose. If cholesterol sulfate enters the cell alongside the glucose, then all the players are available. I conjecture that cholesterol sulfate is the catalyst that seeds the lipid raft. Iron sulfate is then formed by bonding the iron in the heme unit in myoglobin to a sulfate ion provided by cholesterol sulfate. The cholesterol is left behind in the cell wall, thus enriching the newly forming lipid raft with cholesterol. The hydrogen peroxide, provided by the mitochondria upon insulin stimulation, catalyzes the dissolution of glucose by the iron sulfate. The pumped hydrogen can pair up with the reduced sulfur (S<sup>-2</sup>) to form hydrogen sulfide, a gas that can easily diffuse back across the membrane for a repeat cycle. The oxygen that is released from the sulfate radical is picked up by the myoglobin, sequestered inside the molecule for safe travel to the mitochondria. Glucose breakdown products and oxygen are then delivered to the mitochondria to complete the process that ends with water, carbon dioxide, and ATP -- all while keeping the cell's cytoplasmic proteins safe from glucose and oxygen exposure.<br /><br />If I'm right about this role for cholesterol sulfate both in seeding the lipid raft and in providing the sulfate ion, then this process breaks down miserably when cholesterol sulfate is not available. First of all, the lipid raft is not formed. Without the lipid raft, the glucose can not enter the cell. Intense physical exercise can allow glucose to enter the muscle cells even in the absence of insulin [Ojuka2002]. However, this will lead to dangerous exposure of the cell's proteins to glycation (because there is no iron sulfate to degrade the glucose). Glycation interferes with the proteins' ability to perform their jobs, and leaves them more vulnerable to oxidation damage. One of the important affected proteins would be myoglobin: it would no longer be able to effectively carry oxygen to the mitochondria. Furthermore, oxidized myoglobin released into the blood stream by crippled muscle cells leads to painful and crippling rhabdomyolysis, and possible subsequent kidney failure. This explanation accounts for the observation that sulfur deficiency leads to muscle pain and inflammation.Stephaniehttp://www.blogger.com/profile/00273178904990742948noreply@blogger.com3tag:blogger.com,1999:blog-1099974205222264538.post-7566160604190762952010-09-27T16:27:00.001-07:002010-09-27T16:27:24.675-07:00 5. The Metabolc Syndrome The metabolic syndrome is a term used to encapsulate a complex set of markers associated with increased risk to heart disease. The profile includes (1) insulin resistance and dysfunctional glucose metabolism in muscle cells, (2) excess triglycerides in the blood serum, (3) high levels of LDL, particularly small dense LDL, the worst kind, (4) low levels of HDL (the "good" cholesterol) and reduced cholesterol content within the individual HDL particles, (5) elevated blood pressure, and (6) obesity, particularly excess abdominal fat. I have argued previously that this syndrome is brought on by a diet that is high in empty carbohydrates (particularly fructose) and low in fats and cholesterol, along with a poor vitamin D status [Seneff2010]. While I still believe that all of these factors are contributory, I would now add another factor as well: insufficient dietary sulfate.<br /><br />I have described in a <a href= "http://people.csail.mit.edu/seneff/obesity_epidemic_metabolic_syndrome.html"> <font color = "red" >previous essay</font></a>, my interpretation of obesity as being driven by a need for abundant fat cells to convert glucose to fat because the muscle cells are unable to efficiently utilize glucose as fuel. With sulfur deficiency comes the answer as to why muscle cells would be defective in glucose management: they can't come up with enough cholesterol sulfate to seed the lipid raft needed to import the glucose.<br /><br /> An alternative way to ovecome a muscle cell's defective glucose metabolism is to exercise vigorously, so that the generated AMPK (an indicator of energy shortage) induces the GLUT4 to migrate to the membrane even in the absence of insulin [Ojuka2002]. Once the glucose is inside the muscle cell, however, the iron-sulfate mechanism just described is dysfunctional, both because there's no cholesterol sulfate and because there's no hydrogen peroxide. Additionally, with intensive exercise there's also a reduced supply of oxygen, so the glucose must be processed anaerobically in the cytoplasm to produce lactate. The lactate is released into the blood stream and shipped to the heart and brain, both of which are able to use it as fuel. But the cell membrane remains depleted in cholesterol, and this makes it vulnerable to future oxidative damage.<br /><br />Another way to compensate for defective glucose metabolism in the muscle cells is to gain weight. Fat cells must now convert glucose into fat and release it into the blood stream as triglycerides, to fuel the muscle cells. In the context of a low fat diet, sulfur deficiency becomes that much worse a problem. Sulfur deficiency interferes with glucose metabolism, so it's a much healthier choice to simply avoid glucose sources (carbohydrates) in the diet; i.e. to adopt a very low-carb diet. Then the fat in the diet can supply the muscles with fuel, and the fat cells are not burdened with having to store up so much reserve fat. <br /><br />Insulin suppresses the release of fats from fat cells [Scappola1995]. This forces the fat cells to flood the bloodstream with triglycerides when insulin levels are low, i.e., after prolonged periods of fasting, such as overnight. The fat cells must dump enough triglycerides into the bloodstream during fasting periods to fuel the muscles when the dietary supply of carbohydrates keeps insulin levels elevated, and the release of fats from the fat cells is repressed. As the dietary carbs come in, blood sugar levels rise dramatically because the muscle cells can't utilize it.<br /><br />The liver also processes excess glucose into fat, and packages it up into LDL, to further supply fuel to the defective muscle cells. Because the liver is so preoccupied with processing glucose and fructose into LDL, it falls behind on the generation of HDL, the "good" cholesterol. So the result is elevated levels of LDL, triglycerides, and blood sugar, and reduced levels of HDL, four key components of the metabolic syndrome.<br /><br />The chronic presence of excess glucose and fructose in the blood stream leads to a host of problems, all related to glycation damage of blood stream proteins by glucose exposure. One of the key proteins that gets damaged is the apolipoprotein, apoB, that's encased in the membrane of the LDL particles. Damaged apoB inhibits the ability of LDL to efficiently deliver its contents (fat and cholesterol) to the tissues. Fat cells again come to the rescue, by scavenging the broken LDL particles (through a mechanism that does not require apoB to be healthy), taking them apart, and extracting and refurbishing their cholesterol. In order to function properly, the fat cells must have intact ApoE, an antioxidant that cleans up oxidized cholesterol and transports it to the cell membrane for delivery to HDL particles.Stephaniehttp://www.blogger.com/profile/00273178904990742948noreply@blogger.com3tag:blogger.com,1999:blog-1099974205222264538.post-51416593987981205302010-09-27T16:25:00.002-07:002010-09-27T16:26:46.823-07:00 6. Fat Cells, Macrophages and Atherosclerosis While diligently converting glucose to stored fats, the fat cells are awash in glucose, which damages their apoE through glycation [Li1997]. Once their apoE is damaged, they can no longer transport cholesterol to the membrane. Excess cholesterol accumulates inside the fat cells and eventually destroys their ability to synthesize proteins. Concurrently, their cell membrane becomes depleted in cholesterol, because they can no longer deliver it to the membrane [Seneff2010]. A fat cell that has deteriorated to this degree has no choice but to die: it sends out distress signals that call in macrophages. The macrophages essentially consume the dysfunctional fat cell, wrapping their own membrane around the fat cell's membrane that is now barely able to hold its contents inside [Cinti2005].<br /><br />Macrophages are also principle players in the fatty streaks that appear along the sides of major arteries leading to the heart, and are associated with plaque build-up and heart disease. In a fascinating set of experiments, Ma et al. [Ma2008] have shown that the sulfate ion attached to oxidized forms of cholesterol is highly <i>protective</i> against fatty streaks and atherosclerosis. In a set of in-vitro experiments, they demonstrated diametrically opposite reactions from macrophages to 25-hydroxyl cholesterol (25-HC) versus its sulfoconjugate 25-hydroxyl cholesterol <i>sulfate</i> (25-HC3S). Whereas 25-HC present in the medium causes the macrophages to synthesize and store cholesterol and fatty acids, 25-HC3S has the exact opposite effect: it promotes the release of cholesterol to the medium and causes fat stores to shrink. Furthermore, while 25-HC added to the medium led to apoptosis and cell death, 25-HC3S did not. I suggest that the sulfate radical is essential for the process that feeds cholesterol and oxygen to the heart muscle.Stephaniehttp://www.blogger.com/profile/00273178904990742948noreply@blogger.com2tag:blogger.com,1999:blog-1099974205222264538.post-62798439616965527032010-09-27T16:25:00.001-07:002010-09-27T16:25:42.513-07:00 7. Sulfur and Alzheimer's With an aging population, Alzheimer's disease is on the rise, and it has been argued that the rate of increase is disproportionately high compared to the increase in the raw number of elderly people [Waldman2009]. Because of a conviction that the amyloid beta plaque that is a signature of Alzheimer's is also the cause, the pharmaceutical industry has spent hundreds of millions, if not billions, of dollars pursuing drugs that reduce the amount of plaque accumulating in the brain. Thus far, drug trials have been so disappointing that many are beginning to believe that amyloid beta is not the cause after all. Recent drug trials have shown not only no improvement, but actually a further decline in cognitive function, compared to placebo (<a href= "http://www.nytimes.com/2010/08/18/business/18lilly.html"> <font color = "red" >New York Times Article</font></a>). I have argued <a href= "http://people.csail.mit.edu/seneff/alzheimers_statins.html"> <font color = "red" >elsewhere</font></a> that amyloid beta may actually be protective against Alzheimer's, and that problems with glucose metabolism are the true culprit in the disease. <br /><br />Once I began to suspect sulfur deficiency as a major factor in Americans' health, I looked into the relationship between sulfur deficiency and Alzheimer's. Imagine my surprise when I came upon a <a href= "http://www.acu-cell.com/dis-alz.html"> <font color = "red" >web page posted by Ronald Roth</font></a>, which shows a plot of the levels of various minerals in the cells of a typical Alzheimer's patient relative to the normal level. Remarkably, sulfur is almost non-existent in the Alzheimer's patient's profile.<br /><br />To quote directly from that site: "While some drugs or antibiotics may slow, or if it should happen, halt the progression of Alzheimer's disease, sulfur supplementation has the potential of not only preventing, but actually reversing the condition, provided it has not progressed to a stage where much damage has been done to the brain."<br /><br />"One major reason for the increase in Alzheimer's disease over the past years has been the bad reputation eggs have been getting in respect to being a high source of cholesterol, despite the fact of dietary intake of cholesterol having little impact on serum cholesterol - which is now also finally acknowledged by mainstream medicine. In the meantime, a large percentage of the population lost out on an excellent source of sulfur and a host of other essential nutrients by following the nutritional misinformation spread on eggs. Of course, onions and garlic are another rich source of sulfur, but volume-wise, they cannot duplicate the amounts obtained from regularly consuming eggs."<br /><br />Why should sulfur deficiency be so important for the brain? I suspect that the answer lies in the mysterious molecule alpha-synuclein, which shows up alongside amyloid-beta in the plaque, and is also present in the Lewy Bodies that are a signature of Parkinson's disease [Olivares2009]. The alpha-synuclein molecule contains four methionine residues, and all four of the sulfur molecules in the methionine residues are converted to sulfoxides in the presence of oxidizing agents such as hydrogen peroxide [Glaser2005]. Just as in the muscle cells, insulin would cause the mitochondria of neurons to release hydrogen peroxide, which would then allow the alpha-synuclein to take up oxygen, in a way that is very reminiscent of what myoglobin can do in muscle cells. The lack of sufficient sulfur should directly impact the neuron's ability to safely carry oxygen, again paralleling the situation in muscle cells. This would mean that other proteins and fats in the neuron would suffer from oxidative damage, leading ultimately to the neuron's destruction. <br /><br />In my essay on <a href= "http://people.csail.mit.edu/seneff/alzheimers_statins.html"> <font color = "red" >Alzheimer's</font></a>, I argued that biologically pro-active restriction in glucose metabolism in the brain (a so-called type-III diabetes and a precursor to Alzheimer's disease) is triggered by a deficiency in cholesterol in the neuron cell membrane. Again, as in muscle cells, glucose entry depends upon cholesterol-rich lipid rafts, and, when the cell is deficient in cholesterol, the brain goes into a mode of metabolism that prefers other nutrients besides glucose.<br /><br />I suspect that a deficiency in cholesterol would come about if there is insufficient cholesterol sulfate, because cholesterol sulfate likely plays an important role in seeding lipid rafts, while concurrently enriching the cell wall in cholesterol. The cell also develops an insensitivity to insulin, and, as a consequence, anaerobic metabolism becomes favored over aerobic metabolism, reducing the chances for alpha-synuclein to become oxidized. Oxidation actually protects alpha-synuclein from fibrillation, a necessary structural change for the accumulation of Lewy bodies in Parkinson's disease (and likely also Alzheimer's plaque) [Glaser2005]Stephaniehttp://www.blogger.com/profile/00273178904990742948noreply@blogger.com3tag:blogger.com,1999:blog-1099974205222264538.post-52284224300113960092010-09-27T16:22:00.000-07:002010-09-27T16:25:14.330-07:00 8. Is The Skin a Solar-Powered Battery for the Heart? The evidence is quite compelling that sunny places afford protection from heart disease. A study described in [Grimes1996] provides an in depth anaylsis of data from around the world showing an inverse relationship between heart disease rates and sunny climate/low latitude. For instance, the cardiovascular-related death rate for men between the ages of 55 and 64 was 761 per 100,000 men in Belfast, Northern Ireland, but only 175 in Toulouse, France. While the obvious biological factor that would be impacted by sunlight is vitamin D, studies conducted specifically on vitamin D status have been inconclusive, with some even showing a significant <i>increased</i> risk for heart disease with increased intake of <i>vitamin D2</i> supplements [Drolet2003]. <br /><br />I believe, first of all, that the distinction between vitamin D3 and vitamin D3-sulfate really matters, and also that the distinction between vitamin D2 and vitamin D3 really matters. Vitamin D2 is the plant form of the vitamin -- it works similarly to D3 with respect to calcium transport, but it cannot be sulfated. Furthermore, apparently the body is unable to produce vitamin D3 sulfate directly from unsulfated vitamin D3 [Lakdawala1977] (which implies that it produces vitamin D3 sulfate directly from cholesterol sulfate). I am not aware of any other food source besides <i>raw</i> milk that contains vitamin D3 in the sulfated form. So, when studies monitor either vitamin D supplements or vitamin D serum levels, they're not getting at the crucial aspect for heart protection, which I think is the serum level of vitamin D3 <i>sulfate</i>.<br /><br />Furthermore, I believe it is extremely likely that vitamin D3 sulfate is not the only thing that's impacted by greater sun exposure, and maybe not even the most important thing. Given that cholesterol sulfate and vitamin D3 sulfate are very similar in molecular structure, I would imagine that both molecules are produced the same way. And since vitamin D3-sulfate synthesis requires sun exposure, I suspect that cholesterol sulfate synthesis may also exploit the sun's radiation energy.<br /><br />Both cholesterol and sulfur afford protection in the skin from radiation damage to the cell's DNA, the kind of damage that can lead to skin cancer. Cholesterol and sulfur become oxidized upon exposure to the high frequency rays in sunlight, thus acting as antioxidants to "take the heat," so to speak. Oxidation of cholesterol is the first step in the process by which cholesterol transforms itself into vitamin D3. Sulfur dioxide in the air is converted nonenzymatically to the sulfate ion upon sun exposure. This is the process that produces acid rain. The oxidation of sulfide (S<sup>-2</sup>) to sulfate (SO<sub>4</sub><sup>-2</sup>), a strongly endothermic reaction [Hockin2003], converts the sun's energy into chemical energy contained in the sulfur-oxygen bonds, while simultaneously picking up four oxygen molecules. Attaching the sulfate ion to cholesterol or vitamin D3 is an ingenious step, because it makes these molecules water-soluble and therefore easily transportable through the blood stream. <br /><br />Hydrogen sulfide (H<sub>2</sub>S) is consistently found in the blood stream in small amounts. As a gas, it can diffuse into the air from capillaries close to the skin's surface. So it is conceivable that we rely on bacteria in the skin to convert sulfide to sulfate. It would not be the first time that humans have struck up a symbiotic relationship with bacteria. If this is true, then washing the skin with antibiotic soap is a bad idea. Phototrophic bacteria, such as <a href="http://microbewiki.kenyon.edu/index.php/Chlorobium"> <font color = "red" >Chlorobium tepidum</font></a>, that can convert H<sub>2</sub>S to H<sub>2</sub>SO<sub>4</sub> exist in nature [Zerkle2009, Wahlund1991], for example in sulfur hot springs in Yellowstone Park. These highly specialized bacteria can convert the light energy from the sun into chemical energy in the sulfate ion. <br /><br />Another possibility is that we have specialized cells in the skin, possibly the keratinocytes, that are able to exploit sunlight to convert sulfide to sulfate, using a similar phototrophic mechanism to C. tepidum. This seems quite plausible, especially considering that both human keratinocytes and C. tepidum can synthesize an interesting UV-B absorbing cofactor, tetrahydrobioptin. This cofactor is found universally in mammalian cells, and one of its roles is to regulate the synthesis of melanin [Schallreut94], the skin pigment that is associated with a tan and protects the skin from damage by UV-light exposure [Costin2007]. However, tetrahydrobiopsin is very rare in the bacterial kingdom, and C. tepidum is one of the very few bacteria that can synthesize it [Cho99]. <br /><br />Let me summarize at this point where I'm on solid ground and where I'm speculating. It is undisputed that the skin synthesizes cholesterol sulfate in large amounts, and it has been suggested that the skin is the major supplier of cholesterol sulfate to the blood stream [Strott2003]. The skin also synthesizes vitamin D3 sulfate, upon exposure to sunlight. Vitamin D3 is synthesized from cholesterol, with oxysterols (created from sun exposure) as an intermediate step (oxysterols are forms of cholesterol with hydroxyl groups attached at various places in the carbon chain). The body can't synthesize vitamin D3 sulfate from vitamin D3 [Lakdawala1977] so it must be that sulfation happens first, producing cholesterol sulfate or hydroxy-cholesterol sulfate, which is then optionally converted to vitamin D3 sulfate or shipped out "as is."<br /><br />Another highly significant feature of skin cells is that the skin stores sulfate ions attached to molecules that are universally present in the intracellular matrix, such as heparan sulfate, chondroitin sulfate, and keratin sulfate [Milstone1994]. Furthermore, it has been shown that exposure of the melanin producing cells (melanocytes) to molecules containing <i>reduced</i> sulfur (-2) leads to <i>suppression</i> of melanin synthesis [Chu2009], whereas exposure to molecules like chondroitin sulfate that contain <i>oxidized</i> sulfur (+6) leads to enhancement of melanin synthesis [Katz1976]. Melanin is a potent UV-light absorber, and it would compete with reduced sulfur for the opportunity to become oxidized. It is therefore logical that, when sulfur is reduced, melanin synthesis should be suppressed, so that sulfur can absorb the solar energy and convert it to very useful chemical bonds in the sulfate ion.<br /><br />The sulfate would eventually be converted back to sulfide by a muscle cell in the heart or a skeletal muscle (simultaneously recovering the energy to fuel the cell and unlocking the oxygen to support aerobic metabolism of glucose), and the cycle would continually repeat.<br /><br />Why am I spending so much time talking about all of this? Well, if I'm right, then the skin can be viewed as a solar-powered battery for the heart, and that is a remarkable concept. The energy in sunlight is converted into chemical energy in the oxygen-sulfur bonds, and then transported through the blood vessels to the heart and skeletal muscles. The cholesterol sulfate and vitamin D3 sufate are carriers that deliver the energy (and the oxygen) "door-to-door" to the individual heart and skeletal muscle cells.<br /><br />Today's lifestyle, especially in America, severely stresses this system. First of all, most Americans believe that any food containing cholesterol is unhealthy, so the diet is extremely low in cholesterol. Eggs are an excellent source of sulfur, but because of their high cholesterol content we have been advised to eat them sparingly. Secondly, as I discussed previously, natural food plant sources of sulfur are likely to be deficient due to sulfur depletion in the soil. Thirdly, water softeners remove sulfur from our water supply, which would otherwise be a good source. Fourthly, we have been discouraged from eating too much red meat, an excellent source of sulfur-containing amino acids. Finally, we have been instructed by doctors and other authoritarian sources to stay out of the sun and wear high SPF sunscreen whenever we do get sun exposure.<br /><br />Another significant contributor is the high carbohydrate, low fat diet, which leads to excess glucose in the blood stream that glycates LDL particles and renders them ineffective in delivering cholesterol to the tissues. One of those tissues is the skin, so skin becomes further depleted in cholesterol due to glycation damage to LDL.Stephaniehttp://www.blogger.com/profile/00273178904990742948noreply@blogger.com7tag:blogger.com,1999:blog-1099974205222264538.post-50859012303827470982010-09-27T16:09:00.001-07:002010-09-27T16:19:47.972-07:00 9. Sulfur Deficiency and Muscle Wasting DiseasesIn browsing the Web, I recently came upon a remarkable article [Dröge1997] which develops a persuasive theory that low blood serum levels of two sulfur-containing molecules are a characteristic feature of a number of diseases/conditions. All of these diseases are associated with muscle wasting, despite adequate nutrition. The authors have coined the term "low CG syndrome" to represent this observed profile., where "CG" stands for the amino acid "cysteine," and the tripeptide "glutathione," both of which contain a sulfhydryl radical "-S-H" that is essential to their function. Glutathione is synthesized from the amino acids cysteine, glutamate, and glycine, and glutamate deficiency figures into the disease process as well, as I will discuss later.<br /><br />The list of diseases/conditions associated with low CG syndrome is surprising and very revealing: HIV infection, cancer, major injuries, sepsis (blood poisoning), Crohn's disease (irritable bowel syndrome), ulcerative colitis, chronic fatigue syndrome, and athletic over-training. The paper [Drage1997] is dense but beautifully written, and it includes informative diagrams that explain the intricate feedback mechanisms between the liver and the muscles that lead to muscle wasting. <br /><br />This paper fills in some missing holes in my theory, but the authors never suggest that sulfur <i>deficiency</i> might actually be a <i>precursor</i> to the development of low CG syndrome. I think that, particularly with respect to Crohn's disease, chronic fatigue syndrome, and excessive exercise, sulfur deficiency may precede and provoke the muscle wasting phenomenon. The biochemistry involved is complicated, but I will try to explain it in as simple terms as possible.<br /><br />I will use <a href="https://health.google.com/health/ref/Crohn%27s+disease"> <font color = "red" >Crohn's disease</font></a> as my primary focus for discussion: an inflammation of the intestines, associated with a wide range of symptoms, including reduced appetite, low-grade fever, bowel inflammation, diarrhea, skin rashes, mouth sores, and swollen gums. Several of these symptoms suggest problems with the interface between the body and the external world: i.e., a vulnerability to invasive pathogens. I mentioned before that cholesterol sulfate plays a crucial role in the barrier that keeps pathogens from penetrating the skin. It logically plays a similar role everywhere there is an opportunity for bacteria to invade, and certainly a prime opportunity is available at the endothelial barrier in the intestines. Thus, I hypothesize that the intestinal inflammation and low-grade fever are due to an overactive immune system, necessitated by the fact that pathogens have easier access when the endothelial cells are deficient in cholesterol sulfate. The skin rashes and mouth and gum problems are a manifestation of inflammation elsewhere in the barrier.<br /><br />Ordinarily, the liver supplies cholesterol sulfate to the gall bladder, where it is mixed into bile acids, and subsequently released into the digestive system to assist in the digestion of fats. If a person consistently eats a low-fat diet, the amount of cholesterol sulfate delivered to the digestive system from the liver will be reduced. This will logically result in a digestive system that is more vulnerable to invasion by pathogens.<br /><br />The sulfate that's combined with cholesterol in the liver is synthesized from cysteine (one of the two proteins that are deficient in low CG syndome). So insufficient bioavailability of cysteine will lead to a reduced production of cholesterol sulfate by the liver. This will, in turn, make it difficult to digest fats, likely, over time, compelling the person to adhere to a low-fat diet. Whether low-fat diet or sulfur deficiency comes first, the end result is a vulnerability to infective agents in the intestines, with a consequential heightened immune response. <br /><br />[Dröge1997] further discussses how a reduction in the synthesis of sulfate from cysteine in the liver leads to increased compensatory activity in another biological pathway in the liver that converts glutamate to arginine and urea. Glutamate is highly significant because it is produced mainly by the breakdown of amino acids (proteins in the muscles); i.e., by muscle wasting. The muscle cells are triggered to cannibalize themselves in order to provide adequate glutamate to the liver, mainly, in my view, in order to generate enough arginine to replace the role of sulfate in muscle glucose metabolism (i.e., these activities in the liver and muscles are circular and mutually supportive).<br /><br />Arginine is the major source of nitric oxide (NO) and NO is the next best thing for muscle glucose metabolism in the absence of cholesterol sulfate. NO is a poor substitue for SO<sub>4</sub><sup>-2</sup>, but it can function in some of the missing roles. As you will recall, I propose that cholesterol SO<sub>4</sub><sup>-2</sup> accomplishes a number of important things in muscle cells: it delivers oxygen to myoglobin, it supplies cholesterol to the cell membrane, it helps break down glucose, protects the cell's proteins from glycation and oxidation damage, and provides energy to the cell. NO can help in reducing glycation damage, as nitrogen can be reduced from +2 to 0 (whereas sulfur was reduced from +6 to -2). It also provides oxygen, but it is unable to transfer the oxygen directly to myoglobin by binding with the iron molecule, as was the case for sulfate. NO does not supply cholesterol, so cholesterol deficiency remains a problem, leaving the cell's proteins and fats more vulnerable to oxidative damage. Furthermore, NO itself is an oxidizing agent, so myoglobin becomes disabled, due to both oxidation and glycation damage. The muscle cell, therefore, engages in mitochondrial oxidation of glucose at its own peril: better to revert to anaerobic metabolism of glucose to decrease the risk of damage. Anaerobic metabolism of glucose results in a build-up of lactic acid, which, as explained in [Dröge1997] further enhances the need for the liver to metabolize glutamate, thus augmenting the feedback loop.<br /><br />Furthermore, as you'll recall, if I'm right about cholesterol sulfate seeding lipid rafts, then, with a cholesterol sulfate deficiency, the entry of both glucose and fat into the muscle cell are compromised. This situation leaves the cell with little choice but to exploit its internal proteins as fuel, manifested as muscle wasting.<br /><br />In summary, a number of different arguments lead to the hypothesis that sulfur deficiency causes the liver to shift from producing cholesterol sulfate to producing arginine (and subsequently nitric oxide). This leaves the intestines and muscle cells vulnerable to oxidation damage, which can explain both the intestinal inflammation and the muscle wasting associated with Crohn's disease.<br /><br />The immune system depends upon abundant cholesterol to defend against severe stress. I have previously argued that high serum cholesterol is <a href="http://people.csail.mit.edu/seneff/statins_pregnancy_sepsis_cancer_heart_failure.html#do_statins_cause_sepsis"> <font color = "red" >protective against sepsis</font></a>. It is worth repeating here the abstract from [Wilson2003], who studied changes in blood cholesterol levels following trauma, infection, and multiple organ failure:<br /><br />"Hypocholesterolemia is an important observation following trauma. In a study of critically ill trauma patients, mean cholesterol levels were significantly lower (119 ± 44 mg/dl) than expected values (201 ± 17 mg/dl). In patients who died, final cholesterol levels fell by 33% versus a 28% increase in survivors. Cholesterol levels were also adversely affected by infection or organ system dysfunction. Other studies have illustrated the clinical significance of hypocholesterolemia. Because lipoproteins can bind and neutralize lipopolysaccharide, hypocholesterolemia can negatively impact outcome. New therapies directed at increasing low cholesterol levels may become important options for the treatment of sepsis." <br /><br />Thus, many of these conditions/diseases that lead to muscle wasting may do so because cholesterol (and therefore cholesterol sulfate) is depleted from the blood serum. This results in the same feedback loop between the liver and the muscles that I discussed with regard to Crohn's disease. So I think it's plausible that the muscle wasting associated with all of these conditions is caused by this same feedback mechanism.<br /><br />I have discussed the role cysteine plays in providing sulfate to the liver. But what is the role of glutathione, the other sulfur-containing protein that's depleted in low GC syndrome? Muscle cells ordinarily contain significant levels of glutathione, and its depletion leads to mitochondrial damage [Martensson1989]. Patients undergoing surgical trauma have been found to exhibit reduced glutathione levels in their skeletal muscles [Luo1996]. It is tempting to speculate that cholesterol sulfate provides the sulfur needed for glutathione synthesis, so that the deficiency would be explained by the reduced availability of cholesterol following the immune system's heightened response to surgical trauma. Glutathione is a potent antioxidant, so its deficiency will further contribute to dysfunction of the muscle cell's mitochondria, therefore greatly impairing its energy supply. <br /><br />There is a growing awareness that glutathione deficiency may play a role in many diseases. You may want to check out <a href="http://www.amazing-glutathione.com/glutathione-deficiency.html"> <font color = "red" >this Web site</font></a> describing a long list of diseases that may be impacted by glutathione deficiency. Whether the problems arise just due to insufficient supply of the glutathione molecule itself, or whether a more general sulfur deficiency is the root cause, is perhaps hard to say, but provocative nonetheless.Stephaniehttp://www.blogger.com/profile/00273178904990742948noreply@blogger.com6tag:blogger.com,1999:blog-1099974205222264538.post-64865649353281092012010-09-27T16:09:00.000-07:002010-09-27T16:14:36.962-07:00 9. Sulfur Deficiency and Muscle Wasting DiseasesA serious disorder known as <a href="http://www.emedicine.com/ped/TOPIC2117.HTM"> (<font color = "red" >"Smith-Lemli-Opitz"</font>) </a> syndrome (SLOS) is characterized by a genetic defect resulting in an inability to synthesize adequate cholesterol. Most fetuses that are unfortunate to be conceived with this genetic defect don't make it to 16 weeks of gestation before the pregnancy ends in a miscarriage. If they do manage to make it to term, they typically suffer from major brain defects, resulting in autism or other forms of mental retardation [9].Stephaniehttp://www.blogger.com/profile/00273178904990742948noreply@blogger.com3tag:blogger.com,1999:blog-1099974205222264538.post-34742098078975069392010-09-27T16:08:00.000-07:002010-09-27T16:09:48.859-07:00 10. Summary Although sulfur is an essential element in human biology, we hear surprisingly little about sulfur in discussions on health. Sulfur binds strongly with oxygen, and is able to stably carry a charge ranging from +6 to -2, and is therefore very versatile in supporting aerobic metabolism. There is strong evidence that sulfur deficiency plays a role in diseases ranging from Alzheimer's to cancer to heart disease. Particularly intriguing is the relationship between sulfur deficiency and muscle wasting, a signature of end-stage cancer, AIDS, Crohn's disease, and chronic fatigue syndrome.<br /><br />The African rift zone, where humans are believed to have first made their appearance several million years ago, would have been rich with sulfur supplied by active volcanism. It is striking that people living today in places where sulfur is abundantly provided by recent volcanism enjoy a low risk for heart disease and obesity.<br /><br />In my research on sulfur, I was drawn to two mysterious molecules: cholesterol sulfate and vitamin D3 sulfate. Researchers have not yet determined the role that cholesterol sulfate plays in the blood stream, despite the fact that it is ubiquitous there. Research experiments have clearly shown that cholesterol sulfate is protective against heart disease. I have developed a theory proposing that cholesterol sulfate is central to the formation of lipid rafts, which, in turn, are essential for aerobic glucose metabolism. I would predict that deficiencies in cholesterol sulfate lead to severe defects in muscle metabolism, and this includes the heart muscle. My theory would explain the protective role of cholesterol sulfate in heart disease and muscle wasting diseases.<br /><br />I have also argued that cholesterol sulfate delivers oxygen to myoglobin in muscle cells, resulting in safe oxygen transport to the mitochondria. I argue a similar role for alpha-synuclein in the brain. There is a striking relationship between Alzheimer's and sulfur depletion in neurons in the brain. Sulfur plays a key role in protectiing proteins in neurons and muscle cells from oxidative damage, while maintaining adequate oxygen supply to the mitochondria. <br /><br />When muscles become impaired in glucose metabolism due to reduced availability of cholesterol sulfate, proliferating fat cells become involved in converting glucose to fat. This provides an alternative fuel for the muscle cells, and replenishes the cholesterol supply by storing and refurbishing cholesterol extracted from defective LDL. Thin people with cholesterol and sulfur deficiency are vulnerable to a wide range of problems, such as Crohn's disease, chronic fatigue syndrome, and muscle wasting, because fat cells are not available to ameliorate the situation.<br /><br />Cholesterol sulfate in the epithelium protects from invasion of pathogens through the skin, which greatly reduces the burden placed on the immune system. Perhaps the most intriguing possibility presented here is the idea that sulfur provides a way for the skin to become a solar-powered battery: to store the energy from sunlight as chemical energy in the sulfate molecule. This seems like a very sensible and practical scheme, and the biochemistry involved has been demonstrated to work in phototrophic sulfur-metabolizing bacteria found in sulfur hot springs. <br /><br />The skin produces vitamin D3 <i>sulfate</i> upon exposure to sunlight, and the vitamin D3 found in breast milk is also sulfated. In light of these facts, it is quite surprising to me that so little research has been directed towards understanding what role sulfated vitamin D3 plays in the body. It is recently becoming apparent that vitamin D3 promotes a strong immune system and offers protection against cancer, yet how it achieves these benefits is not at all clear. I strongly suspect that it is vitamin D3 sulfate that carries out this aspect of vitamin D3's positive influence.<br /><br />Modern lifestyle practices conspire to induce major deficiencies in cholesterol sulfate and vitamin D3 sulfate. We are encouraged to actively avoid sun exposure and to minimize dietary intake of cholesterol-containing foods. We are encouraged to consume a high-carbohydrate/low-fat diet which, as I have argued previously (Seneff2010), leads to impaired cholesterol uptake in cells. We are told nothing about sulfur, yet many factors, ranging from the Clean Air Act to intensive farming to water softeners, deplete the supply of sulfur in our food and water.<br /><br />Fortunately, correcting these deficiencies at the individual level is easy and straightforward. If you just throw away the sunscreen and eat more eggs, those two steps alone may greatly increase your chances of living a long and healthy life.<br /><br /><br /><br /><br /><a rel="license" href="http://creativecommons.org/licenses/by/3.0/us/"><img alt="Creative Commons License" style="border-width:0" src="http://i.creativecommons.org/l/by/3.0/us/88x31.png" /></a><br /><span xmlns:dc="http://purl.org/dc/elements/1.1/" href="http://purl.org/dc/dcmitype/Text" property="dc:title" rel="dc:type"> Could Sulfur Deficiency be a Contributing Factor to Obesity, Heart Disease, Alzheimer's and Chronic Fatigue Syndrome? </span> by <a xmlns:cc="http://creativecommons.org/ns#" href="http://people.csail.mit.edu/seneff/sulfur.sep1.html" property="cc:attributionName" rel="cc:attributionURL">Stephanie Seneff</a> is licensed under a <a rel="license" href="http://creativecommons.org/licenses/by/3.0/us/">Creative Commons Attribution 3.0 United States License</a>.Stephaniehttp://www.blogger.com/profile/00273178904990742948noreply@blogger.com3