CROSS REFERENCE TO RELATED APPLICATION
Organ Failure and Nutrition
This application is a continuation-in-part of U.S. application Ser. No. 09/002,765, filed Jan. 6, 1998, which is a continuation-in-part of U.S. application Ser. No. 08/826,234 filed Mar. 27, 1997.
- Relationship of Malnutrition to Mitochondrial Function
There are four critical organ systems that are especially likely to fail in aging and critical illness. They are the cardiovascular, central nervous, musculoskeletal and immune systems.
Protein-calorie malnutrition contributes to both skeletal and cardiac1 muscle dysfunction in patients with cardiac failure. Muscle is composed of water, minerals, nitrogen and glycogen2,3. Feeding wasted individuals results in a gain of the multiple elements in lean tissue4 including potassium. Body potassium, has been used as an index of body cell mass5, the metabolically active component of the lean tissue. In contrast to body nitrogen, body potassium responds rapidly to feeding by both oral and intravenous routes6,7,8,9,10. It has been shown that in malnutrition there is a change in muscle membrane potential resulting in reduced intracellular ionic potassium. The reduced cellular potassium cannot be simply corrected by giving potassium but requires restitution of nutrition. The above mentioned observations suggest that cell ion uptake, an energy dependent process, occurs earlier than protein synthesis during nutritional support. This concept has received experimental support by two studies using 31P-NMR which showed that malnutrition was associated with a reduced rate of oxidative phosphorylation, suggesting a mitochondrial abnormality11,12.
Cell energetics are also important for muscle activity and it has been shown13,14,15,16,17,18,19,20,21,22 that skeletal muscle function, including that of the diaphragm, can be rapidly altered by nutrient deprivation and restored by refeeding. Also the changes in muscle function are specific to alterations in the nutritional status and are not influenced by sepsis, trauma, renal failure and steroid administration15,17. Christie and Hill indicated that nutritional support improves muscle, t, including diaphragmatic function before any increase in body protein or body mass20. Windsor and Hill21 demonstrated that the functional effects of nutrition are more important than subnormal body protein as an index of surgical risk. Hanning and her colleagues22 demonstrated the ability of stimulated muscle function as demonstrated by a slow relaxation rate and an altered force-frequency curve to predict the ability of patients with cystic fibrosis to grow as an outcome measure. In contrast, body composition, protein biochemistry, muscle power on an ergometer or use of supplements did not predict growth potential. Among the macronutrients, Castenada et al23 have shown that protein deficiency can profoundly alter muscle function even when energy intake is sufficient to meet requirements.
- SUMMARY OF THE INVENTION
The data given above indicate that it is critical to correct protein-calorie malnutrition, with an emphasis on protein repletion, in order to obtain the maximum functional benefits of administering skeletal muscle specific micronutrients. Current diet supplementing strategies for correcting protein-calorie malnutrition focus on giving supplements of protein and energy (carbohydrates and fats). No supplement to date has addressed the cascading series of metabolic abnormalities that can lead to mitochondrial dysfunction.
We have found that nutrition can be used to prevent or delay the onset of cardiac failure and thereby, promote recovery in disease states affecting the heart. Similar considerations apply to diseases of the other organ systems indicated above.
We have found that the central effect of nutrition in all these systems can be unified into its influence on mitochondrial energetics. That is: inadequate nutitional substrate is a cause of impaired cell energetics. This has led us to invent a composition for the improvement of mitochondrial energetics.
We have shown that in the skeletal muscle protein-calorie malnutrition profoundly reduces mitochondrial oxidative phosphorylation and reduces calcium cycling in cardiac muscle1. We have found that there is profound reduction of respiratory chain complex I, II and IV activity in animals given a protein-calorie deficient diet. In addition, complex I activity is similarly reduced in lymphocyte mitochondria showing that these effects are not cardiac specific but apply to mitochondria in other tissues, and protein feeding rapidly restored the abnormality when it was simply due to protein-energy malnutrition (unpublished data).
- Heart Failure
In addition, certain micronutrients and amino acids also influence mitochondrial function in general. For example, carnitine improves mitochondrial DNA transcription and translation in aged animals24. A specific acyl derivative of carnitine, acetyl-carnitine has been used for mitochondrial DNA synthesis based on findings observed in patients treated with anti-retroviral agents25. Coenzyme Q can alter immune function26 and may protect the central nervous system from injury and neurodegeneration27. On the basis of the above considerations, a nutritional supplement that could maintain or restore mitochondrial function will prevent cardiac failure or aid recovery from cardiac disease. In addition it could also aid in the management of neurodegenerative, musculoskeletal including the muscular abnormality in chronic obstructive lung disease (COPD)16 and immune disorders.
Congestive heart failure has emerged as a major health problem during the past two decades. Its morbidity and mortality have shown a steady increase since 197028; heart failure now affects approximately 1% of the population of the United States and Canada. These data reflect both the aging of our population and the success of modern cardiovascular medicine in converting acute, often previously fatal, cardiac disease into a more chronic process.
The underlying abnormality in congestive heart disease is myocardial dysfunction leading to inadequate blood flow to peripheral tissues. Although there have been considerable advances in our understanding of the pathogenesis of heart failure in recent years, critical questions remain about the evolution of cardiac dysfunction to terminal failure. The importance of elucidating the mechanisms responsible for the evolution of maladaptive hypertrophy to cardiac failure is emphasized by the fact that in spite of our advances, no presently available therapeutic intervention has been shown to substantially improve the long-term survival of patients with dilated cardiomyopathy and congestive heart failure. The underlying heart disease is relentlessly progressive in almost all patients who develop symptoms of overt failure and mortality continues to be unacceptably high; for example, in a recent heart failure trial, SOLVD, 40% of patients in the symptomatic treated group were dead within 4 years29. Heart transplantation appears to be the only prospect to improve long term survival for many patients.
The reason for this dismal outcome despite modern advances lies in the fact that several metabolic abnormalities have been found in the failing myocardium which together as indicated below result in progressive loss of cardiac myocytes (muscle).
There is a progressive accumulation of calcium in the muscle, which in turn results in increased calcium in the mitochondria. The progressive increase in mitochondrial calcium as well as the basic cardiac disease (ischemic, viral, toxic, genetic) decrease myocyte energy production and increase oxidative stress resulting in free radical damage. The combined result of these three processes promotes myocyte dysfunction and death. In addition these processes also influence skeletal muscle and contribute to fatigue and disability.
The modern pharmacological therapy of heart failure has focused on the amelioration of fluid overload and hemodynamic abnormalities and has not addressed the fundamental fact that if there is progressive loss of cardiac muscle then the patient will inevitably succumb. That is: the inexorable myocyte loss by apoptosis that occurs in heart failure is the key factor responsible for myocardial decompensation and the demise of the patient30. Oxidative stress, calcium overload and cellular energy deficiency are well known as principal stimuli for the development of apoptosis.
Among the factors that aggravate myocyte dysfunction there is increasing evidence for the role of nutritional deficiencies both due to reduced intake and to insufficient intake in relation to augmented requirements caused by the underlying disease state, a phenomenon which we will refer to as “conditioned deficiency”. In this situation the recommended daily allowances (RDA) do not apply, as requirements may exceed the standard RDA.
The presence of protein-energy malnutrition has been recognized by surveys of hospitalized patients using anthropometric, biochemical and immunologic measures of nutritional status.
These surveys have indicated that 50-68% of patients with congestive heart failure were significantly malnourished31. The proportion of malnourished heart failure patients has been found to be higher than that of patients with cancer, alcoholism or those with acute infection. The cause of protein-energy malnutrition is due to both reduced intake and increased energy demands. Cardiac failure results in a cascade of metabolic effects such as tissue hypoxia, anorexia, hypermetabolism, weakness, dyspnea and hypomotility of the gastrointestinal tract all leading to poor nutrient intake. Anorexia can be aggravated by unpalatable restrictive diets or by converting enzyme inhibitors or by an excess of diuretics, opiates and digitalis. Characteristics of the disease process such as fatigue and early satiety have all been reported in congestive heart failure patients consuming self-selected diets32,33. These factors lead to compromised food and nutrient intake and subsequently contribute to the poor nutritional status of these cardiac patients. In addition, patients with heart failure have been shown to have significantly increased resting metabolic rates34,35,36,37 possibly due to the increased work of breathing, fever, cytokines or elevated sympathetic nervous system outflow.
The RDA for the vitamins and related micronutrients recommended by federal nutrition authorities in Canada, the United States and Western Europe (e.g. The Canada Food Guide) are obtained through the analysis of deficiency data in otherwise healthy humans and animals. We have found that with the advent of disease, or due to genetic predisposition, specific metabolic pathways in individual organs and the function of some of these systems alter the nutritional requirements causing conditioned deficiency of both macro- (protein including amino acids, carbohydrates and fats) and micronutrients (electrolytes, trace elements, vitamins and special nutrient substances). Certain pharmaceutical agents or treatment strategies also influence these requirements.
- Musculoskeletal System
The above considerations indicate that for heart failure and in other conditions detailed below the nutrient intake is instrumental in determining the evolution of tissue damage—its amelioration or acceleration. RDA data, although suitable for a healthy population, are not necessarily appropriate for patients suffering from certain forms of illness or predisposed to sickness through genetic constitution. There is a need for a nutritional supplement that can be taken by persons suffering from illness or with a genetic predisposition to illness.
- Central Nervous System
Data given above have clearly demonstrated the relationship of protein deficiency and mitochondrial dysfunction in skeletal muscle. In addition several nutrient agents have been shown to improve skeletal muscle performance. These include creatine, carnitine and taurine.
- Immune System
Mitochondrial dysfunction has been noted as an important factor in several neurodegnerative diseases38. A central role for defective mitochondrial energy production, and the resulting increased levels of free radicals, in the pathogenesis of various neurodegenerative diseases is gaining increasing acceptance39.
- THE INTERACTING PATHWAYS RESPONSIBLE FOR MITOCHONDRIAL FUNCTION
Immune dysfunction occurs with aging and there is growing evidence that reduced immunity is related to reduced mitochondrial dysfunction40.
We have found that the critical path in these interactions is the flow of energy substrates into the mitochondria through carnitine, the transfer of electrons through the complexes via CoQ10, and the modulation of the calcium pump by taurine. We consider the constituents of this path, namely Carnitine, CoQ10 and Taurine, to be the core constituents required to promote mitochondrial function. We have found that these compounds act together on this critical path to provide a synergistic effect.
The other constituents of the cascade given in FIG. 1 aid the action of this core by modulating oxidative stress which results from external factors and mitochondrial dysfunction and in turn promotes further dysfunction.
- DETAILS OF ALTERED MITOCHONDRIAL ENERGETICS IN HEART FAILURE
Adequate energy production is essential not only for cellular function but also for long term cell survival. Cellular energy production from nutrients, especially fatty acids need the coordinated action of a number of co-factors. Three factors namely, carnitine (critical for the transport of long chain fatty acid substrate), coenzyme Q10 (a key transducer for mitochondrial oxidative phosphorylation), and taurine (a key modulator of calcium accumulation) are important in promoting normal cell energetics.
The data for mitochondrial energetic dysfunction has been clearly documented in heart failure and therefore the following details will focus on heart failure as a paradigm.
- REGULATING INTRACELLULAR CALCIUM
In heart failure deficiency of carnitine promotes accumulation of toxic long-chain fatty acids; deficiency of CoQ10 alters electron transport and mitochondrial calcium accumulation also occurs, which can be corrected by the action of taurine. From FIG. 1 it can be seen that normalization of any one of the above three constituents alone will not be sufficient to significantly benefit myocardial energy production in the presence of abnormalities the other factors in the myocardial bioenergetic pathway. In addition, from FIG. 1, it can also be seen that the added action of creatine, known to be deficient in cardiac failure and antioxidants to reduce oxidative stress, known to be elevated in cardiac failure, will enhance the action of the three core constituents carnitine, CoQ10 and taurine. For these constituents to be effective in remodelling the heart, the addition of protein is essential in any supplement. These substances can be given as oral replacements to benefit both myocyte function and long-term survival. Details of the deficiencies and altered metabolism of these constituents in cardiac failure and other diseases are given below. However, it has become apparent that this paradigm applies to a number of other diseases; these will be briefly discussed in each section where appropriate.
Metabolism and Physiological Role of Myocardial Taurine
The failing myocardium exhibits an increase in calcium content and impaired movement of intracellular calcium. Impaired uptake of calcium adversely affects diastolic relaxation whereas the kinetics of transsarcolemmal calcium flux and calcium release by the sarcoplasmic reticulum is a principal determinant of systolic function. Chronic intracellular calcium overload ultimately leads to cell death.
Taurine (2-aminoethanesulfonic acid) is a unique amino acid, which lacks a carboxyl group, and as such it does not enter into protein synthesis. Taurine appears to be an important amino acid for the modulation of cellular calcium levels, exhibiting a remarkable biphasic action by increasing or decreasing calcium levels appropriate to the maintenance of cellular calcium homeostasis41,42,43. In the heart, taurine appears to do this by affecting several myocardial membrane Systems41,42,43. It is reported to enhance Ca++-induced Ca++ release from the sarcoplasmic reticulum both directly and through inhibition of the enzyme phospholipid methyl transferase, influencing the phospholipid environment of the ryanodine-sensitive calcium channel. It also modulates cardiac Ca++ and Na+ through the cardiac sarcolemmal Na+-Ca++ exchanger and a taurine/sodium exchanger. Taurine also has antioxidant properties and reacts with a variety of potentially toxic intracellular aldehydes including acetaldehyde and malonlyldialdehyde44,45.
Taurine is found in particularly high concentrations in the heart (15-25mmole/g protein) representing approximately 60% of the free amino acid pool41,46. Plasma levels are approximately 50-80mmol/L. Taurine is not an essential amino acid in humans as it can be synthesized from cysteine or methionine46; however, most taurine in humans is obtained directly through dietary sources, particularly from fish and milk. Biosynthetic capacity is maturation dependent, being almost non-existent in the human fetus and newborn and progressively increasing until adulthood47,48. Taurine uptake by the myocyte is an active process and b-amino acids such as beta-alanine share the transport site; it is saturable at taurine concentrations of 200 mmole46,48. In the heart the transport of taurine, like that of carnitine, can be stimulated by beta-adrenergic agonists or dibutyryl-c-AMP; however, in other tissues the c-GMP pathways seem to be important46. The taurine transporter of all tissues is regulated by the activation of two calcium sensitive enzymes: protein kinase C (which inhibits the transporter) or calmodulin (which stimulates transport)48. This reciprocal regulation of intracellular taurine levels by these two enzymes is consistent with a physiologic role for taurine in the maintenance of intracellular calcium homeostasis.
- Taurine Levels and Taurine Supplementation in Heart Failure
The observation that TNF-α levels49 and soluble TNF receptors50,51 are raised in heart failure suggest increased cytokine activity in this condition. Grimble52 has shown that the requirement for sulfur containing amino acids is increased when TNF-α is infused. Of greater significance is the fact that transsulfuration of dietary methionine to cysteine is reduced and in consequence levels of taurine and lung glutathione fall unless the animals are supplemented with cysteine. These findings suggest that with increased cytokine activity as observed by us51 in severe heart failure, the need for cysteine and taurine will increase. Since cysteine will replenish not only taurine but also glutathione (an important endogenous antioxidant -see ‘Oxidative Stress’ section, below), it may be an important supplement for replenishing both.
Cardiac taurine concentrations are altered in heart disease. Cats have very little taurine biosynthetic capacity and may exhibit a taurine deficient cardiomyopathy53. Prolonged taurine depletion of the myocardium has been shown to decrease contractile force through reduction of myofibrils54. This finding is of interest because increased calcium levels in the myocyte can activate calcium dependant proteinases that in turn can lead to the breakdown and loss of myofibils54. Taurine depletion also renders the heart more susceptible to ischemic injury55. In this context it should be noted that myocardial taurine depletion has been reported following acute ischemic injury56 and cardiovascular bypass surgery57.
In species other than the cat, myocardial hypertrophy and failure is associated with an increase in cardiac taurine concentration41,58. In spite of this increase, orally administered taurine has been shown to have a cAMP-independent positive inotropic effect in animal models of left ventricular dysfunction41. Taurine administration has been shown significantly reduce calcium overload and myocardial damage in a variety of heart failure models including that induced by the calcium paradox, doxorubicin or isoproterenol or in hamster cardiomyopathy41,59,60,61,62; it also has been reported to increase the survival of rabbits with aortic regurgitation63. Taurine may have a beneficial effect on cardiac arrhythmias64 including those associated with digitalis or catecholamine excess65. Studies of taurine administration in humans have been limited. However, in patients suffering from congestive heart failure taurine, given in an oral dose of 1 gram three times per day, has been reported to be extremely well tolerated and to improve both hemodynamic state and functional capacity41,56,67.
- MYOCARDIAL ENERGETICS
Carnitine in Health and Disease
Taurine also appears to function as an osmoregulator and neuromodulator in the brain68. In addition there is evidence that taurine modulates calcium influx and efflux in the brain, increases resistance to hypoxia and reduces seizure activity when administered intraperitoneally. In streptozotocin-induced diabetic rats taurine also appears to protect against the development of renal dysfunction45; cardiac studies have not been performed in this model. On the same lines taurine also protects the kidney and liver against doxorubicin (adriamycin) toxicity69.
L-carnitine, an amino acid derivative (3-hydroxy-4-N-trimethylaminobutyric acid), plays a critical role in this bioenergetic pathway as it is essential for the transport of long-chain fatty acids from the cytoplasm into the sites of beta-oxidation within the mitochondrial matrix.70 The importance of carnitine has been recognized by the observation that carnitine deficiency occurs in several genetically determined metabolic abnormalities71,72 where it is associated with the development of cardiomyopathy and skeletal muscle dysfunction. L-carnitine administration to these patients restored to a great extent cardiac and skeletal muscle function.
Evaluation of carnitine metabolism in several cardiac pathologies has led to the realization that carnitine deficiency may also be acquired and organ selective, a finding of great significance because fatty acid oxidation is a major source of energy for the myocardium. The impaired heart, regardless of the etiology of the dysfunction (including ischemic or non-ischemic dilated cardiomyopathy, coronary, hypertensive, diabetic and valvular heart disease), exhibits a marked depletion (up to 50%) of myocardial carnitine levels (particularly free carnitine) in both animal models and man73,74,75, with evolution of the heart disease. On the other hand, despite low cardiac levels, plasma carnitine levels increased76. This finding makes plasma levels unrepresentative of the levels in the heart.
In addition to promoting the entry of fat into the mitochondria, carnitine binds acyl groups and releases free CoA. These processes benefit the myocyte in two ways, first it removes toxic short chain acyl groups and second maintains sufficient amounts of free CoA for mitochondrial function.
An example of the detoxifying action of carnitine is seen in the ischemic myocardium77,78. In ischemic myocardium or skeletal muscle there is an accumulation of long chain acyl-CoA; these compounds are potentially toxic as they exhibit both detergent-like properties and can impair mitochondrial energy production through the inhibition of a mitochondrial membrane enzyme, adenine nucleotide translocase, which transfers newly synthesized ATP from the inner mitochondria space into the cytoplasm. Carnitine protects the heart (or skeletal muscle) from the accumulation of these metabolic poisons by forming acylcarnitines, which can freely diffuse out of the cell and be eliminated through the urine.
- Carnitine Supplementation and Treatment
Carnitine deficiency has also been observed in patients with chronic renal failure; an improvement in cardiac function following carnitine therapy has been reported for those on hemodialysis79.
Body stores of 1-carnitine are supplied by both diet and via endogenous biosynthesis from trimethyllysine. The concentration of carnitine in normal adult cardiac and skeletal muscle is approximately 8-15 nmol/mg non-collagen protein; plasma levels are approximately 35-50 mmol/L. Thus plasma levels are generally not good measures of tissue concentrations. Under normal conditions approximately 80% of carnitine is free and the remainder complexed as fatty acylcarnitine. A 20-50:1 intracellular to extracellular carnitine gradient is maintained by a sodium-dependent plasma membrane transport system. Carnitine transport can be stimulated by beta-adrenergic agonists or dibutyryl-cAMP.
Following oral administration, peak plasma concentration occurs at 3 hours and decays with a T½ of 3-4 hours; the turnover of endogenous cardiac or skeletal muscle carnitine is likely on the order of several days. The bioavailability of 1-carnitine is limited to approximately 5-20% probably due to clearance by the liver. L- carnitine is well tolerated and no adverse effects have been described.
Acetyl-1-carnitine and proprionyl-1-carnitine are naturally occurring derivatives of 1-carnitine80. Acetyl-1-carnitine has been shown to influence mitochondrial DNA synthesis, is depleted by antiretroviral drugs and is non-toxic when infused intravenously81. The administration of acetyl-1-carnitine has been shown to effectively replace decreased carnitine stores in the brain and heart associated with aging in rats82. Acetyl-1-carnitine has excellent penetration of the CSF and is likely to be of benefit in neurodegenerative conditions such as Alzheimer's disease81,83. Acetyl-1-carnitine has also been shown to be of benefit to peripheral nerve function in experimental diabetes84.
Proprionyl-1-carnitine has also been used for cardiac therapy85,86,87,88. Proprionyl-1-carnitine directly penetrates the cell membrane and has a high affinity for the protein carriers; within the mitochondria the enzyme carnitine acetyl transferase releases 1-carnitine and proprionyl-CoA and the latter is transformed into succinyl CoA which can prime the citric acid cycle and the production of ATP89,90. The proprionyl group appears to stimulate fatty acid oxidation (whereas the acetyl group inhibits this)87.
- Ubiquinone or Coenzyme Q10
Coenzyme Q10 in Health and Diseasereviewed in 99
L-carnitine (3-5 grams) or proprionyl-1-carnitine (1.5-3.0 grams) administration has been shown to result in significant hemodynamic improvement and an overall benefit in the functional capacity of animals and patients with heart failure or myocardial ischemia72,73,79,85,86,,91,92,93,94,95,96,97,98,. Clinical studies report a reduction in cardiac damage, when 1-carnitine is taken from 4-12 weeks following myocardial infarction78,92. It also appears to benefit patients who suffer from skeletal muscle ischemia manifested as intermittent claudication. Circulating levels of tumour necrosis factor, a cytokine that leads to muscle wasting and cardiac dysfunction, correlates with functional class and prognosis of patients with heart failure51; in this context it is of interest that proprionyl-1-carnitine administration tends to normalize the circulating levels of this cytokine in patients with heart failure98.
Coenzyme Q10 or ubiquinone (2,3 dimethoxy-5 methyl-6-decaprenyl benzoquinone) plays a vital role as a rate-limiting carrier for the flow of electrons through complexes I, II and III of the mitochondrial respiratory chain. It is also a major endogenous lipophilic antioxidant and, like vitamin C, can regenerate a-tocopherol, the active form of vitamin E by reducing the a-tocopherol radical. A deficiency of ubiquinone caused by actual loss or through oxidation of the molecule can result in an impairment of energy production. The molecule is sited within the inner mitochondrial membrane but is also associated with the membranes of other intracellular organelles. It is also an important component of circulating LDL particles, protecting LDL from oxidation.
Ubiquinone is actively biosynthesized with the cells. The quinone ring is synthesized from the amino acid tyrosine and the polyisoprenoid side chain is formed through the acetyl CoA-mevalonate pathway. The latter pathway is under the control of the enzyme hydroxy-methylglutaryl coenzyme A reductase (HMGCoA reductase) and is also used for cholesterol synthesis; inhibition of this pathway using HMGCoA reductase inhibitors, drugs which decrease plasma cholesterol, also results in a parallel decrease in plasma ubiquinone100 and may also reduce tissue ubiquinone levels101.
- Ubiquinone Supplementation and Treatment99,105,106,107
Significantly reduced levels of myocardial ubiquinone are found in heart failure in both animal models and man102,103,104. Since the heart depends upon aerobic oxidation for its energy needs, ubiquinone, which is critically necessary for oxidative energy production is very important for cardiac function. The antioxidant properties of ubiquinone would add to this benefit.
Ubiquinone is widespread throughout all food groups and thus body stores may also be partially supplied by diet. The concentration of ubiquinone in normal cardiac muscle is approximately 0.4-0.5 mg/mg dry weight, slightly less in skeletal muscle and 0.6-1.3mg/ml in plasma. Oral absorption is slow and markedly enhanced in the presence of lipid; plasma levels peak at 5-10 hours and decay with a T½ of 34 hours. There is a large hepatic first pass effect so that only about 2-5 % of an oral dose is taken up by the myocardium. The mean steady state level in plasma increases 4-7 fold after 4 days of dosing at 100 mg 3 times daily. Side effects are virtually absent; however, asymptomatic elevations in liver enzymes (LDH, SGOT) have been described with doses of 300 mg/day.
Role of Creatine in Health and Disease
Oral ubiquinone therapy has been shown to result to beneficially affect cardiac dysfunction in a variety of animal paradigms108,109,110. Oral ubiquinone also has been reported to reduce the age-associated decline in mitochondrial respiratory function in rat skeletal muscle111. The controlled trials in patients with heart failure show clinical benefit, reducing symptoms, lessening hospitalization and improving myocardial performance99,,102,103,105,,112,113,114,115.
Creatine phosphate (PCr) is the primary high-energy phosphate reservoir of the heart and skeletal muscle. High-energy phosphate is transferred from PCr to ADP to form ATP through catalysis by creatine kinase116:
Muscle creatine stores are maintained through biosynthesis from endogenous precursors arginine, glycine, and methionine in the liver, pancreas and kidneys, and through the ingestion of meat and fish. The concentration of total creatine in normal adult human myocardium or skeletal muscle is approximately 140 mmol/g protein; creatine phosphate constitutes about 65-80% of the total creatine under aerobic conditions116. Creatine is accumulated by muscle against a large concentration gradient from the blood; the transporter is probably driven by the extracellular/intracellular Na+ electrochemical potential116,117. There is evidence that increased adrenergic drive (a characteristic of heart failure) can decrease myocardial creatine and creatine kinase stores123,124.
Experimental creatine depletion in animals results in structural, metabolic and functional abnormalities in muscle117. Myocardial creatine content and myocardial energetics is reduced in a wide variety of animal paradigms of heart failure116,118,119,123,120. No data is available regarding creatine replacement in these models. However creatine has been shown to attenuate myocardial metabolic stress in rats caused by inhibition of nitric oxide synthesis121.
- Creatine Supplementation and Treatment
Hearts from patients with coronary artery disease, aortic stenosis or heart failure all show a marked reduction in total creatine (up to 50%) with an expected concomitant reduction in creatine phosphate122,123; creatine kinase is also reduced116,117. These reductions interact synergistically decreasing myocardial capacity for ATP synthesis by up to 80%116,117,124. Such an energy deficit has a significantly adverse impact on myocardial function and survival. Similar abnormalities of energy stores and production are seen in the skeletal muscles of patients of heart failure; these play an important role in compromising the functional capacity of these patients125. Recently, it has been reported that the myocardial PCr/ATP ratio may be a better predictor of patient mortality in dilated cardiomyopathy than left ventricular ejection fraction or the patient's functional class126.
The role of creatine supplementation may not be observed in normal cardiac or skeletal muscle under normal levels of performance. However, supplementation has been shown to increase performance in situations where the availability of creatine phosphate is important127,128. Ingestion of 3 g of creatine per day for one month (or 20 g per day for one week) increases muscle creatine content and can improve performance99. Daily turnover of creatine to creatinine for a 70 kg male has been estimated to be approximately 2 g129. Creatine supplements will increase skeletal muscle creatine and increase muscle resistance to fatigue during short-term intense exercise where it reduces lactate accumulation130,131,132. It also appears to be of benefit when given in adequate doses to patients with heart failure where myocardial and perhaps skeletal muscle creatine and creatine phosphate levels are depressed. The administration of a creatine supplement to patients with heart failure did not increase cardiac ejection fraction but significantly increased not only skeletal muscle creatine phosphate but also muscle strength and endurance133 and thus would benefit patient symptom-limited performance.
Thiamine or vitamin B1 status may be compromised in heart failure due to a variety of causes. Thiamine is a water-soluble vitamin which functions as a coenzyme in a variety of enzyme systems especially those related to energy metabolism. Thiamine is stored in very small quantities (approximately 30 mg) with approximately half of the body stores being found in skeletal muscle with the remainder being found in the heart, kidney and nervous tissue including the brain. Since little is stored, thiamine requirements must be met daily. Thiamine requirements are related to daily energy expenditure134 and metabolizable energy intake, especially carbohydrate intake and therefore patients with increased metabolic rates or poor intakes, such as those with heart failure, may be at increased risk for deficiency during acute illness134,135. The Food and Nutrition Board, USA recommends that for adults with energy intakes below 2000 kcal/day that a basal requirement of 1.0 mg thiamine per day be maintained136. The Canadian recommendations support an intake of 0.4 mg/1000 kcal or no less than 0.8 mg/day for adults137. Thiamine intake in patients with heart disease has been examined in only one study using a semi-quantitative food frequency questionnaire focusing on foods high in thiamine138. Nutrient analysis indicated a low overall intake of thiamine of 0.966 mg/day with 33% of patients not meeting the Recommended Dietary Allowance (RDA) for thiamine65. This study also indicated that thiamine-deficient diets were more common among patients treated as out-patients rather than in-patients.
Necropsy studies indicate that thiamine deficiency is underdiagnosed in life135. Classical deficiency signs are often absent or they are not recognized139. Thiamine deficiency results in beri beri, which can have neurological or cardiac effects. The symptoms that are most common are mental confusion, anorexia, muscle weakness ataxia, edema, muscle wasting, tachycardia and an enlarged heart140. Thiamine deficiency leads to several major derangements of the cardiovascular system including peripheral dilatation leading to a high output state, biventricular myocardial failure, retention of sodium and water leading to edema as well as a relative depression of left ventricular function with low ejection fraction140,141. This picture will be masked in preexisting low-output heart failure. Since thiamine deficiency will exacerbate co-existing heart failure, correction of this deficiency through supplementation has the potential to improve cardiac status in patients with congestive heart failure.
In addition to poor dietary intake and increased metabolic utilization, referred to above, patients with heart failure also may be at risk for thiamine deficiency because of their need for diuretics. There is evidence from both animal and human studies that diuretics, especially those which affect the Loop of Henle such as furosemide, cause increased urinary losses of thiamine even in the presence of thiamine deficiency142,143. It appears that furosemide treatment may block the kidneys' ability to adapt thiamine excretion in order to prevent thiamine deficiency. These data demonstrate that patients with heart failure are at increased risk of thiamine deficiency due to a combination of 1) poor intake resulting from anorexia and unpalatable diets, 2) hypermetabolism, and 3) enhanced excretion caused by the concurrent use of diuretics.
There have been a handful of studies which support a high prevalence of thiamine deficiency in both in- and outpatients with congestive heart failure—from 13% to 91% depending on the population studied138,143,144,145. A very high (91%) prevalence of thiamine deficiency has been reported in a group of congestive heart patients on long-term furosemide treatments143. The average dose of furosemide ranged from 80-240 mg/day. Biochemically, the furosemide-treated group had severe thiamine deficiency indicating that doses of this magnitude have significant effects on thiamine status. These investigators undertook a randomized double-blind trial of thiamine supplementation in 30 in-patients with heart failure and on long-term furosemide therapy146. Patients were randomized to receive 200 mg intravenous thiamine or a placebo for seven days after which all subjects were placed on an oral supplement of 200 mg/day of thiamine and followed for an additional six weeks. They saw a significant diuresis with increased excretion of sodium and water within two days of thiamine supplementation in comparison with the placebo group whose excretion remained constant. The mean left ventricular ejection fraction increased significantly after one week of thiamine supplementation but not with the placebo. After six weeks of oral thiamine supplementation left ventricular ejection fraction was increased by 22%. This study demonstrates a significant improvement in left ventricular function as a result of thiamine supplementation in patients with CHF. In addition, improvement in left ventricular function was accompanied by diuresis and increased sodium excretion, which is hypothesized to be one of the major effects of thiamine supplementation.
- REDUCING OXIDATIVE STRESS
Finally, it should also be noted that thiamine deficiency is also commonly present in patients on hemodialysis147, in patients in intensive care units148 and perhaps cognitive impairment in the aged149.
Cells are constantly subjected to interplay between free radical injury and protective mechanisms to prevent or minimize free radical injury. Oxidative stress has been defined as a disturbance in the equilibrium between pro- and anti-oxidative systems. A number of different challenges increase oxidative stress resulting in damage to lipids, proteins, DNA and carbohydrates.
- Role of Oxidative Stress in Cardiac Disease
Dietary Antioxidants—Vitamin E, Vitamin C, Cysteine and Selenium—counteract the effect of free radicals generated by external factors and by mitochondrial dysfunction. Cysteine is the precursor of glutathione, one of the most potent antioxidants present in the cell.
Until recently there has been a reluctance to accept that oxidative stress can be important in the pathogenesis of cardiac disease however, recent investigation suggests that oxidative stress may be a very important contributor to the deterioration of the hypertrophied or failing myocardium.
For example, reactive oxygen species have been shown to be critical components of the apoptosis pathway150; myocyte loss by apoptosis is now thought to be a significant contributor to the inexorable deterioration of the failing myocardium151. The importance of oxidative stress in heart failure is not surprising because a number of factors associated with heart failure, such as increased plasma catecholamines,152 and cardiac sympathetic tone153, microvascular reperfusion injury154,155 cytokine stimulation49,50 and mitochondrial DNA mutations (particularly complex I)156 are known stimuli for free radical production and oxidative stress157,158,159,160. Coenzyme Q10 and taurine (and its precursor cysteine), discussed above are important endogenous antioxidants or antioxidant precursors.
Peroxidative damage has been demonstrated in the hearts of dogs, guinea pigs and rats with heart failure due to pressure or volume overload161,162,163 . Vitamin E administration benefited164 both myocardial structure and function. We have observed decreases in the levels of glutathione peroxidase and a-tocopherol and a concomitant increase in protein oxidation in the myocardium of cardiomyopathic hamsters during the late stages of hamster cardiomyopathy165; an elevation of myocardial free radicals and lipid peroxides have also been demonstrated in this model166. The administration of vitamin E appears to completely normalize these findings163.
Recently, we have also demonstrated a significant increase in the plasma level of lipid peroxides and malonyldialdehyde, markers of oxidative stress, in patients suffering from congestive heart failure51. The increase in oxidative stress was related to the clinical severity of heart failure with the highest levels of lipid peroxidation and malonyldialdehyde being observed in class 3 and class 4 patients. Increased free radical activity is also seen in patients on life support or in intensive care unit settings167. These observations suggest that antioxidant supplements should be important additions to the therapy of heart failure and severely ill patients.
The ability to withstand peroxidative injury is partially dependent on diet. A good dietary intake of the antioxidant vitamins C and E, and trace nutrient minerals such as selenium together with adequate cysteine as a precursor for glutathionc synthesis arc important for protection against free radical injury. Fat intake and composition is also important, the need for vitamin E may be increased by an increased intake of polyunsaturated fatty acids168. It has been suggested that the vitamin E requirement=5.96+0.25(% PUFA kcal+g PUFAs). With the North American diet increasing in PUFA, Diplock has suggested that the current RDA (recommended daily allowances) be increased fourfold169. Dietary antioxidants may reduce the risk of ischemic heart disease and the extent of myocardial infarction170,171,172. Recent reports suggest an increased need for vitamin C in patients with diabetes mellitus173. Finally oxidative stress is also felt to be a major contributor to chronic neurodegenerative disease and the tissue deterioration and immune dysfunction associated with aging174,175,176,177,178,179,180,181,182,183,184.
Correction of Mitochondrial Abnormalities
Other investigators have used certain of these nutrients alone. We have found that a preferred nutritional supplement would replace the constituents given above and influence several metabolic pathways interacting to subserve mitochondrial function. We have found that replacing only one of the core constituents will not correct the connected abnormalities in multiple parts of the myocardial bioenergetic pathway which is deranged in cardiac failure. Rather, our supplement helps to correct the cascading series of metabolic abnormalities in patients with myocardial dysfunction and other diseases mentioned above, rather than merely a problem at a single point in a pathway.
In a preferred embodiment, the invention relates to a method of medical treatment of a disease, disorder or abnormal physical state in a mammal selected from the group consisting of heart disease and functional deterioration associated with ageing, the method comprising administering to a mammal an effective amount of a carrier and a nutritional supplement comprising L-Carnitine or its functional analogue, Coenzyme Q10 (Ubiquinone) or its functional analogue and Taurine or its precursors in a single or divided daily dose.
In one variation, the method includes administering to a mammal an effective amount of a carrier and a nutritional supplement comprising L-Carnitine, Coenzyme Q10 (Ubiquinone) and Taurine or its precursors in a single or divided daily dose. The mammal is preferably one of the group including humans, dogs, cats and horses. The disease, disorder or abnormal physical state may include a disease, disorder or abnormal physical state that is due in whole or in part to aging.
BRIEF DESCRIPTION OF THE DRAWINGS
Another variation of the invention includes a method of increasing neuromuscular or muscular or athletic performance in a mammal, the method including administering to the mammal an effective amount of a carrier and a nutritional supplement comprising L-Carnitine or its functional analogue, Coenzyme Q10 (Ubiquinone) or its functional analogue and Taurine or its precursors in a single or divided daily dose. Another aspect of the method includes administering to the mammal an effective amount of a carrier and a nutritional supplement comprising L-Carnitine, Coenzyme Q10 (Ubiquinone) and Taurine or its precursors in a single or divided daily dose. The mammal is preferably one of the group including humans, dogs, cats and horses. The method and composition improves functional performance and muscular performance. In one embodiment, for an 80 kg man, suitable amounts would include at least about 2.7 g taurine, at least about 2.7 g L-carnitine and at least about 135 mg of CoQ10.
FIG. 1 is a schematic diagram of the interaction of the nutrients included in this invention and their interaction in cell energetics.
FIG. 2 shows electron micrographs of heart specimens from hamsters in Experiment 1. (a) control non-treated heart (b) cocktail treated heart (c) normal hamster heart.
FIG. 3 shows the mean pressure of contraction of hamster hearts (Langendorff Model) in Experiment 1. (a) normal hamster heart (b) control non-treated heart (c) control non treated heart (d) control non-treated heart and cocktail treated heart (e) cocktail treated heart (f) cocktail treated heart.
FIG. 4 shows plasma vitamin E levels as a measure of compliance.
FIG. 5 shows that exercise capacity at 12 weeks with the supplement increased by approximately 30%.
DETAILED DESCRIPTION OF THE INVENTION
FIG. 6(a) shows a preferred embodiment of a supplement of the invention. One packet includes about 125 mL of solution. About 250 mL is the recommended daily dose for a human (preferably for a male of about 80 kg). Variations of this embodiment may be made. For example, many of the compounds indicated in brackets on the chart may be varied, such as to use forms of Vitamin C other than the specific form indicated in brackets. Masses used in variations are preferably at least about those masses listed in the chart. As well, specific changes include using at least about 67.5 mg of Coenzyme Q10, at least about 1.35 g of L-carnitine, at least about 875 mg of creatine and at least about 1.35 g of taurine per pack. Each pack includes about 125 mL of solution (carrier). These amounts are preferably given at 250 mL daily in a single or divided daily dose. (b) shows a variation of the supplement. Amounts greater or less than than the masses (including ranges, where shown) shown in FIGS. 6(a) and (b) may be administered depending on individual need. In another variation of the supplement of the invention, carnitine, taurine, and coenzyme Q10 may be administered alone.
Researchers have not recognized that optimizing mitochondrial function depends upon the synergistic correction of cellular and mitochonidrial energy substrates (FIG. 1) which will lead to improved energetics, reduced oxidative stress and better calcium homeostasis, a synergistic response clinically beneficial to the patient.
The conventional approach has been to try single nutrients. This has led to conflicting results, at best.
The combination of nutrients of this invention addresses what we have found to be an interrelated series of disruptions in cellular metabolism, that are present in heart failure and conditions such as aging, chronic neurodegenerative disease, immune diseases such as AIDS, chronic multisystem disease, chronic lung or renal disease, chronic fatigue syndrome, patients on immunosuppressive drugs post-transplantation, cancer patients on doxorubicin or related drugs, wasting or cachexia from cancer or sepsis and in normal humans wishing better neuromuscular or athletic performance, and thus provides more reliable, effective treatment.
The combination is preferably delivered orally, but other methods of administration such as intravenous administration may be used.
The invention is a nutritional supplement that helps to correct or prevent the cascading series of metabolic abnormalities responsible principally for cardiac disease but will have a similar effect on neuromuscular, central nervous and immune system dysfunction in a wide variety of diseases. Rather than merely addressing problems at particular points in metabolic pathways, the nutritional supplement of the invention uses a holistic approach to restoring and improving function at many points in cell metabolism, for example at multiple points along the mitochondrial bioenergetic pathway. The effectiveness of this nutritional supplement in preventing and correcting myocardial dysfunction has been demonstrated in vivo (Example 1).
This invention relates to a dietary supplement comprising effective amounts of L-Carnitine (or its functional analogues such as Acetyl-Carnitine or Proprionyl-1-Carnitine), Coenzyme Q10 (Ubiquinone or its functional analogues) and Taurine as the minimal number of core constituents essential for the correction of the abnormality in mitochondrial energetics in cardiac failure and the different diseases referred to above. Additional supplementation with Cysteine, Creatine, Vitamin E (RRR-d-alpha-tocopheryl), Vitamin C (ascorbic acid), Selenium, and Thiamin in a high protein nutritional feeding are preferred.
This invention relates to a dietary supplement taken in a high protein formulation such as a dairy based drink, a dehydrated dairy product, soya based drink or dehydrated product, or a nutritional bar which may contain: L-Carnitine 0.5-5 g or its functional analogues such as Acetyl- and Proprionyl-1-Carnitine 3 g, Coenzyme Q10 (Ubiquinone) 30-200 mg (preferably at least about 150 mg) or its functional analogues, Taurine 0.1-3 g. Addition of Cysteine 0.5 gm-1.5 g, Creatine 2.5 g, Vitamin E (RRR-d-alpha-tocopheryl) 600 IU, Vitamin C (ascorbic acid) 1000 mg, Selenium 50 meg, Thiamine 25 mg will aid the action of the core constituents. These doses may vary 25% to 300% for specialized formulations. Coenzyme Q10 (Ubiquinone) preferably is at least about 150 mg.
This formulation ensures a high quality protein to optimize muscle function so as to allow the above nutrients in combination to synergistically interact for the benefit of the patient—that is the effect of all of the ingredients combined will be greater than the sum of the individual parts as they address a cascading series of metabolic abnormalities. There is a core of specific nutrients, which must be combined to be effective, and a larger number for optimal effectiveness. Conversely omission of the core will detract from the overall efficacy of this supplement. In addition to maintaining protein stores, the supplement corrects abnormalities in: (a) myocardial energetics, (b) intracellular calcium and (c) oxidative stress.
- Effectiveness of Nutrient Cocktail In Vivo
This supplement also benefits patients, with or without heart failure, with other conditions in which cellular nutrition, mitochondrial energetics and function are impaired or less than desired and oxidative stress is increased, including but not exclusively for musculoskeletal, immune and disorders of the central nervous system especially those related to aging. Such disorders include neurodegenerative disease, immune diseases, stroke, AIDS, chronic multisystem disease, respiratory muscle fatigue such as chronic obstructive lung disease, lung or renal disease, chronic fatigue syndrome, patients on immunosuppressive drugs, cancer patients treated with drugs such as doxorubicin, wasting, cachexia from cancer or sepsis
- Example 1
In vivo data gained from cardiomyopathic hamsters showed the synergistic effect of the nutritional supplement (Experiment 1). It also showed the feasibility of providing a cocktail of nutrients to cardiomyopathic hamsters and that the mixture positively affected cardiac structure, function and markers of oxidative stress, deterioration of mitochondrial and myofibrillar structures in non-treated animals with improved preservation in treated animals. The results show that there are increased areas of necrosis in non-treated hearts in comparison with treated hearts. These results show that this cocktail of nutrients is effective in preserving myocyte function and structure.
Cardiac Nutrient Cocktail Study
- Method of Delivery
We performed a pilot study in order to determine the feasibility of providing a “cocktail” of nutrients as well as their effect on indices of oxidative stress as well as on myocardial structure and function.
|Composition of Cardiac Cocktail |
| ||L-carnitine ||300 mg/kg/day |
| ||Vitamin E ||147 mg/kg/day |
| ||Vitamin C ||100 mg/kg/day |
| ||Coenzyme Q10 ||15 mg/kg/day |
| ||Thiamine (B1) ||100 mg/kg |
| ||Cysteine ||12 mg/day |
| ||Selenium ||0.05 mg/day (5 mg/kg diet) |
| ||Taurine ||188 mg/day (18.8 g/L) |
| ||Creatine ||100 mg/day (1% diet) |
| || |
- Study Design
The nutrients were delivered in 10 ml of raspberry Jell-O. Water soluble nutrients were added directly to the cooled Jell-O while lipid soluble components were mixed with 15 ml 20% intralipid prior to their addition to the cocktail mixture.
180-day old cardiomyopathic hamsters were started on supplementation after a two-week acclimation period. 18 animals received Jell-O supplemented with nutrients while 18 received the identical Jell-O but without nutrients. The animals received full supplementation for eight weeks, at this time the animals were 278 days of age. At this time, 6 treated, 6 untreated and 6 non-diseased hamsters underwent the Langendorff procedure modified for hamsters following which the hearts were preserved for electron microscopy. The remaining animals were sacrificed and blood, hearts, livers and muscle were collected for biochemistry.
a) Mortality—In the treated group, 2 hamsters died. Of the non-treated group, 4 hamsters died with an additional hamster being moribund at the time of study.
b) Appearance—The treated hamsters remained active and alert with no visible edema. In contrast, two of the non-treated animals became grossly edematous with exceptionally large black livers. The control non-treated animals were less active and appeared less bright.
c) Biochemistry—The heart and plasma were analysed for indices of oxidative stress.
| || |
| || |
| ||Control - Non ||Cocktail |
| ||Treated ||Treated |
| || |
|Heart Glutathione Peroxidase Activity ||113.05 ± 8.20 ||134.34 ± 16.7 |
|Units/min/mg protein |
|Heart Malondialydehyde ug/g wet ||2.00 ± 0.25 ||1.67 ± 0.22 |
|Plasma Malondialydehyde nmol/ml ||0.17 ± 0.009 ||0.14 ± 0.04 |
|Plasma Glutathione Peroxidase Activity ||5.24 ± 0.82 ||6.03 ± 0.51 |
|Units/mm/mg protein |
|Ratio Heart Weight: Body Weight ||0.008 ± 0.0007 ||0.007 ± 0.0006 |
|D. Function ||Non-Treated ||Treated ||Normal Hamster |
|* MeanPressure- ||20.84 ± 5.97 ||44.17 ± 4.55 ||92.50 ± 11.99 |
|Langendorff mmHg |
Electron micrographs (FIGS. 2(a)-c)) show deterioration of mitochondrial and myofibrillar structures in non-treated animals with markedly improved preservation in treated animals. In addition, there are increased areas of necrosis in non-treated hearts in comparison with treated hearts. The results achieved were superior to those projected from individual studies of the ingredients.
e) Recently we have completed another study with the supplement and confirmed all the above results and in addition have shown a complete restoration of systolic contractility as measured by +dP/dT and diastolic relaxation as measured by −dP/dT.
Nutrients have been used, singly or in random combination to treat several conditions including heart disease. We by contrast have found that three defined key components, L-carnitine, coenzyme Q10 and taurine, interact in a way which potentiates the action of each on cell function and energetics. Creatine, thiamine and antioxidants support the action of this core. In addition protein enrichment also aids the maintenance of mitochondrial energetics. We have demonstrated the validity of this claim by preventing the development of heart failure in a genetic animal model normally subject to the inexorable progression of heart failure culminating in death. Other studies using nutrients have not shown comparable benefit.
In another embodiment of the invention, the nutritional supplement is adapted in an amount effective for administration to humans for the purpose of enhancing muscular or athletic performance.
A randomized placebo-controlled safety and efficacy study of a liquid supplement, containing about: 2.7 grains of taurine, 2.7 grams of carnitine, 135 mg coenzyme Q10 plus antioxidant vitamins (vitamin E—400 IU, Vitamin C—250 mg) and 1.75 grams of creatine per 250 milliliters was performed in 33 healthy untrained human volunteers. The supplement was given for 12 weeks at 125 ml twice daily.
There were 2 dropouts in the placebo group between weeks 4 and 12. There were no dropouts in the active supplement group. Exercise capacity, measured as time until exhaustion at 110% of individual VO2 max, was assessed at baseline and at weeks 4 and 12. FIG. 4 shows plasma vitamin E levels as a measure of compliance. FIG. 5 shows the results where exercise capacity at 12 weeks with the supplement increased remarkably by about 30%.
The nutritional supplement of may be adapted (appropriate to body mass and metabolic rate) to be administered to humans and other mammals, such as dogs, cats and horses, in amounts effective for correcting diseases, conditions and infirmities described in this application, including those due to aging. They may also be adapted (appropriate to body mass and metabolic rate) to be administered to humans and other mammals to enhance muscular performance. Mammals tend to have a high and variable metabolic rate. Their requirements per gram of tissue differs from humans. By way of example, a rat has about four times the metabolic rate of a human. The amounts to be administered will also vary with the age of the mammal. The appropriate amount to be administered will be apparent to one skilled in the art.
All publications, patents and patent applications are herein incorporated by reference in their entirety to the same extent as if each individual publication, patent or patent application was specifically and individually indicated to be incorporated by reference in its entirety. U.S. application Ser. No. 09/002,765, filed Jan. 6, 1998 (“Composition for Improvement of Cellular Nutrition and Mitochondrial Energetics”), U.S. application Ser. No. 08/826,234 filed Mar. 27, 1997 and PCT application no. PCV/CA98/00286 filed Mar. 25, 1998 (“Nutritional Composition for Improvements in Cell Energetics”) are incorporated by reference in their entirety.
The present invention has been described in terms of particular embodiments found or proposed by the present inventors to comprise preferred modes for the practice of the invention. It will be appreciated by those of skill in the art that, in light of the present disclosure, numerous modifications and changes can be made in the particular embodiments exemplified without departing from the intended scope of the invention. All such modifications are intended to be included within the scope of the appended claims.
1 O'Brien, P. J., H. Shen, D. Bissonnette and K. N. Jeejeebhoy. Effects of hypocaloric feeding and refeeding on myocardial Ca and ATP cycling in the rat. Molec Cell Biochem 142: 151-161, 1995.
2 Heymsfeld S B, Lichtman S, Baumgartner R N, Wang J, Kamen Y, Aliprants A and Pierson R N. Body composition of humans: comparison of two improved four-compartment models that differ in expense, technical complexity and radiation exposure. Am J Clin Nutr 52:52-58, 1990
3 Roubenoff R and Kehayias J J. The meaning and measurement of lean body mass. Nut Rev 49:165-175, 1991.
4 Hill G L, King R F G J, Smith R C. et al. Multi-element analysis of the living body by neutron activation analysis-application to critically ill patients receiving intravenous nutrition. Br J Surg 66:868-872, 1979.
5 Moore F D, Olesen K H, McMurrey J D, Parker H V, Ball M R, and Boyden C M. The body cell mass and its supporting environment. Philadelphia: W B Saunders 1963.
6 Jeejeebhoy K N, J P Baker, S L Wolman, D E Wesson, B Langer, J E Harrison and K G McNeill. Critical evaluation of the role of clinical assessment and body composition studies in patients with malnutrition and after total parenteral nutrition. Am J Clin Nutr 35:(Suppl.), 1117-1127, 1982.
7 Russell, D Mc R, P J Prendergast, P L Darby, P E Garfinkel, J Whitwell and K N Jeejeebhoy. A comparison between muscle function and body composition in anorexia nervosa: the effect of refeeding. Am J Clin Nutr 38: 229-237, 1983.
8 Collins J P, Oxby C B and Hill G L. Intravenous amino acids and intravenous hyperalimentation as protein-sparing therapy after major surgery-a controlled trial. Lancet 1:788-791, 1978.
9 Almond D J, King R E G J, Burkinshaw L. Potassium depletion in surgical patients: Intracellular cation deficiency is independent of loss of body protein. Clin Nutr. 6:45-50, 1987
10 Almond D J, King R F G J, Burkinshaw L, Laughland A, McMahon M J. Influence of energy source upon body composition in patients receiving intravenous nutrition. JPEN 13:471-477, 1989
11 Pichard C, C Vaughan, R Struk, R L Armstrong and K N Jeejeebhoy. The effect of dietary manipulations (fasting, hypocaloric feeding and subsequent refeeding) on rat muscle energetics as assessed by nuclear magnetic resonance spectroscopy. J Clin Invest 82: 895-901, 1988.
12 Mijan de la Torre A, Madapallimattam A, Cross A, Armstrong R L, Jeejeebhoy K N. Effect of fasting, hypocaloric feeding and refeeding on the energetics of stimulated rat muscle as assessed by nuclear magnetic resonance spectroscopy. J Clin Invest 92:114-121, 1993.
13 Lopes J, D Mc R Russell, J Whitwell and K N Jeejeebhoy. Skeletal muscle function in malnutrition. Am J Clin Nutr 36:602-610, 1982.
14 Russell, D Mc R, L A Leiter, J Whitwell, E B Marliss and K N Jeejeebhoy. Skeletal muscle function during hypocaloric diets and fasting: a comparison with standard nutritional assessment parameters. Am J Clin Nutr 37: 133- 138, 1983.
15 Berkelhammer C H, L A Leiter, K N Jeejeebhoy, A S Detsky, D G Oreopoulos, P R Uldall and J P Baker. Skeletal muscle function in chronic renal failure: an index of nutritional status. Am J Clin Nutr42: 845-854, 1985.
16 Fraser, I M, D Mc R Russell, S Whittaker, N Zamel, R Goldstein, K N Jeejeebhoy and A C Bryan. Skeletal and diaphragmatic muscle function in malnourished chronic obstructive lung disease. Am Rev Respir Dis 129: A269, 1984.
17 Brough W, G Horne, A Blount, M H Irving and K N Jeejeebhoy. Effects of nutrient intake, surgery, sepsis, and long term administration of steroids on muscle function. Br Med J 293: 983-988, 1986.
18 Russell, D Mc R, H L Atwood, J S Whittaker, T Itakura, P M Walker, D A G Mickle and K N Jeejeebhoy. The effect of fasting and hypocaloric diets on the functional and metabolic characteristics of rat gastrocnemius muscle. Clin Sci 67:185-194, 1984.
19 Dureuil B, Viires N, Veber B, Pavlovic D, Pariente R, Desmonts J M, Aubier M. Acute diaphragmatic changes induced by starvation in rats. Am J Clin Nutr 49:738-44, 1989.
20 Christie P M and Hill G L. Effect of intravenous nutrition on nutrition and function in acute attacks of inflammatory bowel disease. Gastroenterology 99:730-736,1990.
21 Windsor J A and Hill G L. Weight loss with physiologic impairment: A basic indicator of surgical risk. Ann Surg 207:290-296,1988.
22 Hanning R M, Blimkie C J R, Bar-Or O, Lands L C, Moss L A and Wilson W M. Relationship among nutritional status and skeletal and respiratory muscle function in cystic fibrosis: does early dietary supplementation make a difference. Am J Clin Nutr 57:580-587, 1993.
23 Castaneda, C., Charnley, J. M., Evans, W. J., and Crim, M. C. Elderly women accommodate to a low-protein diet with losses of body cell mass, muscle function, and immune response. Am J Clin Nutr 62:30-39, 1995
24 Gadaleta M N, Petruzzella V, Daddabbo L, Olivieri C, Fracasso F, Loguercio Polosa P, Cantatore P. Mitochondrial DNA transcription and translation in aged rat. Effect of acetyl-L-carnitine. Ann N Y Acad Sci 717 :150-60, 1994.
25 Famularo G, Moretti S, Marcellini S, Trinchieri V, Tzantzoglou S, Santini G, Longo A, De Simone C. Acetyl-carnitine deficiency in AIDS patients with neurotoxicity on treatment with antiretroviral nucleoside analogues. AIDS 11:185-90, 1997.
26 Folkers K; Wolaniuk A. Research on coenzyme Q10 in clinical medicine and in immunomodulation.Drugs Exp Clin Res 11:539-45,1985.
27 Schulz J B, Matthews R T, Henshaw D R, Beal M F. Neuroprotective strategies for treatmentof lesions produced by mitochondrial toxins: implications for neurodegenerative diseases. Neuroscience 71:1043-8, 1996.
28. Codd, M. B., Sugrue, D. D., Gersh, B. J. et al. Epidemiology of idiopathic dilated and hypertrophic cardiomyopathy: a population-based study in Olmsted County Minnesota, 1975-1984. Circulation 80: 564-572, 1989.
29. Bourassa, M. G., Gurne, O., Bangdiwala, S. I. et al. for the Studies of Left Ventricular Dysfunction SOLVD) Investigators. Natural history and patterns of current practice in heart failure. J Am Coll Cardiol 22(suppl A): 14A-19A, 1993.
30. Sole M. J. Shifting the paradigm for the treatment of dilated cardiomyopathy. Eur Heart J 16(suppl O): 176-179, 1995.
31. Freeman, L., Roubenoff, R. The nutrition implications of cardiac cachexia. Nutr Reviews 52: 340-347, 1994.
32. Rock, L., Leonard, L. B. Nutrition care of cardiac transplant patients. Top Clin Nutr 5: 1-9, 1990.
33. Heymsfield, S. B., Hoff, R. D., Gray T. F., et al. Heart disease. In: J. M. Kinney, K. N Jeejeebhoy, G. L. Hill, O. E. Owen, eds. W. B. Saunders, Philadelphia. 1988, pp 477-509.
34. Pittman, J. G., Cohen, P. The pathogenesis of cardiac cachexia. N Engl J Med 271: 403-409, 1964.
35. Riley, M., Elborn, J. S., McKane, W. R., et al. Resting energy expenditure in chronic heart failure. Clin Sci. 80: 633-639, 1991.
36. Shiihara, H. Pre- and post-operative nutritional assessment of cardiac cachexia. J Jpn Assoc Thorac Surg 39: 183-191, 1991.
37. Poehlman, E. T., Scheffers, J., Gottlieb, S. S., et al. Increased resting metabolic rate in patients with congestive heart failure. Ann Int. Med. 121: 860-862, 1994.
38 Cooper J M. Schapira A H. Mitochondrial dysfunction in neurodegeneration. Journal of Bioenergetics & Biomembranes. 29:175-83, 1997 .
39 Beal M F. Mitochondria, free radicals, and neurodegeneration. Current Opinion in Neurobiology. 6:661-6, 1996.
40 Weindruch R. Caloric restriction and aging Scientific American. 274:46-52, 1996.
41. Azuma, J., Sawamura, A., Awata, N. Usefulness of taurine in chronic congestive heart failure and its prospective application. Jpn Circ J 56: 95-99, 1992.
42. Schaffer, S. W., Ballard, C., Azuma, J. Mechanisms underlying physiological and pharmacological actions of taurine on myocardial calcium transport. In: R. Huxtable, D. V. Michalk, eds. Taurine in Health and Disease. Adv Exp Med Biol 359: 171-180, 1994.
43. Satoh, H. Cardioprotective actions of taurine against intracellular and extracellular calcium-induced effects. In: R. Huxtable, D. V. Michalk, eds. Taurine in Health and Disease. Adv Exp Med Biol 359: 181-96, 1994.
44. Ogasawara, M., Nakamura, T., Koyama, I. et al. Reactivity of taurine with aldehydes and its physiological role. In: R. Huxtable, D. V. Michalk, eds. Taurine in Health and Disease. Adv Exp Med Biol 359: 71-78, 1994
45. Trachtman, H., Sturman, J. A. Taurine and experimental kidney disease, In: R. Huxtable, D. V. Michalk, eds. Taurine in Health and Disease. Adv Exp Med Biol 359:149-157, 1994.
46. Huxtable, R. J., Chubb, J., Azari, J. Physiological and experimental regulation of taurine content in the heart. Fed Proc 39: 2685-2690, 1980.
47. Sturman, J. A. Taurine in development. Physiol Rev 73: 119-147, 1993.
48. Ganapathy, V., Leibach, F. H. Expression and regulation of the taurine transporter in cultured cell lines of human origin. In: R. Huxtable, D. V. Michalk, eds. Taurine in Health and Disease. Adv Exp Med Biol 359: 51-57, 1994.
49. Packer, M. Is tumor necrosis factor an important neurohormonal mechanism in chronic heart failure? Circulation 92: 1379-1382, 1995.
50. Ferrari, R., Bachetti, T., Confortini, R. et al. Tumor necrosis factor soluble receptors in patients with various degrees of congestive heart failure. Circulation 92: 1479-1486, 1995.
51. Geranmaygan, A., Keith, M., Sole, M. J., et al. Increased oxidative stress in patients with congestive heart failure. Submitted
52. Grimble, R. F., Jackson, A. A., Persaud, C., et al. Cysteine and glycine supplementation modulate the metabolic response to tumor necrosis factor alpha in rats fed a low protein diet. J Nutr 122: 2066-2073, 1992.
53. Pion, P. D., Kittleson, M. D., Rogers, Q. R. et al. Myocardial failure in cats associated with low plasma taurine: a reversible cardiomyoapthy. Science 237: 764-768, 1987.
54. Lake, N. Alterations of ventricular contractility and myofibrillar loss in taurine deficient hearts. In: R. Huxtable, D. V. Michalk, eds. Taurine in Health and Disease. Adv Exp Med Biol 359: 335-342, 1994.
55. Schaffer, S. W., Allo, S., Harada, Mozaffari, M. Potentiation of myocardial ischemic injury by drug- induced taurine depletion. In: R. J. Huxtable, F. Franconi, A. Giotti, eds. The Biology of Taurine. Plenum Press, New York. 1987, pp 151-158.
56. Crass III, M. F., Lombardini, J. B. Loss of cardiac muscle taurine after acute left ventricular ischemia. Life Sci 21: 951-958, 1977.
57. Suleiman, M. S., Fernando, H. C., Dihmis, W. C. A loss of taurine and other amino acids from ventricles of patients undergoing bypass surgery. Br Heart J 69: 241-245, 1993.
58. Huxtable, R., Bressle, R. Taurine concentrations in congestive heart failure. Science 184: 1187-1188, 1974.
59. Kramer, J. H., Chovan, J. P. Schaffer, S. W. The effect of taurine on calcium paradox and ischemic heart failure. Am J Physiol 240: H238-H246, 1981.
60. Ohta, H., Azuma, J., Awata, N. et al. Mechanism of the protective action of taurine against isoprenaline induced myocardial damage. Cardiovasc Res 22: 407-413, 1988.
61. Hamaguchi, T. Azuma, J., Awata, N., et al. Reduction of doxorubicin-induced cardiotoxicity in mice by taurine. Res Comm Chem Pathol Pharmacol 59: 21-30,1988.
62. Azari, J., Brumbaugh, P., Barbeau, A., et al. Taurine decreases lesion severity in the hearts of cardiomyopathic Syrian hamsters. Can J Neur Sciences 7: 435-440, 1980.
63.Takihara, K., Azuma, J., Awata, N. Beneficial effect of taurine in rabbits with chronic congestive heart failure. Am Heart J 112: 1278-1284, 1986.
64. Wang, G. X., Duan, J.,Zhou, S., et al. Antiarrhythmic action of taurine. In: J. B. Lombardini ed. Taurine. Plenum Press, New York. 1992, pp 187-192
65. Read, W. O., Welty, J. D. Effect of taurine on epinephrine and digoxin-induced irregularities of the dog heart. J Pharmacol Exp Ther 139: 283-289, 1963.
66. Azuma, J., Sawamura, A., Awata, N. Therapeutic effect of taurine in congestive heart failure: a double-blind crossover trial. Clin Cardiol 8: 276-282, 1985.
67. Zavolovskaia, L. I., Elizarova, E. P., Orlov, V. A. [The clinical efficacy of taufon in the combined treatment of patients with chronic circulatory failure] Eksperimnentalnaia i Klinicheskaia Farmakologiia 58: 29-32, 1995. (in Russian).
68 Oja S. S. Saransaari, P. Taurine as an osmoregulator and neuromodulator in the brain. Metab Brain Dis 11:153-164, 1996.
69 Venkatesan N, Venkatesan O, Karthikeyan J and Armugam V. Protection by taurine against adriamycin induced proteinuria and hyperlipidemia in rats. Proc Soc Exp Biol Med 215:158-164,1997.
70. Hoppel C. The physiological role of carnitine. In: L-carnitine and Its Role in Medicine: From Function to Therapy. R. Ferrari, D. DiMauro, and G. Sherwood, eds. Academic Press, Toronto. 1992, pp5-20
71. Engel, A. G. Carnitine deficiency syndromes and lipid storage myopathies. Ch 57 In: Myology, Basic and Clinical. A. G. Engel and B. Q. Banker eds. McGraw-Hill Book Co., Toronto. 1986, pp1663-1696.
72.Waber, L. J., Valle, D., Neill, C. et al. Carnitine deficiency presenting as familial cardiomyopathy: a treatable defect in carnitine transport. J Ped 101: 700-705, 1982.
73. Kobayashi, A., Masumura, Y., Yamazaki, N. L-carnitine treatment for congestive heart failure; experimental and clinical study. Jpn Circ J 56: 86-94, 1992.
74.Regitz, V., Bossaller, C., Strasser, R. et al. Metabolic alterations in end-stage and less severe heart failure—myocardial carnitine decrease. J Clin Chem Clin Biochem 28: 611-617, 1990.
75. Regitz, V., Shug, A. L., Fleck, E. Defective myocardial carnitine metabolism in congestive heart failure secondary to dilated cardiomyopathy and coronary, hypertensive and valvular heart diseases. Am J Cardiol 65: 755- 760, 1990.
76. Pierpont, M. E. M., Judd, D., Goldenberg, I. F. Myocardial carnitine in end-stage congestive heart failure. Am J Cardiol 64: 56-60, 1989.
77. Fujisawa, S., Kobayashi, A., Hironaka, Y. et al. Effect of 1-carnitine on the cellular distribution of carnitine and its acyl derivatives in the ischemic heart. Jpn Heart J. 33: 693-705, 1992.
78. Jacoba, K. G. C., Abarquez, R. F., Topacio. G. O. et al. Effect of 1-carnitine on the limitation of infarct size in one-month post-myocardial infarction cases: A multicentre, randomized, parallel, placebo- controlled trial. Clin Drug Invest 11: 90-96, 1996.
79. Van Es, A., Henny, F. C., Kooistra, M. P. et al. Amelioration of cardiac function by 1-carnitine administration in patients on haemodialysis. In: G. Guarnieri, G. Panzetta, G. Toigo, eds. Metabolic and Nutritional Abnormalities in Kidney Disease. Vol 98, Contrib Nephrology. Karger, Basel. 1992, pp28-35.
80 Longo A, Bruno G, Curti S, Mancinelli A, Miotto, G. Determination of 1-carnitine, acetyl-1-carnitine and proprionyl-1-carnitine in human plasma by high performance liquid chromatography after pre-column derivatization with 1-aminoanthracene. J Chromatography B 686: 129-139, 1996.
81Bruno G, Scaccianoce S, Bonamini M, Patacchioli F R, Cesarino F, Grassini P, Sorrentino E, Angelucci L, Lenzi G L. Acetyl-L-carnitine in Alzheimer disease: a short-term studio on CSF neurotransmitters and neuropeptides. Alzheimer Dis Assoc Disord 9:128-31,1995.
82 Maccari, F., Arseni, A., Chiodi, P., Ramacci M. T., Angelucci, L. The levels of carnitines in brain and other tissues of rats of different ages: Effect of acetyl-1carnitine administration. Exp Gerentology 25: 127-134, 1990.
83 Pettegrew, J. W., Klunk, W. E., Panchalingam, K., Kanfer, J. N., McClure, R. J. Clinical and neurochemical effects of acetyl-1-carnitine in Alzheimer's disease. Neurobiol Aging 16: 1-4, 1995.
84 Lowitt, S. Malone, J. I. Korthals, J., Benford, S. Acetyl-1-carnitine corrects the altered peripheral nerve function of experimental diabetes. Metabolism 44: 677-680, 1995.
85. Mancini, M. Rengo, F., Lingetti, M. et al. Controlled study on the therapeutic efficacy of proprionyl-1-carnitine in patients with congestive heart failure. Arzneim-Forsch/Drug Res. 42: 1101-1104, 1992.
86. Dhalla, N. S., Dixon, I. M. C., Shah, K. R. et al. Beneficial effects of 1-carnitine and derivatives on heart membranes in experimental diabetes. In: L-carnitine and Its Role in Medicine: From Function to Therapy. R. Ferrari, D. DiMauro, and G. Sherwood, eds. Academic Press, Toronto. 1992, pp 411-426.
87 Paulson, D. J., Traxler, J. Schmidt, M., Noonan, J., Shug, A. L. Protection of the ischemic myocardium by 1-proprionylcarnitine: effects on the recovery of cardiac output after ischemia and reperfusion, carnitine transport, and fatty acid oxidation. Cardiovascular Res 20: 536-541, 1986.
88 Bartels, G. L., Remme, W. J., Holwerda, K. J., Kruijssen, D. A. C. M. Anti-ischemic efficacy of 1-propionylcarnitine—a promising novel metabolic approach to ischaemia? Eur Heart J 17: 414-420, 1996. 89. Hulsmann, W. C. Biochemical profile of proprionyl-1-carnitine. Cardiovasc Drugs Ther 5(suppll): 7-9, 1991.
90 Di Lissa, F. Menabo, R. Barbato, R., Silprandi, N. Contrasting efects of proprionate and proprionyl-1-carnitine on energy linked processes in ischemic hearts. Am J Physiol 267 (Heart Circ Physiol 36:) H455-H461, 1994.
91. Whitmer, J. T. L-carnitine treatment improves cardiac performance and restores high-phosphate pools in cardiomyopathic Syrian hamster. Circ Res 61: 396-408, 1987.
92. Fernandez, C. Profile of long-term 1-carnitine therapy in cardiopathic patients. In: L-carnitine and Its Role in Medicine: From Function to Therapy. R. Ferrari, D. DiMauro, and G. Sherwood, eds. Academic Press, Toronto. 1992, pp337-341.
93 Maresca, P., Corsico, N., Arrigoni-Martelli, E. Mancinelli R., Mannni, E. Proprionyl-1-carnitine improves mechanical performance of papillary muscle from dilated cardiomyopathic hamsters. Ann N Y Acad Sci 752: 207-209, 1995.
94 El Alaoui-Talibi, Z., Guendouz, A., Moravec, M., Moravec J. Control of oxidative metabolism in volume-overloaded rat hearts: effect of proprionyl-1-carnitine Am J Physiol 272(Heart Cic Physiol 41:) H1615-H124, 1997.
95 Bartels, G. L., Remme W. J., Pillay M., Schonfeld, D. H. W., Kruijssen, D. A. C. M. Effects of 1-proprionylcarnitine on ischemia-induced myocardial dysfunction in men with angina pectoris. Am J Cardiol 74: 125-130, 1994.
96 De Guili F, Cargoni, A., Pasini, E., Mazzoletti, A., Confortini, R. Ferrari, R. Effect of proprionyl-1-carnitine on heart and skeletal muscle metabolism during congestive heart failure. In: The Failing Heart. N. S. Dhalla, R. E. Beamish, N. Takeda, M. Nagano eds Lippincot-Raven Publishers Philadelphia 1995, pp 401-411.
97 The Carnitine System: A New Therapeutical Approach to Cardiovascular Diseases. J. W. De Jong, R. Ferrari eds Kluwer Academic Publishers, Boston, 1995, pp 1-393.
98 Bachetti T., Corti, A., Cassani, G., Confortini, R., Mazzoletti, A., Ferrari, R. Cytokines in end stage congestive heart failure: Effect of proprionyl-1-carnitine. Eur Heart J. 15: 1267-1273, 1994.
99. Littaru, G. P. Energy and Defence: Facts and perspectives on Coenzyme Q10 in biology and medicine. Casa Editrice Scientifica Internazionale, Rome. 1995, pp1-91.
100. Folkers, K., Langsjoen, P., Willis, R. et al. Lovastatin decreases coenzyme Q levels in humans. Proc Nat Acad Sci USA 87:8931-8934, 1990.
101. Low, P., Anderson, M., Edlund, C. et al. Effects of menvinolin treatment on tissue dolichol and ubiquinone levels in the rat. Biochem Biophys Acta 1165: 102-109, 1992.
102. Folkers, K. Langsjoen, P., Langsjoen, P. H. Therapy with coenzyme Q10 of patients in heart failure who are eligible or ineligible for a transplant. Biochem Biophys Res Comm 182: 247-253, 1992.
103. Folkers, K. Vadhanavikit, S. Mortensen, S. A. Biochemical rationale and myocardial tissue data on the effective therapy of cardiomyoapthy with coenzyme Q10. Proc Natl Acad Sci USA. 82: 901-904, 1985.
104. Kitamura, N., Yamaguchi, A. Otaki, M. et al. Myocardial tissue level of coenzyme Q10 in patients with cardiac failure. In: Biomedical and Clinical Aspects of Coenzyme Q10 K. Folkers, Y. Yamamura eds. Volume 4. Elsevier Science Publishers, New York. 1984, pp243-253
105. Nylander, M., Weiner, J., Ruokonene, I. et al. Plasma levels of coenzyme Q10 before and after oral supplementation: a bioavailability study. CoQ Res Biol Med 3: 25-32,1995.
106. Mohr, D. Bowry, V. W., Stocker, R. Dietary supplementation with coenzyme Q10 results in increased levels of ubiquinol-10 within circulating lipoproteins and increased resistance of human low-density lipoprotein to the initiation of lipid peroxidation. Biochim Biophys Acta 1126: 247-254, 1992.
107. Greenberg S., Frishman, W. H. Coenzyme Q10: a new drug for cardiovascular disease. J Clin Pharmacol 30: 596-608, 1990.
108. Momomura, S., Serizawa, T., Ohtani, Y. et al. Coenzyme Q10 attenuates the progression of cardiomyopathy in hamsters. Jpn Heart J 32: 101-110, 1991.
109. Guarnieri, C., Muscari, C., Manfroni, S. et al. The effect of treatment with coenzyme Q10 on the mitochondrial function and superoxide radical formation in cardiac muscle hypertrophied by mild aortic stenosis. J Mol Cell Cardiol 19: 63-71, 1987.
110. Azuma, J., Ohta, K. Takihara, K. et al. Effect of coenzyme Q on myocardial injury induced by isoproterenol and by calcium paradox. In: Biomedical and Clinical Aspects of Coenzyme Q10. K. Folkers, Y. Yamamura, eds. Volume 5. Elsevier Science Publishers, New York. 1986, pp213-222.
111. Sugiyama, S. Yamada, K. Ozawa, T. Preservation of mitochondrial respiratory function by coenzyme Q10 in aged rat skeletal muscle. Biochem and Mol Biol Intl 37: 1111-1120, 1995.
112. Morisco, C. Trimarco, B., Condorelli, M. Effect of coenzyme Q10 therapy in patients with congestive heart failure: a long-term multicentre randomized study. Clin Investig 71: S134-S136, 1993.
113. Langsjoen, P. Langsjoen P. H. Folkers, K. Long-term efficacy and safety of coenzyme Q10 therapy for idiopathic dilated cardiomyopathy. Am J Cardiol 65: 521-523, 1990.
114. Langsjoen, P. H., Vadhanavikit, S., Folkers, K. Response of patients in classes III and IV of cardiomyopathy to therapy in a blind and crossover trial with coenzyme Q10. Proc Natl Acad Sci USA 82: 4240-4244, 1985.
115. Langsjoen, H. Langsjoen, P. Langsjoen, P. Willis, R., Folkers, K. Usefulness of coenzyme Q10 in clinical cardiology: a long-term study. Molec Apects Med 15: S165-S175, 1994.
116. Ingwall, J. S. Is cardiac failure a consequence of decreased energy reserve? Circulation 87(suppl VII): VII-58-VII-62, 1993. 117. Wyss, M., Walliman, T. 1-4 creatine metabolism and the consequences of creatine depletion in muscle. Mol Cell Biochem 133/134: 51-66, 1994.
118. Jeejeebhoy, K. N., Sole, M. J. Unpublished
119. Nascimben, L., Friedrich J., Liao, R. et al. Enalapril treatment increases cardiac performance and energy reserve via the creatine kinase reaction in myocardium of Syrian myopathic hamsters with advanced heart failure. Circulation 91: 1824-1833, 1995.
120. Tian, R. Nasciben, L., Kaddurah-Daouk, R., Ingwall, J. S. Depletion of energy reserve via the creatine kinase reaction during the evolution of heart failure in cardiomyopathic hamsters. J Mol Cell Cardiol 28: 755-765, 1996.
121. Constantin-Teodosiu, D., Greenhaff, P. L., Gardiner, S. M. et al. Attenuation by creatine of myocardial metabolic stress in Brattleboro rats caused by chronic inhibition of nitric oxide synthase. Br J Pharmacol 116: 3288-92, 1995.
122. Ingwall, J. S., Kramer, M. F., Fifer, M. A., et al. The creatine kinase system in normal and diseased human myocardium. N Engl J Med 313: 1050-1054, 1985.
123. Nascimben, L., Ingwall, J. S., Pauletto, P., et al. Creatine kinase system in failing and nonfailing human myocardium. Circulation 94: 1894-1901, 1996.
124. Neubauer, S., Krahe, T., Schindler, R. et al. 31P magnetic resonance spectroscopy in dilated cardiomyopathy and coronary artery disease: altered cardiac high energy phosphate metabolism in heart failure. Circulation 86:1810-1818, 1992.
125. Massie, B. M., Conway, M., Rajagopalan, B. et al. Skeletal muscle metabolism during exercise under ischemic conditions in congestive heart failure. Circulation 78: 320-326, 1988.
126. Neubauer, S., Horn, M., Cramer, M., et al. In patients with cardiomyopathy the myocardial phosphocreatine/ATP ratio predicts mortality better than ejection fraction or NYHA class. Circulation 94(suppl I): I-30, 1996 (abstract).
127. Greenhaff, P. L. Creatine and its application as an ergogenic aid. Int J Sport Nutr 5(suppl): S100-S110, 1995.
128. Maughan, R. J. Creatine supplementation and exercise performance. Int J Sport Nutr 5: 94-101, 1995.
129. Hultman, E., Soderland, K., Timmons, J. A., Cerderblad G., Greenhaff, P. L. Muscle creatine loading in men. J Appl Physiol 81: 232-237, 1996.
130. Dawson, B., Cutler, M., Moody, A., et al. Effects of oral creatine loading on single and repeated maximal short sprints. Australian J of Science & Med in Sport 27: 56-71, 1995.
131. Balsom, P. D., Sonderlund, D., Sjodin, B. Skeletal muscle metabolism during short duration high intensity exercise: influence of creatine supplementation. Acta Physiologica Scandinavica 154: 303-310, 1995.
132. Balsom, P. D., Soderlund, K., Ekblom, B. Creatine in humans with special reference to creatine supplementation. Sports Med 18: 268-280, 1994
133. Gordon, A., Hultman, E., Kaijser, L. Creatine supplementation in chronic heart failure increases skeletal muscle creatine phosphate and muscle performance. Cardiovasc Res 30: 413-418, 1995.
134. Sauerblich, H. E., Herman, Y. F. Stevens, C. O. et al. Thiamin requirement of the adult human. Am J Clin Nutr 332: 2237-2248, 1979.
135. O'Keeffe, S. T., Tormey, W. P., Glasgow, R., et al. Thiamine deficiency in hospitalized elderly patients. Gerontology 40: 18-24, 1994.
136. Recommended Dietary Allowances (8th ed). Washington D.C.: Natl Acad Sci-Natl Res Council. 1974.
137. Recommended Nutrient Intakes for Canadians. Ottawa: Ministry of National Health and Welfare. 1983.
138. Brady, J. A., Rock, C. L., Horneffer, M. R. Thiamin status, diuretic medications and the management of congestive heart failure. J Am Diet Assoc 95: 541-544, 1995.
139. Harper, C. G., Giles, M., Finlay-Jones, R. Clinical signs in the Wernicke-Korsakoff complex: a retrospective analysis of 131 cases diagnosed at necropsy. J Neurol Neurosurg Psychiatry 49: 341-345, 1986.
140. McCormick, D. B. Thiamin In: M. E. Shils, V. R. Young,, eds. Modern Nutrition in Health and Disease. Lea & Febiger, Philadelphia, 1988. pp335-361.
141. Leslie, D., Gheorghiade, M. Is there a role for thiamine supplementation in the management of heart failure? Am Heart J 131: 1248-1250, 1996.
142. Yui, Y., Itokawa, Y., Kawai, C. Furosemide induced thiamine deficiency. Cardiovasc Res 14: 537-540, 1980.
143. Seligman, H., Halkin, H., Rauchfleisch, S., et al. Thiamine deficiency in patients with congestive failure receiving long-term furosemide therapy: a pilot study. Am. J. Med. 91: 151-155, 1991.
144. Kwok, T. Falconer-Smith, J. F., Potter, J. F. et al. Thiamine status of elderly patients with cardiac failure. Age Ageing 21: 67-71, 1992.
145. Pfitzenmeyer, P., Guilland, J. C., d'Athis Ph., et al. Thiamine status of elderly patients with cardiac failure including the effects of supplementation. Int J Vit Nutr Res 64: 113-118, 1994.
146. Shimon, I., Almong S., Vered, Z., et al. Improved left ventricular function after thiamine-supplementation in patients with congestive heart failure receiving long-term furosemide therapy. Am J Med 98: 485-490, 1995
147. Descombes, E., Hanck, H. B., Fellay, G. Water soluble vitamins in chronic hemodialysis patients and need for supplementation. Kidney International 43: 1319-1328, 1993.
148. Jeejeebhoy, K. N., Sole, M. J., Mickallis, D. Unpublished
149 Kanofsky J. D. Thiamin status and cognitive impairment in the elderly J Am Coll Nutr 15: 206-222., 1996.
150 McGowan, A. J., Ruiz-Ruiz, M. C., Gurman, A. M. et al. Reactive oxygen intermediates (ROI): common mediator(s) of poly(ADP-ribose) polymerase (PARP) cleavage and apoptosis. FEBS Letters 392: 299-303, 1996.
151 Narula, J., Haider, N., Virmani, R., et al. Apoptosis in myocytes in end-stage heart failure. N Engl J Med 335: 1182-1189, 1996.
152 Cohn, J. N., Levine, T. B., Olivari, M. T., et al. Plasma norepinephrine as a guide to prognosis in patients with congestive heart failure. N Engl J Med 311: 819-823, 1984.
153 Daly, P., Sole, M. J. Myocardial catecholamines and the pathophysiology of heart failure. Circulation 82: 35-43, 1990.
154 Factor, S. M., Minase, T., Cho, S. et al. Microvascular spasm in the cardiomyopathic Syrian hamster: a preventable cause of focal myocardial necrosis. Circulation 66: 342-345, 1982.
155 Sole, M. J., Liu, P. Viral myocarditis: a paradigm for understanding the pathogenesis and treatment of dilated cardiomyopathy. J Am Coll Cardiol 22 [Suppl A]: 99a-105A, 1993.
156 156 Bobba, A., Giannattasio, S., Pucci, A., et al. Characterization of mitochondrial DNA in primary cardiomyopathies. Clin Chim Acta 243: 181-189, 1995.
157 Singal, B. K., Beamish, R. E., Dhalla, N. S. Potential oxidative pathways of catecholamines in the formation of lipid peroxides and genesis of heart disease. Adv Exp Med Biol 161: 391-401, 1983.
158 Flitter, W. D.. Free radicals and myocardial reperfusion injury. Br Med Bull 49: 545-555, 1993.
159 O'Donnell, V. B., Spycher, S., Azzi, A. Involvement of oxidants and oxidant-generating enzyme(s) in tumour-necrosis-factor-a-mediated apoptosis: role for lipoxygenase pathway but not mitochondrial respiratory chain. Biochem J. 310: 133-141, 1995.
160 Cortopassi, G., Wang, E. Modelling the effects of age-related mtDNA mutation accumulation: Complex I deficiency, superoxide and cell death. Biochim Biophys Acta 1271: 171-176, 1995.
161 Singh, N. Dhalla, A. K., Singal, P. K. Oxidative stress and heart failure. Molec Cell Biochem 147: 77-81, 1995.
162 Prasad, K., Gupta, J. B., Kalra, J. et al. Oxidative stress as a mechanism of cardiac failure in chronic volume overload in canine model. J Mol Cell Cardiol 28: 375-385, 1996.
163 Dhalla, A. K., Singal, P. K. Antioxidant changes in hypertrophied and failing guinea pig hearts. Am J Physiol 266: H1280-H1285, 1994.
164 Dhalla, A. K., Hill, M. F., Singal, P. K. Role of oxidative stress in transition of hypertrophy to heart failure. J Am Coll Cardiol 28: 506-514, 1996.
165 Li, R-K., Sole, M. J., Mickle, D. A. G., et al. Vitamin F and oxidative stress in the heart of the cardiomyopathic Syrian hamster. Free Radical Biol Med in press.
166 Fukuchi, T., Kobayashi, A., Kaneko, M., et al. Possible involvement of free radicals and antioxidants in the early stages of the development of cardiomyopathy in the Bio 14.6 Syrian hamster. Jpn Heart J 32: 655-666, 1991.
167 Sheng, Z. Y., Yang, H. M. The concept and diagnosis of multiple systems organ failure. Chin Med J 107: 563-569, 1994.
168 Horwitt, M. K. Supplementation with vitamin E. Am J Clin Nutr 47: 1088-1089, 1988.
169 Diplock, A. T. Dietary supplementation with antioxidants. Is there a case for exceeding the recommended dietary allowance? Free Radical Biol Med 3: 199-201, 1987.
170 Rimm, E. B., Stampfer, M. J. Ascherio, A., et al. Vitamin E consumption and the risk of coronary heart disease in men. N Engl J. Med 328: 1450-1456, 1993.
171 Singh, R. B., Niaz, M. A., Rastogi, S. S., et al. Usefulness of antioxidant vitamins in suspected acute myocardial infarction (the Indian experiment of infarct survival-3). Am J Cardiol 77: 232-236, 1996.
172 Axford-Gately, R. A., Wilson, G. J. Myocardial infarct size reduction by single high dose or repeated low dose vitamin E supplementation in rabbits. Can J Cardiol 9: 94-98, 1993.
173 Will, J. C., Byers, T. Does diabetes mellitus increase the requirement for vitamin C? Nutr Reviews 54: 193-202, 1996.
174 Weindruch, R., Sohal, R. S. Caloric intake and aging. N Engl J Med 337: 986-994, 1997.
175 Stadtman, E. R. Protein oxidation and aging. Science 257: 1220-1224, 1992.
176 Forster, M. J. Dubey, A., Dawson, K. M. et al. Age-related losses of cognitive function and motor skills in mice are associated with oxidative protein damage in the brain. Proc Natl Acad Sci USA 93: 4765-4769, 1996.
177 Williams, L. R. Oxidative stress, age-related neurodegeneration and the potential for neurotrophic treatment. Cerebrovase Brain Met Rev 7: 55-73, 1995.
178 Meydani, S. N., Wu, D., Santos, M. S., Hayek, M. G. Antioxidants and immune response in aged persons: overview of present evidence. Am J Clin Nutr 62(Suppl 6) 1462S-1476S, 1995.
179 Carney, J. M., Starke-Reed, P. E., Oliver, C. N. et al. Reversal of age-related brain protein oxidation, decrease in enzyme activity, and loss in temporal and spatial memory by chronic administration of the spin-trapping compound N-tert-butyl-alpha-phenylnitrone. Proc Natl Acad Sci USA 88: 3633-3666, 1991.
180 Poulin, J. E., Cover, C., Gustafson, M. R., Kay, M. B. Vitamin E prevents oxidative modification of brain and lymphocyte band 3 proteins during aging. Proc Natl Acad Sci USA 93: 5600-5603, 1996.
181 Smith, C. D., Carney, J. M., Starke-Reed, P. E., Oliver, C. N. et al. Excess brain protein oxidation and enzyme dysfunction in normal aging and in Alzheimer disease. Proc Natl Acad Sci USA 88: 10540-10543, 1991.
182 Muscari, C., Giaccari, A., Giordano, E. et al. Role of reactive oxygen species in cardiovascular aging. Mol Cell Biochem 160/161: 159-166, 1996.
183 Joseph, J. A., Denisova, N., Villabos-Molina, R., et al. Oxidative stress and age-related neuronal deficits. Mol Chem Neuropathol 28: 35-40, 1996.
184 Markesbery, W. R. Oxidative stress and Alzheimer's disease. Free Radoc Biol Med 23: 134-147, 1997.