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Remembering A Health Food Pioneer: Willie Pelzer

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Remembering A Health Food Pioneer: Willie Pelzer

Remembering A Health Food Pioneer

Willie Pelzer 

1934-2013

By Lio Perron

The life story of the man affectionately known as the “Granola King” is a compelling one.  Born in a German family from Lemberg, Poland, Withold Pelzer sailed across the Atlantic to chart a new life in North America after the end of the Second World War. Unfazed by his lack of post-secondary education, his rudimentary English and the backlash against German immigrants at the time, the hard-working 17-year-old found work in the sugar beet fields of Lethbridge, Alberta. A few years later, he moved to Saskatchewan to work on a grain farm. In 1961, after a decade in Western Canada, Pelzer and his young bride moved to Ontario to find new opportunities.

Five decades later, through sheer willpower, Pelzer went on to invent granola in his kitchen. He built a sizable fortune and had an unprecedented impact on Canada’s natural food industry with a string of innovative food products. Today, Sunny Crunch Foods Ltd. has a staff of 100 employees producing an impressive range of products, including granola cereal, nutrition bars, meal replacement bars, sports nutrition bars, energy bars, snack bars, protein powders and herbal capsules. The company’s products are sold in Canada and around the world under different labels.  

“It all goes back to grandma’s recipe. She would take oats, sweeten them with honey and served them warm for dessert. Kids just loved it. No artificial preservatives, just whole grain goodness and organic honey,” explains his son Richard Pelzer who has succeeded his father as company president.

The self-made millionaire gambled his life on a simple family recipe in an industry that was struggling to gain acceptance and recognition in the late 1960s. On Sunny Crunch’s website, Pelzer is quoted as saying “Porridge was one of the only foods I knew of that used rolled oats and I wanted to come up with a new kind of food that would make use of rolled oats and other healthy grains in an attractive new way.”

However, the road to riches was a rocky one. “Dominion was the dominant chain back then. He would visit the personal buyer of each store and try to convince them to sell his granola cereal, but the answer was always the same. This is bird seeds, this is not a product for consumers. They’ll never eat this. No thanks, please go away,” says Richard Pelzer.

The Birth of Canada’s Natural Health Product Industry

Despite repeated setbacks, Willie Pelzer took it all in stride. The dream within his heart was too intense for him to quit in the face of temporary defeat. In 1964, Pelzer, Bill Cooper, Walt Zimmerman, Al Smith, Lionel Pasen and other like-minded health food pioneers banded together to create the Canadian Health Food Association. 

“Willie’s talent was getting rid of all the clutter. He saw the basic problem. Very often we would all get lost in the minutiae that doesn’t count. He got rid of all that and got down to the bare facts. It was a pleasure working with him,” explains industry veteran Lionel Pasen, founder of the Toronto-based company, Natural Products Consulting Corp.

The CHFA now represents over 1,000 businesses across Canada. It’s the country’s largest trade association dedicated to natural health and organic products. Members include manufacturers, retailers, wholesalers, distributors, and importers of natural and organic products. These can include foods, vitamin and mineral supplements, herbal products, homeopathics, sports nutrition products, and health and beauty aids.

Since its creation, the CHFA has played a pivotal role in educating the public about its members’ products. It has also successfully lobbied governments to help grow and protect the industry. Today the natural health product industry is worth $3 billion annually to the Canadian economy. However, in the early days the CHFA remained wary of unsolicited government interference. For example, in the 1970s the Ontario government toyed with the idea of restricting the sale of vitamins to pharmacies alone. It’s a move both Pasen and Pelzer were determined to prevent.

“We met with Dr Allan Dyer who was a senior civil servant representing the Ontario government. Willie and I walked out of the meeting saying ‘Dr. Dyer, you’ll request the next meeting with us’. We went down to a restaurant and I said to Willie, what are we going to do?” says Pasen with a chuckle. “We started a letter campaign. Fortunately, there was a postal strike at the time. By the time the strike was over we had collected 40,000 signatures and the government backed off.” 

Willie Pelzer’s life was intricately linked to the Canadian Health Food Association. He presided over the CHFA from 1969 to 1971, and again from 1991 to 1997. “Willie Pelzer assumed the presidency of the CHFA for a second time at a critical juncture in its history. His energy and personal generosity provided a stable financial platform that allowed the Association to grow into the dynamic organization it is today,” says William Gall, past CHFA Board Chair.

By the early 1990s, the industry wanted its own regulation. With the support of health food stores, they launched another letter campaign. The CHFA lobbied federal Health Minister Allan Rock, whose wife was a natural health aficionado. The government accepted all 53 recommendations put forward by the industry and created a new directorate at Health Canada to regulate the industry.

A Recipe for Success

While Pelzer’s commitment to the CHFA was never in doubt, making granola a mainstream food item remained his overriding priority. In the late 1960s and early 1970s health food stores were few and far between. In order to reach the largest number of customers, Pelzer and his wife set their sights on big retailers. 

“We would make stands in grocery stores and shopping malls and handout granola for people to taste and tell them where they can buy it,” explains his wife, Joy Pelzer. These forays into large supermarkets fueled his desire to achieve success despite his inability to capture the imagination of big grocery chain buyers.

Not one to take no for an answer, Pelzer kept going back to meet the same buyers over and over again. 

“One day, he got to the last buyer and of course he said ‘no’ because that’s what they all said,” explains Richard. “As he was walking out of the office, for whatever reason, he got an inspiration and turned around. For the next 20 minutes, he preached a sermon to the buyer. He doesn’t know where it came from, he can’t recall what he said. For 20 minutes, he pointed his finger at the buyer telling him he’s making a mistake. The buyer said, Willie I want you to do a demonstration at my store for one weekend and we’ll take it from there.”

It was a daunting task. If Pelzer had failed to capture people’s imagination, it likely would have been a fatal blow to his dream. Just like a “clutch” athlete, Pelzer had an uncanny ability to perform above expectations in high-pressure situations. When the time came, he loaded the front seat, the back seat and the trunk of his car and drove to the supermarket to set up a booth in the cereal aisle. By the end of the weekend he had sold out. 

In one weekend, Pelzer had won over customers. The growing popularity of granola wasn’t lost on Dominion buyers. One by one, they started ordering his granola cereal in much larger quantities.

Despite their early success, he and Joy kept reinvesting every penny back into the business when they first launched Sunny Crunch Foods Ltd. “We worked day and night,” says Joy. “We would start at eight in the morning and go until two in the morning when we first opened our little factory. We were baking and packaging for delivery the next day. We took jobs cleaning offices to make extra money. We had to do this for about ten years.”

They eventually found success, but it didn’t change Pelzer, according to those closest to him. He remained generous towards the CHFA and gregarious toward his employees. 

“When you make a mistake, bosses usually get upset and it might be cause for dismissal,” says Vasco Rego, who has been plant manager at Sunny Crunch Foods Ltd for the past 36 years. “He saw mistakes as a positive because he felt people usually learn from them.” 

Through his street-smarts, resourcefulness, stubbornness and hard work, Pelzer created a health food giant out of nothing. At the time of his death, he was at the top of the food chain as one of the richest and most successful entrepreneurs in Canada’s health food industry. 

Willie Pelzer passed away at the Mackenzie Health hospital in Richmond Hill on Friday, May 17 with his family at his side. He was 79 years old. IHR

Insomnia

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Insomnia

Role for acupuncture and melatonin

Abstract

“Sleep may be the price you pay so your brain can be plastic the next day,” is an intriguing statement by Cirelli and Tononi (Cirelli 2008). The 2002 Canadian Community Health Survey (CCHS) showed that as many as 18% of participants averaged less than five hours of sleep per night (Shields 2005). Sleep is a vital and recurrent function in daily life. It is thought to be an essential component of good health and hypothesized to have the following functions: energy conservation, memory consolidation, regeneration of substrates, and rest. Lack of sleep can have a negative impact on a person’s career, physical activity level, social interaction, mental aptitude and quality of life. There can be a severe economic burden caused by lack of sleep related to motor vehicle accidents, decreased workplace productivity, absenteeism from commitments and health-care costs from co-morbidities. Melatonin and acupuncture are two natural therapies with large bodies of evidence supporting their use in the management of insomnia.

Features of Sleep and Insomnia

Insomnia is defined as a state of insufficient sleep; it can include difficulties in falling asleep, staying asleep or a combination of both (Balch 2006). Acute insomnia is thought to last less than one to three months on average and consists of difficulty getting to sleep, continuing sleep, and/or sleep is poor quality; resulting in compromised daytime function (AASM 2005). Chronic insomnia can be defined as a failure to get a full night’s sleep on the majority of days in a month. The deleterious impact of insomnia must be considered as an issue on its own and/or as a sign of an underlying major medical diagnoses. There is a plethora of evidence from cognitive, endocrine, neurological, and behavioral disciplines that links insomnia to stress and that attributes insomnia to a state of hyper-arousal and receptor imbalance. Risk factors associated with insomnia include female gender, widowed or single status, low education or income, being unemployed, smoking status, stress, chronic disease, pain, restriction of activity and health dissatisfaction (Sutton 2001).

Sleep is divided into two types: rapid eye movement (REM) and non REM sleep (NREM) with four associated stages, of which the first three are NREM. The four stages of sleep are as follows: Stage one consists of light sleep where one is easily awakened, while stage two consists of theta activity and alpha waves interrupted by k complexes and sleep spindles. Stage three is a deep sleep that consists of slow brain waves, known as delta waves, accompanied by a lack of reaction to environmental stimuli. Stage four consists of the deepest sleep (REM), without major muscle movements (Dement 1975). It is also noted that in this phase, electroencephalogram (EEG) waves are similar to those of an awakened individual.

The Etiology and Physiology of Insomnia

The etiological basis for insomnia includes but is not limited to disturbances in circadian rhythms, stress, genetic conditions, other co-morbid diagnoses, drugs, alcohol, medications and hormonal imbalances. Physiological mechanisms of insomnia include theories that involve the central and autonomic nervous systems (CNS, and ANS respectively). People with insomnia and poor sleep are found to be more alert on daytime alertness tests, such as the multiple sleep latency test (MSLT) (Bonnet 1995, Seidel 1984). They are thought to have altered electroencephalographic (EEG) activity, (beta/gamma) waves at night (Krystal 2002).

Neurological imaging diagnostics such as single photon emission computed tomography (SPECT), positron emission tomography (PET), or proton magnetic resonance spectroscopy (MRS) have shown that insomniacs may have selective hyperactive brain coordinates (Nofzinger 2004, Smith 2002, Winkelman 2008). A highly active ANS that is found in insomniacs coincides with higher metabolic rates, body temperatures, and heart rates (Freedman 1982). There is a reinforced activation of the hypothalamic-pituitaryadrenal (HPA) axis by corticotrophin-releasing hormone (CRH) (Vgontzas 2001). Additionally, high cortisol levels maintain an increased sympathetic tone. Overall, these explanations suggest that therapeutics designed to restore the balance between the ANS and CNS, by mediating sympathetic and parasympathetic tone, could improve insomnia and optimize sleep quality.

Beyond pharmacotherapy, there are a large number of natural therapeutic options available to patients with insomnia such as melatonin, passionflower, 5-hydroxy tryptophan, L-theanine and acupuncture. It has been suggested that benzodiazepine, melatonin and histamine receptor antagonist activity have all been theoretically linked to insomnia. Supplements should be chosen with the associated mechanisms of action in mind, given the variation in pharmacokinetics. In addition to nutritional applications of treatment, eastern therapeutic strategies such as Traditional Chinese Medicine (TCM) methods have also been efficacious.

Eastern Perspectives in Insomnia

TCM etiologies of insomnia include but are not limited to anger, worry, overwork, excessive sexual activity, irregular diet, and a term known as gall bladder timid and residual heat (Maciocia 2008). It is thought that abnormal yin organs are unable to house essence, thereby causing the person to be restless and awake. The state of the mind and the ethereal soul are crucial to proper sleep. Additionally, internal organ disharmony disrupts essence and essence disrupts the mind; essence and Qi are the roots of the mind and without proper rooting of the mind, insomnia could result (Maciocia 2008). Sleep is contingent on normal functioning of the heart/mind, liver/ethereal soul and the kidneys/will power.

TCM theories are based on organ pair and meridian associated diagnoses resulting from either “full-excess” or “empty-deficiency” conditions. The deficiency or excess conditions are a result of imbalance in the connection between the body and the mind. The full conditions are: liver fire blazing, heart fire blazing, phlegm heat harassing the mind, heart Qi stagnation, heart blood stasis, residual heat in the diaphragm and retention of food (Maciocia 2008). The empty conditions are: heart and spleen blood deficiency, heart yin deficiency, heart and kidneys not harmonized, heart and gall bladder deficiency and liver yin deficiency (Maciocia 2008). The intent is to drain in excess conditions and support/ tonify in deficiency.

TCM strategies of acupuncture and herbal medications are individualized depending on what type of excess or deficient condition the person has. For instance, the diagnosis of insomnia as heart fire blazing is treated by needling points such as bladder 15, 44, spleen 6, Ren 15 and Du 19, with a herbal remedy known as XieXin Tang that collectively clears the heart and calms the mind (Maciocia 2008).

Evidence for the Role of Acupuncture in Insomnia

There is a large body of evidence that has shown the efficacy of acupuncture in reducing insomnia by itself or as part of a disease complex. It is effective as an independent therapy or in conjunction with other treatments. One study utilized the Pittsburgh Sleep Quality Index (PSQI) to compare acupuncture to Zolpidem (10mg) in a psychosomatic clinic of 33 patients with primary insomnia. There were significant improvements in both groups: regression analysis showed the results as a baseline PSQI score of 4.13 (p<0.001), the second score 1.32 (p=0.005), and the third 1.49 (p=0.03), confirming that acupuncture was just as effective as the medication (Tu 2012).

One systematic review examined 33 trials with 2293 patients aged 15 to 98 years of age to assess the safety and efficacy of acupuncture for insomnia alone and with other diagnoses, such as stroke, end-stage renal disease, perimenopause, pregnancy and psychiatric diagnoses. They evaluated needle, electro or magnetic acupuncture and pressure. Compared with no treatment (2 studies, 280 participants) or placebo (2 studies, 112 participants), acupressure resulted in more people with improvement in sleep quality (compared to no treatment: OR 13.08, 95% confidence interval (CI) 1.79 to 95.59; compared to placebo: OR 6.62, 95% CI 1.78 to 24.55) (Cheuk 2012). Compared with other treatment alone, acupuncture combined with other treatment was shown to marginally increase the percentage of people with improved sleep quality (13 studies, 883 participants, OR 3.08, 95% CI 1.93 to 4.90) (Cheuk 2012). Overall, needle acupuncture showed the highest efficacy out of all the subgroups. Another systematic review demonstrated through the measurement of subjective sleep outcomes that acupuncture reduced sleep latency, increased sleep and wake ratio (sleep efficiency), resulted in better sleep duration/quality, and minimal insomnia symptoms (Huang 2009).

One interesting, randomized, double blind, placebo controlled trial showed that in postmenopausal women aged 50-67 years old who had insomnia, (as measured by polysomnography exam (PSG) and questionnaires (WHOQOLBREF, Beck Depression Inventory and PSQI)), the acupuncture group had significantly lower scores on the PSQI and improved WHOQOL outcomes after five weeks; confirming that acupuncture improved sleep quality and quality of life in postmenopausal women with insomnia (Hachu 2013). Collectively, the variety of type of studies demonstrate that acupuncture has a measurable improvement on subjective and objective sleep criteria, and that this could potentially be applied to designing individualized treatment strategies for insomnia patients.

Evidence for the Role of Melatonin in Insomnia

Melatonin is a hormone secreted by the pineal gland that regulates circadian rhythm and signals conditions for sleep in the body (Fritz 2009). Administration of exogenous melatonin, typically between 1-5mg under the tongue half an hour before bed, has been shown to decrease sleep latency, increase sleep duration, and increase REM sleep (Brzezinski 2005, Buscemi 2005). Melatonin has also been demonstrated to assist in the discontinuation of benzodiazepine sleeping medications (Garfinkel1999, Kunz 2012). In a randomized controlled trial, 19 of 24 patients who discontinued benzodiazepines using melatonin continued to have good quality sleep six months later (Garkinkel 1999). Another analysis found that after receiving a prescription for melatonin, one third of patients discontinued use of benzodiazepines (Kunz 2012). Melatonin has been studied in several populations including shift workers, patients on benzodiazepines, patients with idiopathic insomnia, patients with schizophrenia, the elderly including patients with Alzheimer’s and Parkinsons, as well as children, including children with neurodevelopment conditions such as Asperger’s, epilepsy, and ADHD (Fritz 2009). Melatonin has an excellent safety profile, with the exception of concurrent use with calcium channel blockers (CCBs), a blood pressure medication; one study found a small decrease in the effect of CCBs when combined with melatonin (Grossman 2006).

Conclusion

Insomnia by itself or in association with other diagnoses can immensely decrease quality of life, as well as be an increased economic burden on the healthcare system and society. Mechanisms of action thought to be targeted are abnormal melatonin production and receptor function, as well as HPA axis imbalance. Melatonin and acupuncture have shown immense potential in insomnia management and are well supported by a large body of evidence. Melatonin has been shown in many studies to improve sleep quality and latency, morning alertness and quality of life at a dose range of 2-4 mg for approximately three weeks. Acupuncture has been shown in several studies to improve sleep efficiency, quality and overall quality of life, and even in some studies enabled patients to reduce medication, especially with individualized protocols. Collectively, these effective natural treatments in addition to targeting the root cause of disease have shown promise in reducing insomnia associated morbidity, and increasing overall wellbeing.

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Garfinkel D, Zisapel N, Wainstein J, Laudon M. Facilitation of benzodiazepine discontinuation by melatonin: a new clinical approach. Arch Intern Med. 1999 Nov 8;159(20):2456-60.

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Huang W, Kutner N, Bliwise DL. A systematic review of the effects of acupuncture in treating insomnia. Sleep Med Rev. 2009 Feb;13(1): 73-104.

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Sports Nutrition

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Sports Nutrition

In the context of lifestyle medicine

In the course of their practice, integrative healthcare practitioners encounter patients who are on an exercise continuum, a continuum that ranges from sedentary adults with good intentions all the way to elite athletes. The rapid expansion of research under the umbrella term ‘sports nutrition’ is relevant not only to the very small percentage of North Americans who might be considered elite athletes, but may also be of use to more ordinary folks. We argue that central research findings related to nutritional influences on performance can be selectively used as a means to support sedentary adults who may be struggling with initiating and maintaining an exercise program. Low motivation and high perceived exertion, associated with a lowered mental outlook, are primary obstacles to exercise adherence. We hypothesize that the same nutritional variables that assist in athletic success, including the Mediterranean diet, plant-based antioxidants such as astaxanthin, cherries and beetjuice, fish oil, branched chain amino acids, and creatine, may be factors with much more broad public health implications in terms of being able to support lifestyle change among the ordinary North American population.

Introduction

There are volumes of international research attesting to the value of regular exercise in the reduction of chronic disease risk, overall mortality, and improvement in mental outlook (Blumenthal 2011). Whereas much has been written concerning the obesity epidemic and the prevalence of sedentary behavior, there are small signs of hope related to participation in moderate intensity physical activity among Canadians. For example, the latest statistics show a 4% increase in the number of physically active Canadians compared to this number in 2003 (Humphreys 2013), and separate data also reflects an increase in time spent in more intense levels of leisure time physical activity (Gilmour 2007). Participation in structured athletics, particularly among females in academic programs, has seen dramatic growth (Shriver 2013). Trends also suggest that North Americans are now more likely to receive physical activity recommendations from healthcare providers – a 40% increase over the last decade (Barnes 2012). Current medical students are more likely to be physically active than the general population, and a high level (69%) of Canadian medical students perceive exercise counseling to be highly relevant to clinical practice (Holtz 2013). Overall, this signals a beginning of a shift in perception, with both the public and the medical profession recognizing the importance of physical activity, and beginning to implement small lifestyle changes.

None of these encouraging signs, however, should be used to gloss over the staggering realities that despite these changes, only 15% of Canadian adults are meeting current guidelines for physical activity (Colley 2011). The same research suggesting that soon-to-be physicians believe exercise counseling to be important also shows that 86% of these graduating medical students consider themselves to be ill-prepared for such counseling: 70% reported no training on interacting with patients concerning exercise (Holtz 2013). The more salient point is that integrative healthcare practitioners are increasingly being called upon to provide exercise counseling to patients on a wide spectrum with regard to their comfort and familiarity with physical activity. This spectrum includes Canadians showing the beginnings of interest in the initiating a more active lifestyle for preventive health or for disease management, as well as those who are engaged in elite athletic endeavors. In between these extremes is the so-called “weekend warrior,” engaged in irregular patterns of sport participation, and/or those involved in regularly structured athletic programs such are recreational hockey. Part of the valued expertise that integrative healthcare practitioners bring to such encounters with this diverse group of patients is their in-depth knowledge of nutrition as it relates to dietary practices as well as nutritional supplementation.

The Relevance of Nutrition

The lessons learned from recent studies in the realm of nutrition in sports performance may, at first glance, seem to be of little importance, even trivial, when discussing serious and potentially life-threatening conditions, such as depression and obesity. However, investigations aimed at outcomes such as hitting the finish line tape faster, increasing endurance time to fatigue, or pushing just a bit more weight in strength training, ultimately provide nutritional insights with broader potential (Maughan 2011). It is becoming clear that nutrition is a variable relevant to all those who are on the exercise/sport continuum. It has the potential to influence motivation to participate in physical activity; to reduce perception of fatigue, a barrier that often impedes subsequent participation; to influence cognitive readiness for exercise; to enhance the enjoyment of the exercise experience; to enhance recovery for the next bout of physical activity; to minimize the risk of injury; and, genetic endowment and training being equal, to influence performance itself.

Although the term sports nutrition is often associated with having a primary role in support of the muscles (readiness for task, repair, anabolic processes etc.), nutritional influences are also of vital importance for motivation and performance, factors that are centrally regulated through the central nervous system (CNS). In sedentary adults, the motivation to engage in physical activity is low, and the normal post-exercise lift in mood is often not experienced. For example, in those with depression and/or obesity (vs. healthy/normal weight controls), motivation is a primary barrier to physical activity (Searle 2011). Among these patients, despite their awareness of the benefits of exercise, there are lower pleasure ratings reported after exercise, and perceived exertion is much higher while energy levels lower (Ekkekakis 2011), and this in turn impairs intent to participate in future physical activity (Weinstein 2010). On the other hand, a more positive perception of the experience of exercise encourages future participation (Annesi 2005, Kwan 2010). Integrative healthcare practitioners can take advantage of recent discoveries in nutritional sciences as a means to help break the cycle of negative affect and associated higher levels of perceived exertion, both of which contribute to a generalized exercise intolerance.

The Mediterranean Diet Example

Before discussing more reductionist investigations of single nutrients, e.g. branch chain amino acids, vitamins, minerals etc., it may be worthwhile to examine the influence of the broad aspects of diet as they related to mental outlook and performance. The Mediterranean (Med) diet provides what may be a gold standard for general support of physical and mental performance (Sofi 2010, 2008). The basis of what is now referred to as the contemporary Med diet has been in place for some 10,000 years (Berry 2011), and although details are sparse, there is certainly evidence that various dietary protocols were part of the training schemes of ancient athletes in the region (Grivetti 1997). In the modern context, there are clear characteristics of adherence to a Med diet vs. those consuming standard North American fare (Box 1).

The benefits of high adherence to a Med diet have been well described, ranging from protection against cognitive decline to reduction in the risk of metabolic syndrome (Sofi 2013, 2010, 2008). However, its association with positive mental outlook and resiliency against depressive symptoms bears mention. In a 5-year prospective study, greater adherence to the Med diet was associated with a 25-30% reduction in the risk of depressive symptoms (Sánchez-Villegas 2009). Moreover, adherence to the Med diet has also been linked to physical performance (Milaneschi 2011). In a recent study, adherence to the Med diet was linked to better objective performance results (measured via selfselected pace and walking speed over a 20m distance) in community-dwelling older adults (Shahar 2012). A recent intervention trial found that a 10-day Med diet significantly increased vigor, alertness and contentment among participants vs. controls (McMillan 2011). These are precisely the mood changes that would work towards undoing the motivational barriers to exercise. Among the many mechanisms whereby the Med diet can influence mental outlook, however its ability to reduce inflammatory markers is likely key. Intentional elevation of inflammatory cytokines in healthy adults has been shown to causes low-grade anxiety, depressive symptoms and mental fatigue (DellaGioia 2013, Reichenberg 2001), while Med diet interventions are known to reduce systemic markers of inflammation (Richard 2013). The Med diet may in this way psychologically augment the experience of exercise.

Specific Med Diet Elements

Among the specific components of the Med diet, fish intake and carotenoid intake (an accepted marker of fruit and vegetable consumption), have been linked to muscle strength and physical performance (Cesari 2004, Robinson 2008, Semba 2007). Mechanistically, there is evidence, although not unequivocal, that omega-3 fatty acid-rich fish oil can limit red blood cell deformability, muscle damage and overall inflammation associated with exercise (Mickleborough 2013). Furthermore, omega-3 fatty acids, and eicosapentaenoic acid (EPA) specifically, can potentially influence mental outlook in ways conducive to motivation (Hegarty 2013). Carotenoids serve as a marker of fruit and vegetable consumption, and their associated array of plant-based antioxidants; there is evidence to suggest that antioxidants may provide benefit in reducing the perception of effort during exercise. For example, compared to controls, heart rate and perceived exertion were reduced, and general fatigue score was decreased, when overweight adults were given 500mg of vitamin C for four weeks (Huck 2013).

Supplementation with the carotenoid astaxanthin for 90 days has been shown to reduce objective markers of muscle cell damage (serum creatine kinase) in elite soccer players (Djordjevic 2012). In another study, 28 days of astaxanthin supplementation (vs. placebo) has been shown to significantly improve cycling performance time among competitive cyclists (Earnest 2011). Other colorful dietary components, cherries for example, have been shown to be helpful in reducing postexercise pain, markers of muscle cell damage and inflammation, as well as a more rapid restoration of muscle strength (Bowtell 2011, Connolly 2006, Howatson 2010).

In addition to omega-3 fatty acids and antioxidants, the Med diet is also very high in dietary nitrate, a compound found in green leafy vegetables and beetroot that can increase blood flow in support of exercise performance. Close to a dozen studies using beetroot juice, nitrate-depleted beetroot juice and/or supplemental nitrate (Lidder 2013) have shown that dietary nitrate can improve exercise performance by increasing the efficiency of oxygen utilization (O2 cost is reduced) and increasing ATP synthesis. Among athletes and recreationally fit adults, beetroot significantly increases time to exhaustion (15%), enhances running velocity and reduces perceived exertion over longdistance running, and improves performance in team-sport exercise (Lansley 2011, Murphy 2012, Wylie 2013). One cup of beetjuice contains 5.5 mmol of nitrate, the equivalent of about 350mg; studies have used supplemental dosage forms containing up to 1500mg nitrate in healthy volunteers (Kapil 2010). These findings, showing more efficient metabolic pathways and lowered perceived exertion in particular, clearly suggest potential benefit in those sedentary adults/children who might contemplate initiating an exercise protocol. Of course, leafy green vegetables also carry significant levels of magnesium (Mg) per serving. Mg plays an important role in enzymatic support of muscular function, and even mild deficiency may compromise muscle performance (Matias 2010). Among athletes, dietary Mg intake is positively associated with various isokinetic strength variables and jumping performance tests, independent of total energy intake (Santos 2011). Mg intake is notoriously low in the general population, and this includes athletes and those with depressive disorders (Santos 2011, Yary 2013). Indeed, low Mg levels have been associated with depressive symptoms, chronic fatigue, and muscular pain. Mg is a versatile nutrient, supporting both muscle and mood; it is one that can help mitigate inflammation and oxidative stress, as well as support normal neurotransmitter functioning (Barbagallo 2009). Again, the relevance of Mg on the continuum, from those struggling to break a sedentary lifestyle to those in which sports performance is part of their identity, seems obvious.

General Considerations – Protein, Carbohydrate, Hydration

Scientific investigations in the realm of sports nutrition, particularly related to performance outcomes, have made it increasingly clear that a needs-based, individualized approach is most appropriate (Maughan 2011). That said, there are general nutritional considerations in supporting the process of exercise adaptation. All forms of exercise increase the rate of protein oxidation relative to the resting state; this translates into an increase in dietary protein demands (Phillips 2012). Of course, increased protein does not imply excess protein – 20 to 25 grams of high-quality mixed protein, or 6g of essential amino acids, consumed before or soon after exercise, is typically more than adequate to support remodeling and adaptive processes in muscle tissue subsequent to exercise (Maughan 2012). Branch chain amino acids (BCAA) have an anabolic effect on human muscle tissue (Borgenvik 2012), and supplemented alone or together with taurine, the BCAAs have been shown to reduce post-exercise muscle soreness (Howatson 2012, Ra 2013). Although not a protein, creatine is a significant nitrogen source, and one that can have value in preventing muscular fatigue (Rawson 2011) and loss of skill under conditions of sleep deprivation at relatively small (2-5g) doses (Cook 2011, Rawson 2011). In keeping with our contention that what is good for the athlete may be good for the sedentary adult seeking motivation, consider that 5g creatine per day has been recently shown to augment the effectiveness of antidepressant medications (Lyoo 2012).

Carbohydrate provides critical fuel for the initiation and maintenance of physical activity, training and/or competition. At lower intensity exercise, the body can take advantage of fat oxidation, however, with increasing intensity (particularly endurance exercise) the demand for carbohydrate utilization grows. Endurance exercise in the fasted state is generally not well tolerated in humans (Maughan 2010), and the increase in perceived effort in those without adequate carbohydrate reserves (glycogen) would certainly not encourage adherence to exercise as medicine (Maughan 2012). For post-exercise recovery efforts, although the science is incomplete, the preponderance of evidence suggests that a 2:1 ratio of carbohydrate to protein supplementation is helpful to restore glycogen reserves (Spaccarotella 2011).

The maintenance of proper hydration is yet another top line consideration in sports nutrition. Progressive loss of water and electrolytes as a result of increasing exercise intensity and duration is itself a cause of fatigue. The initiation of exercise in a dehydrated state has been shown to compromise performance in endurance (Goulet 2012). Again, the cognitive angle of even the mildest forms of dehydration cannot be overlooked. Recent studies in schoolchildren and adults have shown diminished cognitive performance in association with mild dehydration, and cognitive restoration produced by correction to a normal hydrated state (Edmonds 2013, Fadda 2012). Perceived exertion is higher among athletes in states of relatively modest hypohydration (Barr 1999), therefore this is likely to be compounded among those who are sedentary. Maintenance of normal hydration (sodium, potassium, magnesium and calcium) before, during, and after physical activity may go a long way toward encouraging adherence to an exercise program.

Conclusion

Studies derived from the burgeoning area of sports nutrition are not simply of relevance to the very small percentage of North Americans who might be considered elite athletes. IHPs can take advantage of sports nutrition research as a means to support a wide variety of patients/clients, perhaps most importantly those sedentary adults who are struggling with the difficult task of initiating and maintaining an exercise program. Low motivation and high perceived exertion, associated with a lowered mental outlook, are primary obstacles to exercise adherence – the same nutritional variables that might allow an athlete to collect a medal or push an extra rep on a weight bar, are the very factors that might have broad public health implications.

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DHEA Part II

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DHEA Part II
Autoimmunity and Bone Health

Dehydroepiandrosterone (DHEA) is a weak androgen secreted from the adrenal cortex, and an immunologically active hormone. DHEA supplementation has been shown to improve bone density and metabolism in several patient populations, including the elderly, as well as patients with lupus or anorexia. DHEA has also been shown to improve measures of disease activity in patients with lupus and inflammatory bowel disease, and may have the ability to reduce steroid medication requirements. In the contexts of bone health and autoimmunity, dosing of DHEA ranges from 50-200mg daily. Adverse effects appear to be limited to acne and hirsutism in women. This paper reviews findings from nine human trials of DHEA for bone health and nine human trials of DHEA in patients with autoimmune disease.

Introduction

Dehydroepiandrosterone (DHEA), and its sulphated precursor DHEA-S, is a weak adrenal androgen and the most abundant steroid hormone in the body (Andus 2003). Circulating levels peak during the third decade of life (DHEAS 10μM and DHEA 10nM, slightly lower in women) and decline to approximately 20-30% those levels by the seventh decade of life (Hazeldine 2010). DHEA acts as a precursor for other steroid hormones in the body, including androstenedione, estradiol, and testosterone (Papierska 2012, Weiss 2009), and has been shown to possess immune regulatory and anti-inflammatory effects through various pathways, including inhibition of NF-kappaB activation and inhibition of IL-6 and IL-12 secretion via PPAR-alpha (Andus 2003). DHEA has been shown to inhibit Th2 cytokine secretion associated with asthma in vitro and inhibit bronchial hyperreactivity in vivo (Liou 2011). In older human subjects aged 65-75 years with impaired glucose tolerance, DHEA supplementation (50mg/d) for two years not only improved glucose tolerance but also reduced circulating levels of IL-6 and TNFalpha (Weiss 2011). DHEA has also been shown to offset the thymic involution that occurs with glucocorticoid administration in animals (Hazeldine 2010, May 1990).

Unfortunately, DHEA is currently banned in Canada due to historical abuse by professional athletes. Nonetheless, as this remains a promising therapy for applications in female fertility (reviewed in Part I) as well as bone health and autoimmune disease, we hope that the status of DHEA may be revisited by Health Canada in the future. Prasterone is a proprietary, synthetic form of DHEA that is sold as a prescription drug.

Bone Health

DHEA supplementation has been shown to increase bone mineral density (BMD) among older adults, patients with anorexia, as well as SLE patients on prednisone therapy (Table 1). As might be expected, this effect seems to be most marked among those with pre-existing bone loss as opposed to those with normal BMD at baseline, as well as among women compared to men (Papierska 2012, Von Muhlen 2008, Weiss 2009). In a study of older men and women on prednisone and with osteopenia, supplementation with DHEA resulted in increased serum IGF-1 and osteocalcin alongside increased lumbar and femoral BMD (Papierska 2012). Among elderly patients with normal bone density, however, further benefit on BMD from DHEA supplementation appears to be restricted to women, with no significant BMD increase demonstrated among elderly men with normal BMD (Kahn 2002, Von Muhlen 2008, Weiss 2009). The reason for this gender difference is unclear, but may be related to differential hormone metabolism. Indeed, DHEA supplementation among older adults (25-50mg/d) has been shown to increase serum levels of androstenedione, testosterone, and estradiol particularly in women (Papierska 2012, Von Muhlen 2008, Weiss 2009).

Among patients with SLE, DHEA supplementation has also been shown to significantly improve BMD when compared to placebo (Hartkamp 2004, Mease 2005, Sanchez-Guerrero 2008). A randomized placebo controlled double blind study found that DHEA supplementation was also of benefit among young women with anorexia. A dose of 50mg/d was able to restore physiologic DHEA levels within three months, and dosages between 50-300mg were able to improve markers of bone metabolism including osteocalcin and urinary N-telopeptides (Gordon 1999).

Table 1. Human Trials of DHEA for Bone Health
Table 1. Human Trials of DHEA for Bone Health
Table 2. Human Trials of DHEA for Autoimmune Disease: Impact on Disease Activity
Table 2. Human Trials of DHEA for Autoimmune Disease: Impact on Disease Activity

Autoimmune Disease Systemic Lupus Erythematosus

A total of eight human trials, five RCTs and three prospective trials, conducted in patients with SLE report significant improvements in disease activity and/ or corticosteroid requirements associated with supplementation of DHEA at a dosage of 50-200mg daily (See Table 2). The Systemic Lupus Activity Measure (SLAM) and SLE Disease Activity Index (SLEDAI) scoring systems were used to assess impact on disease activity.

Four of five RCTs found significant improvements in measures of disease activity, decreased number of disease flares, or reduced prednisone requirements (Chang 2002, Petri 2004, Petri 2002, van Vollenhoven 1995). Two RCTs demonstrated significant improvements in SLEDAI score with DHEA compared to placebo (Petri 2004, van Vollenhoven 1995). An earlier study by Petri et al found that among patients with active, steroiddependent disease, a significantly greater number of DHEA-treated patients were able to sustain a reduction in steroid dose equal to or greater than 7.5mg/d, defined as responders, compared to placebo (P=0.031). Chang et al found that although there was no significant difference in the SLAM score reductions of patients treated with DHEA or placebo, the number of patients with disease flares was significantly lower in the DHEA group (2002). DHEA-treated patients also had significantly better global assessment scores (p= 0.005) and fewer serious adverse events, most of which were related to SLE disease flares (p= 0.010) (Chang 2002).

The only adverse events reported with DHEA dosages between 50-200mg/d were acne and hirsutism (Barry 1998, Petri 2004, 2002, van Vollenhoven 1998, 1995, 1994).

Inflammatory Bowel Disease

Abnormally lower DHEA/ DHEA-S levels have been described in patients with inflammatory bowel disease, compared to healthy controls (de la Torre 1998, Straub 1998). One study found that among patients with ulcerative colitis, DHEA-S was 1350 nmol/L, and in patients with Crohn’s disease DHEA-S was 1850 nmol/L, while healthy controls had a level almost two-fold higher, 3300 nmol/L (p<0.001 and p<0.01 respectively) (de la Torre 1998).

One prospective trial in patients with chronic, active, refractory IBD found that treatment with 200mg DHEA once daily for approximately two months induced remission in 60% of patients (12 of 20 patients in total; six Crohn’s and six ulcerative colitis) (Andus 2003). Remission was defined as Crohn’s disease activity index <150 or clinical activity index ≤4, which scores eight sign and symptoms of ulcerative colitis, including the number of soft stools; blood in the stools; general wellbeing; abdominal cramps or pain; fever; extraintestinal manifestations; erythrocyte sedimentation rate; and hemoglobin level (Andus 2003).

Subsequent to the publication of this study, the same group of authors published a case report relating the use of DHEA to treat pouchitis (Klebl 2003). A 35-year old women with chronic active pouchitis was treated with 200mg DHEA per day for eight weeks. Her stool frequency dropped from 15-18 per day to eight per day. Eight weeks after her DHEA treatment was discontinued, the numbers of stools increased again to 12-18 per day and her pain returned, indicating that DHEA was likely responsible for her initial improvement, and that ongoing treatment may be required.

Currently, the small number of studies that have examined DHEA supplementation in patients with Sjogren’s syndrome and rheumatoid arthritis have failed to show benefit (Giltay 1998, Hartkamp 2008, Pillemer 2004, Virkki 2010), however this area deserves further study.

Conclusion

DHEA therapy in the treatment of osteopenia and osteoporosis as well as SLE and IBD is supported by human clinical research. Evidence demonstrates that DHEA can increase BMD in the elderly as well as in patients with SLE or anorexia. DHEA may be a disease-modifying agent in patients with SLE and IBD, decreasing disease activity and acting as steroid-sparing agent. Adverse events reported with use of DHEA 50-200mg for up to one year appear to be limited to mild acne and hirsutism. In future it is hoped that this promising agent may once again become available for human therapeutic application in Canada.

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Mease PJ, Ginzler EM, Gluck OS, Schiff M, Goldman A, Greenwald M, Cohen S, Egan R, Quarles BJ, Schwartz KE. Effects of prasterone on bone mineral density in women with systemic lupus erythematosus receiving chronic glucocorticoid therapy. J Rheumatol. 2005 Apr;32(4):616-21.

Papierska L, Rabijewski M, Kasperlik-Załuska A, Zgliczyński W. Effect of DHEA supplementation on serum IGF-1, osteocalcin, and bone mineral density in postmenopausal, glucocorticoid-treated women. Adv Med Sci. 2012 Jun 1;57(1):51-7.

Petri MA, Mease PJ, Merrill JT, Lahita RG, Iannini MJ, Yocum DE, Ginzler EM, Katz RS, Gluck OS, Genovese MC, Van Vollenhoven R, Kalunian KC, Manzi S, Greenwald MW, Buyon JP, Olsen NJ, Schiff MH, Kavanaugh AF, Caldwell JR, Ramsey- Goldman R, St Clair EW, Goldman AL, Egan RM, Polisson RP, Moder KG, Rothfield NF, Spencer RT, Hobbs K, Fessler BJ, Calabrese LH, Moreland LW, Cohen SB, Quarles BJ, Strand V, Gurwith M, Schwartz KE. Effects of prasterone on disease activity and symptoms in women with active systemic lupus erythematosus. Arthritis Rheum. 2004 Sep;50(9):2858-68.

Petri MA, Lahita RG, Van Vollenhoven RF, Merrill JT, Schiff M, Ginzler EM, Strand V, Kunz A, Gorelick KJ, Schwartz KE; GL601 Study Group. Effects of prasterone on corticosteroid requirements of women with systemic lupus erythematosus: a doubleblind, randomized, placebo-controlled trial. Arthritis Rheum. 2002 Jul;46(7):1820-9.

Pillemer SR, Brennan MT, Sankar V, Leakan RA, Smith JA, Grisius M, Ligier S, Radfar L, Kok MR, Kingman A, Fox PC. Pilot clinical trial of dehydroepiandrosterone (DHEA) versus placebo for Sjögren’s syndrome. Arthritis Rheum. 2004 Aug 15;51(4):601-4.

Sánchez-Guerrero J, Fragoso-Loyo HE, Neuwelt CM, Wallace DJ, Ginzler EM, Sherrer YR, McIlwain HH, Freeman PG, Aranow C, Petri MA, Deodhar AA, Blanton E, Manzi S, Kavanaugh A, Lisse JR, Ramsey-Goldman R, McKay JD, Kivitz AJ, Mease PJ, Winkler AE, Kahl LE, Lee AH, Furie RA, Strand CV, Lou L, Ahmed M, Quarles B, Schwartz KE. Effects of prasterone on bone mineral density in women with active systemic lupus erythematosus receiving chronic glucocorticoid therapy. J Rheumatol. 2008 Aug;35(8):1567-75.

Straub RH, Vogl D, Gross V, Lang B, Schölmerich J, Andus T. Association of humoral markers of inflammation and dehydroepiandrosterone sulfate or cortisol serum levels in patients with chronic inflammatory bowel disease. Am J Gastroenterol. 1998 Nov;93(11):2197-202.

vanVollenhoven RF, Park JL, Genovese MC, West JP, McGuire JL. A double-blind, placebo-controlled, clinical trial of dehydroepiandrosterone in severe systemic lupus erythematosus. Lupus. 1999;8(3):181-7.[Abstr]

vanVollenhoven RF, Morabito LM, Engleman EG, McGuire JL. Treatment of systemic lupus erythematosus with dehydroepiandrosterone: 50 patients treated up to 12 months. J Rheumatol. 1998 Feb;25(2):285-9.[Abstr]

vanVollenhoven RF, Engleman EG, McGuire JL. Dehydroepiandrosterone in systemic lupus erythematosus. Results of a double-blind, placebo-controlled, randomized clinical trial. Arthritis Rheum. 1995 Dec;38(12):1826-31.[Abstr]

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Virkki LM, Porola P, Forsblad-d’Elia H, Valtysdottir S, Solovieva SA, Konttinen YT. Dehydroepiandrosterone (DHEA) substitution treatment for severe fatigue in DHEA-deficient patients with primary Sjögren’s syndrome.Arthritis Care Res (Hoboken). 2010 Jan 15;62(1):118-24.

vonMühlen D, Laughlin GA, Kritz-Silverstein D, Bergstrom J, Bettencourt R. Effect of dehydroepiandrosterone supplementation on bone mineral density, bone markers, and body composition in older adults: the DAWN trial. Osteoporos Int. 2008 May;19(5):699-707.

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Acid-Alkaline Balance

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Acid-Alkaline Balance

The Third Dimension of Fruits and Vegetables

The health benefits of a fruit and vegetable based diet are well known, and are most often attributed to their phytonutrient content: vitamins, minerals, flavonoids, phenolic compounds, and fibre. Overlooked in this regard is the role that fruit and vegetable consumption plays in acid-alkaline balance. Fruits and vegetables contain minerals such as potassium and bicarbonate that contribute to the creation of a slightly alkaline state within the body. This is discussed in relation to bone health, cortisol secretion, cancer development and progression, as well as renal function. There is a possibility that supplementation with greens drinks or salts of potassium and/ or bicarbonate can shift the body toward a more alkaline state, and this is supported by preliminary research. Healthcare providers can teach their patients to assess acid-alkaline balance through testing of morning urine; this type of objective testing has the additional benefit of helping patients observe the physiological changes occurring through diet and lifestyle modification, and may help increase long term compliance.

Introduction

In recent years, government and private sector dietetic organizations have stepped up efforts to encourage the North American population to consume greater amounts of fruits and vegetables. The rationale for this effort is quite clear, it is built upon volumes of international research showing that fruits and vegetables offer a layer of protection against a variety of chronic diseases, most notably cardiovascular disease and cancer (Khaw 2008). The health benefits of dietary fruits and vegetables are generally attributed to two major nutritional aspects – antioxidant (vitamin/ mineral and phytochemical) and fibre content. Indeed, when examined independently, dietary antioxidant and fibre content derived from fruits and vegetables are health protective (Mente 2009). However, there is yet a third avenue of health promotion provided by fruits and vegetables – the underappreciated contribution of dietary bases within fruits and vegetables has assumed great importance in the face of our contemporary, acidheavy diet (Welch 2008).

The pH Scale and PRAL

Acid-alkaline states are reflected on the potential of hydrogen (pH) numerical scale. The scale runs from 1 – 14, with 1 being most acidic and 14 most alkaline. Importantly, since this is a logarithmic scale, moving between each full number (i.e. 6 to 7) represents a 10-fold difference; therefore small changes are not insignificant. The pH of our bodily systems is tightly regulated, operating near the middle of the pH scale. In the blood stream, which operates in the narrow range of 7.35-7.45 (LTO 2013), any deviation away from this near-neutral pH may compromise our very survival. Since the bone matrix contains a relatively abundant alkali reserve in the form of calcium and magnesium cations, it may be called upon to buffer the tightly regulated blood pH in the presence of acidic influences. Today, one of the most consistent acidic influences on pH regulatory systems is the western diet (Berkemeyer 2009).

The notion that diet can play a role in the acid-base balance is not a new one; researchers first showed almost a century ago that meats, dairy, eggs and grains can increase the acidity of urine in humans, while dietary fruits and vegetables can have an alkaline influence on the urine (Sherman 1912). The determination of the acid or base potential of a food or beverage in the human body has advanced with a great degree of sophistication since the early days of subjecting the food to combustion and determining if the ash was acid or alkaline. Today a laboratory test known as the Potential Renal Acid Load (PRAL) has emerged as a reliable indicator of the acid or alkaline potential of a food or beverage in the human body, and is being actively studied by Thomas Remer as a convenient way to estimate the net renal acid excretion due to various food (Remer 1995, 2003). The PRAL model takes into account the following factors in determining the relative acidity of a particular food: “mineral and protein composition of [the] foods, the average intestinal absorption rates of the respective nutrients, sulfur metabolism, and urinary excretion of organic acids” (Remer 1995). Foods and beverages that have potential to contribute to the net acid load in the human body, particularly those rich in proteins and sodium, are said to have a high, or more positive PRAL. Conversely, foods and beverages that are abundant in potassium, bicarbonate and alkaline minerals are said to have a lower, or more negative, PRAL score (Remer 1995). In general, fruits and vegetables consistently have a negative PRAL score (more alkaline), while grains, meats, fish, and dairy products have a positive PRAL score (more acidic). Sometimes foods or beverages patients might consider to be ‘acidic’, such as citrus fruits or tomatoes, are actually quite alkaline in the body due to the presence of minerals and bicarbonates.

Tipping the Scale

The contemporary North American diet contains an abundance of acid-forming foods, particularly animal meats, cheeses, grains, soft drinks and processed foods. Our societal deficiency of fruits and vegetables is obvious: less than 40% of Americans consume the minimum recommended five servings of fruits and vegetables according to a the National Health and Nutrition Examination Survey (NHANES) 1999- 2000 survey (Guenther 2006). Statistics such as this demonstrate widespread deficiency of alkaline minerals and bicarbonates in the American diet, with the scale clearly tipped in favour of acidic foods. Using a computational model, researchers have demonstrated that the mean net endogenous acid production (NEAP) projected for preagricultural, hunter-gatherer diets was -88 mEq/d, compared to +48 mEq/d based on the standard American diet as recorded in the NHANES III study (Sebastian 2002). This increased dietary acid load is not without consequence to human health.

Bone Health

The relationship between acid-rich foods – consumed in the absence of buffering alkaline-rich fruits and vegetables – and the development of osteoporosis is of particular interest to scientists. Since blood pH must remain in a narrow (slightly alkaline) range for survival, calcium and magnesium are removed from the bone to buffer, or neutralize a continuously acidic diet; consequently, calcium and magnesium are drawn into the circulation and excreted through the urine (Rylander 2006). Over time this may lead to an appreciable loss of minerals from the bone matrix. Research does indicate that frequent consumption of high PRAL acid-forming foods (cheese, meats, processed grains) and infrequent consumption of potassium and bicarbonate-rich, alkaline-forming foods (fruits and vegetables) is associated with increased urinary calcium and magnesium loss and a greater risk of osteoporosis (Vormann 2008, Wynn 2008) Scientists have also shown that the bone-forming osteoblasts are less active in acidic environments. Even a modest change to a more alkaline diet has been associated with a 50 percent reduction in fracture risk (Lanham-New 2008).

More recent studies have linked an acidic diet to increased body weight, increased waist circumference (Berkemeyer 2009, Remer 2007) and various markers of cardiovascular disease, including elevated cholesterol and hypertension (Murakami 2008). Recently it was reported that adults with metabolic syndrome are likely to have more acidic urine than adults without metabolic syndrome (Maalouf 2007). On the other hand, more alkaline urine is associated with a greater percentage of lean body mass (Dawson- Hughes 2008). In addition, a more alkaline diet rich in potassium was found to improve mental outlook and energy in otherwise healthy adults (Torres 2008).

The Cortisol Connection

Swiss researchers discovered an important physiological change induced by an acidic, fast-food-type diet, a change with enormous implications for human health: consuming an acidic, Western-type diet for nine days was shown to significantly elevate the stress hormone cortisol. When the researchers neutralized the Western diet with bicarbonate supplements, the cortisol levels dropped down to normal (Maurer 2003). Since cortisol increases the propensity to gain abdominal fat, promotes inflammation and oxidative stress, disturbs immune function and mood states (Epel 2009), this research makes pH an important consideration in human health. The lesson in this research may be less about acid foods themselves, and more about the importance of neutralization with our absentee fruits and vegetables.

Cancer Controversy

For years an avoidance of acidic foods has been advocated for reducing cancer risk, malignancy and recurrence. Indeed, the possibility of acidic meats rich in sodium promoting cancer was first hypothesized more than a century ago (Braithwaite 1901). While supporting the protective role of fruits and vegetables in cancer, the American Institute for Cancer Research (AICR) recently scoffed at the notion of any connection between acidic foods and cancer (AICR 2008). Several lines of research indicate that further investigation is required, however, and the preponderance of recent evidence suggests that there may indeed be a connection. First, more alkaline urine has been noted to enhance the excretion of a variety of xenobiotics in a process known as ion trapping at the kidney. In the presence of more alkaline urine, certain environmental toxins and drugs existing as weak acids are more readily excreted and reabsorption by the renal tubules is inhibited (Minich 2007).

Given that alkaline, mineral-rich, drinking water can influence urinary pH, there has been surprisingly little research into the connection between the pH of municipal water and cancer rates. The only studies conducted so far, three separate investigations in Asia, found that rates of esophageal, rectal, and pancreatic cancer mortality rates are all significantly lower in regions where municipal water pH is more alkaline (Yang 1999a, Yang 1999b, Yang 1999c). Use of soft water was associated with a 42% increased risk of esophageal cancer (adjusted OR 1.42, 95% confidence interval 1.22-1.66) (Yang 1999a), 38% increased risk of rectal cancer (AOR 1.38, 95% CI 1.10-1.73) (Yang 1999b), and 39% increased risk of pancreatic cancer (AOR 1.39, 95% CI 1.09–1.76) (Yang 1999c). In addition, Russian researchers reported that among 150 patients recovering from gastric cancer, those consuming alkaline mineral water had approximately 3.8-fold reduced recurrence rates, while 300 breast cancer patients postmastectomy had a 20% reduction in recurrence, though unfortunately further details were unavailable due to language limitations (Vladimirov 2004).

Secondly, the research showing that the typical acid-heavy North American diet can increase cortisol, and that neutralization of the same acidic diet with bicarbonate can reduce cortisol in humans, is also of great relevance to cancer. In animal studies, elevations in cortisol is associated with rapid growth of tumors, and in human depressive disorders, characterized by hypercortisolemia, a more rapid breast cancer progression has been noted in 19 out of 24 studies (Palesh 2007). Various aspects of the immune system are compromised with chronic cortisol elevation (Epel 2009). It is also true that a shift toward a more acidic environment diminishes the functionality of natural killer cells and promotes the production of inflammatory cytokines (Kellum 2004, Lardner 2001).

The most convincing evidence comes from direct studies of the pH of cancer cells and their surrounding environments. A number of recent studies using pH sensitive magnetic resonance imaging contrast agents and microelectrodes have consistently shown that the extracellular pH of tissue surrounding tumors is significantly lower than that of healthy tissues (Bellone 2013, Estrella 2013). A low pH microenvironment is now known to increase the growth and invasiveness of tumor cells in vivo. Bathing tumor cells in an acidic pH (6.7, typical of the extracellular pH of tumors) increases invasiveness by 4-fold (Silva 2009).

Discounting test-tube studies in cancer and acidbase balance, the AICR stated that “altering the cell environment of the human body to create a lessacidic, less-cancer-friendly environment is virtually impossible” (2008). Yet, emerging studies are at odds with this claim. In recent animal investigations oral bicarbonate has been shown to increase the extracellular pH of tumors, subsequently reducing the in vivo number and size of tumor metastases. Ultimately the reductions in metastases after oral bicarbonate led to increased survival rates of the animals with tumors (Robey 2009). Importantly, the chronic administration of bicarbonate produced no change in the tightly regulated blood pH, indicating that the results were based on a local buffering phenomenon via the oral alkaline solution. This provides more than mere speculation that the reverse would also be true, i.e. that chronic consumption of oral diet-derived acids can influence the interstitial fluid of primary and/or metastatic tumors in the absence of significant changes to blood pH. While much more research is needed, for now the only myth concerning acid-alkaline diet and cancer is that which states there is no relationship between the two.

Chronic Kidney Disease

A fourth area within medicine that is being studied with respect to acid-base balance is chronic kidney disease, though in this case the population has already progressed from subacute acidosis (Kovesdy 2012). Nonetheless, it is of relevance that in this context, supplementing with alkali may be able to slow the loss of renal function. A study by de Brito-Ashurst et al found that supplementing 134 patients with reduced baseline creatinine clearance (15–30 mL/min) with oral sodium bicarbonate tablets 600mg three times daily, titrated to achieve target serum bicarbonate levels of ≥23 mEq/L, resulted in a slower slope of progression for creatinine clearance and fewer incidences of rapid loss of kidney function and end stage renal disease (defined as creatinine clearance <10 mL/min), compared to usual care (2009). Other studies have shown similar findings, with slower decline in eGFR and decreases in urinary endothelin-1, albuminuria, and tubular injury markers (Kovesdy 2012, Phisitkul 2010).

What to Do?

When a daily diet with a high net acid load becomes the rule rather than the exception, it sets the stage for what has been described as a chronic low-grade level of metabolic acidosis. This, in turn, may be influencing a number of different health outcomes. Therefore, in the clinical setting it is important to establish a baseline of acid-alkaline status in the body. Urinary pH has been established as a reliable surrogate marker for the state of acid-alkaline balance in the human body (Welch 2008), and while 24-hour urinary pH is the gold standard, it may be worthwhile to have patients measure the first morning urine after an overnight fast for 7-10 days. It may also be prudent to have a discussion with patients concerning the meaning of the pH scale itself, reminding them that small changes in the numerical value are of significant importance. For patients the measurement of pH is an inexpensive investment; commercially available litmus paper provides an accurate assessment of pH, equal to that of the dipsticks used by clinicians (Desai 2008).

The obvious clinical priority is to provide the standard advice to consume an abundance and broad variety of fruits and vegetables. Concomitant reduction of meats, dairy and grains will help reduce the net acid load. Reframing the advice away from fibre and antioxidants, moving in the direction of pH may provide a novel stimulus to comply with the healthy change, particularly when provided in the context of a simple urinary test that may enhance goal-oriented motivation. Supplementation is also an option. Recently researchers from the University of Toronto showed that a commercially available supplement with dehydrated plant foods (greens+) can influence urinary pH. The pH of first-morning urine was made significantly more alkaline in otherwise healthy adults after 14 days, particularly among those with more acidic urine pre-supplementation (Berardi 2008). It has also been shown that supplementation with potassium citrate and bicarbonate can influence urinary pH in the direction of alkalinity (Minich 2007).

While the science of net dietary acid load has advanced in recent years, the clinical utility of addressing the acidbase balance is not a recent phenomenon. The notion that frequent consumption of acid-heavy foods can promote inflammation is also not a new concept, nor is the use of diet and bicarbonate supplements to address health. In the 1920s dermatologists reported that alkaline ash foods and bicarbonates were helpful in lowering skin inflammation – “In nearly all cases, active inflammatory processes cease and the eruption rapidly clears when the urine is rendered alkaline” (Ormsby 1920). The acidbase connection to human health requires much more study, since for now there are still more questions than answers. Hopefully in the near future we will have a greater understanding of the mechanisms surrounding the net dietary acid load and its influence on cortisol, interstitial pH, detoxification and our immune systems. The collateral health benefits provide more than enough justification for an emphasis on potassium, bicarbonate, and mineral-rich fruits and vegetables.

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AICR 2008 e-newsletter. May 2008. http://www.aicr.org/site/News2? page=NewsArticle&id=13441. Accessed 24 March 2013.

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Berardi JM, Logan AC, Rao AV. Plant-based dietary supplement increases urinary pH. Journal of the International Society of Sports Medicine. 2008;5:20.

Berkemeyer S. Acid-base balance and weight gain: are there crucial links via protein and organic acids in understanding obesity? Med Hypotheses. 2009 Sep;73(3):347-56.

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de Brito-Ashurst I, Varagunam M, Raftery MJ, Yaqoob MM. Bicarbonate supplementation slows progression of CKD and improves nutritional status. J Am Soc Nephrol. 2009 Sep;20(9):2075-84.

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Estrella V, Chen T, Lloyd M, Wojtkowiak J, Cornnell HH, Ibrahim- Hashim A, Bailey K, Balagurunathan Y, Rothberg JM, Sloane BF, Johnson J, Gatenby RA, Gillies RJ. Acidity generated by the tumor microenvironment drives local invasion. Cancer Res. 2013 Mar 1;73(5):1524-35.

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The Novel Catecholamine- Regulated Protein 40

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The Novel Catecholamine- Regulated Protein 40:

A crucial protein for understanding and diagnosing neurological disorders?

Neurological disorders represent one of the leading global causes of disability. They include both age-related neurodegenerative disorders such as Parkinson’s disease (PD), as well as neurodevelopmental disorders including schizophrenia (SCZ). In the past several decades, researchers have made significant progress in unravelling the pathological mechanisms of these disorders. This has been accelerated by the merging of various scientific disciplines and an enhanced interaction between basic, applied, and translational research in what is now widely known as the field of neuroscience. This integrative field has allowed for researchers to study diseases from different perspectives and to use different strategies for discovery, understanding or treatment. Our recent review on molecular chaperones illustrated how studying this specific set of proteins and their roles in abnormal protein folding, oxidative stress, and mitochondrial function may reveal their greater impact on a range of central nervous system disorders. Specifically, we discussed the diverse roles of a novel molecular chaperone protein called Catecholamine-regulated protein 40 (CRP40; 40kDa) that was discovered in our laboratory. We presented compelling evidence indicating that CRP40 is a molecular chaperone that functions in maintaining proper protein folding and may be involved in dopamine signalling. Further, we found that CRP40 is dysregulated in human post-mortem SCZ and PD brain samples. In part two of our two-part review, we present our newest evidence for the genetic dysregulation of CRP40 in the blood of live SCZ and PD patients. These compelling findings link CRP40 to central nervous system disorders and add significant value to the important global task of identifying valid, reliable, and simple biomarkers for definitive and early diagnosis of these debilitating disorders.

CRP40: A novel molecular chaperone with implications in neurological diseases In part one of our two-part review on molecular chaperones as potential targets in neurological diseases (Lubarda 2013a), we outlined the roles of heat shock proteins in diseases characterized by abnormal protein folding. We presented research done by our laboratory at McMaster University, led by Dr. Joseph Gabriele, on the novel molecular chaperone Catecholamine-regulated protein 40 (CRP40). CRP40 was discovered during investigation of proteins with dopamine (DA) binding abilities and purported to be involved in the DA signalling cascade through its interactions with DA and colocalization with tyrosine hydroxylase, the critical enzyme in DA synthesis (Gabriele 2009). CRP40 was found to belong to a family of heat shock proteins involved in protein folding and maintenance of mitochondrial function and homeostasis — conditions that are impaired in various neurological disease states. It was discovered that CRP40 is an alternative splice variant of the 70kDa heat shock protein, mortalin, which has been linked to Parkinson’s disease (PD) through the following evidence: mortalin is expressed from a putative PD locus on chromosome 5; concentrations of mortalin are reduced in post-mortem PD patients; mutations in the mortalin gene, in regions from which CRP40 is expressed, have been found in German and Spanish PD patients—these mutations have been associated with impairments in mitochondrial function and increased neuronal susceptibility to oxidative damage (Burbulla 2010).

In part one of our review on CRP40, we also revealed evidence for the possible involvement of this protein in neurological disorders such as schizophrenia (SCZ). CRP40 is dysregulated in post-mortem SCZ brain samples. Further investigation showed that CRP40 is unique by being expressed solely in the central nervous system and blood, unlike mortalin, which is expressed ubiquitously. It is well known that a number of brain proteins involved in neurotransmission are also found in the circulating blood system; therefore, certain blood cells may be considered “sentinel tissues” that could reflect levels of these neuropeptides in the brain. Thus, we set out to verify this hypothesis and investigate whether CRP40 levels in the blood mirror the reductions found in post-mortem disease subjects.

We designed methods to measure CRP40 mRNA levels using the versatile technique of real-time reversetranscriptase polymerase chain reaction (RT-PCR) in the blood of live PD and SCZ patients. CRP40 was dysregulated in both of these cases in comparison to healthy, normal controls and negative controls (Alzheimer’s disease, stroke) (Groleau 2013, Lubarda 2013b). These findings point to the possibility of distinct roles of CRP40 in central nervous system disorders. In this review, we discuss the implications of CRP40 in SCZ and PD, and present evidence that may advance our understanding of these complex disorders and open up novel avenues for investigation of heat shock proteins as potential biomarkers.

CRP40: Implications in Schizophrenia

The pathogenesis of SCZ, though still incompletely understood, is instigated by oxidative stress and mitochondrial dysfunction (Bitanihirwe 2011, Ciobica 2011, Martins-de-Souza 2011, Wood 2009, Yao 2001, Yao 2011). Oxidative stress is caused by reactive oxygen species (ROS), natural by-products of mitochondrial energy metabolism. If ROS are not controlled, they can react with important cellular components, leading to irreparable damage like neural degeneration.

Exacerbated ROS production in conjunction with dysfunctional energy metabolism has been observed in human tissue samples from SCZ patients (brain tissue, blood, and cerebrospinal fluid) (Prabakaran 2007, Schwarz 2008). The neurodegeneration observed in SCZ has been associated with genetic variations of heat shock proteins (HSPs) with specific implications in oxidative stress and apoptosis including 70kDA HSPs (HSP70) (Kim 2008, Schon 2003). Since Mortalin and its splice variant, CRP40, show significant homology to HSP70, they have emerged as proteins of interest to researchers studying SCZ.

Many studies done over the last decade have revealed CRP40 as a key player in SCZ. Using human post-mortem SCZ samples obtained from the Stanley Foundation Neuropathology Consortium, western blot analysis revealed a significant decrease in the protein expression of CRP40 in the ventral striatum of SCZ patients compared to healthy subjects (Gabriele 2005, Gabriele 2010a). Non-medicated schizophrenic patients showed an even greater reduction in CRP40 expression than medicated schizophrenics when compared to controls (Gabriele 2005). The effect of antipsychotic drug use on CRP40/mortalin mRNA levels in schizophrenic post-mortem samples was also investigated using human SCZ dorsolateral prefrontal cortex brain samples (Gabriele 2010a). CRP40/mortalin mRNA expression was assessed via real-time RT-PCR, revealing a positive correlation between lifetime antipsychotic drug use and increased CRP40/mortalin mRNA expression in SCZ patients (Gabriele 2010a).

In further studies, CRP40/mortalin were underexpressed by gene knockdown in the medial prefrontal cortex in rats (Gabriele 2010b). Sensorimotor gating was measured by prepulse inhibition (PPI), which assesses the startle reflex response to acoustic stimuli. Deficits in PPI are typically observed in patients diagnosed with SCZ. Significant deficits in PPI were found in the antisense oligodeoxynucleotide treated group when compared to the control group (Gabriele 2010b).

Importantly, it has been observed that haloperidol, a DA receptor agonist and antipsychotic drug used regularly in the treatment of SCZ, causes significant upregulation of CRP40 at the striatum, but not in the frontal cortex (Gabriele 2007, Gabriele 2009, Gabriele 2010a, Sharan 2001). Haloperidol blocks binding of DA to D2 receptors, causing increased free, unbound DA. This excess DA causes CRP40 upregulation (Gabriele 2007, Gabriele 2010a, Sharan 2000). Similarly, amphetamine, a DA agonist used in a rat model of SCZ, causes significant upregulation of CRP40 at the striatum (Gabriele 2002). These findings support the hypothesis that anti-psychotic drugs and psychostimulants like cocaine affect CRP40 protein levels in the mesocorticolimbic brain regions and further reinforce its involvement in these psychotic disorders (Gabriele 2007, Sharan 2001, Sharan 2003).

Most recently, CRP40 has been found dysregulated in human blood samples of patients with SCZ (Groleau 2013). CRP40/mortalin mRNA was analyzed in white blood cells of first episode schizophrenia subjects, chronic/treated schizophrenia subjects and healthy controls. Significant reductions in CRP40/ mortalin mRNA were found among first episode schizophrenia subjects and chronic schizophrenia subjects compared to healthy controls (Groleau 2013). These results suggest a possible functional role of CRP40 in the pathogenesis of schizophrenia, as well as the potential for future diagnostics based on the CRP40 protein.

CRP40: Implications in Parkinson’s disease

Parkinson’s disease (PD) is a movement disorder characterized by progressive degeneration of dopaminergic neurons in the substantia nigra pars compacta (SNc) (Lang 1998, Schapira1999). The resulting loss of DA neurotransmission is associated with impairments in motor and locomotion (Alberts 1965). Researchers have linked mitochondrial dysfunction and oxidative stress to PD pathophysiology (Lang 1998, Schapira 1999). In the SNc, free DA is oxidized to ROS known as Quinones (Stokes 1999). Since natural oxidation of DA leads to the formation of this potent oxidant species, it is of particular interest to PD research. As a protective measure, the human brain employs molecular chaperone proteins to defend against oxidative stressors (Becker 1994, Kaul 2007). PD patients show impaired function of key mitochondrial proteins, many of which are molecular chaperones (Becker 1994, Kaul 2007). Many have reported the possibility of combined effects of increased oxidative stress, impaired mitochondrial function, and dysregulation of molecular chaperone proteins in the pathogenesis of this movement-related disease.

Recent work has exposed the likely role of the CRP40 chaperone in the pathology of Parkinson’s disease. Genetic analysis of a population of PD patients found two previously unidentified missense mutations of Mortalin/ CRP40 in some patients with PD that were completely missing in any control subject (De Mena 2009). Interestingly, mutations in the mortalin gene found in PD patients are located within the C-terminal domain of the protein, from which CRP40 is expressed (Burbulla 2010, Sklar 2004). Since mortalin is involved in mitochondrial health, a mutation of this calibre could be responsible for the cellular stress that leads to degeneration like that seen in PD. Mortalin has been found to interact with PD-implicated proteins such as DJ-1 and α-synuclein (Jin 2006, Jin 2007). A missense mutation in Mortalin, like those described above, would certainly have a negative effect on these important protein interactions (De Mena 2009, Jin 2006, Jin 2007). Among studies using human post-mortem brain samples, Mortalin/ CRP40 were found to be underexpressed in patients with PD compared to controls (Jin 2006, Shi 2008). Specifically, there is a quantitative decrease in Mortalin/ CRP40 expression with progression of the disease (Shi 2008). In a study using rats with intrastriatal injections of 6-hydroxydopamine and reserpine treatment (models of PD), results showed decreased levels of CRP40 in the striatum (Modi 1996).

Most recently, CRP40 has been found dysregulated in human blood samples of patients with PD (Lubarda 2013b). CRP40 mRNA was analyzed in platelets of Parkinson’s disease subjects and healthy controls. Differences were observed between health states with reduced CRP40 expression among PD subjects compared to controls (Lubarda 2013). These results suggest that blood levels of CRP40 are congruent with post-mortem findings showing CRP40 is genetically dysregulated in PD, and that CRP40 exhibits potential as a biomarker and in future diagnostics for Parkinson’s disease.

CRP40: A Future Diagnostic Solution for Neurological Disorders?

The challenges of diagnosing neurological disorders are well known. There is a lack of valid and reliable biomarkers for early detection, specifically those that could monitor disease progression and efficacy of treatments. For instance, diagnosis of SCZ is currently based on clinical criteria and despite extensive research, biomarkers are still lacking (Tandon 2008). Our findings point to the possibility of utilizing CRP40 as a blood biomarker for SCZ. However, more extensive research is required to explore the pathophysiological heterogeneity that is observed in SCZ and to determine whether CRP40 can be used to explain some of the neurobiological processes and clinical expression of the intermediate phenotypes of this complex disease.

Similar strategies can be applied to studying PD as well. The notion of PD as a motor disorder has been altered to encompass several non-motor features, and prodromal symptoms such as hyposmia, rapideye- movement behaviour disorder, constipation, and depression (Siderowf 2012). There is now a paradigm shift in research that focuses on diagnosing PD in prodromal stages; however, biomarkers are sparse (Siderowf 2012). Currently, several of the available biomarkers for PD are based on DA transporter imaging, but the technologies are highly costly, somewhat invasive, and not yet widely available (Siderowf 2012). Further, DA imaging techniques may not be ideal for prodromal detection as the development of DA deficiency may be preceded by other events, such as aberrations in protein folding (Siderowf 2012). For these reasons, investigation of molecular chaperone proteins that regulate protein homeostasis, such as CRP40, could possibly be used to diagnose diseases in early stages. We are currently conducting a large-scale study with funding from the Quebec Consortium for Drug Discovery, in association with AstraZeneca, Merck and Pfizer, to validate the potential use of CRP40 as a biomarker for PD. This initiative could lead to the development of highly specific, sensitive, and more affordable diagnostic tests. As well, this research may herald possibilities for the development of disease-modifying therapies based on restoration of function of essential heat shock proteins.

References

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Bitanihirwe BKY, Woo TUW. Oxidative stress in schizophrenia: an integrated approach. Neuroscience and biobehavioral rev.2011; 35(3):878-893.

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Ciobica A, Padurariu M, Dobrin I, Stefanescu C, Dobrin R.Oxidative stress in schizophrenia – focusing on the main markers. Psychiatria Danubina. 2011;23(3):237 245.

De Mena L, Coto E, Sánchez-Ferrero E, Ribacoba R, Guisasola LM, Salvador C, Blázquez M, Alvarez V. Mutational screening of the mortalin gene (HSPA9) in Parkinson’s disease. J Neural Transm. 2009;116(10):1289-1293.

D’Souza SM, Brown IR. Constitutive expression of heat shock proteins Hsp90, Hsc70, Hsp70 and Hsp60 in neural and non-neural tissues of the rat during postnatal development.Cell Stress Chaperon. 1998;3(3):188-199.

Gabriele JP, Chong VZ, Pontoriero GF, Mishra RK. Decreased expression of a 40-kDa catecholamine-regulated protein in the ventral striatum of schizophrenic brain specimens from the Stanley Foundation Neuropathology Consortium. Schizophrenia Res.2005;74(1):111-119.

Gabriele JP, Pontoriero GF, Thomas N, Ferro MA, Mahadevan G, MacCrimmon DJ, Pristupa ZB, Mishra RK. Antipsychotic drug use is correlated with CRP40/ mortalin-2 mRNA expression in the dorsolateral prefrontal cortex of human postmortem brain specimens. Schizophrenia Res. 2010a;119(1-3):228-231.

Gabriele N, Pontoriero GF, Thomas N, Shethwala SK, Pristupa ZB, Gabriele JP. Knockdown of mortalin-2 within the medial prefrontal cortex impairs normal sensorimotor gating. Synapse. 2010b; 64(11):808-813.

Gabriele J, Pontoriero GF, Thomas N, Thomson C, Skoblenick K, Pristupa ZB, Mishra RK. Cloning, characterization, and functional studies of a human 40-kDa catecholamine-regulated protein: implications in central nervous system disorders. Cell Stress Chaperon. 2009;14(6):555-567.

Gabriele JP, Rajaram M, Zhang B, Sharma S, Mishra RK. Modulation of a 40-kDa catecholamine-regulated protein following D-amphetamine treatment in discrete brain regions. Eur. J. Pharmacol. 2002; 453(1):13-19.

Gabriele J, Thomas N, N-Marandi S, Mishra R. Differential modulation of a 40 kDa catecholamine regulated protein in the core and shell subcompartments of the nucleus accumbens following chronic quinpirole and haloperidol administration in the rat. Synapse. 2007;61(10):835-842.

Groleau SE, Lubarda J, Thomas N, Ferro MA, Pristupa ZB, Mishra RK, Gabriele JP. Human blood analysis reveals differences in gene expression of catecholamine-regulated protein 40 (CRP40) in schizophrenia. Schizophr Res. 2013;143(1):203-206.

Jin J, Hulette C, Wang Y, Zhang T, Pan C, Wadhwa R, et al.Proteomic identification of a stress protein, Mortalin/mthsp70/ GRP75: relevance to Parkinson disease. Mol Cell Proteomics. 2006; 5(7):1193-1204.

Jin J, Li GJ, Davis J, Zhu D, Wang Y, Pan C, Zhang J. Identification of novel proteins associated with both alpha-synuclein and DJ-1. Mol Cell Proteomics. 2007;6(5):845-859.

Kaul SC, Deocaris CC, Wadhwa R. Three faces of Mortalin: a housekeeper, guardian and killer. Exp Gerontol.2007;42(4):263-724.

Kim JJ, Mandelli L, Lim S, Lim HK, Kwon OJ, Pae CU, Serretti A, Nimgaonkar VL, Paik IH, Jun TY. Association analysis of heat shock protein 70 gene polymorphisms in schizophrenia. Eur. Arch. Psychiatry Clin Neurosci.2008;258(4):239-244.

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Lubarda J, Groleau S, Thomas N, Pristupa ZB, Mishra RK, Gabriele JP. Molecular chaperones as 21st century solutions for neurodegenerative disorders. A look at catecholamine- regulated protein 40 (CRP40). Integrated Healthcare Practitioners. 2013a;6(1):65-70.

Lubarda J, Groleau SE, Thomas N, Ferro MA, MishraRK, GabrieleJP. Dysregulation of novel catecholamine-regulated protein 40 (CRP40) in Parkinson’s disease patients. Movement Disord. 2013b [Accepted January 18; MDF-13-0044].

Modi PI, Kashyap A, Nair VD, Ross GM, Fu M, Savelli JE, Marcotte ER, Barlas C, Mishra RK. Modulation of brain catecholamine absorbing proteins by dopaminergic agents. Eur J Pharmacol. 1996;299(1-3):213-220.

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Schapira AH. Neurobiology and treatment of Parkinson’s disease. Trends Pharmacol Sci.2009;30(1):41-47.

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Schwarz E, Prabakaran S, Whitfield P, Major H, Leweke FM, Koethe D, McKenna P, Bahn S. High throughput lipidomic profiling of schizophrenia and bipolar disorder brain tissue reveals alterations of free fatty acids, phosphatidylcholines, and ceramides. J Proteome Res. 2008;7(10):4266-4277.

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Sharan N, Nair VD, Mishra RK. Modulation of a 40-kDa catecholamine regulated protein by dopamine receptor antagonists. Eur. J. Pharmacol. 2001;413(1):73-79.

Shi M, Jin J, Wang Y, Beyer RP, Kitsou E, Albin RL, Gearing M, Pan C, Zhang J. Mortalin: a protein associated with progression of Parkinson disease? J Neuropathol Exp Neurol. 2008;67(2):117-124.

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Stokes AH, Hastings TG, Vrana KE. Cytotoxic and genotoxic potential of dopamine. J Neurosci Res.1999;55(6):659-65.

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Wood SJ, Yücel M, Pantelis C, Berk M. Neurobiology of schizophrenia spectrum disorders: the role of oxidative stress. Annuals of the Academy of Medicine, Singapore. 2009;38(5):396.

Yao JK, Keshavan MS. Antioxidants, redox signaling, and pathophysiology in schizophrenia: an integrative view. Antiox. & Redox Sig.2011;15(7):2011-2035.

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Estrogen metabolism

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Estrogen metabolism

Advances in prediction of disease risk

Since the late 1990’s researchers have been using immunoassays to measure certain estrogen metabolites (2- and 16-hydroxyestrogens) in urine, in an attempt to gauge future risk of breast and prostate cancer, and many practitioners have used these immunoassay measurements in their practices for the past decade. A 2011 meta-analysis examined the body of literature on this topic and questions its validity. This article reviews the conclusions of the meta-analysis and briefly discusses future directions for research, using the newer, LCMSMS technology.

In the last several decades there has been increased recognition that elevated levels of estrogens are causally related to breast cancer (Ziegler 2010). This has led to increased interest in how estrogens are broken down in the body including how various nutritional deficiencies and genetic tendencies relevant to cancer may affect the pattern of urine estrogen metabolites.

Our understanding of estrogen metabolism has been largely shaped by the measurement technologies available. Past measurement techniques have included immunoassays (RIA and ELISA) as well as gas chromato graphic-mass spectrometric (GCMS) methods (Stanczyk 2007). The latest methodology uses liquid chromatography-tandem mass spectrometry (LCMSMS), which is said to provide improved accuracy and specificity (Falk 2008). Certain long-held ideas about estrogen metabolism now need to be revisited in light of improvements in our ability to measure estrogen metabolites.

This review summarizes what we know about the interpretation of urine estrogen metabolite measurements in light of the most recent research and technological developments. In short, it appears that the traditional urine estrogen metabolite ratio measured by ELISA is of less utility for gauging risk of breast cancer than previously thought. It is also apparent that LCMSMS measurements of the urine estrogen metabolites cannot be interpreted in the same way as the older ELISA data. There is increased interest in lesser-studied metabolites such as 4-hydroxyestrone, and 2-methoxyestrone for assessment of risk of estrogen-sensitive cancers, and the development of newer methodologies is sure to usher in further advances in estrogen metabolism research.

2:16 ratio Around 1997, the Estramet™ ELISA estrogen metabolite kit was introduced as a means to measure the ratio of 2-hydroxylated estrogens to 16-hydroxylated estrone. This ratio is often referred to as the “2/16 ratio” or “Estrogen Metabolism Ratio (EMR)”. Based on literature suggesting that 16-hydroxyestrone was carcinogenic and that 2-hydroxyestrogens were protective or at least cancer-neutral, the ratio was deemed to be relevant for breast cancer risk assessment (Lord 2008). A lower ratio (lower than normal 2-hydroxyestrogens) was thus thought to reflect an excess of “bad” estrone (16OHE1) relative to the good/benign estrogens. Numerous therapies have been shown to increase this ratio, mostly by upregulating the enzymes that make 2-hydroxyestrogens (Lord 2002). The rationale for increasing the EMR was to decrease future risk of breast cancer, although this has never been prospectively or retrospectively studied.

Like many immunoassays, the Estramet™ kit lacks specificity, which is evidenced by the fact that the antibody ostensibly directed toward 2-hydroxyestrone cross-reacts with other 2-hydroxylated estrogens (IBL Estramet™ 2008). In other words, two individuals may have the same value for total 2-hydroxyestrogens, but the proportional values of 2-hydroxyestrone (2OHE1), 2-hydroxyestradiol (2OHE2) and 2-hydroxyestriol (2OHE3) may be quite different for each. This makes the interpretation of data generated by the ELISA kit somewhat problematic.

When the Estramet™ kit was introduced in the late 1990’s, it was used in several published prospective and population-based case control studies on the risk of breast cancer. Based on these studies, practitioners understood that an Estramet™ ratio of less than approximately two was associated with increased breast cancer risk in both pre- and postmenopausal women not currently using estrogen therapy (Kabat 1997, Meilahn 1998, Muti 2000). The increased risk of breast cancer comparing the highest to lowest quantiles of the EMR was estimated to be between 15 and 30%, with weak statistical significance (Obi 2011).

More recent literature suggests a need to reevaluate these assumptions. A 2011 meta-analysis by Obi et al reviewed all the literature on the EMR specifically for non-estrogen users and concluded: “For the highest compared with the lowest quantile of urinary EMR, non-significant associations suggested at best a weak protective effect in premenopausal but not in postmenopausal breast cancer (range of odds ratios: 0.50–0.75 for premenopausal and 0.71–1.31 for postmenopausal)” (Obi 2011). In other words, there is no evidence that a higher EMR protects postmenopausal women from breast cancer, and there is weak evidence of a protective effect in premenopausal women. For each of the studies included in Obi’s metaanalysis, a risk reduction or risk increase was calculated by comparing the highest EMRs to the lowest EMRs. Only one of these studies, conducted in China, showed a significant decrease in breast cancer risk. Other studies looking at North American or European populations produced results that clustered around a relative risk of 1 – an indication of no effect on breast cancer risk (Obi 2011). Hence, the notion that the EMR (measured by immunoassay) is associated with breast cancer risk is not on a robust statistical foundation.

The EMR is on even shakier ground when viewed through the lens of estrogensupplementing women. A prospective casecontrol analysis authored by Wellejus (2005) looked at the predictive value of the EMR (as measured by Estramet™) for breast cancer risk in postmenopausal women using estradiolbased HRT. Unlike the no-effect or weakly protective effect of the higher EMR on breast cancer risk in non-estrogen users, Wellejus found that higher EMRs were associated with an increased risk of estrogen receptor positive breast cancer in women supplementing with estradiol-based hormone therapy. The authors found that this increased risk was tied to increased levels of 2-hydroxyestrogens and was independent of 16-hydroxyestrone (16OHE1); they commented that presentation of the metabolites as a ratio is probably misleading in postmenopausal women supplementing with estradiol (Wellejus 2005).

Assuming that there is value in knowing the EMR, should we try to use the urinary levels of specific 2-hydroxylated estrogen metabolites (2OHE1 and 2OHE2) more accurately measured via LCMSMS to calculate an approximation of the immunoassay method EMR? If so, how should laboratories determine the correct threshold?

One laboratory addresses this issue by measuring estrogen metabolite levels both ways: via LCMSMS and via immunoassay, reporting the individual LCMSMS numbers for hydroxyestrones and hydroxyestradiol and reporting the EMR based on immunoassay numbers (Genova Diagnostics references I 2013, Genova Diagnostics II 2013). Another laboratory uses LCMSMS to measure all metabolites, and derives an approximation of the EMR from the following: (2OHE2 + 2OHE1)/16OHE1. The threshold for this LCMSMS ratio was chosen to divide the sample population in the same fashion as did the immunoassay threshold ratio (Metametrix 2013). Ideally, the two thresholds would numerically match, but in practice the LCMSMS threshold is higher. This reflects the fact that the two methodologies are not measuring exactly the same things. In order to properly resolve the issue, new prospective studies would have to be performed using LCMSMS methodology.

An important consideration when working with ratios is their potential to shift focus away from the component numbers. For example, elevated 2-hydroxylated estrogen metabolites are a problem regardless of their level in relation to 16OHE1 (as noted by Wellejus). Conversely if all estrogen metabolite levels are very low, is an abnormal ratio really relevant? Likely not.

EMR in Males

There are several case-control retrospective studies looking at a possible connection between prostate cancer and the EMR (Barba 2009). In general, these studies suggest a weak association between a low EMR and prostate cancer risk. Muti, for example, found that men in the highest tertile for EMR had an 39% lower risk of prostate cancer (OR of 0.61) compared to the lowest tertile, but the confidence interval was wide and crossed 1 (no effect) (Muti 2002). As is the case for women and breast cancer, there are no controlled trials that show a higher EMR protects against prostate cancer, nor any that show that a therapeutic increase in the EMR will lower the risk of prostate cancer. In short, given the current state of research, it appears the EMR is no more valuable in males than it is in females.

In summary, it is this author’s opinion that there is minimal value in reporting the EMR or an approximation thereof in either women or men. There is likely more value in measuring individual metabolite levels. For example, when estrogens are being supplemented (particularly oral estradiol), elevation of “2-hydroxyestrogens” appears to be associated with an increased risk of breast cancer (Wellejus 2005). This leads naturally to the question of whether there is value in examining other estrogen metabolites, or in examining metabolites in different ways.

Hydroxyestrone vs Hydroxyestradiol None of the previously discussed immunoassay studies address the issue of the relative proportions of 2-hydroxyestrone and 2-hydroxyestradiol metabolites. This may be of particular importance when 2-hydroxyestrogens are high. Using an immunoassay like the Estramet™, it is impossible to know if elevated 2-hydroxyestrogens are a result of elevated 2OHE1, 2OHE2 or both because the individual metabolites are indistinguishable to the antibodies. It is therefore worth asking the question: are the relative amounts of each type of estrogen important in the overall total?

To answer that question, it is important to know the following: the various estrogens can exist in either estrone or estradiol form. The enzyme that converts estradiol to its estrone form is 17-hydroxysteroid dehydrogenase Type II. Estrogen pairs that regularly undergo conversion include estrone/estradiol, 2-hydroxyestrone/2- hydroxyestradiol, 4-hydroxyestrone/4-hydroxyestradiol and others (Kitawaki 2000).

Rocky Mountain Analytical has reviewed the partitioning of 2-hydroxyestrogens measured via LCMSMS in first morning urine in a population of women (including women with both recent and remote diagnoses of breast cancer. The data below are presented without regard to breast cancer status. The results are summarized below.

The estrone metabolite appears to be favoured over the estradiol metabolite as the ratio of 2OHE1 to 2OHE2 exceeds 50% across all groups. The lower percentage of 2OHE1 in post-menopause relative to pre-menopause probably reflects differential expression of 17-hydroxysteroid dehydrogenase Type II. Progesterone is known to induce this enzyme (Kitawaki 2000), which means the higher proportion of 2OHE1 relative to 2OHE2 in pre-menopause is likely a consequence of higher progesterone levels found in premenopausal women.

Research published by Huang in late 2011 examined 16 different estrogen metabolites in the urine of postmenopausal women with breast cancer versus healthy controls. The breast cancer patients were found to have higher levels of both 2- and 4-hydroxyestradiol compared to healthy controls. The proportions between estrones and estradiols could only be estimated, as their data did not report 2- and 4-hydroxyestrones separately. Their research does however support the notion that although estrogens are supposed to be able to freely interconvert between the estrone and estradiol forms, the equilibria lie more toward the estradiol side in breast cancer patients (Huang 2012).

The point of the preceding discussion is that it is probably time to move away from some of the older concepts about estrogen metabolism and breast cancer, and start mining the specific (and copious) data provided by LCMSMS technology.

Methoxyestrogens

Further metabolism of hydroxyestrogens involves conversion of a hydroxyl (-OH) group to a methoxy group (-OCH3). Hence there is a family of methoxyestrogens including 2-methoxyestradiol, 2-methoxyestrone, 4-methoxyestradiol and 4-methoxyestrone, which mirror the hydroxyestrogens (Obi 2011).

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There is some literature indicating that methoxyestrogens may be relevant for assessing risk of breast cancer. For example, the study by Huang, mentioned earlier noted lower levels of 2-methoxyestradiol (2-MeOE2) in breast cancer patients compared to controls (Huang 2012). In particular 2-MeOE2, the downstream metabolite of 2-hydroxyestradiol, is thought to mitigate against breast cancer by a variety of mechanisms including anti-angiogenesis and cell cycle regulation (Mueck 2010). This molecule is the focus of active research as a potential treatment for breast cancer.

Huang et al speculated in their paper: “Interestingly, 2-MeOE2 levels, with a p-value of 0.038, were significantly reduced in breast cancer cases. It is speculated that methylation of catechol estrogens might prevent reactive quinine formation as well as weaken binding to the estrogen receptor, both of which should be beneficial for decreased breast cancer risk. Because methylation plays an important role in the protection against rapid tumor growth, 2-MeOE2 could be presumed a potential anticancer biomarker for breast cancer” (Huang 2012).

Conversion of hydroxyestrogen to methoxyestrogen is a transmethylation reaction requiring an adequate supply of methyl donors and magnesium (enzyme cofactor) as well as sufficient catechol orthomethyltransferase (COMT) enzyme activity (Cavalieri 2011). Reduced levels of methoxyestrogens relative to their parent hydroxyestrogens may therefore reflect nutritional factors (insufficient levels of folic acid, B12, magnesium, betaine, SAMe, and/or MSM) plus genetic factors (homo- or heterozygosity for hypofunctioning COMT alleles) (Cavalieri 2011). Hypofunctioning mutations of COMT have been noted in various populations of women with breast cancer (Yager 2000). To date, no prospective trials looking at the predictive value of low levels of 2-methoxyestrogens in relation to breast cancer have been reported.

Once again from the data presented by Huang, the ratio of methoxyestrogen/parent hydroxyestrogen was calculated for postmenopausal breast cancer patients versus healthy postmenopausal controls. In general, the ratios became smaller (closer to zero) for breast cancer patients compared to controls, possibly indicating that there was less methoxylation of parent hydroxyestrogens in breast cancer patients, and supporting the notion that it may be worthwhile to measure and track these various parameters (Huang 2012).

4-Hydroxyestrone

At the time that 16-hydroxyestrone was being put forth as a “bad estrogen”, 4-hydroxyestrone was also emerging as an important metabolite in relation to breast cancer. On balance, evidence is fairly compelling to support a role for 4-hydroxyestrone in breast cancer causation. An extensive monograph on this topic was published in 2000 (Jefcoate 2000). Ercole Cavalieri, a principal researcher in this field, has also published an excellent review (Cavalieri 2002).

The proposed carcinogenic action of 4OHE1 is as follows: after conversion to a quinone form, 4-hydroxyestrone may irreversibly bind to DNA, resulting in removal of purine bases (Cavalieri-1997, 2002). Depurination of oncogenic stretches of DNA may be a very early event in the genesis of breast cancer (Cavalieri 2000).

By cycling back and forth between the semiquinone and quinone forms, 4-hydroxyestrone also generates free radicals, with resultant DNA damage and lipid peroxidation (Davis 1997, Cavalieri 1997).

Gaikwad et al demonstrated that women with untreated breast cancer had the highest levels of purine adducts with 4-hydroxyestrone compared to healthy controls and women at high risk of breast cancer. Conversely, the patients with cancer had the lowest levels of thiol conjugates (including glutathione conjugates) of 4-hydroxyestrone (Gaikwad 2009). Conjugation with glutathione and other thiol-containing molecules prevents depurinating adduct formation (Cavalieri 2011). Low levels of thiol conjugates would imply less protection. Breast cancer patients also had the lowest levels of 4-methoxyestrone, which is another mechanism whereby 4-hydroxyestrone is “detoxified”. However, there is still some controversy regarding the utility of urinary 4OHE1 (as conjugate) levels for assessing breast cancer risk. Gaikwad presented data indicating lower levels of 4-hydroxyestrone in breast cancer patients compared to controls. Rocky Mountain Analytical data shows the opposite, with highest levels of 4-hydroxyestrone appearing in breast cancer patients. This apparent conflict may be a result of different testing methodologies. In any case, the mass balance of estrogen in the 4-hydroxyestrone ‘branch’ of the estrogen metabolism tree appears to be an exciting avenue for future research.

Closing Remarks

The relationship between breast cancer and estrogen metabolites continues to be an active area of research. Emphasis needs to shift away from the 2-16 ratio and toward estrogen metabolite profiles which are more closely linked to breast cancer. Further research may also be directed towards analytes that inform on DNA damage and glutathione deficiency. This is a fascinating area of Women’s Health in which analysis of urine estrogen metabolites via LCMSMS has much to offer.

References

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Social Media changing the behaviour of health care practitioners

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A recent study analyzed the factors that influence the adoption and use of social media by physicians to share medical information. The authors developed a survey that was emailed to 1695 practicing oncologists and primary care physicians in the United States. Responses from 485 physicians were analyzed. Overall, almost one in four physicians used social media daily or many times daily to scan or explore medical information and 14% contributed new information through social media. Physicians largely had favourable attitudes toward the use of social media and most perceived social media to be beneficial, engaging, and a good way to get high-quality information. A majority of the physicians also identified that social media enabled them to care for patients more effectively and improve the quality of patient care they delivered. The main factors influencing physician’s usage or share information was perceived ease of use and usefulness.

New drug targets aggressive Non-Hodgkin’s Lymphoma

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An international research team led by two laboratories from Weill Cornell Medical College have developed a new experimental drug to target an aggressive form of lymphoma, called activated B-cell diffuse large B-cell lymphoma (ABC-DLBCL). An experimental small molecule agent called MI-2 irreversibly inactivates a key protein, MALT1, that is responsible for driving the growth and survival of lymphoma cells. In their study, the researchers showed that the MI-2 inactivates any MALT1 it comes into contact with and also does so without any apparent toxicity in animal models.

Breast milk probiotic launched in Australia: world first

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Biosearch Life’s Hereditum probiotic strain Lactobacillus fermentum Lc40 is being sold in sachet form and it is marketed for pregnant and breastfeeding mothers. Studies have shown that certain strains of probiotics such as this one
can improve the immune system of the mother and infant. In addition, it can prolong breastfeeding by reducing the incidence of mastitis, which the World Health Organization ranks as the number one reason mother’s cease breastfeeding.The probiotic is being sold under the name Qiara and is being endorsed by some professionals as an alternative to antibiotics that have proven ineffective in some situations due to drug resistance in breastfeeding women.