Osteoporosis, aging, tissue renewal, and product science


The incidence of osteoporosis, like obesity, has been increasing in recent decades. The number of hip
fractures in many countries has doubled in the last 30 or 40 years (Bergstrom, et al., 2009). An exception
to that trend was Australia in the period between 2001 and 2006, where the annual incidence of hip fractures
in women over 60 years old decreased by 28.3%. During those years, the number of prescriptions for "hormone
replacement therapy" decreased by 54.6%, and the number of prescriptions for bisphosphonate increased by
245%. The publication of the Women's Health Initiative results in 2002 (showing that the Prem-Pro treatment
caused breast cancer, heart attacks, and dementia), led to a great decrease in the use of estrogen
treatments everywhere. After the FDA approved estrogen's use in 1972 for the prevention of osteoporosis the
number of women using it increased greatly, and by 1994, 44% of women in the US had used it. After the WHI
results were published, the number of prescriptions for "HRT" fell by more than half, and following that
decrease in estrogen sales, the incidence of breast cancer decreased by 9% in women between the ages of 50
and 54.With the incidence of hip fractures increasing while the percentage of women using estrogen was
increasing, it seems likely that there is something wrong with the theory that osteoporosis is caused by an
estrogen deficiency. That theory was derived from the theory that menopause was the consequence of ovarian
failure, resulting from the failure to ovulate and produce estrogen when the supply of eggs was depleted.
The theory was never more than an ideological preference, but the estrogen industry saw it as an opportunity
to create a huge market.There are many studies that seem to imply that the greater incidence of osteoporotic
fractures among women is the result of their exposure to estrogen during their reproductive years. This
would be analogous to the understanding that it is the cumulative exposure to estrogen that ages the nerves
in the hypothalamus that control the cyclic release of the gonadotropic hormones, causing the
menopause.. . . the nature of science itself changed around the middle of the last century, becoming
product and disease oriented, so that now relatively few people are continuing to study bones
objectively.Animal studies show that estrogen stunts growth, including bone growth. The high
estrogen levels in girls' teen years and early twenties accounts for the fact that women's bones are lighter
than men's. In rat studies, treatment with estrogen was found to enlarge the space between the jawbone and
the teeth, which is a factor in periodontal disease (Elzay, 1964). Teeth are very similar to bones, so it's
interesting that treating male or female rats with estrogen increases their incidence of tooth decay, and
removing their gonads was found to decrease the incidence (Muhler and Shafer, 1952). Supplementing them with
thyroid hormone decreased the incidence of cavities in both males and females (Bixler, et al., 1957).One of
the "estrogen receptors" appears to actively contribute to bone loss (Windahl, et al., 1999, 2001). Studies
in dogs following the removal of their ovaries found that there was an increase of bone remodeling and bone
formation rate in the first month, followed by a few months of slowed bone formation, but that by 10 months
after the surgery the bones had returned to their normal remodeling rate, and that "at no time was a
significant reduction in bone volume detected" (Boyce, et al., 1990). With the removal of the ovaries, the
production of progesterone as well as estrogen is affected, but the adrenal glands and other tissues can
produce those hormones.Until the influence of the estrogen industry overwhelmed it, ordinary science was
studying bone development in comprehensive ways, understanding its biological roles and the influences of
the environment on it. But the nature of science itself changed around the middle of the last century,
becoming product and disease oriented, so that now relatively few people are continuing to study bones
objectively.The outstanding physical-chemical property of bone is that it is a reservoir-buffer of carbon
dioxide, able to bind huge amounts of the gas into its structure.When carbon dioxide increases in the
bloodstream it is at first absorbed rapidly by the bones, and if the blood level of CO2 is kept high day
after day, the rate of absorption gradually slows down, but in experiments that have continued for several
weeks the bones were still slowly absorbing more carbon dioxide; the absorption curve seems to be
asymptotic. When people move to or from high altitudes, their bones appear to continue adapting to the
different gas pressures for years. A reduction of atmospheric pressure (which allows the tissues to retain
more carbon dioxide) helps to reduce the calcium loss caused by immobility (Litovka and Berezovs'ka, 2003;
Berezovs'kyi, et al., 2000), and promotes the healing of damaged bone (Bouletreau, et al., 2002). Ultrasound
treatment, which accelerates bone healing, stimulates processes similar to reduced oxygen supply (Tang, et
al., 2007). The mineral in newly formed bone is calcium carbonate, and this is gradually changed to include
a large amount of calcium phosphate. Besides forming part of the mineral, carbon dioxide is also
incorporated into a protein (in a process requiring vitamin K), in a process that causes this protein,
osteocalcin, to bind calcium. The osteocalcin protein is firmly bonded to a collagen molecule. Collagen
forms about 30% of the mass of bone; several percent of the bone consists of other organic molecules,
including osteocalcin, and the rest of the mass of the bone consists of mineral.Thyroid hormone is essential
for forming carbon dioxide. In the early 1940s, experimental rabbits were fed their standard diet, with the
addition of 1% desiccated thyroid gland, which would be equivalent to about 150 grains of Armour thyroid for
a person. They became extremely hypermetabolic, and couldn't eat enough to meet their nutritional needs for
growth and tissue maintenance. When they died, all of their tissues weighed much less than those of animals
that hadn't received the toxic dose of thyroid, except for their bones, which were larger than normal.
Experiments with the thin skull bones of mice have shown that the active thyroid hormone, T3, increases the
formation of bone. To increase cellular respiration and carbon dioxide production, T3 increases the activity
of the enzyme cytochrome oxidase, which uses copper as a co-factor. Increased thyroid activity increases the
absorption of copper from foods.There is an inherited condition in humans, called osteopetrosis or marble
bone disease, caused by lack of a carbonic anhydrase enzyme, which causes them to retain a very high level
of carbon dioxide in their tissues. Using a chemical that inhibits carbonic anhydrase, such as the diuretic
acetazolamide, a similar condition can be produced in animals. Acetazolamide inhibits the bone resorbing
actions of parathyroid hormone, including lactic acid formation and the release of the lysosomal enzyme,
beta-glucuronidase (Hall and Kenny, 1987). While lactic acidosis causes bone loss, acidosis caused by
increased carbonic acid doesn't; low bicarbonate in the body fluids seems to remove carbonate from the bone
(Bushinsky, et al., 1993), and also mineral phosphates (Bushinsky, et al., 2003). The parathyroid hormone,
which removes calcium from bone, causes lactic acid to be formed by bone cells (Nijweide, et al., 1981;
Lafeber, et al., 1986). Lactic acid produced by intense exercise causes calcium loss from bone (Ashizawa, et
al., 1997), and sodium bicarbonate increases calcium retention by bone. Vitamin K2 (Yamaguchi, et al., 2003)
blocks the removal of calcium from bone caused by parathyroid hormone and prostaglandin E2, by completely
blocking their stimulation of lactic acid production by bone tissues. Aspirin, which, like vitamin K,
supports cell respiration and inhibits lactic acid formation, also favors bone calcification. Vitamin K2
stimulates the formation of two important bone proteins, osteocalcin and osteonectin (Bunyaratavej, et al.,
2009), and reduces the activity of estrogen by oxidizing estradiol (Otsuka, et al, 2005).The formation of
eggshell, which is mostly calcium carbonate, is analogous to the early stage of bone formation. In hot
weather, when chickens pant and lower their carbon dioxide, they form thin shells. A sodium bicarbonate
supplement improves the quality of the eggshell (Balnave and Muheereza, 1997; Makled and Charles, 1987).
Chickens that habitually lay eggs with thinner shells have lower blood bicarbonate than those that lay thick
shelled eggs (Wideman and Buss, 1985). One of the arguments for stopping the sale of DDT in the US was that
it was threatening to cause extinction of various species of bird because it caused them to lay eggs with
very weak shells. Several other synthetic estrogenic substances, ethynylestradiol, lindane, PCBs, cause
eggshell thinning, partly by altering carbonic anhydrase activity (Holm, et al, 2006). Estrogen and
serotonin activate carbonic anhydrase in some tissues, progesterone tends to inhibit it. DDE, a metabolite
of DDT, reduces medullary bone formation in birds (Oestricher, et al., 1971) and bone mineral density in men
(Glynn, et al., 2000). Among its estrogenic effects, DDE increases prolactin (Watson, et al., 2007); one
form of DDT inhibits progesterone synthesis and increases estrogen (Wojtowics, et al., 2007)In youth, the
mineralization of the collagen framework is slightly lower than in maturity, and the bones are more
flexible. With aging, the mineralization increases progressively, and the proportion of collagen decreases
slightly, and the bones become increasingly brittle. (Rogers, et al., 1952; Mbuyi-Muamba, et al., 1987).
Collagen is a major part of the extracellular substance everywhere in the body, and its concentration
increases with aging in the non-calcified tissues. There is considerable renewal and modification of
collagen, as new molecules are formed and old molecules broken down, but its average structure changes with
aging, becomes less soluble and more rigid, as the result of chemical cross-links formed between molecules.
These cross-links are involved in regulating the differentiation of bone cells (Turecek, et al., 2008).
Recently (August 2, 2011), Deasey et al., have published evidence showing that cross-linking is required for
bone mineralization (2011).The outstanding physical-chemical property of bone is that it is a
reservoir-buffer of carbon dioxide, able to bind huge amounts of the gas into its structure.Around 1950, Fritz Verzar began studying the changes of collagen that occur with aging, and his work led to
the "collagen theory of aging." He showed that older, stiffer, less elastic tendons have a higher "melting"
or contracting temperature than young tendons. (This effect is responsible for the curling of a piece of
meat when it is frying.) Verzar and his colleagues investigated the effects of hormonal treatments on the
aging of rat collagen, especially in their tail tendons. They found that estrogen treatment increased the
stiffness and the melting temperature of collagenous tissues. While estrogen increased the cross-linking
with aging, removing the pituitary gland was found to retard the aging. Later, the cross-linking enzymes
transglutaminase and lysyl oxidase, which are induced by estrogen, were found to be a major factor in the
cross-linking of collagen and other molecules.When estrogen was found to age the connective tissues, it was
assumed that continual breeding during an animal's life-time, greatly increasing the total exposure of the
tissues to estrogen, would increase the aged rigidity of the connective tissues, but these animals were
found to have less rigid tissues. During pregnancy other hormones, especially progesterone, were also
increased, and it was suggested that this reversed the effects of aging and estrogen. Since most people had
believed that frequent pregnancies would cause a woman to age more rapidly, a large survey of records was
done, to compare the longevity of women with the number of pregnancies. It was found, in the very extensive
Hungarian records, that life-span was increased in proportion to the number of pregnancies.Despite these
very interesting results in the 1950s and 1960s, the growing influence of the estrogen industry changed the
direction of aging research, favoring the belief that decreasing estrogen accelerated the deterioration of
tissues in aging, and the popularity of Denham Harman's "free radical theory of aging" led many people to
assume that random reactions produced by lipid peroxidation were responsible for most of the cross-linking,
and that theory was gradually replaced by the "glycation" theory of aging, in which sugar molecules break
down and form the cross-links, by random, non-enzymic processes. Estrogen's role in aging was completely
by-passed.The meat industry is interested in reducing the toughness of meat, by influencing the nature of
the collagen in muscle. Castrated animals were found to produce meat that was tenderer than that of intact
males. When castrated animals were treated with testosterone, the amount of collagen was increased, making
the meat tougher. But when dihydrotestoserone, which can't be converted to estrogen was used, the meat
didn't become tough. Treatment with estrogen produced the same increase of collagen as treatment with
testosterone, showing that testosterone's effect was mainly the result of its conversion to estrogen
(Miller, et al., 1990).In the 1960s and '70s the estrogen industry was looking for ways to build on the
knowledge that in puberty estrogen is responsible for accelerating the calcification of the growth plate at
the ends of the long bones, and to find a rationale for selling estrogen to all women concerned with the
problems of aging. As bone metabolism was investigated, two kinds of cell were found to be active in
constantly remodeling the bone structure: Osteoclasts (breaking it down), and osteoblasts (building new
bone). Estrogen was found to slow the actions of the osteoclasts, so the idea that it would delay
osteoporosis became the basis for a huge new marketing campaign. Slowing bone metabolism became the focus.
Although estrogen was known to increase prolactin, and prolactin was known to accelerate bone loss, nearly
all publications began to focus on substances in the blood or urine that corresponded to the rate of bone
turnover, with the implication that increasing bone turnover would correspond to a net loss of bone.This was
the context in which, during the 1980s, articles about thyroxine's role in causing osteoporosis began to
appear. The thyroid hormone supports bone renewal, and increases indicators of bone breakdown in the blood
and urine. If estrogen's use was to be justified by slowing bone turnover, then the effects of thyroid,
accelerating bone turnover, should be interpreted as evidence of bone destruction.A basic problem with many
of the publications on thyroid and bone loss was that they were talking about an unphysiological medical
practice (prescribing the pre-hormone, thyroxine), which frequently failed to improve thyroid function, and
could even make it worse, by lowering the amount of T3 in the tissues.Later, it was noticed that high TSH
was associated with the signs of lower bone turnover. TSH rises when there is less thyroid hormone, but
(after the recombinant TSH became available for medical use) a few publications argued that it was the TSH
itself, rather than the absence of thyroid hormone, that was "protecting" the bones (lowering the evidence
of bone turnover). The doctrine that had been developed to support estrogen therapy was now used to oppose
thyroid therapy. Keeping the TSH high would slow bone turnover. Working in this cultural context, genetic
engineers at Amgen identified a protein that inhibited the formation of osteoclast cells, and slowed bone
metabolism. It was suggested that it was responsible for estrogen's suppression of the osteoclasts, and many
publications appeared showing that it was increased by estrogen. It was named "osteoprotegerin," meaning
"the bone protecting protein." Prolactin increases osteoprotegerin (OPG), reducing bone resorption just as
estrogen does. Serotonin also increases OPG, and it turns out that OPG is elevated in all of the
pathological conditions associated with high serotonin, including cancer, pulmonary artery hypertension,
vascular calcification, and even bone loss.While Arthur Everitt, Verzar, and others were studying the
effects of the rat's pituitary (and other glands) on collagen, W. D. Denckla investigated the effects of
reproductive hormones and pituitary removal in a wide variety of animals, including fish and mollusks. He
had noticed that reproduction in various species (e.g., salmon) was quickly followed by rapid aging and
death. Removing the pituitary gland (or its equivalent) and providing thyroid hormone, he found that animals
lacking the pituitary lived much longer than intact animals, and maintained a high metabolic rate. Making
extracts of pituitary glands, he found a fraction (closely related to prolactin and growth hormone) that
suppressed tissue oxygen consumption, and accelerated the degenerative changes of aging.Aging, estrogen,
cortisol, and a variety of stresses, including radiation and lipid peroxidation, chemically alter collagen,
producing cross-links that increase its rigidity, and affect the way it binds minerals. The cross-linking
enzymes induced by estrogen are involved in the normal maturation of bone collagen, and at puberty when
estrogen increases, bone growth is slowed, as the cross-linking and mineralization are accelerated. With
aging and the accumulation of heavy metals and polyunsaturated fats, random oxidative processes increase the
cross-linking. In bones, the relatively large masses of cartilage absorb oxygen and nutrients slowly, so
internally the amount of oxygen is very limited, about 1/5 as much as at the surface, and this low oxygen
tension is an important factor in regulating growth, differentiation, cross-linking, and calcification,
maintaining bone integrity. But in blood vessels the connective tissues are abundantly supplied with oxygen
and nutrients; this is normally a factor regulating the production of collagen and its cross-linking, and
preventing calcification. When the factors promoting collagen synthesis and maturation are increased
systemically, with aging and stress, the excess cross-linking slows the biological renewal process in bones,
but in blood vessels the same processes creating excess cross-linking initiate a calcification process,
involving the various factors that in youth are responsible for normal maturation of bone.Prolactin, like
estrogen, interferes with thyroid function and oxygen consumption (Wade, et al., 1986; Strizhkov, 1991;
Spatling, et al., 1982). Many years ago, repeated lactation was considered to cause osteoporosis and loss of
teeth, and prolactin, which mobilizes calcium from bones for the production of milk, was recognized as an
important factor in bone loss. Drugs that increase prolactin were found to cause osteoporosis. In the 40
years since the drug industry began its intense promotion of estrogen to prevent and treat osteoporosis,
there has been very little attention to the fact that estrogen increases prolactin, which contributes to
osteoporosis, but some people (e.g., Horner, 2009) have noticed that oral contraceptives and menopausal
hormone treatments have damaged the bones of the inner ear, causing otosclerosis and impaired hearing, and
have suggested that prolactin mediates the effect.A few years ago, the "serotonin reuptake inhibitor"
antidepressants, already known to increase prolactin by increasing the effects of serotonin, were found to
be causing osteoporosis after prolonged use. Estrogen increases serotonin, which besides promoting the
secretion of prolactin, also stimulates the production of parathyroid hormone and cortisol, both of which
remove calcium from bone, and contribute to the calcification of blood vessels. The association between
weakened bones and hardened arteries is now widely recognized, but researchers are being careful to avoid
investigating any mechanisms that could affect sales of important drug products, especially estrogen and
antidepressants.Following the recognition that the SSRI drugs were causing osteoporosis, it was discovered
that the serotonin produced in the intestine causes bone loss, and that inhibiting intestinal serotonin
synthesis would stop bone loss and produce a bone building anabolic effect (Inose, et al., 2011). One group
that had been concentrating on the interactions of genes commented that, recognizing the effects of
intestinal serotonin, they had suddenly become aware of "whole organism physiology" (Karsenty and Gershon,
2011).In previous newsletters I have talked about the ability of intestinal irritation and the associated
increase of serotonin to cause headaches, asthma, coughing, heart and blood vessel disease, muscular
dystrophy, flu-like symptoms, arthritis, inflammation of muscles and nerves, depression, and inflammatory
brain diseases. With the new recognition that serotonin is a basic cause of osteoporosis, intestinal health
becomes a major issue in aging research.The protein that inhibits intestinal formation of serotonin is the
low density lipoprotein receptor-related protein. This seems likely to have something to do with the fact
that "low" HDL is associated with better bones. A low level of LDL is associated with increased vertebral
fractures (Kaji, et al., 2010).Cartilage synthesis and turnover are highest at night. It is inhibited by
metabolic acidosis (increased lactic acid), but not by respiratory acidosis (CO2) (Bushinsky, 1995). Since
most calcium is lost from bone during the night (Eastell, et al., 1992; even in children: DeSanto, et al.,
1988) in association with the nocturnal rise of the catabolic substances, such as free fatty acids,
cortisol, prolactin, PTH, and adrenalin, things which minimize the nocturnal stress can decrease the bone
turnover. These include calcium (Blumsohn, et al., 1994) and sugar. Catabolic substances and processes
increase with aging, especially at night. Babies grow most during the night when bone turnover is high, and
even a daytime nap accelerates collagen turnover (Lutchman, et al., 1998). Discussions about whether a
certain person's osteoporosis is "menopausal osteoporosis" or "senile osteoporosis" have neglected the
possibility that osteoporosis doesn't begin in either menopause or old age, but that it is the result of
life-long developmental processes that interact with all the factors that are involved in aging. The fact
that the collagen content of old bone is lower than in young bone (as a percentage of bone weight) shows
that the problem in osteoporosis isn't a lack of calcification, it's a deficiency of tissue renewal,
parallel to sarcopenia, the decrease of muscle mass with aging. Systemically decreased tissue renewal would
account for the association of bone loss with other processes such as male baldness (Morton, et al., 2007)
and Alzheimer's disease (Zhou, et al., 2011, Duthie, et al., 2011).A high level of respiratory energy
production that characterizes young life is needed for tissue renewal. The accumulation of factors that
impair mitochondrial respiration leads to increasing production of stress factors, that are needed for
survival when the organism isn't able to simply produce energetic new tissue as needed. Continually
resorting to these substances progressively reshapes the organism, but the investment in short-term
survival, without eliminating the problematic factors, tends to exacerbate the basic energy problem. This
seems to be the reason that Denckla's animals, deprived of their pituitary glands, but provided with thyroid
hormone, lived so long: they weren't able to mobilize the multiple defenses that reduce the mitochondria's
respiratory energy production. Several things that the geneticists would never be able to fit into their
schemes of "bone regulatory molecules" such as OPG, growth hormone, parathyroid hormone, and estrogen, fit
neatly with the idea that bone health is maintained by respiratory energy and tissue renewal, under the
influence of thyroid hormone. For example, adrenaline, which is increased by stress, aging, and
hypothyroidism (and in many cases by estrogen), causes bone loss. Even the bone loss caused by immobility
can be blocked by an adrenaline blocker such as propranolol. (The stress of immobility also famously
increases serotonin.) Adrenaline tends to decrease carbon dioxide and increase lactic acid, and it strongly
increases parathyroid hormone (Ljunhgall S, et al., 1984). Calcium activates mitochondrial respiration, and
lowers adrenaline (Luft, et al., 1988), parathyroid hormone (Ohgitani, et al., 1997), and prolactin (Kruse
and Kracht, 1981). Copper, which is the co-factor for the cytochrome C oxidase enzyme, activated by thyroid,
is essential for bone formation and maintenance, and is consistently deficient in osteoporosis. Thyroid
hormone increases the body's ability to assimilate copper. Aspirin, which stimulates bone formation, has
other thyroid-like actions, including activation of mitochondrial respiration and energy production, with an
increase of cytochrome C oxidase (Cai, et al., 1996), and it lowers serotonin (Shen, et al., 2011). It also
apparently protects against calcification of the soft tissues, (Vasudev, et al., 2000), though there has
been surprisingly little investigation of that. "Aspirin can promote trabecular bone remodeling, improve
three-dimensional structure of trabecular bone and increase bone density of cancellous in osteoporotic rats
by stimulating bone formation. It may become a new drug for the treatment of osteoporosis" (Chen, et al.,
2011).A wide range of inflammatory mediators that accelerate inflammation and bone loss also inhibit thyroid
function. People who ate more polyunsaturated fat, which inhibits thyroid and oxidative metabolism, were
several times more likely to have osteoporotic fractures (that is, essentially spontaneous fractures) than
people who ate the least (Martinez-Ramirez, et al., 2007). Arachidonic acid stimulates prolactin secretion,
and prolactin acts on the thyroid gland to decrease its activity, and on other tissues to increase their
glycolysis (with lactate production), while decreasing oxidative metabolism (Spatling, et al., 1982;
Strizhkov, 1991). Living at high altitude, which strengthens bones, increases thyroid activity and decreases
prolactin (Richalet, et al., 2010) and parathyroid hormone (Khan, et al., 1996). It lowers free fatty acids,
which lower bone mass by reducing bone formation and increasing bone resorption (Chen, et al., 2010). In
menopausal women, polyunsaturated fatty acids and even monounsaturated fats are associated with bone loss,
fruit and vegetable consumption protects against bone loss (Macdonald, et al., 2004).While it's very
interesting that the drug propranol which blocks adrenaline, and drugs that block serotonin formation, have
bone protective and restorative effects, they also have undesirable side effects. Food choices that optimize
oxidative metabolism are the safest, as well as the most economical, way to approach the problem of
osteoporosis and other degenerative changes. A person can easily perceive changes in appetite, quality of
sleep, changes in skin, hair, and mood, etc., but blood tests could be used to confirm that the right
choices were being made. Tests for vitamin D, parathyroid hormone, free fatty acids, and CO2/bicarbonate, as
well as the hormones, can be helpful, if a person isn't sure whether their diet, sunlight exposure, and
thyroid supplementation is adequate. The popular medical understanding of the organism is based on a
mechanistic view of causality, in which genes have a central role, causing things to develop and function in
certain ways, and that hormones and drugs can cause genes to increase or decrease their activity. Genes that
build bones can be activated by one substance, and genes that tear down bones can be inhibited by another
substance. The "osteoprotegerin" story illustrates the problem with that kind of thinking. Vernadsky's
description of an organism as a "whirlwind of atoms" is probably a better way to think of how "causality"
works. The moving air in a whirlwind forms a self-intensifying system, with the motion reducing the
pressure, causing more air to be drawn into the system. The atoms moving in coordination aren't acting as
separate things, but as parts in a larger thing. The way in which increased metabolism in the bones acts
favorably on the metabolism of kidneys, blood vessels, lungs, liver, digestive system, etc., which in turn
favors the bones' renewal, is analogous to the tendency of a whirlwind to intensify as long as there is a
source of energy. The intensity of oxidative metabolism is the basic factor that permits continuing
coordination of activity, and the harmonious renewal of all the components of the organism.
<h3>REFERENCES</h3>Ann Nutr Aliment. 1975;29(4):305-12. [Effects of administering
diets with starch or sucrose basis on certain parameters of calcium metabolism in the young, growing
rat]. Artus M. (Sucrose maintains calcium homeostasis in vitamin D deficient bones.)J Appl
Physiol. 1997 Oct;83(4):1159-63. A bout of resistance exercise increases urinary calcium
independently of osteoclastic activation in men. Ashizawa N, Fujimura R, Tokuyama K, Suzuki
M.Poult Sci. 1997 Apr;76(4):588-93. Improving eggshell quality at high temperatures with dietary
sodium bicarbonate. Balnave D, Muheereza SK.Fiziol Zh. 2000;46(1):10-6. [The effect of
measured hypoxia on the development of situational osteopenia]. Berezovs'kyi VIa, Litovka IH,
Chaka OH.Plast Reconstr Surg 109(7):2384-97, 2002. Hypoxia and VEGF up-regulate BMP-2 mRNA and
protein expression in microvascular endothelial cells: implications for fracture healing.Bouletreau PJ, Warren SM, Spector JA, Peled ZM, Gerrets RP, Greenwald JA, Longaker MT.J Med Assoc Thai.
2009 Sep; 92 Suppl5:S1-3. The role of vitamin K2 on osteoblastic functions by using stem cell model.
Bunyaratavej N, Sila-Asna M, Bunyaratavej A.J Med Assoc Thai. 2009 Sep;92 Suppl5:S4-6. Highly recommended dose of MK4 for osteoporosis. Bunyaratavej N, Kittimanon N, Jitivirai T,
Tongthongthip B. The recommended dose of Menatretenone is 45 mg three times a day; however the compliant in
daily practice is not convenient. This study shows the twice dose per day is inferior to the recommended
dose. This study used the level of Gla protein in osteocalcin as a parameter for the comparison. The mean of
three-time dose a day is 11.27 nanogram per milliliter while the mean of the other group is 6.07 nanogram
per milliliter after the three-month treatment.Curr Opin Nephrol Hypertens. 1993 Jul;2(4):588-96. Effects of metabolic and respiratory acidosis on bone. Bushinsky DA, Ori Y. Am J Physiol. 1995
Jan;268(1 Pt 1):C80-8. Stimulated osteoclastic and suppressed osteoblastic activity in metabolic but not
respiratory acidosis. Bushinsky DA.J Bone Miner Res. 1993 Jan;8(1):93-102. Physicochemical effects
of acidosis on bone calcium flux and surface ion composition. Bushinsky DA, Wolbach W, Sessler
NE, Mogilevsky R, Levi-Setti R.Am J Physiol Renal Physiol. 2003 Sep;285(3):F532-9. Chronic
acidosis-induced alteration in bone bicarbonate and phosphate. Bushinsky DA, Smith SB, Gavrilov
KL, Gavrilov LF, Li J, Levi-Setti R.Acta Orthop. 2009 Oct;80(5):520-4. The hip fracture incidence
curve is shifting to the right.Bergstrom U, Jonsson H, Gustafson Y, Pettersson U, Stenlund H,
Svensson O.J Dent Res 1957 36:571, The relationship between the histology of the thyroid and the
salivary glands and the incidence of dental caries in the rat, Bixler D, Muhler JC, Shafer WB.J
Bone Miner Res. 1990 Sep;5(9):947-53. Sequential histomorphometric changes in cancellous bone from
ovariohysterectomized dogs. Boyce RW, Franks AF, Jankowsky ML, Orcutt CM, Piacquadio AM, White
JM, Bevan JA.J Clin Endocrinol Metab. 1994 Sep;79(3):730-5. The effect of calcium supplementation on
the circadian rhythm of bone resorption. Blumsohn A, Herrington K, Hannon RA, Shao P, Eyre DR,
Eastell R. Ann Nutr Metab. 2002;46(2):80-7. Urinary collagen cross-links as biochemical markers of
growth: an evaluation of biological variables. Branca F, Valtuena S, Golden MH, Robins S.Arch
Biochem Biophys. 1996 Jan 1;325(1):107-12. Thyromimetic action of the peroxisome proliferators
clofibrate, perfluorooctanoic acid, and acetylsalicylic acid includes changes in mRNA levels for certain
genes involved in mitochondrial biogenesis. Cai Y, Nelson BD, Li R, Luciakova K, dePierre JW.
"Clofibrate, perfluorooctanoic acid, and acetylsalicylic acid all increased the mRNA levels for the
mitochondrial-encoded respiratory-chain components cytochrome c oxidase subunit I and NADH dehydrogenase
subunit I." PLoS One. 2010 Oct 28;5(10):e13704. Obesity reduces bone density associated with
activation of PPARγ and suppression of Wnt/B-catenin in rapidly growing male rats.
Chen JR, Lazarenko OP, Wu X, Tong Y, Blackburn ML, Shankar K, Badger TM, Ronis MJ.Zhonghua Yi Xue Za Zhi.
2011 Apr 5;91(13):925-9. [Effect of aspirin administration for the treatment of osteoporosis in
ovariectomized rat model].
Chen ZW, Wu ZX, Sang HX, Qin GL, Wang LS, Feng J, Wang J, Li XJ, Wang JC, Zhang D.Arch Fr Pediatr.
1987 Dec;44(10):839-41. [Osteocalcin in children of short stature and its nocturnal
variations]. Colle M, Ruffie A, Ruedas E, Chebbo M. "In younger children, Gla-P levels were
significantly lower in patients with growth retardation, when compared with normal children." Amino Acids.
2011 Aug 2. Characterization of the transglutaminase gene family in zebrafish and in vivo analysis
of transglutaminase-dependent bone mineralization. Deasey S, Grichenko O, Du S, Nurminskaya
M.Miner Electrolyte Metab. 1988;14(4):235-9. Circadian rhythm with acrophase at night for urinary
excretion of calcium and magnesium in childhood: population-based data of the cimitile study in southern
Italy. DeSanto NG, DiIorio B, Capasso G, Capodicasa G, Giordano DR, Aulisio M, Paduano C,
Stamler J.QJM. 2011 Jul 18. Non-psychiatric comorbidity associated with Alzheimer's disease. Duthie A, Chew D, Soiza RL.Clin Sci (Lond). 1992 Sep;83(3):375-82. Nyctohemeral changes in bone
turnover assessed by serum bone Gla-protein concentration and urinary deoxypyridinoline excretion:
effects of growth and ageing. Eastell R, Simmons PS, Colwell A, Assiri AM, Burritt MF, Russell
RG, Riggs BL.J. Dental Res. 1964, 43(3), 331.345. Effects of simultaneous administration of estrogen
and parathyroid extract upon teeth, periodontium, and long bones of growing albino mice. Elzay
RP. "There appeared to be more osteoclasts per unit area in the group that received estrogen than in the
group that did not."Osteoporos Int. 2000;11(12):1036-42. Organochlorines and bone mineral density in
Swedish men from the general population. Glynn AW, Michaelsson K, Lind PM, Wolk A, Aune M,
Atuma S, Darnerud PO, Mallmin H.J Biol Chem. 2010 Jun 18;285(25):19561-71. Activation of
glucose-6-phosphate dehydrogenase promotes acute hypoxic pulmonary artery contraction. Gupte
RS, Rawat DK, Chettimada S, Cioffi DL, Wolin MS, Gerthoffer WT, McMurtry IF, Gupte SA. Calcif Tissue Int.
1987 Apr;40(4):212-8. Role of carbonic anhydrase in bone resorption: effect of acetazolamide on
basal and parathyroid hormone-induced bone metabolism. Hall GE, Kenny AD.Environ Toxicol Chem.
2006 Oct;25(10):2787-93.
Embryonic exposure to o,p'-DDT causes eggshell thinning and altered shell gland carbonic anhydrase
expression in the domestic hen. Holm L, Blomqvist A, Brandt I, Brunstrom B, Ridderstrale Y,
Berg C.Hear Res. 2009 Jun;252(1-2):56-60. The effect of sex hormones on bone metabolism of the otic
capsule--an overview. Horner KC.J Bone Miner Res. May 23, 2011:439. Efficacy of
serotonin inhibition in mouse models of bone loss. Inose H, Zhou B, Yadav VK, Guo XE, Karsenty
G, Ducy P.Chem Res Toxicol. 1999 May;12(5):429-36. Tryptamine-4,5-dione, a putative endotoxic
metabolite of the superoxide-mediated oxidation of serotonin, is a mitochondrial toxin: possible
implications in neurodegenerative brain disorders. Jiang XR, Wrona MZ, Dryhurst G.J Pak Med
Assoc. 1996 Jun;46(6):128-31. Changes in plasma electrolytes during acclimatization at high
altitude. Khan DA, Aslam M, Khan ZU.Exp Clin Endocrinol Diabetes. 2010 Jun;118(6):371-6.
Low density lipoprotein-cholesterol levels affect vertebral fracture risk in female patients with
primary hyperparathyroidism.Kaji H, Hisa I, Inoue Y, Sugimoto T. "In conclusion, the present
study demonstrated that lower serum LDL-Chol levels were related to vertebral fracture risk independent of
renal function, age, body size, bone metabolism parameters and the severity of the disease in pHPT
women."Neurochem Int. 2008 Feb;52(3):432-7. Epub 2007 Aug 11. Changes in regional long-term
oxidative metabolism induced by partial serotonergic denervation and chronic variable stress in rat
brain. Kanarik M, Matrov D, Koiv K, Eller M, Tonissaar M, Harro J.Gastroenterology. 2011
Aug;141(2):439-42. The importance of the gastrointestinal tract in the control of bone mass
accrual. Karsenty G, Gershon MD.Brain Res. 2008 Jul 24;1221:93-7.
Increased electrical and metabolic activity in the dorsal raphe nucleus of Parkinsonian rats.
Kaya AH, Vlamings R, Tan S, Lim LW, Magill PJ, Steinbusch HW, Visser-Vandewalle V, Sharp T, Temel
Y.Am J Physiol. 1984 May;246(5 Pt 1):E458-62. Arachidonic acid mobilizes calcium and stimulates
prolactin secretion from GH3 cells. Kolesnick RN, Musacchio I, Thaw C, Gershengorn MC.Bone.
2011 Jul 28. FoxO1, the transcriptional chief of staff of energy metabolism. Kousteni
S.Acta Endocrinol (Copenh). 1981 Nov;98(3):339-44. Inhibitory effect of calcium on serum
prolactin. Kruse K, Kracht U.Endocrinology. 1986 Nov;119(5):2249-55. Parathyroid
hormone-like effects of rainbow trout Stannius products on bone resorption of embryonic mouse calvaria
in vitro. Lafeber FP, Schaefer HI, Herrmann-Erlee MP, Wendelaar Bonga SE.Fiziol Zh.
2003;49(2):58-65. [Oxygen deprivation as the initiator of osteogenesis in hypokinesia].
Litovka IH, Berezovs'ka OP. Exp Clin Endocrinol. 1984 Dec;84(3):313-8. Effects of epinephrine and
norepinephrine on serum parathyroid hormone and calcium in normal subjects. Ljunhgall S,
Akerstrom G, Benson L, Hetta J, Rudberg C, Wide L.Ann Nutr Aliment. 1975;29(4):313-9. [Comparative
study of the effect of free and combined glucose and fructose on the absorption and retention of
calcium]. Lorinet A. (Glucose and starch are poor for calcium absorption and retention, lactose
is good, fructose and inulin are intermediate.)Proc Soc Exp Biol Med. 1988 Apr;187(4):474-81.
Effect of high calcium diet on magnesium, catecholamines, and blood pressure of stroke-prone
spontaneously hypertensive rats. Luft FC, Ganten U, Meyer D, Steinberg H, Gless KH, Unger T,
Ganten D.Exp Clin Endocrinol Diabetes. 1998;106(1):51-6. Longitudinal study of urinary
hydroxy-pyridinium cross-links and growth in healthy infants: higher values with breastfeeding and after
daytime sleep. Lutchman EC, Hardwick TA, Biener R, Chowdhury HA, Trout JR, Shapses SA.Am J Clin
Nutr. 2004 Jan;79(1):155-65. Nutritional associations with bone loss during the menopausal
transition: evidence of a beneficial effect of calcium, alcohol, and fruit and vegetable nutrients and
of a detrimental effect of fatty acids. Macdonald HM, New SA, Golden MH, Campbell MK, Reid
DM.Poult Sci. 1987 Apr;66(4):705-12.
Eggshell quality as influenced by sodium bicarbonate, calcium source, and photoperiod. Makled
MN, Charles OW.Eur J Clin Nutr. 2007 Sep;61(9):1114-20. Dietary fat intake and the risk of
osteoporotic fractures in the elderly.
Martinez-Ramirez MJ, Palma S, Martinez-Gonzalez MA, Delgado-Martinez AD, de la Fuente C,
Delgado-Rodriguez M.Clin Biochem. 1987 Jun;20(3):221-4. Studies on EDTA extracts and collagenase
digests from osteoporotic cancellous bone of the femoral head. Mbuyi-Muamba JM, Gevers G,
Dequeker J.J Anim Sci 1990, 68:1044-1048, Intramuscular collagen and serum hydroxyproline as related
to implanted testosterone, dihydrotestosterone and estradiol-17 beta in growing wethers, Miller
LF, Judge MD, Schanbacker BD.J Aging Health. 2007 Apr;19(2):275-85. Premature graying, balding, and
low bone mineral density in older women and men: the Rancho Bernardo study. Morton DJ,
Kritz-Silverstein D, Riley DJ, Barrett-Connor EL, Wingard DL.J. Dental Res 31: 798, 1952, The
effects of orchiectomy and ovariectomy on dental caries in immature rats, Muhler JC and Shafer
WG.Calcif Tissue Int. 1981;33(5):529-40. Biochemical and histological studies on various bone cell
preparations. Nijweide PJ, van der Plas A, Scherft JP.Gastroenterology. 2007 Aug;133(2):608-18.
Epub 2007 May 21. Serotonin mediates oxidative stress and mitochondrial toxicity in a murine model
of nonalcoholic steatohepatitis. Nocito A, Dahm F, Jochum W, Jang JH, Georgiev P, Bader M,
Renner EL, Clavien PA.Nature. 1971 Feb 19;229(5286):571. DDE reduces medullary bone formation in
birds.Oestreicher MI, Shuman DH, Wurster CF.Nihon Ronen Igakkai Zasshi. 1997
Sep;34(9):743-7.
[Effects of calcium supplementation using AAACa or milk on nocturnal bone resorption in young
women]. Ohgitani S, Fujii Y, Fujita T.Life Sci. 2005 Apr 8;76(21):2473-82.
Vitamin K2 binds 17beta-hydroxysteroid dehydrogenase 4 and modulates estrogen metabolism.
Otsuka M, Kato N, Ichimura T, Abe S, Tanaka Y, Taniguchi H, Hoshida Y, Moriyama M, Wang Y, Shao RX, Narayan
D, Muroyama R, Kanai F, Kawabe T, Isobe T, Omata M.Osteoporos Int. 1995;5(6):472-7. Circadian rhythm
in type I collagen formation in postmenopausal women with and without osteopenia. Pedersen BJ,
Schlemmer A, Rosenquist C, Hassager C, Christiansen C.Acta Physiol Scand. 1997 Oct;161(2):161-9. Serotonin inhibition of 1-methylxanthine metabolism parallels its vasoconstrictor activity and inhibition
of oxygen uptake in perfused rat hindlimb.
Rattigan S, Appleby GJ, Miller KA, Steen JT, Dora KA, Colquhoun EQ, Clark MG.Am J Physiol Regul
Integr Comp Physiol. 2010 Dec;299(6):R1685-92. Effects of high-altitude hypoxia on the hormonal
response to hypothalamic factors. Richalet JP, Letournel M, Souberbielle JC.Biochem. J. vol.
50:537-542, 1952, Studies on the skeletal tissues. 2. The collagen content of bones from rabbits,
oxen and humans. Rogers HJ, Weidmann SM, Parkinson A. Int J Clin Pharmacol Res.
2003;23(2-3):83-92. Eggshell calcium in the prevention and treatment of osteoporosis.
Rovensky J, Stancikova M, Masaryk P, Svik K, Istok R.Br J Nutr. 2002 Mar;87(3):267-75.
Positive effects of a chicken eggshell powder-enriched vitamin-mineral supplement on femoral neck bone
mineral density in healthy late post-menopausal Dutch women. Schaafsma A, van Doormaal JJ,
Muskiet FA, Hofstede GJ, Pakan I, van der Veer E.Poult Sci. 2000 Dec;79(12):1833-8. Mineral, amino
acid, and hormonal composition of chicken eggshell powder and the evaluation of its use in human
nutrition. Schaafsma A, Pakan I, Hofstede GJ, Muskiet FA, Van Der Veer E, De Vries PJ.Ukr
Biokhim Zh. 1990 Nov-Dec;62(6):93-7.
[Effect of inflammatory mediators on respiration in rat liver mitochondria].
Semenov VL.Cell Biochem Biophys. 2011 Sep;61(1):23-31. Aspirin attenuates pulmonary
arterial hypertension in rats by reducing plasma 5-hydroxytryptamine levels. Shen L, Shen J, Pu
J, He B. Arch Gynecol. 1982;231(4):263-7.
Influence of prolactin on metabolism and energy production in perfused corpus luteum bearing bovine
ovaries. Spatling L, Stahler E, Vilmar W, Ullrich J, Buchholz R.Probl Endokrinol (Mosk). 1991
Sep-Oct;37(5):54-8. [Metabolism of thyroid gland cells as affected by prolactin and
emotional-physical stress]. Strizhkov VV.J Biol Chem 282(35):25406-15, 2007.
Ultrasound induces hypoxia-inducible factor-1 activation and inducible nitric-oxide synthase expression
through the integrin/integrin-linked kinase/Akt/mammalian target of rapamycin pathway in
osteoblasts. Tang CH, Lu DY, Tan TW, Fu WM, Yang RS.Calcif Tissue Int. 2008 May;82(5):392-400.
Collagen cross-linking influences osteoblastic differentiation. Turecek C, Fratzl-Zelman N,
Rumpler M, Buchinger B, Spitzer S, Zoehrer R, Durchschlag E, Klaushofer K, Paschalis EP, Varga F.Artif
Organs. 2000 Feb;24(2):129-36.
Synergistic effect of released aspirin/heparin for preventing bovine pericardial calcification.
Vasudev SC, Chandy T, Sharma CP, Mohanty M, Umasankar PR.Am J Physiol. 1986 May;250(5 Pt 2):R845-50. Energy balance and brown adipose tissue thermogenesis during pregnancy in Syrian hamsters.
Wade GN, Jennings G, Trayhurn P.Expert Opin Investig Drugs. 2005 Mar;14(3):251-64. Parathyroid hormone and leptin--new peptides, expanding clinical prospects. Whitfield JF. Leptin,
a member of the cytokine superfamily has a PTH-like osteogenic activity and may even partly mediate PTH
action. But leptin has two drawbacks that cloud its therapeutic future. First, apart from directly
stimulating osteoblastic cells, it targets cells in the hypothalamic ventromedial nuclei and through them it
reduces oestrogenic activity by promoting osteoblast-suppressing adrenergic activity. Second, it stimulates
vascular and heart valve ossification, which leads to such events as heart failure and diabetic limb
amputations.Poult Sci. 1985 May; 64(5):1015-9.
Arterial blood gas, pH, and bicarbonate values in laying hens selected for thick or thin eggshell
production. Wideman RF Jr, Buss EG.J Clin Invest. 1999 Oct; 104(7):895-901. Increased
cortical bone mineral content but unchanged trabecular bone mineral density in female ERbeta(-/-)
mice. Windahl SH, Vidal O, Andersson G, Gustafsson JA, Ohlsson C.J Bone Miner Res. 2001
Aug;16(8):1388-98. Female estrogen receptor beta-/- mice are partially protected against age-related
trabecular bone loss.
Windahl SH, Hollberg K, Vidal O, Gustafsson JA, Ohlsson C, Andersson G.J Physiol Pharmacol. 2007
Dec;58(4):873-85. DDT- and DDE-induced disruption of ovarian steroidogenesis in prepubertal porcine
ovarian follicles: a possible interaction with the main steroidogenic enzymes and estrogen receptor
beta.
Wojtowicz AK, Kajta M, Gregoraszczuk EL.Steroids. 2007 Feb;72(2):124-34. Xenoestrogens are
potent activators of nongenomic estrogenic responses. Watson CS, Bulayeva NN, Wozniak AL, Alyea
RA.Mol Cell Biochem. 2003 Mar;245(1-2):115-20. Inhibitory effect of menaquinone-7 (vitamin K2) on
the bone-resorbing factors-induced bone resorption in elderly female rat femoral tissues in
vitro. Yamaguchi M, Uchiyama S, Tsukamoto Y.Eur J Endocrinol. 2011 Jun;164(6):1035-41. Response of biochemical markers of bone turnover to oral glucose load in diseases that affect bone
metabolism.
Yavropoulou MP, Tomos K, Tsekmekidou X, Anastasiou O, Zebekakis P, Karamouzis M,
Gotzamani-Psarrakou A, Chassapopoulou E, Chalkia P, Yovos JG.Exp Clin Endocrinol. 1984 Dec;84(3):294-8.
Hypercalcaemia and calcitonin inhibit prolactin secretion. Zofkova I, Nedvidkova J.J
Alzheimers Dis. 2011;24(1):101-8. Association between bone mineral density and the risk of
Alzheimer's disease. Zhou R, Deng J, Zhang M, Zhou HD, Wang YJ.


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